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Abstract: Octopamine plays an important role in mediating reward signals in olfactory learning and memory formation in insect. However, its target molecules and signaling pathways are still unknown. In this study, we investigated the effects of octopamine on the voltage-activated Ca2+ channels expressed in native Kenyon cells isolated from the mushroom body of the cricket (Gryllus bimaculatus) brain. The cell-attached patch clamp recordings with 100mM Ba2+ outside showed the presence of dihydropyridine (DHP) sensitive L-type Ca2+ channels with a single channel conductance of approximately 21±2pS (n =12). The open probability (NPo) of single Ca2+ channel currents decreased by about 29±7% (n =6) by bath application of 10μM octopamine. Octopamine-induced decrease in Po was imitated by bath application of 8-Br-cAMP, a membrane-permeable cAMP analog. Pre-treatment of Kenyon cells with the octopamine receptor antagonist phentolamine blocked the inhibitory effect of octopamine on Ca2+ channels. Pre-treatment of Kenyon cells with H-89, a selective inhibitor of cAMP-dependent protein kinase (PKA) attenuated the inhibitory effect of bath applied octopamine on Ca2+ channels. These results indicate that DHP-sensitive L-type Ca2+ channel is a target protein for octopamine and its modulation is mediated via cAMP and PKA-dependent signaling pathways in freshly isolated Kenyon cell in the cricket G. bimaculatus. [Copyright &y& Elsevier]

Abstract: This study compares velar ascent and morphological factors affecting velopharyngeal function between patients with repaired cleft palate and noncleft controls from early childhood to puberty. Lateral cephalograms obtained at rest and during blowing from 61 patients with repaired unilateral cleft lip and palate (cleft group) and 82 noncleft controls (control group) were divided into four developmental stages according to age and were studied cross-sectionally. Indices of nasopharyngeal area were derived from a coordinate system and landmarks on lateral cephalograms. The cleft group had lesser velar ascent, more posterosuperior position of the posterior maxilla, shorter velar length, and lesser pharyngeal depth than did the control group. There was a strong correlation between the vertical position of the posterior maxilla and the pharyngeal depth in the cleft group. Discriminant analysis revealed that the cleft group could be discriminated from the control group primarily on the basis of pharyngeal depth, velar length, and velar ascent. Our results suggest that the posterosuperior position of the posterior maxilla in patients with repaired cleft palate, resulting in reduced pharyngeal depth, represents an effort to facilitate velopharyngeal closure by means of shorter velar length and lesser velar ascent. [Copyright &y& Elsevier]

We studied in vitro hormonal activity of bisphenol A diglycidyl ether (BADGE) and bisphenol F diglycidyl ether (BFDGE), which are used as a material of interior coating for food cans. We also examined related compounds such as 2,2-bis[4-(3-chloro-2-hydroxypropoxy)phenyl]propane (BADGE.2HCl), and bis[4-(3-chloro-2-hydroxypropoxy)phenyl]methane (BFDGE.2HCl) etc. For this purpose, we constructed two stably transfected CHO-K1 cell lines (AR-EcoScreen for androgenic activity and c-luc for cell toxicity evaluation). One stably expresses luciferase with induction of androgen. The other stably expresses luciferase without androgen induction. Also, we have determined the androgenic and anti-androgenic effects of the test chemicals by reporter gene assay with these cell lines. None of the chemicals tested by this assay exhibited androgen agonistic activity. However, BADGE.2HCl and BFDGE.2HCl had the conspicuous antagonistic activity for androgen. These compounds had a high binding affinity for androgen receptor. Furthermore, these two compounds did not show the estrogenic activity in vitro assays. On the contrary, bisphenol A and bisphenol F exhibited anti-androgenic activity in vitro in addition to the estrogenic activity. These results suggest that these chlorohydroxy compounds of BADGE and BFDGE act as androgen antagonist through the process of binding to androgen receptor. [Copyright &y& Elsevier]

A case of follicular dendritic cell tumor (FDC tumor) arised in the aro-pharyngeal region is reported in a 16-year-old Japanese boy. He had a swelling in the right retromolar trigone and soft palate. Clinical examination disclosed a 25 mm×30 mm, elastic hard, ulcerated mass. CT scans showed a low density lesion in the right medial parapharyngeal area. The biopsy specimen revealed a low-grade malignant tumor. The patient underwent a tumorectomy which was extended to include the adjacent palatine tonsil. The tumor cells, with a clear cytoplasm and oval nucleus, were immunoreactive for S-100 (N/A), CD 21 (1F8), fascin (55K-2) and FDC (CNA42). The fascicular sheet pattern arrangement of the tumor cells was partly characteristic. The final diagnosis was an follicular dendritic cell tumor. Three courses of adjuvant chemoradiotherapy were administered postoperatively. Clinical features, pathological diagnosis and immunohistochemical markers for distinguishing FDC tumor are reviewed. [Copyright &y& Elsevier]

We orally administered polyphenone-60 (P-60), green tea extract catechins, in the diet (0, 1.25 and 5%) to male rats for 2, 4 and 8 weeks initiated at 5 weeks old. It was found that a 5% dose to male rats for 2–8 weeks induced goiters and decreased weights of the body, testis and prostate gland. Endocrinologically, elevating plasma thyroid stimulating hormone (TSH), luteinizing hormone (LH) and testosterone levels and decreasing tri-iodothyronine (T3) and thyroxine (T4) levels were induced by this treatment. We also found that P-60 as a whole and some of its constituents exhibited inhibitory effects on human placental aromatase activity by in vitro assay. The concentration of P-60 that required producing 50% inhibition of the aromatase activity (IC50 value) was 28 μg/ml. The IC50 values of (−)-catechin gallate (Cg), (−)-epigallocatechin (EGC), (−)-epigallocatechin gallate (EGCg) and (−)-gallocatechin gallate (GCg) were 5.5×10−6, 1.0×10−4, 6.0×10−5 and 1.5×10−5 m, respectively. (−)- Epicatechin gallate (ECg) at 1.0×10−4 m produced 20% inhibition. (−)-Epicatechin (EC) and (+)-catechin (CT) exhibited no effects on aromatase activity. The endocrinological changes observed in vivo were in conformity with antithyroid effects and aromatase inhibition effects of P-60 and its constituents. [ABSTRACT FROM AUTHOR]

Abstract: We developed PCR-based methods, confronting two pair primers (PCR-CTPP) and sequence-specific primers with a PCR-positive control (PCR-SSPPC) to analyze SNPs at nucleotide position 59 to reflect α-1, 4-fucosyltransferase (Le transferase) activity, which were analyzed by ABI PRISM® 3100 genetic analyzer. We compared 4 kinds of PCR-based methods, PCR restriction fragment-length polymorphism (PCR-RFLP), PCR sequence-specific-primers (PCR-SSP), PCR-CTPP and PCR-SSPPC. We found that all of these methods could determine Lewis genotyping correctly. The frequencies of Le and le alleles were 67.2% and 32.8% respectively. Both PCR-CTPP and PCR-SSPPC for Lewis genotyping are simple, reliable and applicable for forensic and clinical investigation. [Copyright &y& Elsevier]

Abstract: This paper reports effective PCR-based methods, such as sequence-specific primers with a positive control (PCR-SSPPC) and confronting two pairs of primers (PCR-CTPP) for forensic ABO groupings using fragment analysis by ABI PRISM® 3100 genetic analyzer. The method allows the well-established base changes at four nucleotide positions 261, 796, 802, and 803 to be assayed, so that reliable group prediction is established by the presence of three representative alleles such as A, B, and O. As a result, all common alleles in the database were correctly determined by our methods. [Copyright &y& Elsevier]

TH01 typing was performed by semi-nested PCR using a very low concentration of DNA (TH01 type=7–11) as a template. With 10 pg of template DNA, the probabilities of obtaining both of two alleles, only allele 7, only allele 11, or no PCR product, were 23.1%, 15.4%, 38.5% or 23.1%, respectively. Our findings suggest that these differences were found because the initial proportions of alleles 7 and 11 in the PCR tube were not equal. However, the allele distribution of TH01 types 7 and 11 using single cells considered to contain equal amount of the two alleles was similar to that obtained using a very low concentration of DNA. These results indicate that correct genotypes could not be obtained by using equal amount of both alleles, when the concentration of DNA was very low. To obtain a correct genotype, it was necessary to use 100 pg of DNA as the template for PCR amplification. [Copyright &y& Elsevier]

We examined urine and blood DNA obtained from bladder cancer patients with commercially available STR kits such as those for TH01, TPOX and CSF1PO loci. These loci were amplified by PCR, and polymorphic alleles were visualized as bands on gels. The results were compared with each paired DNA. Microsatellite alternation was observed in 5 out of 52 (9.6%) urine DNAs from the patients with bladder cancer. These loss of heterozygosity (LOH) and microsatellite instability (MSI) were observed such as the combination of an intense and a faint band. Complete LOH causing incorrect typing results was not observed. We were able to avoid incorrect typing results when using urine DNA under careful observation. [Copyright &y& Elsevier]