Intramolecular [π4s + π4s] photocycloaddition of carbon- and nitrogen-bridged [32](1,4)naphthalenophanes.

[32](1,4)Naphthalenophanes, bearing carbon-bridge chains (syn- and anti-NPs) and nitrogen-bridge chains (syn- and anti-ANPs), were synthesized, and their X-ray structures and photoreactions were investigated. The intramolecular separation distance between the naphthalene cores for ANPs was shorter than that for NPs, suggesting that intramolecular interactions between the naphthalene rings  were more efficient for ANPs compared to NPs. Upon photoirradiation at 300 nm, anti-NP, syn-ANP and anti-ANP produced the corresponding intramolecular [π4s + π4s] cycloadducts, whereas syn-NP gave an unidentified complex product mixture. Quantum yields for the photo-consumption (ΦPC) of NPs and ANPs were evaluated to quantitatively compare their photoreactivity. The ΦPC values of ANPs were approximately two-fold higher than those of ANPs.Noteworthily, the ΦPC value of syn-ANP was estimated to be unity. Based on these results we discuss the effects of the alignments of the naphthalene cores (anti vs. syn) and the bridging elements (C-bridge vs. N-bridge) on the photoreaction efficiencies of [32](1,4)naphthalenophanes.

Vanadium-Containing Keggin-Type Polyoxometalates, [VM12O40]3- and [VVM11O40]4- (M = Mo, W): Structural Characterization and Voltammetric, NMR, and EPR Studies Related to Electrochemical Reduction at Framework and Central Vanadium Sites.

Vanadium is accommodated in both the framework (VoutV) and central positions (VinV) in the Keggin-type polyoxometalates (POMs) [VinVVoutVM11O40]4- (M = Mo, W; VinVVoutVM11) and in the central position in [VinVM12O40]3- (VinVM12). The structures of the VinVVoutVM11 class have been determined by X-ray crystallography and compared to those of VinVM12 reported previously. A major feature of interest with POMs is their capacity for very extensive reduction, particularly when protonation accompanies the electron transfer step. With VinVVoutVM11 and VinVM12 POMs, knowledge as to whether reduction occurs at V or M sites and the concomitant dependence on acidity has been obtained. Frozen solution EPR spectra obtained following bulk electrolysis showed that the one-electron reduction of VinVMo12 occurs at the molybdenum framework site to give VinVMoVMo11. In contrast, EPR spectra of one-electron reduced VinVW12 at <30 K are consistent with the electron being accommodated on the central V atom in a tetrahedral environment to give VinIVW12. In the case of VinVVoutVM11, the initial reduction occurs at the framework VoutV site to give VinVVoutIVM11. The second electron is delocalized over the Mo framework in two-electron reduced VinVVoutIVMoVMo10, whereas it is accommodated on the central V site in VinIVVoutIVW11. The distance between VinIV and VoutIV in VinIVVoutIVW11 estimated as 3.5 ± 0.2 Å from analysis of the EPR spectrum is consistent with that obtained in VinVVoutVW11 from crystallographic data. Simulations of the cyclic voltammograms as a function of CF3SO3H acid concentration for the initial reduction processes provide excellent agreement with experimental data obtained in acetonitrile (0.10 M [n-Bu4N][PF6]) and allowed acid association constants to be estimated and compared with the literature values available for [XVoutVM11O40]n- (X = S (n = 3), P and As (n= 4); M = Mo, W). The interpretation of the voltammetric data is supported by 51V NMR measurements on the oxidized VV forms of the POMs.

Semiconductor-metal transition in Bi2Se3 caused by impurity doping.

Doping a typical topological insulator, Bi2Se3, with Ag impurity causes a semiconductor-metal (S-M) transition at 35 K. To deepen the understanding of this phenomenon, structural and transport properties of Ag-doped Bi2Se3 were studied. Single-crystal X-ray diffraction (SC-XRD) showed no structural transitions but slight shrinkage of the lattice, indicating no structural origin of the transition. To better understand electronic properties of Ag-doped Bi2Se3, extended analyses of Hall effect and electric-field effect were carried out. Hall effect measurements revealed that the reduction of resistance was accompanied by increases in not only carrier density but carrier mobility. The field-effect mobility is different for positive and negative gate voltages, indicating that the EF is located at around the bottom of the bulk conduction band (BCB) and that the carrier mobility in the bulk is larger than that at the bottom surface at all temperatures. The pinning of the EF at the BCB is found to be a key issue to induce the S-M transition, because the transition can be caused by depinning of the EF or the crossover between the bulk and the top surface transport.

Superconductivity of topological insulator Sb2Te3-ySeyunder pressure.

The crystal structures of Sb2Te3-ySey(y= 0.6 andy= 1.2) at 0-24 GPa were investigated by synchrotron x-ray diffraction. The stoichiometry of Sb2Te3-ySeyused in this study was determined to be Sb2Te2.19(9)Se0.7(2)fory= 0.6 and Sb2Te1.7(1)Se1.3(3)fory= 1.2, on the basis of energy-dispersive x-ray spectroscopy. The sample of Sb2Te2.19(9)Se0.7(2)showed a structural phase transition from a rhombohedral structure (space group No. 166,R3¯m) (phase I) to a monoclinic structure (space group No. 12,C2/m) (phase II), with increasing pressure up to ∼9 GPa. A new structural phase (phase II') emerged at 17.7 GPa, a monoclinic structure with the space groupC2/c(No. 15). Finally, a 9/10-fold monoclinic structure (space group No. 12,C2/m) (phase III) was observed at 21.8 GPa. In contrast, the sample of Sb2Te1.7(1)Se1.3(3)provided only phase I (space group No. 166,R3¯m) and phase II (space group No. 12,C2/m), showing one structural phase transition from 0-19.5 GPa. These samples were not superconductors at ambient pressure, but superconductivity suddenly appeared with increasing pressure. Superconductivity with superconducting transition temperatures (Tc's) of 2 and 4 K was observed above 6 GPa in phase I of Sb2Te2.19(9)Se0.7(2). In this sample, theTcvalues of 6 and 9 K were observed in phase II and phase II' or III of Sb2Te2.19(9)Se0.7(2), respectively. Superconductivity withTc's of 4 and 5 K suddenly emerged in Sb2Te1.7(1)Se1.3(3)at 13.6 GPa, which corresponds to phase II, and it evolved to 6.0 K under further increased pressure. ATcvalue of 9 K was finally found above 15 GPa. The magnetic field dependence ofTcin phase II of Sb2Te2.19(9)Se0.7(2)and Sb2Te1.7(1)Se1.3(3)followed ap-wave polar model, suggesting topologically nontrivial superconductivity.

A cyclic tetrasaccharide, cycloisomaltotetraose, was enzymatically produced from dextran and its crystal structure was determined.

Two bacterial strains isolated from soil, namely Agreia sp. D1110 and Microbacterium trichothecenolyticum D2006, were found to produce a novel oligosaccharide. The oligosaccharide was enzymatically produced from dextran using the culture supernatant of Agreia sp. D1110 or M. trichothecenolyticum D2006. LC-MS and NMR analysis identified the novel oligosaccharide as cyclo-{→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→6)-α-d-Glcp-(1→}, which was named cycloisomaltotetraose, and abbreviated as CI4. CI4 was subsequently crystalized and its X-ray crystallographic structure was determined. CI4 crystals were shown to be pentahydrate, with the CI4 molecules in the crystal structure displaying a unique 3D structure, in which two glucosyl residues in the molecule were facing each other. This unique 3D structure was quite different from the 3D structure of known cyclic tetrasaccharides. This is the first report of CI4 molecules and their unique crystal structure.

Metal-substituted tungstosulfates with Keggin structure: synthesis and characterization.

Simple synthetic procedures for accessing novel metal-substituted tungstosulfates [SMW11O39]4- with Keggin-type structures were developed based on the reaction of metal ions (M = Mn2+, Co2+, Ni2+, and Cu2+) with lacunary tungstosulfate, [SW11O39]6-, which was obtained by treating [SW12O40]2- with a weak base in acetone. All metal-substituted tungstosulfates were characterized by elemental analysis, X-ray crystallography, ESI-MS, IR, Raman, UV-Vis and cyclic voltammetry analyses.

Fermi level tuning of Ag-doped Bi2Se3 topological insulator.

The temperature dependence of the resistivity (ρ) of Ag-doped Bi2Se3 (AgxBi2-xSe3) shows insulating behavior above 35 K, but below 35 K, ρ suddenly decreases with decreasing temperature, in contrast to the metallic behavior for non-doped Bi2Se3 at 1.5-300 K. This significant change in transport properties from metallic behavior clearly shows that the Ag doping of Bi2Se3 can effectively tune the Fermi level downward. The Hall effect measurement shows that carrier is still electron in AgxBi2-xSe3 and the electron density changes with temperature to reasonably explain the transport properties. Furthermore, the positive gating of AgxBi2-xSe3 provides metallic behavior that is similar to that of non-doped Bi2Se3, indicating a successful upward tuning of the Fermi level.

Structural features of a bacterial cyclic α-maltosyl-(1→6)-maltose (CMM) hydrolase critical for CMM recognition and hydrolysis.

Cyclic α-maltosyl-(1→6)-maltose (CMM, cyclo-{→6)-α-d-Glcp-(1→4)-α-d-Glcp-(1→6)-α-d-Glcp-(1→4)-α-d-Glcp-(1→})is a cyclic glucotetrasaccharide with alternating α-1,4 and α-1,6 linkages. CMM is composed of two maltose units and is one of the smallest cyclic glucooligosaccharides. Although CMM is resistant to usual amylases, it is efficiently hydrolyzed by CMM hydrolase (CMMase), belonging to subfamily 20 of glycoside hydrolase family 13 (GH13_20). Here, we determined the ligand-free crystal structure of CMMase from the soil-associated bacterium Arthrobacter globiformis and its structures in complex with maltose, panose, and CMM to elucidate the structural basis of substrate recognition by CMMase. The structures disclosed that although the monomer structure consists of three domains commonly adopted by GH13 and other α-amylase-related enzymes, CMMase forms a unique wing-like dimer structure. The complex structure with CMM revealed four specific subsites, namely -3', -2, -1, and +1'. We also observed that the bound CMM molecule adopts a low-energy conformer compared with the X-ray structure of a single CMM crystal, also determined here. Comparison of the CMMase active site with those in other enzymes of the GH13_20 family revealed that three regions forming the wall of the cleft, denoted PYF (Pro-203/Tyr-204/Phe-205), CS (Cys-163/Ser-164), and Y (Tyr-168), are present only in CMMase and are involved in CMM recognition. Combinations of multiple substitutions in these regions markedly decreased the activity toward CMM, indicating that the specificity for this cyclic tetrasaccharide is supported by the entire shape of the pocket. In summary, our work uncovers the mechanistic basis for the highly specific interactions of CMMase with its substrate CMM.

Preparation of Preyssler-type Phosphotungstate with One Central Potassium Cation and Potassium Cation Migration into the Preyssler Molecule to form Di-Potassium-Encapsulated Derivative.

A mono-potassium cation-encapsulated Preyssler-type phosphotungstate, [P5W30O110K]14- (1), was prepared as a potassium salt, K14[P5W30O110K] (1a), by heating mono-bismuth- or mono-calcium-encapsulated Preyssler-type phosphotungstates (K12[P5W30O110Bi(H2O)] or K13[P5W30O110Ca(H2O)]) in acetate buffer. Characterization of the potassium salt 1a by single-crystal X-ray structure analysis, 31P and 183W nuclear magnetic resonance (NMR) spectroscopy, Fourier transform infrared spectroscopy, high-resolution electrospray ionization mass spectroscopy, and elemental analysis revealed that one potassium cation is encapsulated in the central cavity of the Preyssler-type phosphotungstate molecule with a formal D 5h symmetry. Density functional theory calculations have confirmed that the potassium cation prefers the central position of the cavity over a side position, in which no water molecules are coordinated to the encapsulated potassium cation. 31P NMR and cyclic voltammetry analyses revealed the rapid protonation-deprotonation of the oxygens in the cavity compared to that of other Preyssler-type compounds. Heating of 1a in the solid state afforded a di-K+-encapsulated compound, K13[P5W30O110K2] (2a), indicating that a potassium counter-cation is introduced in one of the side cavities, concomitantly displacing the internal potassium ion from the center to a second side cavity, thus providing a new method to encapsulate an additional cation in Preyssler compounds.

Azulene-Fused Linear Polycyclic Aromatic Hydrocarbons with Small Bandgap, High Stability, and Reversible Stimuli Responsiveness.

Azulene-fused polycyclic aromatic hydrocarbons (PAHs) were synthesized from commercially available azulene in four steps. The resulting azulene conjugates exhibited significantly narrow HOMO-LUMO bandgaps with high air stability, confirmed by photophysical study. Introduction of azulene also enabled the unique reversible stimuli-responsiveness even with the weak acid and base, which can potentially control the degree of conjugation and optoelectronic properties by simple acid-base and redox processes.

Chiral Macrocyclic Organocatalysts for Kinetic Resolution of Disubstituted Epoxides with Carbon Dioxide.

Among chiral macrocycles 1 synthesized, 1m with the 3,5-bis(trifluoromethyl)phenylethynyl group was the best organocatalyst for the enantioselective synthesis of cyclic carbonates from disubstituted or monosubstituted epoxides and CO2. The X-ray crystal structure of 1m revealed a well-defined chiral cavity with multiple hydrogen-bonding sites that is suitable for the enantioselective activation of epoxides. A catalytic cycle proposed was supported by DFT calculations.

Visualization of Protonation/Deprotonation of Active Pharmaceutical Ingredient in Solid State by Vapor Phase Amine-Selective Alkyne Tagging and Raman Imaging.

Here, we report a simple and direct method to visualize the protonation/deprotonation of an amine active pharmaceutical ingredient (API) in the solid state using a solid-vapor reaction with propargyl bromide and Raman imaging for the assessment of the API during the manufacturing process of solid formulations. An alkyne tagging occurred on the free form of solid haloperidol by the vapor phase reaction, and a distinct Raman signal of alkyne was detected. Alkyne signal monitoring by Raman imaging enabled us to visualize the distribution of the free-form haloperidol in a solid formulation. On the other hand, haloperidol hydrochloride did not react with propargyl bromide in the solid-vapor reaction, and the alkyne signal was not observed. Using the difference in reactivity, the protonation/deprotonation of the amine API in the solid state could be visualized. As an example of application, we tried to visually assess the protonation/deprotonation state when the free-form haloperidol was ground with acids using the solid-vapor reaction and Raman imaging and found that haloperidol was partially protonated when ground with 2 equivalents of hydrogen chloride. Furthermore, we demonstrated the relationship between the degree of protonation and the amount of water added as a medium for grinding haloperidol with succinic acid.

Voltammetric and Spectroscopic Studies of α- and ß-[PW12O40]3- Polyoxometalates in Neutral and Acidic Media: Structural Characterization as Their [(n-Bu4N)3][PW12O40] Salts.

The structure of the Keggin-type ß-[PW12O40]3- (PW12) polyoxometalate, with n-Bu4N+ as the countercation, has been determined for the first time by single-crystal X-ray analysis and compared to data obtained from a new determination of the structure of the α-PW12 isomer, having the same countercation. Analysis of cyclic voltammograms obtained in CH3CN (0.1 M [n-Bu4N][PF6]) reveals that the reversible potential for the ß-PW12 isomer always remains ca. 100 mV more positive than that of the α-PW12 isomer on addition of the acid CF3SO3H. Simulations of the cyclic voltammetry as a function of acid concentration over the range 0-5 mM mimic experimental data exceptionally well. These simulation-experiment comparisons provide access to reversible potentials and acidity constants associated with α and ß fully oxidized and one- and two-electron reduced systems and also explain how the two well-resolved one-electron W(VI)/W(V) processes converge into a single two-electron process if sufficient acid is present. 183W NMR spectra of the oxidized forms of the PW12 isomers are acid dependent and in the case of ß-PW12 imply that the bridging oxygens between the WI and WII units are preferentially protonated in acidic media. EPR data on frozen solutions of one-electron reduced ß-[PWVWVI11O40]4- indicate that either the WI or the WIII unit in ß-PW12 is reduced in the ß-[PWVI12O40]3-/ß-[PWVWVI11O40]4- process. In the absence of acid, reversible potentials obtained from the α- and ß-isomers of PW12 and [SiW12O40]4- exhibit a linear relationship with solvent properties such as Lewis acidity, acceptor number, and polarity index.

Encapsulation of Two Potassium Cations in Preyssler-Type Phosphotungstates: Preparation, Structural Characterization, Thermal Stability, Activity as an Acid Catalyst, and HAADF-STEM Images.

Dipotassium cation (K+)-encapsulated Preyssler-type phosphotungstate, [P5W30O110K2]13-, was prepared by heating monobismuth (Bi3+)-encapsulated Preyssler-type phosphotungstate, [P5W30O110Bi(H2O)]12-, in acetate buffer in the presence of an excess amount of potassium cations. Characterization of the isolated potassium salt, K13[P5W30O110K2] (1a), and its acid form, H13[P5W30O110K2] (1b), by single crystal X-ray structure analysis, 31P and 183W nuclear magnetic resonance (NMR), Fourier transform infrared (FT-IR) spectroscopy, cyclic voltammetry (CV), high-resolution electrospray ionization mass spectroscopy (HR-ESI-MS), and elemental analysis revealed that two potassium cations are encapsulated in the Preyssler-type phosphotungstate molecule with formal D5h symmetry, which is the first example of a Preyssler-type compound with two encapsulated cations. Incorporation of two potassium cations enhances the thermal stability of the potassium salt, and the acid form shows catalytic activity for hydration of ethyl acetate. Packing of the Preyssler-type molecules was observed by high-resolution high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM).

Emergence of superconductivity in (NH3)yMxMoSe2 (M: Li, Na and K).

We report syntheses of new superconducting metal-doped MoSe2 materials (MxMoSe2). The superconducting MxMoSe2 samples were prepared using a liquid NH3 technique, and can be represented as '(NH3)yMxMoSe2'. The Tcs of these materials were approximately 5.0 K, independent of x and the specific metal atom. X-ray diffraction patterns of (NH3)yNaxMoSe2 were recorded using polycrystalline powders. An increase in lattice constant c showed that the Na atom was intercalated between MoSe2 layers. The x-independence of c was observed in (NH3)yNaxMoSe2, indicating the formation of a stoichiometric compound in the entire x range, which is consistent with the x-independence of Tc. A metallic edge of the Fermi level was observed in the photoemission spectrum at 30 K, demonstrating its metallic character in the normal state. Doping of MoSe2 with Li and K also yielded superconductivity. Thus, MoSe2 is a promising material for designing new superconductors, as are other transition metal dichalcogenides.

Reversible near-infrared/blue mechanofluorochromism of aminobenzopyranoxanthene.

Mechanochromic organic molecules (MOMs) that exhibit a large difference of fluorescence wavelength between two states have important potential applications, but few such compounds are known. Here, we report a new MOM, cis-ABPX01(0), which shows switchable near-IR and blue fluorescence responses. Detailed spectrophotometric and single-crystal X-ray analyses revealed that the near-IR fluorescence is attributable to fluorescence from slip-stacked dimeric structures in crystals, while the blue fluorescence is attributable to fluorescence from the monomer. Switching between the two is achieved by dynamic structural interconversion between the two molecular packing arrangements in response to mechanical grinding and solvent vapor-fuming.

Thyminate(2-)-bridged cyclic tetranuclear rhodium(III) complexes formed by a template of a sodium, calcium or lanthanoid ion.

In the thyminate(2-)-bridged tetranuclear Cp*Rh(III) complexes incorporating a Na(+), Ca(2+) or Ln(3+) cation, homochiral aggregation of four Rh(III) centres was achieved to form metallacalix[4]arene-type clusters. The thyminate(2-) bridged two Rh(III) and the third metal ion with a µ3-1κN(1):2κ(2)N(3),O(2):3κO(2) mode in the Na(+) and Ca(2+) complexes, while in the Ln(3+) analogues it exhibited a different bridging mode, µ3-1κN(1):2κ(2)N(3),O(4):3κO(2).

Associations between individual-level social capital and self-rated health or depression among elderly men and women.

OBJECTIVES: Recent studies have investigated the relationship between social capital and health, although relatively few studies have evaluated the role of gender in this relationship. This study's aim was to investigate whether individual-level social capital in communities is associated with the self-reported health (self-rated health and depression) of urban elderly women and men living at home, even after adjusting for some confounding factors, and whether there is a gender difference in the association between social capital and health. METHODS: A self-administered questionnaire was mailed to 2,400 people aged between 65 and 79 who were not receiving long-term care insurance services in City A (Tokyo). Indicators of social capital, as represented by cognitive social capital (trust in neighbors and reciprocity) and structural social capital (group membership in local associations), were tested with two self-reported health indicators: perceived health and depression. The associations between social capital and perceptions of health were analyzed for men and women respectively using multivariate logistic regression analysis with adjustments made for age, economic status, educational status, presence of chronic disorders, functional capability levels, marital status, and duration of residence. RESULTS: The number of valid responses was 1,776 (men: n=887, 71.2 ± 4.0 years; women: n=889, 70.9 ± 3.9 years; mean ± SD) with a response rate of 74.5%. Lower levels of cognitive social capital (civic mistrust) were associated with an odds ratio for poorer self-rated health of 1.58 (95% CI [1.07-2.34], P=.022) at the individual level in men. In women, lack of reciprocity (OR=1.63, 95% CI [1.10-2.41], P=.014) was associated with poorer self-rated health. Civic mistrust and lack of reciprocity were associated with depression in men and women. Lack of group membership in local associations was associated with self-rated health (OR=1.68, 95% CI [1.16-2.44], P=.007) and depression (OR=2.24, 95% CI [1.49-3.38], P<.001) in women. CONCLUSION: Civic mistrust was associated with poorer self-rated health and a lack of reciprocity was associated with depression in men. In women, lack of reciprocity was associated with poorer self-rated health and depression, and lack of group membership in local associations was associated with self-rated health and depression. These results lead to the conclusion that there were noticeable gender differences in the relationship between the social capital and self-rated health of the elderly. A longitudinal study should be conducted to clarify the causal relationship between social capital and perceived health.

[Structural homology between streptolysin O (SLO) produced by streptococcus pyogenes and SLO-like protein produced by non-pathogenic streptococci and cross-reactivity of antibody against SLO-like protein to SLO].

Nine clones of non-pathogenic streptococci were isolated from the pharynges of seven healthy subjects, and grown on sheep blood agar plates with a hemolysis or gamma hemolysis, then cultured in LB broth for 16 hrs. Purified streptolysin O (SLO) purchased from Sigma Chemical Co. (Sigma-SLO), SLO antigen as a latex agglutination reagent from A company (A-SLO) and supernatants from four culture media were electrophoresed on 12% SDS-polyacrylamide gel and transferred to PVDF membranes. Immunological analyses of antibodies against SLO in healthy sera and proteins in culture medium demonstrated that healthy sera contained an antibody recognizing Sigma-SLO, A-SLO and a protein of the same size as SLO (SLO-like protein) in culture medium. These findings suggest that healthy subjects develop an antibody directed against SLO-like protein produced by non-pathogenic streptococci, and that this antibody cross-reacts with Sigma-SLO and A-SLO. Using DNA from Streptococcus pyogenes and non-pathogenic streptococci, the SLO gene and SLO-like protein gene were analyzed by direct sequencing with oligonucleotide primers designed to cover no. 74 to approximately 1900 of the SLO gene. There were three different bases resulting in amino acid substitution between the SLO gene and SLO-like protein gene, namely 101Lys (AAA) of SLO to Asn (AAT), 175Met (ATG) to Arg (AGG) and 185Asp (GAT) to Asn (AAT). Remaining 560 residues of 563 amino acids constituting SLO-like protein were homologous to SLO. Non-pathogenic streptococci on the pharynges of healthy subjects produce an SLO-like protein composed of amino acids similar to those of SLO, which induces an antibody against this SLO-like protein in serum. It is likely that an antibody against SLO-like protein in healthy sera cross-reacts with SLO and causes a pseudo-positive reaction on ASO measurement by the latex agglutination method using SLO antigen.

[Immunological analysis of pseudo-positive reaction in healthy adults at anti-streptolysin O measurement by latex agglutination method].

Streptolysin O (SLO) is a toxic immunogenic protein produced by Streptococcus pyogenes (S. pyogenes). The latex agglutination photometric immunoassay with latex coated by SLO (latex agglutination method) has been most widely used for determination of antibody to SLO (ASO). We measured ASO levels by latex agglutination method in serum specimens collected from 159 healthy individuals and eight patients with S. pyogenes, who had a positive S. pyogenes culture from throat swabs. A significant frequency(about 15%) had positive ASO levels (> 200 unit/ml) in healthy individuals aged < 20 (47 individuals) and 20-29 (80 individuals), respectively, however, none of 30 or over (32 individuals) were positive. The SLO specimen purchased from Sigma Chemical Co. showed at least three protein bands on SDS-PAGE; one was considered SLO protein because it had a molecular weight of 64 kD and of the remaining proteins, one had a molecular weight higher and the other had a molecular weight lower than 64 kD. Serum antibodies among the healthy adults to Sigma-SLO specimen consisted of IgG class in the majority, with little IgM class. Immunoblotting analysis revealed that serum antibodies of the patients recognized the 64-kD protein (SLO), and serum antibodies of the healthy adults recognized different proteins from SLO. It appeared that many of the healthy adults with a positive ASO level had antibodies different from ASO, resulting in pseudo-positive ASO values when ASO was measured by latex agglutination method using SLO specimen containing non-specific proteins. Improvement of commercial kits using this method is required.