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Neutrophil extracellular traps (NETs) play important roles in host immunity, as there is increasing evidence of their contribution to the progression of several types of cancers even though their role in colorectal cancers (CRCs) remains unclear. To investigate the clinical relevance of NETs in CRCs, we examined the expression of citrullinated histone H3 using immunohistochemistry and preoperative serum myeloperoxidase–DNA complexes in CRC patients using an enzyme-linked immunosorbent assay. High expression of intratumoral or systemic NETs was found to correlate with poor relapse-free survival (RFS), for which it is an independent prognostic factor. In vitro investigations of CRC cells (HCT116, HT29) revealed that NETs did not affect their proliferation but did promote the migration of CRC cells mediated by neutrophil elastase (NE) released during NETosis to increase extracellular signal-regulated kinase (ERK) activity. In vivo experiments using nude mice (KSN/slc) revealed that NE inhibition suppressed liver metastases in CRC cells, although it did not affect the growth of subcutaneously implanted tumors. Taken together, these results suggest that NET formation correlates with poor prognoses of patients with CRC and that the inhibition of NE could be a potential therapy for CRC metastases.

Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (CRS + HIPEC) is the active treatment for peritoneal carcinomatosis of appendiceal origin. However, surgical management is sometimes difficult in patients with a high-tumor burden. A high-tumor burden was defined as a peritoneal cancer index (PCI) ≥ 28. Among 49 patients receiving CRS + HIPEC, 29 had a PCI ≥ 28. Complete cytoreduction (CC-0/1) was achieved in 20 of the 29 patients with a PCI ≥ 28 and in all 20 patients with a PCI

Rationale: Cystic lymphangiomas are uncommon congenital malformations that originate from lymphatic channels. Lymphangiomas frequently appear in the head, neck, and axillary regions of children. Abdominal cystic lymphangiomas are extremely rare, having a reported incidence of 1 in 20,000 to 250,000. Patient concerns: A 50-year-old female patient was admitted to our hospital with a cough that had persisted for several weeks. Abdominal ultrasonography incidentally revealed a multilocular cystic lesion in the lesser curvature of the stomach. Diagnosis: Preoperative findings indicated that the lesion was cystic lymphangioma. However, the possibility of a pancreatic tumor could not be completely excluded. Interventions: Laparoscopy revealed a multilocular cyst in the lesser curvature of the stomach. The gastrocolic ligament was divided, and the body and tail of the pancreas was exposed in the omental bursa, showing that the cystic lesion was not derived from the pancreas but from the lesser omentum. Although it was located directly beside the left gastric artery, the cyst was enucleated and totally resected laparoscopically without sacrificing the artery. Outcomes: The cystic lesion was histopathologically diagnosed as an abdominal cystic lymphangioma originating from the lesser omentum. The patient was discharged on the postoperative day 4 without complications. Lessons: Preoperative imaging cannot completely distinguish abdominal cystic lymphangiomas from other types of cystic tumors. Because cystic lymphangiomas have the potential to grow, invade vital structures, and develop life-threatening complications, laparoscopic assessment followed by total resection is considered a useful treatment strategy for peripancreatic cystic lesions.

Tributyltin (TBT) has long been recognized as a major environmental pollutant that can cause significant damage to the cellular functions as well as disruption of endocrine homeostasis. TBT induces apoptosis accompanied by production of reactive oxygen species (ROS) in mammalian and yeast cells. We observed that the budding yeast cells exposed to this compound at low concentrations exhibited cell growth arrest, but not cell death. Flow cytometric analysis of yeast cells without synchronization and morphological assessment of cells synchronized at M phase by nocodazole treatment indicated that TBT-exposed Saccharomyces cerevisiae cells were arrested at G1 phase of the cell cycle. This arrest was recovered by the addition of N-acetylcysteine, suggesting the involvement of ROS production by TBT. This is the first study to evaluate the action of TBT on cell cycle events.

Objective: To evaluate whether or not the liquid-based procedure (LBP) for endometrial cytology is as worthwhile for endometrial phasing as conventional slides. Materials and Methods: The subjects were 81 women who underwent endometrial cytology and were defined as negative. The specimens obtained by either Endocyte® or Masubuchi aspiration tube® were processed first with the conventional procedure and then with LBP using TACAS™. Results: (1) The number of subjects diagnosed by the conventional method as having proliferative, mid-, middle-secretory and late-secretory and atrophic phases was 40, 11, 10, 0 and 20, respectively. The rate of agreement with those using LBP was 87.7%. (2) Incidences of large clusters, ductal clusters, palisade arrangement, uneven staining and dirty mucous background detected were significantly higher with the conventional method, whereas with LBP clean background, inconspicuous bonding of cells, scattered solitary glandular cells, clear well-stained cytoplasm and cell compactness were higher. (3) Especially in the proliferative phase, clusters tended to be smaller and lose their architectural structures, and scattered solitary columnar cells were present. (4) Cells in the mid-phase tended to have loose contact and to mimic other phases. Conclusions: Cytodiagnosis of endometrial phasing prepared with LBP is feasible to perform when some modifications are implemented.

本研究は，離乳期の間食内容を調べ，さらに10ヶ月児の間食内容と3歳児の間食内容との関連を縦断的に検討することを目的とした。静岡県伊東市保健センターにおいて，平成12年度から15年度に出生した子ども（1313名）を対象に実施した。10ヶ月児と3歳児健康診査の問診票分を利用した。10ヶ月児では，約10％の者が「アイス」，「ビスケット等甘い菓子」を摂取しており，「スナック菓子」は約4％の者が摂取していた。10ヶ月児の間食内容10種をクラスター分析した結果，「アイス」，「スナック菓子」など5種類と「果物」や「ヨーグルト」（砂糖なし）など5種類に分かれた。そこで，前者を摂取している者を「菓子類あり群」とし，摂取していない者を「菓子類なし群」とした。この2群と3歳児の間食との関連を検討した結果，「菓子類あり群」の方が「菓子類なし群」よりも3歳児における間食の総数が多く，「アイス」，「スナック菓子」，「あめ」，「ガム」，「チョコレート」，「菓子パン」を与えると回答する割合が高かった。本研究は，日本で始めて離乳期の間食内容を示すとともに10ヶ月児の間食内容と3歳児の間食内容は関連があることを示した。今後，さらに離乳期の間食について調査を行っていく必要があると考えられる。（オンラインのみ掲載）

Amino acid analysis of Saccharomyces cerevisiae cells indicated that neutral amino acids such as glycine and alanine were probably excluded from the vacuoles, and that vacuolar H(+)-ATPase (V-ATPase) was involved in the vacuolar compartmentalization of these amino acids. We found that vacuolar membrane vesicles export neutral amino acids in an ATP-dependent manner. This is important in identifying vacuolar transporters for neutral amino acids.

Objective: To evaluate a fully automated processing system (TACAS™ Pro) for liquid-based procedures (LBPs). Methods and Materials: Materials were 3,483 and additionally 502 specimens that were taken at Kanagawa Health Service Association. Specimens obtained with a Cervex-Brush® were first smeared to glass slides using one side of the brush and then processed to TACAS Pro. Results: (1) The microscopy watching time per normal case was 3.65 ± 0.85 min in the conventional procedure, whereas in the LBP it was 1.95 ± 0.60 min, and the latter reduced workload to 53%. (2) The handling time of TACAS Pro per day was 2 h and 25.8 min. The workload at a laboratory offset it and revealed the work saving to be 63.8%. (3) Unsatisfactory rates were 0% in the conventional procedure, whereas in the LBP it was 1.88% at first. The latter rate decreased to 0.5% after system improvement. (4) Specimens which may disturb microscopy analysis were found in 1.06%, including 3 cases of possible carry-over of cells to the following slides. An additional study with the revised system confirmed no carry-over. (5) Incidences of abnormal cytology were consistent between the two methods. Conclusions: The revised automated processing system TACAS Pro is a feasible and useful LBP and reduces the workload of cytology laboratories.

Basic amino acids (lysine, histidine and arginine) accumulated in Saccharomyces cerevisiae vacuoles should be mobilized to cytosolic nitrogen metabolism under starvation. We found that the decrease of vacuolar basic amino acids in response to nitrogen starvation was impaired by the deletion of AVT4 gene encoding a vacuolar transporter. In addition, overexpression of AVT4 reduced the accumulation of basic amino acids in vacuoles under nutrient-rich condition. In contrast to AVT4, the deletion and overexpression of AVT3, which encodes the closest homologue of Avt4p, did not affect the contents of vacuolar basic amino acids. Consistent with these, arginine uptake into vacuolar membrane vesicles was decreased by Avt4p-, but not by Avt3p-overproduction, whereas various neutral amino acids were excreted from vacuolar membrane vesicles in a manner dependent on either Avt4p or Avt3p. These results suggest that Avt4p is a vacuolar amino acid exporter involving in the recycling of basic amino acids.

Tributyltin (TBT) has long been recognized as a major environmental pollutant that can cause significant damage to the cellular functions as well as disruption of endocrine homeostasis. TBT induces apoptosis accompanied by production of reactive oxygen species (ROS) in mammalian and yeast cells. We observed that the budding yeast cells exposed to this compound at low concentrations exhibited cell growth arrest, but not cell death. Flow cytometric analysis of yeast cells without synchronization and morphological assessment of cells synchronized at M phase by nocodazole treatment indicated that TBT-exposed Saccharomyces cerevisiae cells were arrested at G1 phase of the cell cycle. This arrest was recovered by the addition of N-acetylcysteine, suggesting the involvement of ROS production by TBT. This is the first study to evaluate the action of TBT on cell cycle events.

A vacuolar membrane protein, Vba2p of Schizosaccharomyces pombe, is involved in basic amino acid uptake by intact cells. Here we found evidence that Vba2p mediated ATP-dependent lysine uptake by vacuolar membrane vesicles of Saccharomyces cerevisiae. Vba2p was also responsible for quinidine sensitivity, and the addition of lysine improved cell growth on quinidine-containing media. These findings should be useful for further characterization of Vba2p.

キョウチクトウアブラムシの京都における生活環およびその天敵昆虫群構成を明らかにするため,京都市内2か所のキョウチクトウで1990年から3年間サンプリングおよび発生状況調査をおこない,次の結果を得た。1) キョウチクトウアブラムシは,京都において不完全生活環を全うできる。有翅虫は5月上旬に分散をはじめ,その後6月上旬,8月上旬,9月にそれぞれ発生のピークがみられた。秋にキョウチクトウが剪定され,新梢が伸長した場合にはその個体群は冬期にも高密度で推移したが,新梢が伸長しない場合には低密度で推移し,死滅することもあった。2月から3月の厳寒期には,胎生雌は増殖を停止し,その体色は鮮やかな黄色から乳濁色に変化した。越冬虫は3月末から増殖を再開し,4月末に最初の有翅虫が出現した。2) 本種の天敵昆虫として,次の15種を確認した。テントウムシ3種,ヒラタアブ4種,アブラコバエ1種,クサカゲロウ3種,ヒメカゲロウ1種,アブラバチ2種,ツヤコバチ1種。5月下旬から9月中旬までは,ダンダラテントウ(幼虫・成虫)の個体数が最も多く,その捕食が,アブラムシ個体群に対する重要な抑制要因であると考えられた。9月下旬以降はヒラタアブ(幼虫)や捕食寄生バチ類が活動したが,アブラムシの発生量を低下させることはできなかった。キョウチクトウアブラムシの毒性が,その天敵昆虫に及ぼす影響について考察した。

Objective: To evaluate the usefulness of a new liquid-based cytological procedure in a population screening program for cervix cancer. Subjects and Methods: Subjects were 1,000 women who underwent primary screening at the Kanagawa Health Service Association. The cytological specimens obtained by either cotton stick and Cytobrush® or Cervex-Brush® were processed using the Thinlayer Advanced Cytology Assay System (TACAS™), following the manufacturer’s instructions. Results: (1) Cells were evenly distributed on specimens and stained evenly; (2) shrinkage of cells was 5% based on measurement of the nuclear diameters of granulocytes in comparison with those of the conventional procedure; (3) incidences of cells that occupied the whole area, 1/20≤, 1/4≤, 1/4> of the observation fields were 58.8, 26.2, 12.0 and 3.0%, respectively; (4) number of the squamous cells in cases with 1/4> was Conclusion: TACAS is a feasible and useful cytological procedure.

The fission yeast Schizosaccharomyces pombe has a homolog of the budding yeast Atg22p, which is involved in spore formation (Mukaiyama H. et al., Microbiology, 155, 3816–3826 (2009)). GFP-tagged Atg22p in the fission yeast was localized to the vacuolar membrane. Upon disruption of atg22, the amino acid levels of the cellular fraction as well as the vacuolar fraction decreased. The uptake of several amino acids, such as lysine, histidine, and arginine, was impaired in atg22Δ cells. S. pombe Atg22p plays an important role in the compartmentalization of amino acids.

A recent study filling the gap in the genome sequence in the left arm of chromosome 2 of Schizosaccharomyces pombe revealed a homolog of budding yeast Vba2p, a vacuolar transporter of basic amino acids. GFP-tagged Vba2p in fission yeast was localized to the vacuolar membrane. Upon disruption of vba2, the uptake of several amino acids, including lysine, histidine, and arginine, was impaired. A transient increase in lysine uptake under nitrogen starvation was lowered by this mutation. These findings suggest that Vba2p is involved in basic amino acid transport in S. pombe under diverse conditions.

Tributyltin chloride (TBT), an environmental pollutant, is toxic to a variety of eukaryotic and prokaryotic organisms. Although it has been reported that TBT induces apoptotic cell death in mammalian, the action of TBT on eukaryotic microorganisms has not yet been fully investigated. In this study we examined the mechanism involved in cell death caused by TBT exposure in Saccharomyces cerevisiae. The median lethal concentration of TBT was 10 microM for the parent strain BY4741 and 3 microM for the pdr5Delta mutant defective in a major multidrug transporter, respectively. Fluorescence microscopic observations revealed nuclear condensation and chromatin fragmentation in cells treated with TBT indicating that cells underwent an apoptosis-like cell dearth. TBT-induced cell death was suppressed by deletion of the yca1 gene encoding a homologue of the mammalian caspase. In parallel, reactive oxygen species (ROS) were produced by TBT. These results suggest that TBT induces apoptosis-like cell death in yeast via an Yca1p-dependent pathway possibly downstream of the ROS production. This is the first report on TBT-induced apoptotic cell death in yeast.

We have previously reported that the combination of murine recombinant interferon beta (Mu-rIFN beta) with murine recombinant interferon gamma (Mu-rIFN gamma) provided greater inhibition of tumor growth than did each one alone in MethA-bearing mice. In the present study the effect of addition of human recombinant interleukin-2 (Hu-rIL-2) to the combination of Mu-rIFN beta with Mu-rIFN gamma on tumor growth in BALB/c mice bearing syngeneic MethA fibrosarcoma was examined. Low doses of Hu-rIL-2 (5 x 10(3) U or 5 x 10(4) U at 3-day intervals) showed no antitumor activity, while a high dose of Hu-rIL-2 (5 x 10(5) U) showed profound growth inhibition. The administration of IL-2 (ranging between 5 x 10(3) U and 5 x 10(5) U) in addition to the combination of IFN beta and IFN gamma showed more augmented antitumor effects in a dose-dependent manner. Furthermore, the simultaneous administration of IL-2, IFN beta and IFN gamma had more effective therapeutic activity, compared with the sequential administration of interferons and IL-2. These findings indicated that IL-2 in combination with IFN beta and gamma was effective for cancer treatment.

The purpose of this study was to determine the anchorage potential of titanium implants (Branemark; 3.75 × 7 mm) with the use of a sectional arch wire technique for orthodontic mesiodistal tooth movement, as assessed by the osseointegration of implants and tooth movement. Two implants were surgically placed in healed mandibular extraction sites of the second and third premolars on each side in 4 adult male beagle dogs. The implants were surgically uncovered 18 weeks later, and second-stage abutments with soldered edgewise tubes were attached. Segmented edgewise rectangular archwires (0.017 × 0.025 inch) with a T-loop or an L-loop were placed between the implants and the fourth premolars on both sides as the anchorage unit. One segment in each dog served as a loaded side, and the archwire was calibrated to produce 200 g of lateral force on the fourth premolar. The contralateral segment served as an unloaded side and was not subjected to orthodontic force. Sectional wires were activated biweekly 24, 28, 28, and 32 weeks, respectively, depending on the magnitude and the appearance of mesial tipping movement of the fourth premolar. After mandibular impressions were taken to measure the distance between the first molar and the fourth premolar, the animals were euthanized and dissected mandibles were prepared. The specimens were then embedded in polyester resin and cut to take backscattered electron images. On the basis of these images, the percentage of peri-implant bone volume was calculated and defined as an index of osseointegration. The differences between the initial and final fourth premolar to first molar distances varied (7.40, 8.85, 10.50, and 3.30 mm) on the loaded side, whereas the unloaded side showed no movement. Not only was there no statistical difference in the percent of peri-implant bone volume between the loaded and unloaded sides, but there was also no statistical difference between the compression and tension sides in both loaded and unloaded implants, which suggests that the implants maintained rigid osseointegration. In conclusion, the present study demonstrated that endosseous titanium implants can function as anchors for long-term orthodontic mesiodistal movement. (Am J Orthod Dentofacial Orthop 2000;118:601-7)

Tributyltin chloride (TBT), an environmental pollutant, is toxic to a variety of eukaryotic and prokaryotic organisms. Some members of F-ATP synthase (F-ATPase)/vacuolar type ATPase (V-ATPase) superfamily have been identified as the molecular target of this compound. TBT inhibited the activities of H(+)-transporting or Na(+)-transporting F-ATPase as well as H(+)-transporting V-ATPase originated from various organisms. However, the sensitivity to TBT of Na(+)-transporting V-ATPase has not been investigated. We examined the effect of TBT on Na(+)-transporting V-ATPase from an eubacterium Enterococus hirae. The ATP hydrolytic activity of E. hirae V-ATPase in purified form as well as in membrane-bound form was little inhibited by less than 10 microM TBT; IC50 for TBT inhibition of purified enzyme was estimated to be about 35 microM. Active sodium transport by E. hirae cells, indicating the in vivo activity of this V-ATPase, was not inhibited by 20 microM TBT. By contrast, IC50 of H(+)-transporting V-ATPase of the vacuolar membrane vesicles from Saccharomyces cerevisiae was about 0.2 microM. E. hirae V-ATPase is thus extremely less sensitive to TBT.