A case of ovarian small cell carcinoma of the pulmonary type that was observed as it developed.

INTRODUCTION: In the case reported here, the authors observed ovarian small cell carcinoma of the pulmonary type as it developed. CASE: The patient was a 48-year-old woman who underwent a hysterectomy for CIN3 in 2007. A year later, the woman underwent screening for ovarian cancer. A gradually growing ovarian mass was noted. This mass was found to be a mixed tumor. This mixed tumor grew to 36 mm in size, and six months later it had enlarged to 119 mm. After surgery, the tumor was pathologically diagnosed as an ovarian small cell carcinoma of the pulmonary type with a neuroendocrine nature that was positive for CD56 and synaptophysin. Postoperatively, the patient received six courses of combined therapy with irinotecan and cisplatin (CPT-P therapy), and the patient has survived disease- free for over two years. CONCLUSION: Findings suggested that ovarian small cell carcinoma of the pulmonary type is a type I ovarian malignancy that develops through an adenoma-carcinoma sequence.

Aureusidin synthase: a polyphenol oxidase homolog responsible for flower coloration.

Aurones are plant flavonoids that provide yellow color to the flowers of some popular ornamental plants, such as snapdragon and cosmos. In this study, we have identified an enzyme responsible for the synthesis of aurone from chalcones in the yellow snapdragon flower. The enzyme (aureusidin synthase) is a 39-kilodalton, copper-containing glycoprotein catalyzing the hydroxylation and/or oxidative cyclization of the precursor chalcones, 2',4',6',4-tetrahydroxychalcone and 2',4',6',3,4-pentahydroxychalcone. The complementary DNA encoding aureusidin synthase is expressed in the petals of aurone-containing varieties. DNA sequence analysis revealed that aureusidin synthase belongs to the plant polyphenol oxidase family, providing an unequivocal example of the function of the polyphenol oxidase homolog in plants, i.e., flower coloration.

Locally administered low-dose alendronate increases bone mineral density during distraction osteogenesis in a rabbit model.

We investigated the effect of locally administered bisphosphonate on distraction osteogenesis in a rabbit model and evaluated its systemic effect. An osteotomy on the right tibia followed by distraction for four weeks was performed on 47 immature rabbits. They were divided into seven equal groups, with each group receiving a different treatment regime. Saline and three types of dosage of alendronate (low, 0.75 microg/kg; mid, 7.5 microg/kg and high 75 microg/kg) were given by systemic injection in four groups, and saline and two dosages (low and mild) were delivered by local injection to the distraction gap in the remaining three groups. The injections were performed five times weekly during the period of distraction. After nine weeks the animals were killed and image analysis and mechanical testing were performed on the distracted right tibiae and the left tibiae which served as a control group. The local low-dose alendronate group showed a mean increase in bone mineral density of 124.3 mg/cm(3) over the local saline group (analysis of variance, p < 0.05) without any adverse effect on the left control tibiae. The findings indicate that the administration of local low-dose alendronate could be an effective pharmacological means of improving bone formation in distraction osteogenesis.

Pathway of taurolipid B formation from exogenous taurolipid A by Tetrahymena thermophila.

When Tetrahymena thermophila was incubated with taurolipid A isolated from T. pyriformis NT-1, the exogenously added taurolipid A was deacylated rapidly, and taurolipid B content in the cells was increased. The deacylated taurolipid A (lysotaurolipid A) content reached a maximum early on during incubation, and then declined. Taurolipid B and lysotaurolipid B contents in the cells were increased continuously during the incubation. These observations suggest that lysotaurolipid A was an intermediate for taurolipid B formation. When cells were incubated with lysotaurolipid A, newly formed lysotaurolipid B and taurolipid B were observed. Furthermore, when cells were incubated with lysotaurolipid B, only taurolipid B was newly formed. In contrast, newly formed lysotaurolipid B was observed when cells were incubated with exogenous taurolipid B. From the results, we have postulated the biosynthetic pathway of taurolipid B from exogenous taurolipid A in cells of Thermophila.

Isolation and structural characterization of a diacyltaurolipid in cells of Tetrahymena mimbres.

A diacyltaurolipid was found in cells of Tetrahymena mimbres (formerly Tetrahymenas pyriformis NT-1). The lipid accounted for about 0.05% of the total lipids of the cells, and was composed of taurine (1 equiv.), 3,7,13-trihydroxystearic acid (1 equiv.) and non-hydroxy fatty acids (2 equiv.). By mild alkaline hydrolysis, non-hydroxy fatty acids (2 equiv.) and 2-(3,7,13-trihydroxyoctadecanoyl)aminoethanesulfonic acid were obtained. For determination of the positions of the ester bonds of the lipid, the free hydroxy group of the intact lipid was converted to a ketone, and the esterified hydroxy groups were acetylated after methanolic hydrochloric acid hydrolysis. By mass and nuclear magnetic resonance spectrometry, the structure of the derivative was identified as methyl 3,7-diacetoxy-13-oxooctadecanoate. From these results, the diacyltaurolipid isolated from T. mimbres was identified as 2-(3,7-diacyloxy-13-hydroxyoctadecanoyl)aminoethanesulfonic acid (7-acyltaurolipid A). This structure suggests that the diacyltaurolipid may be formed by acylation of taurolipid A.

Identification of pentahydroxystearic acid-containing taurolipid (taurolipid C) isolated from Tetrahymena thermophila.

A pentahydroxystearic acid-containing taurolipid (taurolipid C) was found in cells of Tetrahymena thermophila. The lipid accounted for about 1.2% of the total taurolipids of the cells. The lipid was subjected to mild alkaline and methanolic hydrochloric acid hydrolyses, and the structures of the hydrolyses products were identified by mass and nuclear magnetic resonance spectrometries, as taurine, non-hydroxy fatty acid and 2,3,7,12,13-pentahydroxystearic acd, a novel fatty acid. The NMR spectra of the intact and acetylated lipid showed that the carboxyl group of the pentahydrxyostearic acid was combined with the amino group of taurine, and the hydroxy group at C-3 was esterified with non-hydroxy fatty acids. From these results, the pentahydroxystearic acid-containing taurolipid (taurolipid C) isolated from T. thermophila was identified as 2-(3-acyloxy-2,7,12,13-tetrahydroxyoctadecanoylamino)ethanesulf oni c acid.

Topological analysis of an RND family transporter, MexD of Pseudomonas aeruginosa.

The membrane topology of a resistance-nodulation-division (RND) family transporter, MexD of Pseudomonas aeruginosa, was determined. Although it had been predicted previously that most RND proteins contain 12 transmembrane helices, three independent computer programs used in the present study predicted that MexD possessed 11, 14 or 17 transmembrane segments. To investigate the topology of MexD more thoroughly, 25 MexD-PhoA (alkaline phosphatase) and 18 MexD-Bla (beta-lactamase) fusion plasmids were constructed and analyzed. The resulting topological model had just 12 transmembrane helices and two periplasmic loops of about 300 residues between helices 1 and 2 and helices 7 and 8. It is therefore proposed that the N- and C-termini are located in the cytoplasm and the predicted orientation is consistent with the 'positive-inside rule'. This topological model can be applied to other RND proteins.

Specificity analysis and mechanism of aurone synthesis catalyzed by aureusidin synthase, a polyphenol oxidase homolog responsible for flower coloration.

Aureusidin synthase, which plays a key role in the yellow coloration of snapdragon flowers, is a homolog of plant polyphenol oxidase (PPO). The enzyme specifically acted on chalcones with a 4-monohydroxy or 3,4-dihydroxy B-ring to produce aurones, for whose production the oxidative cyclization of chalcones must be preceded by 3-oxygenation. However, it exhibited virtually no PPO activity toward non-chalcone phenolics. The enzyme was competitively inhibited by phenylthiourea, a specific PPO inhibitor. These results led us to propose a mechanism of aurone synthesis by aureusidin synthase on the basis of known PPO-catalyzed reactions and conclude that the enzyme is a chalcone-specific PPO specialized for aurone biosynthesis.

Molecular characterization of tobacco sulfite reductase: enzyme purification, gene cloning, and gene expression analysis.

A cDNA clone, NtSiR1, that encodes the precursor of ferredoxin-dependent sulfite reductase (Fd-SiR) has been isolated from a cDNA library of tobacco (Nicotiana tabacum cv. SR1). The identity of the cDNA was established by comparison of the purified protein and the predicted structure with the nucleotide sequence. The amino terminus of the purified enzyme was Thr62 of the precursor protein, and the mature region of NtSiR1 consisted of 632 amino acids. Tobacco Fd-SiR is 82, 77, and 48% identical with Fd-SiRs from Zea mays, Arabidopsis thaliana, and a cyanobacterium, respectively. Significant similarity was also found with Escherichia coli NADPH-SiR in the region involved in ligation of siroheme and the [4Fe-4S] cluster. On Northern blot analysis, a transcript of NtSiR1 was detected in leaves, stems, roots, and petals in similar amounts. We also isolated a genomic SiR clone named gNtSiR1. It consists of 8 exons and 7 introns. Genomic Southern blot analysis indicated that at least two SiR genes are present in the tobacco genome.

Ptilomycalin A, a novel Na+, K(+)- or Ca2(+)-ATPase inhibitor, competitively interacts with ATP at its binding site.

Ptilomycalin A inhibited the brain Na+, K(+)-ATPase and Ca2(+)-ATPase from skeletal sarcoplasmic reticulum with an IC50 value of 2 microM and 10 microM, respectively. Kinetic analysis of the inhibitory effects of ptilomycalin A suggests that the inhibition of Na+, K(+)-ATPase is a competitive-, an uncompetitive- and an anticompetitive-type with respect to ATP, Na+ and K+, respectively. The inhibition of Ca2(+)-ATPase by ptilomycalin A is a competitive- or an uncompetitive-type with respect to ATP or Ca2+, respectively. These results suggest that ptilomycalin A interacts with ATP at the ATP binding site of Na+, K(+)-ATPase or Ca2(+)-ATPase. Ptilomycalin A has become a useful biochemical tool for clarifying the ATP binding site in both enzymes.

Purification, characterization, and application of an acid urease from Arthrobacter mobilis.

It has been shown that urea in fermented beverages and foods can serve as a precursor of ethylcarbamate, a potential carcinogen, and acid urease is an effective agent for removing urea in such products. We describe herein the purification and characterization of a novel acid urease from Arthrobacter mobilis SAM 0752 and show its unique application for the removal of urea from fermented beverages using the Japanese rice wine, sake, as an example. The purified acid urease showed an optimum pH for activity at pH 4.2. The enzyme exhibited an apparent K(m) for urea of 3.0 mM and a Vmax of 2370 mumol of urea per mg and min at 37 degrees C and pH 4.2. Gel permeation chromatographic and sodium dodecyl sulfate gel electrophoretic analyses showed that the enzyme has an apparent native molecular weight (M(r)) of 290,000 and consisted of three types of subunit proteins (M(r), 67,000, 16,600, 14,100) denoted by alpha, beta, and gamma. The most probable stoichiometry of the subunits was estimated to be alpha: beta: gamma = 1:1:1, suggesting the enzyme subunit structure of (alpha beta gamma)3. The enzyme also existed as an aggregated form with an M(r) of 580,000. The purified enzyme contained 2 g-atom of nickel per alpha beta gamma unit of the enzyme. Enzyme activity was inhibited by acetohydroxamic acid, HgCl2, and CuCl2. The isoelectric point of the native enzyme was estimated by gel electrofocusing to be 6.8. Urea (50 ppm), which was exogenously added to sake (pH 4.4, 17 +/- 1% (v/v) ethanol), was completely decomposed by incubation with the enzyme (0.09 U ml-1) at 15 degrees C for 13 days. The enzyme was unstable at temperatures higher than 65 degrees C and pHs lower than 4, and was completely inactivated under the conditions of a pasteurization step involved in the traditional sake-making processes. These results indicate that the enzyme is applicable to the elimination of urea in fermented beverages with minimal modification to the conventional process.

Phenylglycine methyl ester, a useful tool for absolute configuration determination of various chiral carboxylic acids

A new chiral anisotropic reagent, phenylglycine methyl ester (PGME), developed for the elucidation of the absolute configuration of chiral alpha,alpha-disubstituted acetic acids, has turned out to be applicable to other substituted carboxylic acids, such as chiral alpha-hydroxy-, alpha-alkoxy-, and alpha-acyloxy-alpha, alpha-disubstituted acetic acids, as well as to chiral beta, beta-disubstituted propionic acids. Because a carboxylic moiety is convertible from other functional groups, e.g., ozonolysis of an olefin and oxidative cleavage of a glycol, the present findings can expand the utility of the PGME method to the absolute configuration determination of various types of organic compounds, even those which initially lack oxygen functions. Several examples of the combination of chemical reactions and the PGME method are described.

High IL-6 synthesis in cultured fibroblasts isolated from radicular cysts.

OBJECTIVE: Inflammatory cytokines have been reported to be related with inflammation and expansion of jaw cysts. In this study, to examine the relationship between radicular cysts and inflammatory cytokines, it was found that there was notable unique evidence on cytokine synthesis from fibroblasts isolated from radicular cysts. METHODS: The expression of such cytokines, namely, interleukin-1beta, IL-1beta, IL-6, IL-8, IL-10, tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), transforming growth factor-beta1 (TGF-beta1), and granulocyte-macrophage colony-stimulating (GM-CSF) mRNA, in nine radicular cysts was examined and compared with that detected in six specimens of healthy gingival mucosa. Furthermore, separating all fibroblasts from their respective radicular cysts, healthy gingival mucosa, and healthy periodontal ligaments, these fibroblast groups were cultured without stimulators and a supernatant for each was obtained to analyse IL-1beta, IL-6, IL-8, TNF-alpha, and IFN-gamma by ELISA. RESULTS: Differences between radicular cysts and healthy gingival mucosa were not clearly shown by the expression of cytokine mRNA. Analysing inflammatory cytokine synthesis in fibroblast groups from these three kinds of tissues, surprisingly, the levels of IL-6 mRNA and protein were recognised to be higher in fibroblasts of radicular cysts than in those of control tissues by ELISA and a real-time RT-PCR. Significant differences in the cultured supernatants of these fibroblast groups were not recognised in the release of IL-1beta, IL-8, TNF-alpha, and IFN-gamma by ELISA. CONCLUSIONS: From these results, it was suggested that fibroblasts inducing IL-6 production might play important roles in the expansion of radicular cysts. It is considered that fibroblasts around radicular cysts may lead to high IL-6 synthesis over time in chronic inflammation.

Graft incorporation within the tibial bone tunnel after anterior cruciate ligament reconstruction with bone-patellar tendon-bone autograft.

We described histologic changes in patellar tendon autografts that occur over time within the tibial tunnel in specimens harvested from patients undergoing revision anterior cruciate ligament reconstruction. Ten patients, averaging 21.2 years of age, were divided into two groups based on the time period between their original and revision surgery: early revision (less than 1 year, four patients) and late revision (more than 1 year, six patients). Among the early revision group, the tendon within the tunnel showed increased cellularity and random collagen bundles. A specimen from the shortest early revision case revealed a normal original bone-tendon junction, whereas others showed an obscured structure. Between the tendon and the tunnel wall, granulation tissue was seen and the bone-tendon junction was still immature. In the late revision group, the tendon appeared similar to normal ligament. The original bone-tendon junction was not seen, and the tendon continued completely to the tunnel wall with Sharpey-like fibers. Observations in the early revision group suggest that tendon remodeling and bone-tendon integration continue for at least several months after transplantation. The original bone-tendon junction appears to have shifted to the proximal patellar tendon-tunnel wall junction with time. These findings are in agreement with prior animal studies.

[Clinical evaluation of Dumon stent for tracheobronchial stenosis due to advanced esophageal carcinoma].

We inserted Dumon stent to 13 patients with tracheobronchial stenosis due to advanced or recurrent esophageal carcinoma. Severe dyspnea was improved in 11 patients except for 2 patients with bilateral recurrent nerve palsy. 3 cases who had radiation therapy or chemotherapy lived over 150 days. We inserted esophageal stent in 4 cases. 2 patients died due to hemoptysis after 156 days and 35 days. We conclude that Dumon stent is one of the useful treatments in order to improve quality of life and prognosis of advanced or recurrent esophageal carcinoma.

[Comprehension of synesthetic expressions: cross-modal modifications of sense adjectives].

This study examined (a) acceptability of phrases whose modifier adjective and head noun refer to different sense modalities, and (b) such cross-modal adjective properties by means of similarity judgment of these phrases. In Experiment 1, 58 subjects rated 60 sense adjectives as to their acceptability in modifying nine modality-denoting nouns (touch, taste, smell; color, sound; memory, mood, idea, personality). The phrases were rated highly acceptable when the adjectives denoting lower-modal qualities were combined with the nouns denoting higher-modal contents (e.g., soft sound, sweet mood, bright memory). In Experiment 2, two groups of 30 subjects judged the similarity of the highly acceptable phrases for each of the nine modality-denoting nouns. Multidimensional scaling of these data revealed two- and three-dimensional configurations of the adjectives for each modality-denoting noun. Multiple regression analysis indicated that two independently rated properties, pleasantness and intensity, provided a satisfactory interpretation of each configuration. Canonical correlation analysis indicated that most of the first- and second-order canonical correlations among the nine three-dimensional configurations were significant. These results suggest that there is a common dimension of sense adjectives across different modalities.

[Effects of personality, cognitive, and situational variables on risk taking behavior].

This study examined the effects of individual difference and situation on risk taking behavior. In Experiment 1, 115 undergraduates completed a questionnaire of personality (sensation seeking, optimism, etc.) and their risk taking behavior, risk perception, and anxiety in eight situations: personal, social, gain-loss, and loss situations. Results indicated an effect of personality on risk taking behavior in personal gain-loss situations (sports and life event), which was mediated by perceived risk controllability. In Experiment 2, 137 undergraduates completed a questionnaire of personality, cognitive variables (risk perception, own competence, and perceived cost and benefit), and risk taking behavior in personal gain-loss situations (sports, life event, and gambling). Results of covariance structure analysis showed that perceived risk controllability affected the relationship between the variables. For instance, risk significance and perceived cost and benefit mediated the effect of 'controllability with skills' on risk taking behavior in the controllable situation (e.g., sports). Similarly, competence and risk perception mediated the effect of 'uncontrollable luck factors' in the chance situation (e.g., life event).

Time-dependent gene expression and immunohistochemical analysis of the injured anterior cruciate ligament.

OBJECTIVES: This study aimed to investigate time-dependent gene expression of injured human anterior cruciate ligament (ACL), and to evaluate the histological changes of the ACL remnant in terms of cellular characterisation. METHODS: Injured human ACL tissues were harvested from 105 patients undergoing primary ACL reconstruction and divided into four phases based on the period from injury to surgery. Phase I was < three weeks, phase II was three to eight weeks, phase III was eight to 20 weeks, and phase IV was ≥ 21 weeks. Gene expressions of these tissues were analysed in each phase by quantitative real-time polymerase chain reaction using selected markers (collagen types 1 and 3, biglycan, decorin, α-smooth muscle actin, IL-6, TGF-ß1, MMP-1, MMP-2 and TIMP-1). Immunohistochemical staining was also performed using primary antibodies against CD68, CD55, Stat3 and phosphorylated-Stat3 (P-Stat3). RESULTS: Expression of IL-6 was mainly seen in phases I, II and III, collagen type 1 in phase II, MMP-1, 2 in phase III, and decorin, TGF-ß1 and α-smooth muscle actin in phase IV. Histologically, degradation and scar formation were seen in the ACL remnant after phase III. The numbers of CD55 and P-Stat3 positive cells were elevated from phase II to phase III. CONCLUSIONS: Elevated cell numbers including P-Stat3 positive cells were not related to collagens but to MMPs' expressions.