Temperature dependence on bandgap of semiconductor photocatalysts.

Light irradiation onto a semiconductor generates heat; however, its electronic structure under high temperature has not yet been well investigated. In this study, we have carefully examined the temperature dependence on the bandgap of simple metal oxides, which are well-known photocatalysts, i.e., TiO2, CeO2, Nb2O5, SnO2 Ta2O5, WO3, ZnO, and ZrO2, using operando UV-visible spectroscopy under controlled temperature (from room temperature to 500 °C). Consequently, a linear decrease in bandgap was seen as a function of temperature with a different slope for each semiconductor. We found that the slope was dependent on the bonding distance between metal and oxygen. This finding is essential to develop a photocatalyst used under the condition involving photo-thermal effect.

Anti-desmocollin autoantibodies in nonclassical pemphigus.

BACKGROUND: Despite the established pathogenic role of anti-desmoglein (Dsg) antibodies in classical pemphigus, the significance of autoantibodies to another desmosomal cadherin, desmocollin (Dsc) is at present unknown. No consistent immunoassay for immunoglobulin (Ig) G autoantibodies to Dscs has been developed. OBJECTIVES: The aim of this study was to develop reliable assays to detect anti-Dsc autoantibodies. METHODS: We expressed soluble recombinant proteins (RPs) of human Dsc1-3 in mammalian cells and examined sera of various types of pemphigus, including 79 paraneoplastic pemphigus (PNP) sera, by novel enzyme-linked immunosorbent assays (ELISAs) using the RPs. We also performed ELISAs of Dsc baculoproteins and used the complementary DNA (cDNA) transfection method, and compared the results with those of mammalian ELISAs. RESULTS: Through mammalian ELISAs, IgG autoantibodies to Dsc1, Dsc2 and Dsc3 were detected in 16.5%, 36.7% and 59.5% of PNP sera, respectively, and considerable numbers of pemphigus herpetiformis (PH) and pemphigus vegetans (PVeg) sera reacted strongly with Dsc1 and Dsc3. Mammalian ELISAs were highly specific and more sensitive than baculoprotein ELISAs or the cDNA transfection method. Several Dsc-positive sera, particularly PH sera, showed no reactivity with Dsgs. The reactivity of PNP serum and PVeg serum with Dscs was not abolished by pre-absorption with Dsg RPs. CONCLUSIONS: The results of these novel ELISAs indicated that IgG anti-Dsc autoantibodies were frequently detected and potentially pathogenic in nonclassical pemphigus.

Evaluation of collaborative tuberculosis and human immunodeficiency virus activities in Phnom Penh, Cambodia.

SETTING: Phnom Penh, Cambodia. OBJECTIVES: 1) To monitor the number of tuberculosis (TB) patients undergoing human immunodeficiency (HIV) testing during TB treatment and trends of referral of TB-HIV patients to HIV services following the appointment of TB-HIV coordinators in TB wards, and 2) to investigate factors that influence undesirable TB treatment outcomes. DESIGN: Retrospective descriptive study based on a review of patient records and interviews with programme staff. RESULTS: Eighty-six per cent of newly registered TB patients underwent HIV testing. Most of the TB-HIV patients were referred to HIV services. Using logistic regression analysis, the risk of an undesirable treatment outcome in extra-pulmonary TB was significantly lower than in smear-positive pulmonary TB. Interviews revealed that patients in poor clinical condition at the start of TB treatment or who faced social problems, such as homelessness or foreign nationality, were at considerable risk for undesirable TB treatment outcomes. CONCLUSION: The appointment of TB-HIV coordinators to TB wards resulted in better HIV testing uptake and referral to HIV care and treatment services. To save TB-HIV patients' lives, it is important to continue this kind of study over a longer term to monitor these activities and to identify high-risk patients.

Analysis of hepatitis B surface antigen mutations in Mongolia: molecular epidemiology and implications for mass vaccination.

Although the potential significance of hepatitis B surface antigen (HBsAg) mutants for failure of immunization has been studied in some endemic countries, whether the "a" determinant variants are responsible for vaccine failure in Mongolia remains unknown. Fifty-nine HBsAg-positive children (age: 8.8 +/- 0.9 years) who had been observed during the nationwide survey of vaccinated cohorts conducted in 2004 were subjected to molecular analyses of hepatitis B virus (HBV). Partial S gene sequences encoding amino acids (aa) 40-171 of HBsAg were determined in 57 children (96.6%) who had detectable HBV DNA. Phylogenetic analysis of the S gene sequences revealed that genotype D accounted for 93.0% and genotype A for 5.3%. Only one child (1.7%) had HBVs of genotypes A and D. HBsAg mutations were found in 17 (29.8%) children ranging from 1 to 4 aa per subject (mean +/- SD, 1.6 +/- 0.9 aa). Pro127Thr and Thr118Ala were the most common substitutions, which occurred in 6 (10.5%) and 3 (5.3%) subjects, respectively; none had Gly145Arg. There were no significant associations in the prevalence of HBsAg mutations with age, sex, residential area, or vaccination status against hepatitis B. Analysis of the deduced amino acid sequence of the entire preS1/preS2/S gene revealed that eight genotype D isolates and one genotype A isolate were quite similar to previously-reported wild-type isolates, suggesting that they are essentially wild-type, but not vaccine-induced mutants. In conclusion, the results demonstrate that hepatitis B surface gene mutants do not play a significant role in vaccination failure in Mongolia.

Correlation coefficients between the dioxin levels in mother's milk and the distances to the nearest waste incinerator which was the largest source of dioxins from each mother's place of residence in Tokyo, Japan.

BACKGROUND: To observe the relationship between the PCDD/F and Co-PCB levels in samples of human breast milk and nearby waste incinerators in Tokyo, Japan. METHODS: Breast milk was taken from 240 mothers residing in Tokyo, Japan to measure and analyze the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs; 14 congeners), polychlorinated dibenzofurans (PCDFs; 15 congeners), and coplanar polychlorinated biphenyls (Co-PCBs; 12 congeners) contained in the fat. Individual milk samples (about 50 ml) were obtained from the mothers 30 days after delivery, between the months of June and September in 1999 and 2000. A map of Tokyo was used to measure the distances between each mother's place of residence and the closest public and industrial waste incinerators. RESULTS: The distances to the nearest waste incinerators bore no apparent correlations with the congeners of PCDD/Fs and Co-PCBs. The distances were also uncorrelated with the mean toxic equivalent quantities (TEQs) of PCDD/Fs (the sum of PCDDs and PCDFs), Co-PCBs, and the total PCDD/Fs and Co-PCBs. CONCLUSIONS: Although waste incinerators were the largest source of dioxins in Japan at the time of the study, the dioxins levels of mother's milk bore no apparent relationships with the distances between the mothers' domiciles and the nearest waste incinerators. In this study, several meaningful factors were not taken into account, namely, the wind direction, the level of dioxin emitted from each incinerator, the level of environmental pollution of dioxins, and the average time the mothers stayed at home each day. A full understanding of these points awaits future studies.

Kawasaki disease in parents and children.

AIM: To estimate the probability that the parents of patients with Kawasaki disease also had a history of the same disease. METHODS: Self-reported parents' histories of Kawasaki disease were collected from data of the 16th nationwide survey of the disease conducted in Japan from January 1999 to December 2000. The incidence of Kawasaki disease was calculated by using data reported in all 16 nationwide surveys and live births in the Japanese vital statistics. The expected number of parents with a history of Kawasaki disease in the general population, which was calculated by using the assumed number of parents in the vital statistics and the incidence of this disease, was compared with the observed number. RESULTS: Among 14,163 parent pairs of patients with Kawasaki disease, 33 parents (25 mothers and 8 fathers) had a history of the disease. The number of parents expected to have a history of Kawasaki disease was 16.1 (8.4 mothers and 7.7 fathers). From a Poisson distribution, the probability of the observed number was less than 0.001 among parents or mothers. The prevalence of a recurrence of Kawasaki disease and incidences involving siblings of patients whose parents had a history of the disease were five or six times higher than those of all patients who were reported in the 16th survey. CONCLUSION: When compared with parents in the general population, the probability of a history of Kawasaki disease was significantly higher in those parents whose children suffered from the same disease. This suggests that, epidemiologically, a genetic predisposition to Kawasaki disease may be implicated in its occurrence.

[Ventilatory failure due to the limitation of chest movement in a case of FPS].

A 72-year-old woman presented with cervicothoracal skin lesions mimicked to scleroderma and muscular atrophy in 1996. Because of the elevation of serum creatinine kinase (CK), muscular biopsy was performed at another institution. Under the diagnosis of polymyositis, she was treated with corticosteroid. Despite of the decrease in serum CK levels by corticosteroid therapy, skin lesions and mascular dystrophy gradually worsened to extend to the regions of major pectoral, paravertebral, and femoral muscles. In 1997, she was admitted to our hospital because of dyspnea. On admission, the limitation of the chest movement was obvious and she developed respiratory arrest due to CO2 narcosis. The femoral magnetic resonance image (MRI) showed increased signal intensity of subcutaneous tissues and fascia on T2-weighted image. The block biopsy specimens obtained from the cervical lesion revealed fibrotic thickness and chronic inflammation of subcutaneous septa, fascia, and perimysium. She was treated by mechanical ventilation and cimetidine and weekly methotrexate were added to the corticosteroid therapy because of the diagnosis of FPS. Thereafter, the skin and muscular lesions as well as the MRI findings were improved. The concept of FPS was proposed by Naschitz et al. This condition is pathologically characterized by cicatrizing fascitis, septal and lobular panniculitis, and perimysial fibrosis and peripheral blood and tissue eosinophilia is not important for diagnosis. FPS includes classical eosinophilic fascitis but is also associated with several disorders such as malignancy. This case is suggestive of the therapeutic consideration of FPS in terms of the response to cimetidine and MTX.

Methotrexate (MTX) inhibits osteoblastic differentiation in vitro: possible mechanism of MTX osteopathy.

OBJECTIVE: To clarify the mechanism of impaired bone formation during low dose methotrexate (MTX) therapy. METHODS: The in vitro effects of MTX on the function and differentiation of osteoblastic cells were investigated using (1) a mouse osteogenic cell line (MC3T3-E1) with the capacity to differentiate into osteoblastic or osteocytes, (2) a human osteoblastic osteosarcoma cell line (SaOS-2) with a mature osteoblastic phenotype, and (3) mouse bone marrow stromal cells containing osteoblast precursors. Osteoblast function was assessed by measuring the cellular activity of alkaline phosphatase (ALP) and the mineralization capacity of cultures. RESULTS: MTX suppressed ALP activity dose-dependently in growing MC3T3-E1 cells, but proliferation of these cells was only inhibited by a high concentration of MTX. In contrast, inhibition of ALP activity in MC3T3-E1 cells of mature osteoblastic phenotype was only observed with 10(-8) M and 10(-7) M MTX, and proliferation was not influenced. ALP activity and the proliferation of SaOS-2 cells were not inhibited by MTX, even when growing cells were treated. However, both ALP activity and formation of calcified nodules in bone marrow stromal cell cultures were significantly suppressed by MTX at concentrations between l0(-10) and 10(-7) M. CONCLUSION: These results suggest that MTX suppresses bone formation by inhibiting the differentiation of early osteoblastic cells.

Elevation of serum hepatic aminotransferases during treatment of rheumatoid arthritis with low-dose methotrexate. Risk factors and response to folic acid.

Sixty-six rheumatoid arthritis (RA) patients were analyzed retrospectively to assess the incidence and risk factors for elevation of serum hepatic aminotransferases during methotrexate (MTX) therapy. The effect of folate supplementation on serum ALT and RA activity was evaluated prospectively in 14 patients who showed a sustained high serum level of ALT. The frequency of elevation of serum AST or ALT was 4-5 times greater than in patients taking other DMARDs. Multivariate linear regression analysis demonstrated that elevation of ALT was independently associated with sex (female), obesity, baseline ALT, MTX dose, and gastrointestinal side effects. Folate supplementation caused ALT levels to decrease in all patients within 3 months. Eleven patients showed no change of RA activity, but 3 patients dropped out of the study because of the exacerbation of RA. These results suggest that careful monitoring of serum hepatic aminotransferases is necessary in patients with predisposing factors, especially those receiving more than 0.15 mg/kg of MTX weekly. Folate supplementation can reverse the sustained elevation of ALT, but might cause exacerbation of RA in some patients.

Conserved T cell receptor complementarity-determining region 3 of ocular T cells in mice with experimental autoimmune uveoretinitis.

PURPOSE: To characterize T cells infiltrating into the ocular tissues of mice with experimental autoimmune uveoretinitis (EAU) immunized with interphotoreceptor retinoid-binding protein (IRBP). METHODS: The T cell receptor (TCR) repertoire on T cells obtained from ocular lesions of EAU mice was analyzed by reverse transcriptase-polymerase chain reaction. The clonotype of the T cells was examined by the single-strand conformation polymorphism (SSCP) method, followed by sequence analysis of the TCR beta-chain complementarity-determining region 3 (CDR3). RESULTS: The repertoire of the TCR BV gene in T cells from inflamed lesions was heterogeneous. SSCP analysis showed accumulation of multiple T cells specifically in ocular tissues of EAU mice, suggesting that these cells were expanded by an antigen-driven stimulation. Junctional sequence analysis demonstrated the presence of highly conserved amino acid sequence motifs (AGTGG, AGD) in CDR3 of BV2-positive T cells. CONCLUSIONS: Our findings suggest that T cells infiltrating into ocular lesions of EAU mice recognize restricted T cell epitopes of IRBP, resulting in autoimmune uveoretinitis.

Hypoplastic left heart syndrome: duration of survival without surgical intervention.

BACKGROUND: Mortality rate for heart transplantation for patients with hypoplastic left heart syndrome (HLHS) has improved, but there is a considerable wait until a suitable donor is available. Thus it is important to examine the duration of survival and risk factors for early death in patients with HLHS who did not undergo surgical intervention. METHODS AND RESULTS: Twenty-six consecutive patients were studied retrospectively. Duration of survival and the 14 following variables were investigated: date of birth, body weight at birth, cardiothoracic ratio, ascending aorta diameter, interatrial communication size, coarctation of the aorta, tricuspid regurgitation, anatomic subtype (patency) of mitral and aortic valve, arterial blood gas findings (pH, PaO 2, SaO 2, PaCO2, base excess), and ST depression in the electrocardiogram. Twenty patients survived <60 days (group A) and 6 patients survived beyond 60 days (group B). The duration of survival (mean [SD]) was 60 (151) days overall (1 patient is currently alive at 783 days). The long-term survivors (beyond 60 days) increased significantly after 1991 (P <.05). Coarctation of the aorta was a significant risk of early death (<60 days) (P <.05). Interatrial communication size was significantly smaller in group B than in group A (P <.05). The mean pH and base excess were significantly lower in group A than in group B. The other 9 variables showed no significant difference between the 2 groups. CONCLUSIONS: There was a significant correlation of long-term survival with stabilized ductal blood flow without coarctation of the aorta, adequate restriction of interatrial communication without severe hypoxemia, and no metabolic acidosis.

Magnetic Effects on Electrolyte Solutions in Pulse and Alternating Fields.

The contribution of the pulse and alternating magnetic fields on the magnetic effects is examined and compared with that of the static field, using a rotational device by which the pulse and alternating fields are able to be applied to the stationary sample. The followings are found: (i) the substantial time required to reach the maximum magnetic effect in the pulse and alternating fields is much smaller than the time in the static field, (ii) the magnetic effect does depend on the frequency of magnetic field, and (iii) the pulse and alternating magnetic fields make the quasi-stable structure more stable than the static field. The results are discussed and compared with the magnetic effects in flow loops reported elsewhere. Copyright 1999 Academic Press.

Characterization of physical entrapment and chemical conjugation of adriamycin in polymeric micelles and their design for in vivo delivery to a solid tumor.

An anticancer drug adriamycin (ADR) was incorporated into polymeric micelles forming from poly(ethylene glycol)-poly(aspartic acid) block copolymer by chemical conjugation and physical entrapment. Structural stability of the polymeric micelles was found to be dependent on both the contents of chemically conjugated and physically entrapped ADR. The polymeric micelle with high contents of the chemically conjugated ADR and the physically entrapped ADR expressed very high in vivo antitumor activity against murine C 26 tumor, while the polymeric micelle with only the chemically conjugated ADR showed negligible in vivo activity. This indicates that the physically entrapped ADR played a major role in antitumor activity in vivo. For the polymeric micelle with the high ADR contents, it was found that a dimer of adriamycin molecules formed and that this dimer was physically entrapped in the inner core of the micelle as well as intact ADR.

[Osteoporosis in rheumatoid arthritis].

Patients with rheumatoid arthritis (RA) develop both periarticular and generalized osteoporosis. Periarticular osteopenia in appendicular bones occurs early in the course of RA and is one of the earliest radiological signs of RA. An uncoupled state in bone resorption-formation linkage, contributes to the development of periarticular osteopenia and it might be mediated through an increased productions of cytokines and prostaglandins by synovium and bone marrow. Accordingly, early suppression of rheumatoid synovitis is necessary for the prevention of periarticular osteopenia. Generalized osteoporosis is also common in RA and leads to increased risk of fractures. Generalized osteoporosis considered to be multifactorial and factors contributing to lumbar osteoporosis might be different from those to loss of appendicular bones, such as femur and radius. Corticosteroids and menopausal state are important risk factors for lumbar osteoporosis. Rheumatoid activity and reduced physical activity are also important determinants. According to the previous studies, however, the influence of functional impairment is more prominent in the femoral BMD compared to spinal BMD. In addition to control of RA and maintenance of physical activity, hormone replacement therapy (HRT) and bisphosphonate are possible agents for the treatment of osteoporosis in RA patients, especially postmenopausal women.

[Clinical relapse of rheumatoid arthritis (escape phenomenon) during low-dose methotrexate therapy].

OBJECTIVE: To clarify the incidence and background of clinical relapse (escape phenomenon) during low-dose methotrexate therapy for rheumatoid arthritis. METHODS: Seventy one patients with rheumatoid arthritis (RA) were analyzed. They were started on therapy with methotrexate (MTX) between April 1, 1991 and May 30, 1995. Among them, 60 patients showed clinical improvement within 6 months after the start of the therapy and were subjected to the analysis for clinical relapse (escape phenomenon). RESULTS: Twelve patients showed an initial improvement followed by a relapse with increased serum CRP and number of painful joints despite the MTX therapy was continued. Two types of the relapses were seen; (1) early, escape (relapse after an initial brief improvement) in 7 patients, and (2) late escape (relapse after a long-term improvement with MTX therapy) in 5 patients. The early escape was seen at 9.0 +/- 0.7 months after the start of therapy while the late escape was seen at 23.3 +/- 4.8 months. Patients with both types of escape phenomenon had the longer duration of the disease and more advanced stage. There was no relationship between clinical relapse and age, baseline RA activity, MTX dose, or concurrent use of corticosteroids and other disease modifying anti-rheumatic drugs. The efficacy of MTX for RA was restored by increasing dose of MTX in 11 patients. CONCLUSION: These results suggest that clinical relapse is not rare in RA patients during low-dose methotrexate therapy, but could be improved by increasing dose.

Ionized magnesium level in whole blood of healthy Japanese children.

Magnesium (Mg) catalyzes or activates more than 300 enzymes in the body, and it plays a pivotal role in the metabolism of carbohydrates, fats and proteins. Until recently Mg has only been measured as a total substance concentration; however, it is the free ionized form of magnesium (iMg2+) that is physiologically active. In 1992, a novel ion selective electrode (ISE) utilizing a neutral carrier-based membrane for assessment of iMg2+ in whole blood, plasma or serum was designed and it was used to measure the iMg2+ level in whole blood of 160 healthy Japanese children. There was a significant positive correlation between serum total magnesium (SMg) and iMg2+. Percent ionized magnesium (iMg2+/SMg) was 58.3 +/- 4.1% and increased with growth (age and bodyweight). The level of iMg2+ was 1.29 +/- 0.08 mg/dL (range, 1.09-1.51) ¿0.535 +/- 0.033 mmol/L (range, 0.452-0.627)¿ and was constant irrespective of growth. On the other hand, SMg, serum total calcium (SCa) and ionized calcium (iCa2+) decreased with growth. Thus only iMg2+ level was constant irrespective of growth. It is suspected that the buffer action of protein binding Mg may keep the iMg2+ level constant, because iMg2+ plays an important part in the activation of many enzymes.

Short-term low dose methotrexate ameliorates abnormal bone metabolism and bone loss in adjuvant induced arthritis.

OBJECTIVE: To clarify whether short term weekly methotrexate (MTX) therapy aggravates bone abnormalities in adjuvant induced arthritis (AIA) or improves them through its antiarthritic effect. METHODS: Bone metabolism and bone mineral density (BMD) were studied in 6 groups of Lewis rats: (1) normal controls, (2) rats given MTX 0.3 mg/kg weekly, (3) rats given MTX 3 mg/kg weekly, (4) AIA rats, (5) AIA rats given MTX 0.3 mg/kg weekly, and (6) AIA rats given MTX 3 mg/kg weekly. Osteogenic activity was determined from serum osteocalcin levels and number of marrow fibroblast colony forming units (osteogenic precursor cells). Bone resorptive activity was assayed by detecting osteoclast-like cells and pit formation in bone marrow cultures. RESULTS: In control rats, MTX (3 mg/kg weekly) suppressed osteogenic activity, as shown by low serum osteocalcin levels and decreased growth of marrow fibroblast colony forming units. Osteoclast-like cells and pit formation in bone marrow cultures from control rats were increased by MTX, but BMD was unchanged. In rats with AIA, MTX (3 mg/kg) suppressed arthritis and restored the decreased osteogenic activity of bone marrow cells, and reduced their increased bone resorptive activity. These changes resulted in a significant increase of periarticular BMD in the femur. CONCLUSION: Low dose weekly MTX therapy had a favorable effect on abnormal bone metabolism and osteopenia in rats with AIA.

Rapid enlargement of cardiac rhabdomyoma during corticotropin therapy for infantile spasms.

OBJECTIVE: To investigate the influence of corticotropin therapy on cardiac rhabdomyoma. DESIGN: Analysis of data from echocardiography performed on in-patients. PATIENTS: Six patients with rhabdomyoma who were admitted to the authors' medical centre with either convulsion (five cases) or prematurity (one case) between 1985 and 1995. Five had tuberous sclerosis. INTERVENTION: Size of cardiac tumours of each patient was measured by echocardiography, and volume index was calculated as the ratio of the tumour volume to its initial volume. MAIN RESULT: Increase in size of some of the tumours was found during corticotropin therapy on follow-up echocardiography. Maximum volume indexes of tumours in the case of patients (n = 4) who did not receive corticotropin therapy was 1.2 to 3.7, whereas those of patients (n = 2) who received therapy was 9.1 to 12; one of the latter patients died. CONCLUSION: Corticotropin may contribute to the enlargement of cardiac rhabdomyoma. The size of cardiac rhabdomyomas must be carefully followed when patients are treated with corticotropin.

Long-term follow up of abdominal aortic aneurysm complicating Kawasaki disease: comparison of the effectiveness of different imaging methods.

A 2 month old boy with Kawasaki disease developed the rare complication of abdominal aortic aneurysm (AAA). He was followed up over 7 years by ultrasonography (2D-ECHO) with and without Doppler flow evaluation, angiography and computed tomography. Calcification was noted 33 months after the onset of the disease but the aneurysm did not decrease in size. 2D-ECHO was adequate for evaluating the size of an aneurysm, but did not show thrombus formation or calcification. Doppler flow studies did not show abnormally high blood flow velocities either at the inflow or outlet of aneurysms, which are indicative of stenosis. On the other hand an abdominal aortogram provided information regarding the luminal shape and abnormalities in flow pattern suggestive of thrombus formation. Fluoroscopy cannot demonstrate calcification in the early stages. Computed tomography (CT) was the imaging method of choice for the evaluation of obstructive or calcific changes; an organizing thrombus was clearly demonstrated and early detection of calcification was possible.

Primary role of NADH formed by glucose dehydrogenase in ATP provision at the early stage of spore germination in Bacillus megaterium QM B1551.

Metabolic events involved in energy metabolism were studied in order to evaluate the ATP-forming ability of Bacillus megaterium QM B1551 spores at the very early stage of germination. When heat-activated spores were germinated on glucose as a sole substrate, its oxidation into gluconate (catalyzed by glucose dehydrogenase, EC 1.1.1.47), the accompanying NADH formation, oxygen uptake, and RNA synthesis were initiated immediately after germination, even when anaerobic breakdown of 3-phosphoglycerate (an ATP source for spores) and the subsequent glucose metabolism via the phosphorylating pathway were impaired by potassium fluoride (KF). In contrast, fructose metabolism and the accompanying metabolic events did not begin until a few minutes after triggering of germination, and those events were entirely abolished by KF, indicating that fructose metabolism is initiated exclusively via its phosphorylation by the ATP derived from endogenous 3-phosphoglycerate. Thus those results provided further evidence for our previous proposal (Otani et al (1987) Microbiol. Immunol. 31: 967-974; Sano et al (1988) Biochem. Biophys. Res. Commun. 151: 48-52) that the first molecules of ATP in germinating spores can be efficiently generated via aerobic oxidation of NADH, which is formed by glucose dehydrogenase. Fluorescence monitoring of NADH in germinating spores also supported this conclusion.