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

), blaCTX-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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A full account is given of the chemical behavior observed for 7, -dialkyladeninium salts (16). On treatment with boiling 1N aqueous NaOH for 60min, 16a, , d, (X=I), 16c (X=Br), and 16 (X=ClO₄) rearranged to isomeric N⁶, 7-dialkyladenines (21a-) in 50-91% yields. Treatment of the salts with 0.N aqueous at room temperature for 30-min or with Amberlite CG-400 (OH^-) in H₂O at room temperature gave the ring-opened derivatives a- (in the trans-formamide form) in 56-83% yields, and rate constants for the ring-opening reactions of 16a, , d-g (X=ClO₄) and 16c (X=Br) leading to a-g were determined in H₂O at pH .84 and ionic strength 0.50 at 25°C. Cyclization of a with NaH in AcNMe₂ at room temperature or with boiling 1N aqueous NaOH produced 21a in 84% or 72% yield, respectively.In solution, the trans-formamides seemed to transform slowly into the cis-formamides 23, attaining equilibria. The existence of such an equilibrium in D₂O or Me₂SO-d₆ at 25°C or in H₂O at pH .84 and ionic strength 0.50 at 25°C was kinetically confirmed in the case of a, and the mechanism of the rearrangement of 16 to 21 through is discussed on the basis of the above kinetic results and Deslongchamps' theory of stereoelectronic control. On treatment with NaBH₄ in MeOH at room temperature, 16a (X=I) furnished the 7, 8-dihydro derivative 28 (84% yield), which slowly decomposed in H₂O at 60°C to give a in 49% yield.The 7, -dialkyladeninium salts (16) were found to be obtainable from N'-alkoxy-1-alkyl--formamidoimidazole-4-carboxamidines () through an alternative synthetic route : Alkylations of with alkyl halides in HCONMe₂ in the absence of base, followed by hydrogenolysis of the N'-alkoxy group and cyclizatio (or vice versa) produced 16 in acceptable yields. In order to interpret the proton nuclear magnetic resonance spectrum of a, the 2-deuterated species 26 was also synthesized from 24 via 25 and 27.

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Map-based cloning techniques are widely used to identify the mutated gene(s) in plant, because chemical agents, such as EMS, are generally used to induce mutations. However, map-based cloning was used to be time consuming to identify the mutated gene(s).

Whole genome sequence information has already been opened in Arabidopsis, we established a rapid and high throughput mapping technique, ARAMAP system, based on the information of single nucleotide polymorphism (SNP). To demonstrate the accuracy of this system, we performed SNP-based cloning for a floral internode mutant that we isolated from T-DNA tagging line. Although this mutant is well linked with T-DNA insertion, by using this system we could map to the same locus within two days in a region between 8.559Mb and 13.13Mb of a chromosome. We finished setting of functional markers on the five chromosomes of Arabidopsis in a interval of 0.26Mb to be able to fine mapping of mutant.

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We screened the differentiation-inducing activities of 39 mushroom extracts from Akita prefecture, Japan, on the mouse osteoblastic cell line, MCT-1. Sixteen phosphate buffered saline (PBS), 8 boiled PBS, 14 ethanol and 12 methanol extracts induced alkaline phosphatase (ALP) activities, an indicator of MCT-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-,,6β-triol (1). 1 induced ALP activities of MCT-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 MCT-1 cells by 1, ergosterol (2), ergocalciferol (), cholesta-,,6β-triol (4), 7-dehydrocholesterol () and cholecalciferol (6) were tested. The enzyme activities of MCT-1 cells were increased .0-fold by 10 μM 1 and 2.4-fold by 10 μM 4. However, 2, , and 6 did not induce MCT-1 cell ALP activity at 0.1—10 μM. These results suggested that the OH groups at C- and/or C-6 of 1 and 4 played an important role in their differentiation-inducing activities on MCT-1 cells. Furthermore, 1 suppressed induction of MCT-1 cell apoptosis by serum starvation.

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Reactions of nor-diterpenoid dilactones isolated from Podocarpus plants have been investigated. The 7: 8,: 11-dienolide type [C] dilactones, the least available but biologically the most active members, are mainly derived from naturally abundant dilactone components of α-pyrone type [A] and 7: 8-epoxy-: 11-enolide type [] for total-synthetic and biological purposes. From nagilactone (), for example, a combined sequence of hydrogenolysis of 7: 8-epoxide group and dehydration gives nagilactone (7), β-hydroxynagilactone (8) and 2: -dehydronagilactone (19), which have been found from natural sources. From nagilactone A (1), photo-hydration and concomitant double bond migration of its acetate results in a lactol (), which is reduced to 1β-hydroxynagilactone (), another natural type [C] dilactone. Derivation of the type [A] or [] from others is not successful. Some attempts for conversion of the ring A functional groups have also been studied.

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Six new cembranoids sarcophytol P (), R (4), S (), K (8), (11), and T (13) were isolated from the soft coral Sarcophyton glaucum. Sarcophytol P () was shown to be the 20-hydroxy derivative of the major component sarcophytol A (1), and afforded the cyclization product 6 in CHCl_ at room temperature, in a same way as in 1. Sarcophytols R (4) and S () were correlated to 1, by conversion of its 7R,8R and 7S,8S epoxide derivatives. Sarcophytol K(8) was a 13, 14-dihydroxycembranoid having a 1,Z-diene moiety. The absolute configuration of 8 and its 1Z,- and 1Z,Z-isomers sarcophytol (2) and J (10) were determined by circular dichroism study of their bis-p-dimethylaminobenzoate derivatives. Sarcophytols (11) and T (13) were 1- and 1,Z-isomers of 1. Compound 11 showed characteristic broadening of the ^1H-NMR chemical shifts, due to the restricted conformational interconversion. Using the three cembranoids sarcophytols (11, 1,), N (15, 1Z,Z) and T (13,1,Z), spontaneous autoxidation-cyclization, in CHCl_,was examined, in order to compare the stereochemical course of the reaction with that of 1 (1Z,), which affords trans-fused bicyclo[..0]tetradecene systems. The 1,Z-isomer 13 gave the same cyclization product 18, as in the reaction of 1, even though it is isomeric at C-1,. The 1Z,Z isomer 15 gave 19, in consequence of the reversed geometry at C- of 15. The 1,-isomer (11) gave the bicyclic product , having an antipodal fusion as compared with 19. This was confirmed by PCC oxidation of 19 and , giving enantiomeric ketones 23 and 24 respectively. The 1(14)-epoxide 26 was shown to be the immediate precursor of , and acounted for the inversion of the geometry at C-1 of the cyclization product. Compound 26 is isomeric with the epoxide 17 derived from 1. The epoxide 17 is the postulated precursor in the conversion of 13 to 18. It is noteworthy that the mode of antipodal fusion of the cyclopentane rings, in 18 and , was controlled by the chirality of the epoxy rings. The C-14 hydroxyl group participates in the transannular cyclization, but was found not to be the requisite functional group for the reaction. Similar treatment of cembrene C (30), the parent hydrocarbon of 1, also reacted in CHCl_ giving the bicyclic product 32.

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Four new sterols, , 6α-epoxy-(, 24R)-ergosta-8(14), -diene-, 7β-diol (1), (, 24R)-ergosta-8, -diene-, , 6β, 7α-tetrol (2), (, 24R)-ergosta-7, (11), -triene-, , 6β-triol () and , , 6β-trihydroxy-(, 24R)-ergost--en-7-one (4), have been isolated from the fruit bodies of Grifola frondosa (FR.) S. . GRAY (Polyporaceae)together with fourteen known ones (-18), of which two ( and 6) are reported for the first time from a natural source. The structures of these compounds were elucidated on the basis of spectral data.

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A new glucuronide, (R)-, 16β, , 26-tetrahydroxycholest--ene -O-α-L-rhamnopyranosyl-(1→2)-β-D-gluc-uronopyranoside (1), was isolated from the aerial parts of Chinese Solanum lyratum THUNB. The structure of this new steroidal glucuronide was deduced by spectroscopic means and the identity of its aglycone was substantiated by comparison with an authentic sample derived from diosgenin acetate.

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Three new azaphilone derivatives designated falconensins (), (6), and G (7) were isolated from the mycelium o Emericella falconensis, along with falconensins A (1), (2), C (), D (4), and H (8), and three hopane-type triterpenes, zeorin (), hepane-7β, -diol (10), and hopane-6α, 7β, -triol (11). The structures of -7 were confirmed by spectroscopic investigation and chemical correlations. The absolute stereochemistry of falconensins A (1) to G(7) was also established.

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We reported a case of basal cell nevus syndrome (BCNS) . A 12-year old boy visited us for the examination of numerous skin pits on the palms. He showed hypertelorism, mandibular cyst, calcification of the falx, and mental retardation. Clinical diagnosis of BCNS was made by these findings. Examination of other family members did not reveal palmar pits or any other signs of BCNS. Skin biopsy from the palmar pits showed reduced thickness of the stratum corneum and the presence of basal cell carcinoma in the dermis.
Chromosome-analysis of peripheral leukocytes revealed the deletion of q21.-.1. His parents did not show any abnormalities in chromosome-analysis. Very few cases of BCNS have been reported that have basal cell carcinoma on the palms. [Skin Cancer (Japan) 2000; 15: 27-30]

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Three monoclonal antibodies (MoAb), 1 , 1 G and 4, were produced by fusion of spleen cells of a BALB/c mouse immunized with Ca - cell from human squamous cell carcinoma of gingiva and murine myeloma cells SP 2/o.They were screened by 51Cr release assay of complement dependent cytotoxicity.Their isotypes were all IgM which recognized the cell surface antigens.Five MoAbs, 1 11, N 2, N 4, N 18 and 4 , were prepared against NA cell from human tongue squamous cell carcinoma by the same procedure, but they were screened by radioimmunoassay using 125I-protein A.All MoAbs had isotypes IgG and only four recognized their antigen in the cytoplasm.N 4 recognized the antigen existing in both cytoplasm and cell membrane.
The reactivity of each MoAb to various cells and tissues was analyzed by immunostaining method.They were separable into three groups by the cell specificity, i..the first group including 1 , 1 G , and 4 reacted to all squamous cell carcinoma cells tested but not to the normal diploid fibroblast WI 38.The second group comprising of 1 11, N 2, N 4 and N 18 was reactable to both squamous cell carcinoma cells and WI 38.The third group of 4 reacted to only Ca - used as the immunogen.From the study using paraffin embedded tissues including oral cancers and normal tissues, it was revealed that 1 and 1 G stained the cell membrane of the cancer nest in squamous cell carcinoma and the prickle cell layer in the oral mucosa.The second group of MoAb, 1 11, N 2, N 4 and N 18, stained both parenchymal and stroma cells simultaneously. 4, interestingly, showed different reactivity between individuals, namely in a positive case it stained not only the normal and malignant cell of squamous epithelial origin but also endothelial cells and erythrocytes in the stroma.However, in a negative case it did not stain any type of cell, suggesting that 4 may recognize an antigen related to the blood group.

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