We screened the differentiation-inducing activities of 39 mushroom extracts from Akita prefecture, Japan, on the mouse osteoblastic cell line, MC3T3-1. Sixteen phosphate buffered saline (PBS), 8 boiled PBS, 14 ethanol and 12 methanol extracts induced alkaline phosphatase (ALP) activities, an indicator of MC3T3-1 cell differentiation. The enzyme activities were markedly induced by extracts of Tricholoma auratum, and we isolated the active compound from methanol extracts of this mushroom. Physical data for the isolated active compound were identical to those for (,24R)-ergosta-7,-diene-3β,,-triol (1). 1 induced ALP activities of MC3T3-1 cells and promoted cell proliferation. To investigate the relationships between the chemical structure and differentiation-inducing activity of the compound, ALP-inducing activities of MC3T3-1 cells by 1, ergosterol (2), ergocalciferol (3), cholesta-3β,,-triol (4), 7-dehydrocholesterol () and cholecalciferol () were tested. The enzyme activities of MC3T3-1 cells were increased 3.0-fold by 10 μM 1 and 2.4-fold by 10 μM 4. However, 2, 3, and did not induce MC3T3-1 cell ALP activity at 0.1—10 μM. These results suggested that the OH groups at C- and/or C- of 1 and 4 played an important role in their differentiation-inducing activities on MC3T3-1 cells. Furthermore, 1 suppressed induction of MC3T3-1 cell apoptosis by serum starvation.

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The ability of Climacodon septentrionalis to immobilize and kill a mycophagous nematode (Aphelenchoides sp.) in vitro is described for the first time. Two isolates produced droplets (20–45 μm in diameter) that formed at the apices of tall, stalked, and branching secretory cells (700–1,500 μm tall). On 2% modified malt extract agar, nematodes became enveloped in the droplets, which restricted their ability to move and resulted in complete immobilization and death within several hours of contact. The rate of decomposition of the nematodes varied considerably, with most individuals persisting for weeks whereas others were degraded within several days and appeared to be colonized by dense hyphal growth. This study provides the first documentation of a non-agaricoid fungus producing secretory cells that are able to immobilize nematodes.

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The effects of the pressure of ambient gas and sample temperature on photo-acoustic displacement (PAD) were studied quantitatively using an extremely sensitive laser interferometric probe. For silicon, the PAD signal measured at atmospheric pressure increased by about 18% as compared to that obtained in a vacuum, and varied by less than 0.7% for a change in pressure of % around 1 atm. This effect was attributed to the decrease in refractive index of the gas adjacent to the sample. Temperature rise caused PAD to increase proportionally, and for GaAs, PAD varied 0.4% per degree. The variation of PAD with temperature was accurately explained by the temperature dependence of thermal expansion coefficient and thermal conductivity of the sample.

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Some existing analytical methods which have been used in geotechnical engineering are modified to handle the shear strength reduction with time due to softening in fissured, over-consolidated clays and mudstones. These are limit equilibrium analysis of slope stability, calculation of earth pressure and calculation of stress and displacement around a tunnel. The softening effect is incorporated in these analyses using the failure criterion and the time-dependent strength parameters proposed previously by the authors. Examples are presented which emphasize the importance of appreciating the softening effect in analyses and further illustrate that the effect of shear strength reduction with time due to softening could easily be captured with existing analytical methods.

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The gamma-ray spectrum and the conversion-electron spectrum are measured in the decay of the ^116mIn activity. Directional correlations are also measured for 818–1293 and 1097–1293 keV cascades. The M1-2 mixing ratio δ, and the 0-2 mixing ratio μ_k are obtained for 818.7 keV +→2₁⁺ transition to be δ=1.52−

0.22

+0.26, and μ_k≤

6

.1×10⁻⁴ respectively. The

E

2 branching ratio

B

(

E

2;

2₂

+→0_g⁺):

B

(

E

2;

2₂

+→2₁⁺):

B

(

E

2;

2₂

+→0₁⁺) is determined to be 0.0158:1.0:

5

.86.

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Brassinolide analogues, (R, 23R, 24R)-2α, 3α, , 23-tetrahydroxy--homo-7-oxa--ergostan--one (24-epibrassinolide) (10) and (S, 23S, 24R)-2α, 3α, , 23-tetrahydroxy--homo-7-oxa--ergostan--one (), were synthesized from brassicasterol (3a) in five steps and with ca. 20% overall yield. The key steps are the direct formation of (, 24R)-3α, -cyclo--ergost--en--one (4) from brassicasterol mesylate (3), the acid-catalyzed rearrangement of 4 to (, 24R)--ergosta-2, -dien--one (), and the Baeyer-Villiger oxidation of the tetrahydroxy -ergostan--ones 7 and 8.

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The gamma-ray spectrum and the conversion-electron spectrum are measured in the decay of the Tb¹⁵² activity. The L-subshell electron lines of the 586.7 keV +→2₁⁺ transition are separated by the ion-free beta-ray spectrometer. The M1-2 mixing ratio, δ², and the dimensionless ratio X of 0-2 mixing of the 586.7 keV transition are obtained to be δ²≤0.16 and 0.034≤X≤0.038, respectively. The 2 branching ratio (2; +→0_g⁺): (2; +→2₁⁺): (2; +→0₁⁺) is determined to be 0.023 : 1.0 : ..

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The phase diagrams of the Ni-Mo- and Ni-W- ternary systems in the region of less than 50mol% were constructed by thermodynamic calculation, based on the data obtained by thermodynamic measurement of the related materials. We found three ternary eutectic points and three or two ternary peritecto-eutectic points as follows:
:L (1365K, 71.mol%Ni-.0mol%Mo-.mol%)=(Ni)++
:L (1355K, 62.mol%Ni-2.mol%Mo-30.mol%)=++
:L (1445K, 42.0mol%Ni-30.mol%Mo-10.3mol%)=(Ni)+NiMo+
P₁:L (1812K, 34.mol%Ni-42.3mol%Mo-.8mol%)+MoB=+
P₂:L (1633K, 42.3mol%Ni-40.4mol%Mo-17.3mol%)+Mo=+
P₃:L (1812K, 53.mol%Ni-33.7mol%Mo-12.8mol%)+Mo=NiMo+
:L (1622K, 51.0mol%Ni-31.mol%W-17.4mol%)=(Ni)+W+
:L (1260K, 71.0mol%Ni-7.0mol%W-.0mol%)=(Ni)++
:L (1291K, 65.4mol%Ni-4.8mol%W-29.8mol%)=++
P₁:L (2115K, 23.8mol%Ni-43.1mol%W-33.1mol%)+WB=+
P₂:L (1657K, 48.mol%Ni-33.1mol%W-18.0mol%)+=W+
The calculated phase diagrams are expected to be useful for the development of new Ni-based heat-, corrosion- or wear-resistance alloys.

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The solubility and supersolubility of L-phenylalanine in aqueous solutions is firstly reported in this work. At temperatures from 273.15 to 343.15 K, both solubility and supersolubility in water generally increase with the temperature, and the average metastable zone width is about 8.3°C. At the temperature of 298.15 K, the solubility remarkably decreases with an increase in the concentration of ethanol, which indicates that ethanol can be used as an antisolvent for the precipitation and crystallization of L-phenylalanine. When sodium chloride is added to the solution, the solubility of L-phenylalanine increases with the concentration of sodium chloride.

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Monoclonal antibodies (MAbs) specific to the -kDa coat protein (CP) of Broad bean wilt virus 2 (BBWV-2) were produced. The -kDa CP of the IA isolate expressed in

E

. coli was used to immunize mice and three independent hybridoma cell lines were screened. In western blot analyses, the MAbs (MAb3-, MAb-12A and MAb-2A) reacted to the bacterially expressed -kDa CP of the IA isolate and most BBWV-2 isolates, but not to some other BBWV-2 isolates and other plant viruses. In immuno-electron microscopy, MAb3- and MAb-2A reacted with BBWV-2 particles to the 10-

5

dilution.

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We purified a sterol with antitumor activity from the edible mushroom Sarcodon aspratus (BERK.) S. ITO and identified it as ,8α-epidioxy--ergosta-,(11),-trien-3β-ol (,11-dehydroergosterol peroxide ((11)-DHEP)). Purified (11)-DHEP was a more effective inhibitor of HL60 leukemia cell growth and stronger apoptosis-inducer than ,8α-epidioxy--ergosta-,-dien-3β-ol (ergosterol peroxide (EP)) that we had previously identified as an apoptosis inducer. Moreover, (11)-DHEP selectively suppressed the growth of HT29 human colon adenocarcinoma cells but not WI38 normal human fibroblasts. After d incubation of HT29 with 7 μM (11)-DHEP, the number of S phase cells decreased from 23 to 15% of total diploid cells and 17% became hypodiploid. Expression of the cyclin-dependent kinase inhibitor 1A (p21, WAF1, Cip1) (CDKN1A), which has been shown to cause cell cycle arrest and apoptosis in HT29 cells, was induced by (11)-DHEP. These results suggest that (11)-DHEP inhibits HT29 cell growth by inducing CDKN1A expression, thus causing cell cycle arrest and apoptosis.

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This study aimed to determine the prevalence of fecal carriage of extended spectrum β-lactamase (ESBL) and/or plasmidic AmpC β-lactamase (pAmpC) producing Escherichia coli among dogs (n=428) in Turkey. Polymerase chain reaction (PCR) and sequencing were used to characterize genes encoding β-lactamase and plasmid mediated quinolone resistance (PMQR). Antimicrobial susceptibility testing and PCRs for virulence genes and phylogenetic groups were also performed. Cefotaxime resistant

. coli isolates were detected in 95 (.2%) of the swab samples. Sequencing analysis results showed occurrence of various β-lactamase genes: bla_CTX-M-15 (62), blaTEM-

1b

(42), bla_CMY-2 (

22

), bla_CTX-M-3 (16), bla_CTX-M-1 (15), bla_OXA-1 (

9

) and bla_SHV-12 (3) alone or in combination. The most frequently encountered phylogenetic group was group A₁ (35.8%), followed by group D₂ (

22

.1%),

B

1 (15.8%), D₁ (

9

.

5

%), A₀ (7.4%),

B2₂

(

5

.3%) and

B2₃

(4.2%), respectively. PMQR genes, aac(

6

’)-Ib-cr, qnrS1 and qnrB10 were detected in 25.3, 10.

5

and 1.1% of the isolates, respectively. While all isolates were susceptible to imipenem and amikacin, resistance rates to non-β-lactam antibiotics ranged from 20.0% for tobramycin to 56.8% for tetracycline. The virulence genes were only detected in 34 (36.2%) of the isolates and this isolates carried single or various combination of virulence genes of iucD, papC, papE, f17a-A and eaeA. Four isolates were identified as human virulent pandemic CTX-M-15 producing

E

. coli clone O25

b

:ST131/

B

2. To the best of our knowledge, this is the first study to show fecal carriage of ESBL/pAmpC type β-lactamase producing

E

. coli isolates among dogs in Turkey.

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Melithasterols A-D (1-4), isolated from a gorgoinan coral Melithaea ocracea of Okinawa, were shown to be cholestane-, 24-methyl--dehydrocholestane-, -dehydrocholestane-, and 24-methylenecholestane-derivatives. respectively, having a common 3β, 7α -dihydroxy-α, α-epoxy-Δ^8 steroid nucleus. This was confirmed by direct comparison with the authentic sample prepared by lead tetra-acetate oxydation of cholest--ene- 3β, α, 8α-triol 3-monoacetate. Twelve new polyhydroxysterols were isolated from an Alcyonium sp. (7,8,), and Sclerophytum sp. (14,16,19,20,21,a,23,26,28) soft corals collected off the coasts of the Andaman and Nicobar Islands in the Indian Ocean. All the Sclerophytum sp. soft corals contained 17a or its 25-deacetyl derivative. Compounds 7 to were identified as 24-methylenecholest--ene- 3β, 16β-diol-3-0-α-L fucoside (7) and its 7β-(8) and 7α-hydroxy () derivatives. Compound 14 was shown to be 24S-24-methylcholest--ene-3β,25ξ,26-triol and was correlated to the known compound 15 by α, β-glycolation. Compound 16 was shown to be β, α-isomer of 17, correlating to 17 by PCC oxidation followed by dehydration. Compound 19 was identified with 25-deacetyl derivative of the known compound lobosterol (19a) by hydrolysis of authentic lobosterol. Compounds 20 and 21 were shown to be 7-dehydro-(20) and -dehydro-(21) derivative of 17a and 17, respectively, by comparisons of their spectral data with reference compounds. Compounds a and 23 were 24S-methylcholestane-3β, α 25-triol--one 25-monoacetate (a) and its 25-deacetoxyl derivative (23). Partial PCC oxidation of 17a afforded a. Compounds 26 (andamansterol) and 28 (nicobarsterol) were shown to be gorgost--ene-3β, α. 11α, 21-tetrol and novel secosteroid, 11,21-cyclo--homo-11-oxa-, 11-secoergostane-3β, α, 12ξ-triol--one, respectively, by spectral analyses (H-HCOSY, HMQC, HMBC).

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