Search

Lipomyces starkeyi is an oil-producing yeast that can produce triacylglycerol (TAG) from glycerol as a carbon source. The TAG was mainly produced after nitrogen depletion alongside reduced cell proliferation. To obtain clues for enhancing the TAG production, cell metabolism during the TAG-producing phase was characterized by metabolomics with 13C labeling. The turnover analysis showed that the time constants of intermediates from glycerol to pyruvate (Pyr) were large, whereas those of tricarboxylic acid (TCA) cycle intermediates were much smaller than that of Pyr. Surprisingly, the time constants of intermediates in gluconeogenesis and the pentose phosphate (PP) pathway were large, suggesting that a large amount of the uptaken glycerol was metabolized via the PP pathway. To synthesize fatty acids that make up TAG from acetyl-CoA (AcCoA), 14 molecules of nicotinamide adenine dinucleotide phosphate (NADPH) per C16 fatty acid molecule are required. Because the oxidative PP pathway generates NADPH, this pathway would contribute to supply NADPH for fatty acid synthesis. To confirm that the oxidative PP pathway can supply the NADPH required for TAG production, flux analysis was conducted based on the measured specific rates and mass balances. Flux analysis revealed that the NADPH necessary for TAG production was supplied by metabolizing 48.2% of the uptaken glycerol through gluconeogenesis and the PP pathway. This result was consistent with the result of the 13C-labeling experiment. Furthermore, comparison of the actual flux distribution with the ideal flux distribution for TAG production suggested that it is necessary to flow more dihydroxyacetonephosphate (DHAP) through gluconeogenesis to improve TAG yield.

Skeletal muscle is an important organ for controlling the development of type 2 diabetes. We discovered Panax notoginseng roots as a candidate to improve hyperglycemia through in vitro muscle cells screening test. Saponins are considered as the active ingredients of ginseng. However, in the body, saponins are converted to dammarane-type triterpenes, which may account for the anti-hyperglycemic activity. We developed a method for producing a dammarane-type triterpene extract (DTE) from Panax notoginseng roots and investigated the extract’s potential anti-hyperglycemic activity. We found that DTE had stronger suppressive activity on blood glucose levels than the saponin extract (SE) did in KK-Ay mice. Additionally, DTE improved oral glucose tolerance, insulin sensitivity, glucose uptake, and Akt phosphorylation in skeletal muscle. These results suggest that DTE is a promising agent for controlling hyperglycemia by enhancing glucose uptake in skeletal muscle.

Lipomyces starkeyi is an oil-producing yeast that can produce triacylglycerol (TAG) from glycerol as a carbon source. The TAG was mainly produced after nitrogen depletion alongside reduced cell proliferation. To obtain clues for enhancing the TAG production, cell metabolism during the TAG-producing phase was characterized by metabolomics with

Using a monoclonal immunoradiometrical assay, we measured the concentrations of the specific epitope M3 of tissue polypeptide antigen (TPA), namely, tissue polypeptide specific antigen (TPS), in serum and amniotic fluid obtained from normal, healthy, nonpregnant and pregnant Japanese women (NHNJW and NHPJW). The cut-off value of the serum TPS level was set at 130 Ul, based on the mean+2 standard deviations in the NHNJW. The serial measurement of serum TPS levels demonstrated the significant emporal elevation of TPS level near the time of ovula-tion (±3 days) in the ovulatory women. In the NHPJW, the mean serum TPS level and the positivity rate (>130 U/l) increased with the advance of gestation, reaching 183.7 U/l and 66.7%, respectively, in the third trimester. Maternal serum levels were much elevated just before (mean: 982.3 U/l) and after (mean: 824.6 U/l) delivery, and were reduced to about one-fourth by 3 days. In addition, markedly high TPS concentrations were found in the retroplacental blood (one case: 9,076 U/l) and the amniotic fluid (mean: 11,650 U/l). The serum TPS level correlated well (r=0.871) with the serum TPA level in the TPS range of 2-2,000 U/l, while a poor correlation (r=0.273) was found in the TPS range of 2–500 U/l. The present study thus obtained fundamental data on TPS in Japanese women. The fluctuation and change of serum TPS levels during the menstrual cycle and in pregnancy should be taken into consideration when we apply TPS as a tumor marker for women.

目的 : 日母分類にてクラス III a と判定された標本を用いて, 日母分類とベセスダシステム 2001 の比較検討を行った.方法 : 扁平上皮系の異常にて日母分類クラス III a と判定された 120 例の細胞標本を, ベセスダシステム 2001 の基準に従って評価を行った. 細胞学的特徴, 組織診結果について検討した.成績 : 対象 120 例のうち適正標本は 117 例で NILM が 28 例, ASC-US が 28 例, ASC-H が 2 例, LSIL が 47 例, HSIL が 12 例であった. 日母分類クラス III a の細胞学的特徴は核腫大とクロマチン増量の頻度が高く, CIN2 以上を有意に検出する所見は核腫大であった.結論 : 日母分類クラス III a は, ベセスダシステム 2001 にて評価すると幅広い推定病変を含んでいることが明らかになった. 表層から中層細胞の異常を認める細胞標本において, 核腫大所見は CIN2 以上の検出に有用である可能性が示唆された.

Objective. The aim of this study was to evaluate precisely the differences in the spectra of human papillomavirus (HPV) types detected by different generic primer pairs commonly used for detection of this extraordinarily heterogeneous virus. Methods. Three sets of polymerase chain reaction (PCR) primers for the L1 open reading frame (ORF) and two sets for E6/E7 ORFs were used to detect HPVs in DNAs from 107 cervical tissues, including 77 cervical neoplasias. HPV types were determined by analysis of restriction fragment length polymorphisms (RFLPs) and nucleotide sequencing. Results. A high overall detection rate of HPV in cervical neoplasias (76/77, 98.7%) was achieved by polymerase chain reaction (PCR) amplification with multiple sets of generic primers, while the detection rate for each individual primer pair varied from 48/77 (62%) to 70/77 (91%). Only in 34 of 77 cases (44%) were HPV DNAs positive for all sets of primer pairs. Further determination of HPV types by RFLPs and nucleotide sequencing showed inconsistencies between the PCR primer pairs used. Conclusion. Our study revealed that the HPV detection rate is critically affected by the choice of PCR primers, and that appropriate use of combinations of generic PCR primer sets followed by RFLP analyses is both necessary and sufficient for typing most HPVs in cervical lesions. More precise methods such as sequencing would be necessary in only a few cases.

We describe an improved copper ferrocyanide-based method for cytochemical detection of glucose-6-phosphate dehydrogenase (G6PD), which was used to localize the enzyme within the ultrastructure of rat hepatocytes and adrenocortical cells. With this method, glutaraldehyde fixation and the addition of exogenous electron carriers (for example, phenazine methosulfate) to the cytochemical reaction medium were essential. Copper ferrocyanide reaction product showing the distribution of G6PD was readily recognized at the light microscopic level as Hatchett's brown staining and at the electron microscopic level as electron-dense deposits. Within stained regions, enzyme cytochemical G6PD activity was found to be associated with ribosome-like structures. Because G6PD is a soluble, cytosolic enzyme, its displacement or extraction may occur during conventional fixation. We, therefore, combined a rapid-freezing technique with G6PD enzyme cytochemistry. The resultant rapid-freezing enzyme cytochemistry enabled us to show the subcellular distribution of G6PD in a more life-like state; the localization of G6PD in rapidly frozen cells was in substantial agreement with that in conventionally fixed cells.

Background. Teratomas of the greater omentum are rare. In most cases, they are composed of mature tissue elements and behave clinically in a benign fashion. Case. A 62-year-old woman with a history of bilateral salpingo-oophorectomy due to ectopic pregnancy was found to have multiple cystic teratomas of the omentum. There was a histologic evidence of a sequential transition from benign through borderline to obviously invasive adenocarcinoma within the lining of the cysts as well as peritoneal dissemination. Conclusion. We defined the present case as malignant transformation of a mature teratoma, but have no evidence whatsoever for the origin of multiple mature teratomas. Further detailed analyses with more elaborated procedures on many relevant reports would be required to establish the origin and biologic behavior of teratomas of the omentum.

今回われわれは子宮頸部生検組織および手術材料にて子宮頸部上皮内新生物Cervical IntraepithelialNeoplasia (CIN) I～IIIと診断された130症例を用いin situ hybridization (ISH) を実施し, human papillomavirus (HPV) の検索を行い以下の結果を得た.内訳はCINI31例, CINII43例, CIN III56例である.HybridizationはVira TypeTM Human Papillomavirus TissueHybridization Kit (LTI, 東レ) を用いた.使用したDNAプローブはHPV6/11, 16/18, 31/33/35型のビオチン標識混合プローブである.1.CIN全体におけるHPV-DNAの検出頻度は69.5%(91/130) であった.また, 各病変の陽性率はCIN I58.1%(18/31), CIN II74.4%(32/43) CIN III73.3%(41/56) であった.2.CIN異型度とISHの陽性所見はCIN Iで表層集中型 (Type S), CIN IIで全層不均等分布型 (Type A1), CIN IIIで全層均等分布型 (Type A2) がそれぞれ最も高い頻度を示した.3.Koilocytosisの出現頻度はHPV-DNA陰性例に比べ, 陽性例に頻度が高かった.Dysker-atosis, Parakeratosisの出現頻度は陽性例と陰性例で大差を認めなかった.

凍結融解過程におけるストロー内の牛卵子の封入位置と透明帯損傷の関係について検討した。垂直にしたストロー内では卵子の位置が綿栓上,液体の凹面,溶液中となるように吸入し,水平にしたストロー内では,卵子を中央のグリセロール液層内に吸入し,ストロー壁に付着するように設定した。各ストローは,室温から-5°Cまで毎分1°Cの速度で冷却し,-5°Cで5分間保持後植氷を行ないさらに同温度で10分間保持したのち,毎分0.3°Cの速度で-30°Cまで冷却し,液体窒素中へ浸漬した。これらのストローを,+37°C温湯中に浸漬して融解し,卵子の透明帯損傷の割合を実体顕微鏡下で検査した。ストローを垂直にしてウシ卵子を綿栓上,液体の凹面,溶液中に位置させて凍結融解した試験では,透明帯の損傷割合がそれぞれ42.6%(86/202),21.6%(22/102),22.0%(22/100)となり綿栓上で有意(P

To evaluate drug-induced neurotoxicity with combination consecutive low-dose CDDP (CLD-CDDP) and weekly paclitaxel in comparison with TJ therapy. Neurotoxicity was judged according to accumulative scores on questionnaires.Weekly TP therapy (CDDP: 10 mg/m2, days 1-7, paclitaxel: 60-80 mg/m2, days 1, 8, 15) and TJ therapy (paclitaxel: 170-180 mg/m2, 3 hours, carboplatin AUC 4-5) were given to 24 subjects in 57 courses and 19 subjects in 30 courses as previously reported, respectively.1. Significant differences were found in numbness accumulative scores (p = 0.011) between weekly TP and TJ therapy, but not in pain accumulative scores (p = 0.926) between weekly TP and TJ therapy. 2. Peak pain at days 3-4 of paclitaxel that appeared during TJ therapy disappeared during weekly TP therapy, while numbness persisted to a lesser extent with weekly TP therapy than with TJ therapy. 3. Numbness was observed to accumulate in some subjects but not in others during weekly TP therapy.The patients' subjective symptoms were able to be objectively evaluated with the questionnaires. Weekly paclitaxel was observed to reduce neurotoxicity, which might be reinforced in combination with CDDP.

Cyclic guanosine monophosphate (cGMP)-phosphodiesterase (PDE) in the outer segment of vertebrate retinal rod cells is one of key enzymes mediating phototransduction. We report here on light-induced PDE activity in intact rat retinal rod cells processed by rapid-freezing enzyme cytochemistry, a new morphological technique that is a combination of rapid-freezing, freeze-substitution fixation, and subsequent enzyme cytochemistry. This technique quickly immobilizes and preserves both enzyme activity and the cell ultrastructure in a state approximating living conditions; consequently, it has proved useful for cytochemical detection of light-induced PDE activity. Using this technique we observed that the catalytic site of PDE molecules in rapid-frozen outer segments was predominantly located in the extradiscal spaces; PDE activity was significantly greater in light than in darkness; and illumination elicited marked increases in PDE activity in dark-adapted cells. Light-induced PDE activity was first cytochemically detected after 3 s of illumination and reached a peak within 5 s, at which time it was in virtually the same as that seen in fully light adapted cells. In addition, cytochemical guanosine triphosphatase (GTPase) activity in dark-adapted cells, as well as corresponding PDE activity, increased in a time-dependent manner with illumination duration; this acceleration in GTPase activity closely paralleled the PDE activity. Thus, our results suggest, in part, the existence of reciprocal regulation of PDE and activated transducin alpha subunit, thereby modulating light adaptation in rod cells.

2種の絨毛癌培養細胞 (BeWoおよびHCCM-5株) の形態と, methotrexate (MTX) によるDNA, RNA, 蛋白合成, 細胞生存率および形態変化をin vitroで観察した.両細胞ともMTXに濃度依存性にDNA, RNA, 蛋白合成, 細胞生存率が低下した.MTX 10-6M 48時間処理後のDNA合成, RNA合成, 蛋白合成, 細胞生存率はBeWo細胞ではそれぞれ無処理対照の61, 27, 30, 87%, HCCM-5細胞では42, 23, 30, 66%であった.同条件で両細胞に細胞質の淡染化, 細胞質内空胞および顆粒の出現, 核の増大と淡染化, 核不整形化, 核小体の増大と1個の核内に出現する核小体数の減少を認め, HCCM-5細胞により著明であった.上記形態変化はMTXによるDNA, RNA, 蛋白合成の抑制により出現したと考えられた.

Co(II)TPP 1 - catalysed isomerization of a series of electronegative substituted quadricyclanes( 1 ) to the corresponding norbornadiene( 2 ) was found to proceed via radicophilic attack of the metal to 1 .

組織学的に子宮頸管炎と診断された161例の生検組織に酵素抗体法PAP (peroxidase antiperoxidase) を応用し, 組織学的にC.trachomatisの検出を試みた.1.子宮頸管炎161例中52例にPAP陽性所見を認めた.その頻度は32.3%であった.2.年齢別頻度は～30歳, 26.7%, 31～40歳33.3%, 41～50歳, 33.3%, 51～60歳, 35.3%, 61歳～, 16.7%, と各年代にわたり, 30%近くの感染を認めた.3.感染細胞は円柱上皮が最も多く, 扁平上皮化生, 扁平上皮の順であった.4.感染細胞におけるPAP陽性所見は, 円柱上皮細胞の核に隣接した一側または, 周囲に認められ, 特に腺腔側に出現するものが圧倒的に多く注目された.また, 細胞診で認められる円形様の封入体は少数であった.5.一部の症例に螢光抗体法を行い, PAP陽性部位に一致して, 陽性所見を認めた.

New series of platinum complexes of cyclopropenylidenes of the types of PtX2(CP)2 and trans-PtX(PBu3)2(CP) have been synthesized, where CP is di-t-butylcyclopropenylidene (BCP) or bis(diisopropylamino)cyclopropenylidene (ACP). The 13C-NMR chemical shifts, and 13C−195Pt coupling constants (1JPtC) for the complexes are discussed in comparison with those values derived from closely related series of compounds, trans-PtCl(PR3)2L; L CH3, C6H5 and CCBu-t. An excellent linear relationship through the origin was obtained between 1JPtC and the formal ‘s’ % character of the carbon directly bonded to Pt for the series trans-PtCl(PR3)2L in which the PtC bond is regarded as a pure σ-linkage, whereas 1JPtC deviates largely from this relationship when pπdπ bonding interaction possibly exists in the PtC bond. The NMR data suggest the strong nmr trans-influence of the cyclopropenylidenes and that in the PtCP bond the σ-interaction is appreciable but the π-interaction is negligible.

子宮体部横紋筋肉腫の1症例についてその臨床経過, 細胞診所見, 各種特殊染色を用いた病理組織学的所見, および組織レベルの細胞核DNA量の測定を行い比較検討した.1) 本症例は術前に細胞診, 組織診によって肉腫と診断し, 術後, 摘出物の検索によって中胚葉性混合腫瘍のうち, 横紋筋肉腫成分のみからなるきわめてまれな症例と確認された.2) 細胞診では壊死性背景のなかに, 大小不同の著しく, 核膜の著明な紡錘型から卵円形の非上皮性悪性腫瘍由来の細胞を強く示唆する悪性細胞が認められた.3) H-E染色では横紋筋肉腫細胞と横紋筋芽細胞とみられる2種類の細胞を認めた.またVG, MA, PAS染色などの特殊染色より, 横紋を有する細胞, PAS陽性細胞などが確認できた.4) 組織レベルのDNA定量では, 2倍体から12倍体に及ぶ, 漸減性の大きなモードがヒストグラム上に認められ, DNA量からも, 横紋筋肉腫細胞の多形性が裏付けられた.

New palladium complexes of cyclopropenylidenes of the type of trans-Cl2Pd(CP)(Py-X) have been synthesized, where CP is di-t-butylcyclopropenylidene or bis(diisopropylamino)cyclopropenylidene, and Py-X is 4-substituted pyridines (X = H, CH3, CN). The complexes revealed high catalytic activity toward the cycloreversion of quadricyclane (Q) to norbornadiene (NBD). Based on a detailed kinetic study, it is concluded that liberation of Py-X is prerequisite for the metal to interact with Q, The rate of the reaction obeys a rate expression d[NBD]/dt = kcat[cat] 1 2 [Q]. The apparent catalytic activity (kcat) can be related to the catalytic activity (k*) of the coordinatively unsaturated species Cl2Pd(CP) and the dissociation constant (K) for pyndine liberation by kcat= k*K 1 2 . The values k* and K are discussed in relation to the nature of ligands.

Antitumor effects of hematoporphyrin derivative (HpD) plus argon dye laser were examined using various human gynecologic tumor cells in vitro. Irradiation alone with argon dye laser showed no effect on the DNA synthesis of cells. Treatment of the cells with HpD inhibited the DNA synthesis depending on the concentration and the exposure time. Photoradiation by argon dye laser following treatment of the cells with HpD 40 μg/ml for 2 h killed more than 80% of cells when the cells were irradiated for more than 3 min. Prominent degenerative changes of the cytoplasm and the nucleus appeared within 1 h after photoradiation. These changes were confirmed by morphology and DNA histogram. There were no differences of sensitivity to photoradiation among the three histologically different kinds of cells lines used.

Endometrial carcinoma was diagnosed in a premenopausal woman suffering with systemic lupus erythematosus. She had received both predonisolone and an im-munosuppressive agent for more than 10 years. Anovulatory cycles persisted during drug administration, along with dysfunctional uterine bleeding. The serum estrogen: progesterone ratio was high. Repeated endometrial biopsies revealed a progression of change from benign proliferation to cystic hyperplasia, adenomatous hyperplasia, atypical hyperplasia and invasive adenocarcinoma. These clinical data suggest that a result of long-term unopposed endogenous estrogen can have been the cause of the endometrial carcinoma.