Search

Background: Superficial temporal artery (direct) and encephalomyosynangiosis (indirect) revascularization may develop after combined bypass surgery in pediatric patients with moyamoya disease (MMD). However, arterial development varies widely among patients, and the underlying mechanisms remain unknown. Objectives: We evaluated the relationship between the development of donor arteries after bypass surgery in pediatric patients with MMD and the MMD-susceptibility gene variant c.14576G>A of ring finger protein (RNF) 213. Methods: The data of pediatric patients with MMD (age RNF213 c.14576G>A status was examined. Results: Forty-eight hemispheres from 28 pediatric patients with MMD were examined. Three hemispheres belonged to patients with the AA genotype; 33 to patients with the AG genotype (AA/AG group); and 12 to patients with the GG genotype (GG group; wild type). The CCRs for the DTA were significantly higher in patients with RNF213 variant (AA/AG group; 2.5 ± 0.1) than in the GG group (2.0 ± 0.2) (p = 0.03), whereas the CCRs for the STA were significantly higher in the GG (1.6 ± 0.1) than in the AA/AG group (1.3 ± 0.6) (p = 0.02). There was no significant difference in the CCRs for the MMA and basilar artery between the groups. Other factors, including sex, age, and MRA grading, were not associated with the development of specific bypass development. Conclusions: The extent of collateral development associated with direct or indirect bypass was found to differ between the genotypes of the RNF213 c.14576G>A associated with pediatric MMD. This genetic variant correlates with the development of the disease and affects revascularization after bypass surgery in pediatric patients with MMD.

A chemoselectivity switchable microflow reaction was developed to generate reactive and unstable intermediates. The switchable chemoselectivity of this reaction enables a selection for one of two different intermediates, an aryllithium or a benzyl lithium, at will from the same starting material. Starting from bromo-substituted styrenes, the aryllithium intermediates were converted to the substituted styrenes, whereas the benzyl lithium intermediates were engaged in an anionic polymerization. These chemoselectivity-switchable reactions can be integrated to produce polymers that cannot be formed during typical polymerization reactions.

In vertebrates, newly emerging transformed cells are often apically extruded from epithelial layers through cell competition with surrounding normal epithelial cells. However, the underlying molecular mechanism remains elusive. Here, using phospho-SILAC screening, we show that phosphorylation of AHNAK2 is elevated in normal cells neighboring RasV12 cells soon after the induction of RasV12 expression, which is mediated by calcium-dependent protein kinase C. In addition, transient upsurges of intracellular calcium, which we call calcium sparks, frequently occur in normal cells neighboring RasV12 cells, which are mediated by mechanosensitive calcium channel TRPC1 upon membrane stretching. Calcium sparks then enhance cell movements of both normal and RasV12 cells through phosphorylation of AHNAK2 and promote apical extrusion. Moreover, comparable calcium sparks positively regulate apical extrusion of RasV12-transformed cells in zebrafish larvae as well. Hence, calcium sparks play a crucial role in the elimination of transformed cells at the early phase of cell competition.

The external quenching method based on flow microreactors allows the generation and use of short-lived fluoro-substituted methyllithium reagents, such as fluoromethyllithium, fluoroiodomethyllithium, and fluoroiodostannylmethyllithium. Highly chemoselective reactions have been developed, opening new opportunities in the synthesis of fluorinated molecules using fluorinated organometallics.

Summary: p53 is a tumor suppressor protein, and its missense mutations are frequently found in human cancers. During the multi-step progression of cancer, p53 mutations generally accumulate at the mid or late stage, but not in the early stage, and the underlying mechanism is still unclear. In this study, using mammalian cell culture and mouse ex vivo systems, we demonstrate that when p53R273H- or p53R175H-expressing cells are surrounded by normal epithelial cells, mutant p53 cells undergo necroptosis and are basally extruded from the epithelial monolayer. When mutant p53 cells alone are present, cell death does not occur, indicating that necroptosis results from cell competition with the surrounding normal cells. Furthermore, when p53R273H mutation occurs within RasV12-transformed epithelia, cell death is strongly suppressed and most of the p53R273H-expressing cells remain intact. These results suggest that the order of oncogenic mutations in cancer development could be dictated by cell competition. : Watanabe et al. demonstrate that mutant p53 cells undergo necroptosis and are eliminated from epithelia via cell competition with surrounding normal cells. Moreover, the eradication of mutant p53 cells is suppressed within RasV12-transformed epithelia, suggesting that the order of oncogenic mutations in cancer development could be dictated by cell competition. Keywords: cell competition, p53, necroptosis, p53R273H, p53R175H, RasV12, multi-step cancer progression, MDCK cells, mouse intestinal organoids

CeO2-supported Ru (Ru/CeO2) worked as an effective and reusable heterogeneous catalyst for the selective dissociation of the C-N bond in amides, particularly primary amides, with H2 in water solvent at low reaction temperature of 333 K, and high yields of the corresponding alcohols were obtained from primary amides.

At the initial stage of carcinogenesis, newly emerging transformed cells are often eliminated from epithelial layers via cell competition with the surrounding normal cells. For instance, when surrounded by normal cells, oncoprotein RasV12-transformed cells are extruded into the apical lumen of epithelia. During cancer development, multiple oncogenic mutations accumulate within epithelial tissues. However, it remains elusive whether and how cell competition is also involved in this process. In this study, using a mammalian cell culture model system, we have investigated what happens upon the consecutive mutations of Ras and tumor suppressor protein Scribble. When Ras mutation occurs under the Scribble-knockdown background, apical extrusion of Scribble/Ras double-mutant cells is strongly diminished. In addition, at the boundary with Scribble/Ras cells, Scribble-knockdown cells frequently undergo apoptosis and are actively engulfed by the neighboring Scribble/Ras cells. The comparable apoptosis and engulfment phenotypes are also observed in Drosophila epithelial tissues between Scribble/Ras double-mutant and Scribble single-mutant cells. Furthermore, mitochondrial membrane potential is enhanced in Scribble/Ras cells, causing the increased mitochondrial reactive oxygen species (ROS). Suppression of mitochondrial membrane potential or ROS production diminishes apoptosis and engulfment of the surrounding Scribble-knockdown cells, indicating that mitochondrial metabolism plays a key role in the competitive interaction between double- and single-mutant cells. Moreover, mTOR (mechanistic target of rapamycin kinase) acts downstream of these processes. These results imply that sequential oncogenic mutations can profoundly influence cell competition, a transition from loser to winner. Further studies would open new avenues for cell competition-based cancer treatment, thereby blocking clonal expansion of more malignant populations within tumors.

Recent studies have revealed that newly emerging transformed cells are often apically extruded from epithelial tissues. During this process, normal epithelial cells can recognize and actively eliminate transformed cells, a process called epithelial defence against cancer (EDAC). Here, we show that mitochondrial membrane potential is diminished in RasV12-transformed cells when they are surrounded by normal cells. In addition, glucose uptake is elevated, leading to higher lactate production. The mitochondrial dysfunction is driven by upregulation of pyruvate dehydrogenase kinase 4 (PDK4), which positively regulates elimination of RasV12-transformed cells. Furthermore, EDAC from the surrounding normal cells, involving filamin, drives the Warburg-effect-like metabolic alteration. Moreover, using a cell-competition mouse model, we demonstrate that PDK-mediated metabolic changes promote the elimination of RasV12-transformed cells from intestinal epithelia. These data indicate that non-cell-autonomous metabolic modulation is a crucial regulator for cell competition, shedding light on the unexplored events at the initial stage of carcinogenesis.

Despite the fact that gut microbiota is closely associated with obesity, few studies have focused on the influences of paraprobiotics as food ingredients on both obesity prevention and the gut microbial community. In this study, we evaluated the effects of fragmented Lactobacillus amylovorus CP1563 (CP1563) as a paraprobiotic for obesity prevention and investigated its effects on the gut microbial community in pre-obese subjects. One hundred sixty-nine healthy subjects with a body mass index from 25.0 to 29.9 kg/m2 ingested beverages with or without the fragmented CP1563 containing 10-hydroxyoctadecanoic acid (10-HOA) for 12 weeks. The changes in abdominal, total, visceral, and subcutaneous fatty areas were significantly lower in the CP1563-10-HOA group than in the placebo group at 12 weeks. Furthermore, 16S rRNA gene sequencing of fecal DNA revealed that the changes in the abundances of the genera Roseburia and Lachnospiraceae;g were significantly greater in the CP1563-10-HOA group than in the placebo group, and the changes in the abundances of the genus Collinsella was significantly smaller in the CP1563-10HOA group than in the placebo group. Our results showed that continuous ingestion of the fragmented CP1563 containing 10-HOA reduced abdominal body fat and affected the gut microbial community in pre-obese healthy subjects. Our findings may contribute to the understanding of the relationship between the anti-obesity effect of paraprobiotics and gut microbiota.

Obesity is a risk factor of postoperative complications. We experienced 2 extremely obese patients:a 32-year-old male with coronary artery disease and a 75-year-old female with aortic valve stenosis. Their initial body weights were 133 kg and 88.5 kg, respectively, and their initial body mass indexes (BMIs) were both 41. Their BMIs were reduced to 35.5 and 35, respectively, after preoperative weight reduction. Off-pump coronary artery bypass grafting and aortic valve replacement were performed, respectively. After surgery, the non-invasive positive pressure ventilation( NPPV) support was effective, and their postoperative courses were uneventful. Preoperative weight reduction and NPPV are useful for extremely obese patients who undergo cardiac surgery.

Summary At the early stage of cancer development, oncogenic mutations often cause multilayered epithelial structures. However, the underlying molecular mechanism still remains enigmatic. By performing a series of screenings targeting plasma membrane proteins, we have found that collagen XVII (COL17A1) and CD44 accumulate in RasV12-, Src-, or ErbB2-transformed epithelial cells. In addition, the expression of COL17A1 and CD44 is also regulated by cell density and upon apical cell extrusion. We further demonstrate that the expression of COL17A1 and CD44 is profoundly upregulated at the upper layers of multilayered, transformed epithelia in vitro and in vivo. The accumulated COL17A1 and CD44 suppress mitochondrial membrane potential and reactive oxygen species (ROS) production. The diminished intracellular ROS level then promotes resistance against ferroptosis-mediated cell death upon cell extrusion, thereby positively regulating the formation of multilayered structures. To further understand the functional role of COL17A1, we performed comprehensive metabolome analysis and compared intracellular metabolites between RasV12 and COL17A1-knockout RasV12 cells. The data imply that COL17A1 regulates the metabolic pathway from the GABA shunt to mitochondrial complex I through succinate, thereby suppressing the ROS production. Moreover, we demonstrate that CD44 regulates membrane accumulation of COL17A1 in multilayered structures. These results suggest that CD44 and COL17A1 are crucial regulators for the clonal expansion of transformed cells within multilayered epithelia, thus being potential targets for early diagnosis and preventive treatment for precancerous lesions.

Normal epithelial cells have an ability to sense and actively eliminate neighboring transformed cells, a process called epithelial defense against cancer (EDAC). Exogenous S1P plays a crucial role in EDAC; the S1P–S1PR2 pathway regulates Rho–Rho kinase–filamin in the surrounding normal cells, promoting apical extrusion of RasV12-transformed cells from epithelia., At the initial step of carcinogenesis, transformation occurs in single cells within epithelia, where the newly emerging transformed cells are surrounded by normal epithelial cells. A recent study revealed that normal epithelial cells have an ability to sense and actively eliminate the neighboring transformed cells, a process named epithelial defense against cancer (EDAC). However, the molecular mechanism of this tumor-suppressive activity is largely unknown. In this study, we investigated a role for the sphingosine-1-phosphate (S1P)–S1P receptor 2 (S1PR2) pathway in EDAC. First, we show that addition of the S1PR2 inhibitor significantly suppresses apical extrusion of RasV12-transformed cells that are surrounded by normal cells. In addition, knockdown of S1PR2 in normal cells induces the same effect, indicating that S1PR2 in the surrounding normal cells plays a positive role in the apical elimination of the transformed cells. Of importance, not endogenous S1P but exogenous S1P is involved in this process. By using FRET analyses, we demonstrate that S1PR2 mediates Rho activation in normal cells neighboring RasV12-transformed cells, thereby promoting accumulation of filamin, a crucial regulator of EDAC. Collectively these data indicate that S1P is a key extrinsic factor that affects the outcome of cell competition between normal and transformed epithelial cells.

Recent studies have revealed that newly emerging transformed cells are often eliminated from epithelia via cell competition with the surrounding normal epithelial cells. However, it remains unknown whether and how soluble factors are involved in this cancer preventive phenomenon. By performing stable isotope labeling with amino acids in cell culture (SILAC)-based quantitative mass spectrometric analyses, we have identified ADAM-like Decysin-1 (ADAMDEC1) as a soluble protein whose expression is upregulated in the mix culture of normal and RasV12-transformed epithelial cells. Expression of ADAMDEC1 is elevated in normal epithelial cells co-cultured with RasV12 cells. Knockdown of ADAMDEC1 in the surrounding normal cells substantially suppresses apical extrusion of RasV12 cells, suggesting that ADAMDEC1 secreted by normal cells positively regulate the elimination of the neighboring transformed cells. In addition, we show that the metalloproteinase activity of ADAMDEC1 is dispensable for the regulation of apical extrusion. Furthermore, ADAMDEC1 facilitates the accumulation of filamin, a crucial regulator of Epithelial Defense Against Cancer (EDAC), in normal cells at the interface with RasV12 cells. This is the first report demonstrating that an epithelial intrinsic soluble factor is involved in cell competition in mammals.

CeO2-supported Ru (Ru/CeO2) worked as an effective and reusable heterogeneous catalyst for the selective dissociation of the C-N bond in amides, particularly primary amides, with H2 in water solvent at low reaction temperature of 333 K, and high yields of the corresponding alcohols were obtained from primary amides.

At the initial stage of carcinogenesis, transformation occurs in single cells within the epithelium. Recent studies have revealed that the newly emerging transformed cells are often apically eliminated from epithelial tissues. However, the underlying molecular mechanisms of this cancer preventive phenomenon still remain elusive. In this study, we first demonstrate that myosin-II accumulates in Src-transformed cells when they are surrounded by normal epithelial cells. Knock-down of the heavy chains of myosin-II substantially diminishes apical extrusion of Src cells, suggesting that accumulated myosin-II positively regulates the apical elimination of transformed cells. Furthermore, we have identified β-spectrin as a myosin-II-binding protein under the coculture of normal and Src-transformed epithelial cells. β-spectrin is also accumulated in Src cells that are surrounded by normal cells, and the β-spectrin accumulation is regulated by myosin-II. Moreover, knock-down of β-spectrin significantly suppresses apical extrusion of Src cells. Collectively, these results indicate that accumulation of the myosin-II-spectrin complex plays a positive role in apical extrusion of Src-transformed epithelial cells. Further elucidation of the molecular mechanisms of apical extrusion would lead to the establishment of a novel type of cancer preventive medicine.

Nitrous oxide (N2O) is an important greenhouse gas that can be emitted from wastewater treatment plants (WWTPs). Such emissions are reportedly process specific and related to operational parameters. This study was conducted to clarify spatial and daily variations of N2O in a full-scale activated sludge anoxic/oxic process that consisted of an anoxic tank and three oxic tanks (oxic-1, oxic-2 and oxic-3), all of which except the final sedimentation tank were fully covered. Higher dissolved N2O (D-N2O) loading and gaseous N2O (G-N2O) emissions were observed for oxic-3 than for the anoxic, oxic-1, and oxic-2 tanks, implying that there was higher N2O production potential via nitrification in the latter stage of the oxic tank. Moreover, the sudden decrease in dissolved oxygen concentration after the peak was found to lead to abrupt production of D-N2O at oxic-3 in the anoxic/oxic process. The increases in AOB amoA, AOB nirK and the following AOB norB gene transcripts at the end of the oxic-2 tank suggested that nitrifier denitrification occurred to produce N2O under low dissolved oxygen conditions when the N2O peak was observed. Additionally, the much lower transcription levels of the two nosZ genes suggested lower N2O consumption. The N2O emission factors ranged from 0.087% to 0.302%, and lower N2O emission factors were observed during summer.

Recent studies have revealed that newly emerging RasV12-transformed cells are often apically extruded from the epithelial layer. During this cancer preventive process, cytoskeletal proteins plectin and Epithelial Protein Lost In Neoplasm (EPLIN) are accumulated in RasV12 cells that are surrounded by normal cells, which positively regulate the apical elimination of transformed cells. However, the downstream regulators of the plectin-EPLIN complex remain to be identified. In this study, we have found that paxillin binds to EPLIN specifically in the mix culture of normal and RasV12-transformed cells. In addition, paxillin is accumulated in RasV12 cells surrounded by normal cells. Paxillin, plectin and EPLIN mutually influence their non-cell-autonomous accumulation, and paxillin plays a crucial role in apical extrusion of RasV12 cells. We also demonstrate that in RasV12 cells surrounded by normal cells, acetylated tubulin is accumulated. Furthermore, acetylation of tubulin is promoted by paxillin that suppresses the activity of histone deacetylase (HDAC) 6. Collectively, these results indicate that in concert with plectin and EPLIN, paxillin positively regulates apical extrusion of RasV12-transformed cells by promoting microtubule acetylation. This study shed light on the unexplored events occurring at the initial stage of carcinogenesis and would potentially lead to a novel type of cancer preventive medicine.

Several lines of evidence have revealed that newly emerging transformed cells are often eliminated from the epithelium, though the underlying molecular mechanisms of this cancer preventive phenomenon still remain elusive. In this study, using mammalian cell culture systems we have identified plectin, a versatile cytoskeletal linker protein, as a novel regulator for apical extrusion of RasV12-transformed cells. Plectin is accumulated in RasV12 cells when they are surrounded by normal epithelial cells. Similarly, cytoskeletal proteins tubulin, keratin, and Epithelial Protein Lost In Neoplasm (EPLIN) are also accumulated in the transformed cells surrounded by normal cells. Knockdown or functional disruption of one of these molecules diminishes the accumulation of the others, indicating that the accumulation process of the individual protein mutually depends on each other. Furthermore, plectin-knockdown attenuates caveolin-1 (Cav-1) enrichment and PKA activity in RasV12 cells and profoundly suppresses the apical extrusion. These results indicate that the plectin-microtubules-EPLIN complex positively regulates apical elimination of RasV12-transformed cells from the epithelium in a coordinated fashion. Further development of this study would open a new avenue for cancer preventive medicine.

Newly emerging transformed cells are often eliminated from epithelial tissues. Recent studies have revealed that this cancer-preventive process involves the interaction with the surrounding normal epithelial cells; however, the molecular mechanisms underlying this phenomenon remain largely unknown. In this study, using mammalian cell culture and zebrafish embryo systems, we have elucidated the functional involvement of endocytosis in the elimination of RasV12-transformed cells. First, we show that Rab5, a crucial regulator of endocytosis, is accumulated in RasV12-transformed cells that are surrounded by normal epithelial cells, which is accompanied by up-regulation of clathrin-dependent endocytosis. Addition of chlorpromazine or coexpression of a dominant-negative mutant of Rab5 suppresses apical extrusion of RasV12 cells from the epithelium. We also show in zebrafish embryos that Rab5 plays an important role in the elimination of transformed cells from the enveloping layer epithelium. In addition, Rab5-mediated endocytosis of E-cadherin is enhanced at the boundary between normal and RasV12 cells. Rab5 functions upstream of epithelial protein lost in neoplasm (EPLIN), which plays a positive role in apical extrusion of RasV12 cells by regulating protein kinase A. Furthermore, we have revealed that epithelial defense against cancer (EDAC) from normal epithelial cells substantially impacts on Rab5 accumulation in the neighboring transformed cells. This report demonstrates that Rab5-mediated endocytosis is a crucial regulator for the competitive interaction between normal and transformed epithelial cells in mammals.

Background Although many patients complaining of chest pain visit the emergency department, very few are diagnosed with spontaneous pneumomediastinum (SPM). We present the management of 20 patients with SPM. Methods We retrospectively analyzed the clinical features, past history, chest X-rays (CXRs), 64-slice helical computed tomography (CT) images, and clinical course of 20 patients with SPM (19 men and 1 woman) who visited the emergency department of the Tokyo Metropolitan Bokutoh Hospital between 2005 and 2010. Results SPM predominantly resulted from physical exertion during such activities as sports and weight lifting (8 patients). The most common complaint was chest pain (15 patients), followed by dysphagia (10 patients), and dyspnea (8 patients). Subcutaneous emphysema was detected in 9 patients. The mean body mass index of the patients was 20.8±1.4kg/m 2 . Although CXR findings of pneumomediastinum were absent in 5 patients, CT showed findings of the Macklin effect in all patients—interstitial gas was observed in the perihilar area in all patients and in the peripheral lung area in 9 patients (45%). Ten patients were hospitalized. The others received ambulatory care in the form of analgesics without antibiotics. All patients fully recovered without complications. Conclusions Our results showed that 64-slice helical CT is more reliable than CXRs for diagnosing SPM. Moreover, some patients with SPM can be treated without hospitalization, thus decreasing medical expenses for these patients.

In Drosophila, normal and transformed cells compete with each other for survival in a process called cell competition. However, it is not known whether comparable phenomena also occur in mammals. Scribble is a tumor suppressor protein in Drosophila and mammals. In this study we examine the interface between normal and Scribble-knockdown epithelial cells using Madin–Darby Canine Kidney (MDCK) cells expressing Scribble short hairpin RNA (shRNA) in a tetracycline-inducible manner. We observe that Scribble-knockdown cells undergo apoptosis and are apically extruded from the epithelium when surrounded by normal cells. Apoptosis does not occur when Scribble-knockdown cells are cultured alone, suggesting that the presence of surrounding normal cells induces the cell death. We also show that death of Scribble-knockdown cells occurs independently of apical extrusion. Finally, we demonstrate that apoptosis of Scribble-knockdown cells depends on activation of p38 mitogen-activated protein kinase (MAPK). This is the first demonstration that an oncogenic transformation within an epithelium induces cell competition in a mammalian cell culture system.

Zirconium was loaded onto orange waste, a cheap and available agricultural waste in Japan, to investigate the feasibility of its utilization for phosphorus recovery from secondary effluent and side-stream liquid, which contain 5.9 and 68.2 mg/dm3 phosphorus, respectively. The phosphorus removal from side-stream liquid by using zirconium-loaded saponified orange waste (Zr-SOW) gel increased with an increasing solid/liquid ratio, and it was found that Zr-SOW gel showed better performance than zirconium ferrite. The prepared adsorbent was effective for phosphorus removal and exhibited a reasonably high adsorption capacity, twice than that of zirconium ferrite. The secondary effluent was treated in a column packed with Zr-SOW gel, and an dynamic adsorption capacity of 1.3 mol-P/kg was attained. The adsorbed phosphorus from the column was successfully eluted as a concentrated form by using a small amount of 0.2 M NaOH. Throughout the elution process, zirconium was not leaked from the adsorption gel.

The prognostic factors following aortic root reconstruction were studied in 19 patients including 13 with annuloaortic ectasia (AAE) and 6 without AAE (non-AAE). The preoperative diagnosis of six non-AAE patients was a dissecting aneurysm in five of the patients and supravalvular aortic stenosis associated with stenosis of the right coronary ostia in one patients. In the AAE group, the Bentall's method was initially selected in 11 patients and the Cabrol's method in the remaining 2 patients. In the non-AAE group, ascending aortic replacement was performed in 4 patients, patch plasty of the ascending aorta in 1 patient, and entry closure in the other patient. In this group, aorto-coronary bypass grafting using a saphenous vein graft was performed in 4 patients, ostioplasty of the right coronary artery (RCA) in 1, and the Bentall's method in 1. During the postoperative acute phase, one AAE patient died of acute myocardial infarction 3 days after surgery; the remaining 18 patients survived. In the follow-up study, 3 patients died of cardiac events which included two cardiac failures and one arrhythmia. The preoperative left ventricular diameter in the end-diastolic phase (LVDd) of 2 AAE patients who died of cardiac failure was 80 mm or larger and the left ventricular function remained unchanged after surgery. One non-AAE patient who underwent RCA ostioplasty suddenly died of arrhythmia. Postanastomotic leakage around the left coronary ostia associated with the patent Cabrol's trick occurred in 1 AAE patient and mitral valve regurgitation occurred in the other non-AAE patient. Reoperation using Cabrol's procedure and mitral valve replacement were performed for these 2 patients, respectively. Preoperative low cardiac function and large LVDd may influence the late results in AAE patients, therefore, earlier operations should be recommended.

We investigated the efficacy of University of Wisconsin (UW) solution for its ability to preserve hearts for 12 hours as compared with the results with modified Collins (MC) solution. After orthotopic transplantation, hearts stored in the UW solution (68%) showed significantly (p

We evaluated the availability of original “sandwich plasty” for the treatment of functional mitral regurgitation (FMR) associated with ischemic heart disease (IHD) and aortic valve disease (AVD). Forty-three patients were reviewed, including 27 IHD patients and 16 AVD patients. Preoperatively severe FMR was detected in 14 patients, moderate FMR in 26, and mild FMR in 3. The papillary muscle heads of anterior leaflets and posterior leaflets were approximated using Teflon-pledgeted 3-0 Ticron sutures at anterolateral and posteromedial commissural portions. After surgery, residual moderate FMR was observed in 1 patient and mild FMR in 3 patients. Tenting height of the mitral valve significantly decreased. FMR free rates 2 years after surgery were 93% among IHD patients and 83% in AVD patients. “Sandwich plasty” was simple and effective for the treatment of functional FMR caused by tethering effects due to left ventricular dilatation.

下水処理プラントからの二次処理水，返流水中のリンと下水汚泥焼却灰から硫酸や塩酸溶液により抽出したリンを，ミカン搾汁残渣を起源とした吸着剤（Zr-SOJR）によって吸着性を検討した．二次処理水と返流水のリン濃度は5.9mg/Lと68.2mg/Lであった．返流水のリン吸着率はバッチ実験において液固比の増加とともに増加した．その吸着能力はジルコニウムフェライトより高い値となった．二次処理水の吸着では，カラム通水実験により通水量が吸着剤容量の300倍以上で破過に達した．焼却灰からのリン抽出は酸溶媒の使用により効率的に行え，Zr-SOJRで吸着できることを示した．

Differentiation of mate recognition systems is one of the important steps for speciation in animals. For some insects, a contact sex pheromone present on the cuticular surface is indispensable in discriminating reproductive partners. In Callosobruchus species (Coleoptera: Chrysomelidae: Bruchinae), contact sex pheromones have been found in two species, Callosobruchus chinensis (L.) and Callosobruchus maculatus (Fabricius). It was suggested, however, that these two species lacked the ability to discriminate their conspecific and/or heterosexual partners. To elucidate this inconsistency, we verified the existence of contact sex pheromones from two other species, Callosobruchus rhodesianus (Pic) and Callosobruchus analis (Fabricius). As a result, unlike C. chinensis and C. maculatus, the males of C. rhodesianus and C. analis were able to discriminate their heterosexual partners. Comparing cross-copulation behavior, i.e., copulation behavior between two species, against these four species indicated that the mate recognition specificities were quite different. Males of C. rhodesianus and C. analis had highly species-specific mating behavior, whereas males of C. chinensis and C. maculatus were much less specific. These results indicate that variation in mate recognition can arise even among congeneric species living in a sympatric environment, and this variation might have arisen during species differentiation. Based on our results in combination with previous reports on interspecific competition, we suggest that the observed asymmetric cross-copulation behavior might be, at least partially, an adaptation for surviving interspecific competition.

We initiated an original papillary muscle head approximation procedure, commonly known as Sandwich plasty, for the treatment of ischemic mitral regurgitation (MR). In this study, we evaluated the appropriateness of this procedure for functional MR associated with aortic valve disease. Fifteen patients who had undergone Sandwich plasty combined with aortic valve surgery were included in this study. The mean age of the patients was 69 years. Predominant aortic valve diseases were regurgitation in 8 patients and stenosis in 7 patients. Aortic valve replacement was performed in 14 patients and David surgery in one. The mitral valve was approached through the aortic annulus in 9 patients (the transaortic group). Six other patients with mitral valve annulus of 30 mm or larger underwent concomitant mitral ring-annuloplasty through a left atrial incision (the LA group). The tenting height of the mitral valve and left ventricle diastolic diameter significantly decreased after surgery in both groups. After surgery, residual moderate or mild MR was detected in two patients in the transaortic group. In the LA group, residual mitral regurgitation was not detected. In the follow-up study, prominent MR occurred in two patients in the transaortic group. The MR free rate two years after surgery was 83% in the total patient population. Sandwich plasty was simple and effective in the treatment of functional mitral regurgitation combined with aortic valve surgery. A transaortic approach is effective in obviating a separate left atriotomy and reducing operation time.

心臓手術後急性期における短時間作用型β1 遮断薬・塩酸ランジオロール（オノアクト®）の有効性と安全な投与方法を検討した．心臓大血管手術後の成人10症例を対象とした．年齢は平均66（53～81）歳で男女比は9対1であった．主疾患は弁膜症7例，虚血性心疾患，胸部大動脈瘤，大動脈解離が各1例であった．頻脈の内訳は上室性（心房細粗動，上室性頻拍）が6例，心室性（心室性頻拍）が4例であった．頻脈性不整脈に対して，上室性ではIa，Ic群抗不整脈薬を，心室性ではIb群抗不整脈薬を第1選択として投与し，無効例でオノアクトを投与した．オノアクトの投与は初期負荷投与は行わず，静脈内少量持続投与とした．投与量は，導入量が平均0.018 mg/kg/分で，効果発現後は平均0.01 mg/kg/分で維持した．臨床効果：10例中9例で有効であった．上室性では，心房細動の4例中3例が洞調律に復帰し，1例で心房細動が持続したが心拍数の安定が得られた．心房粗動の1例では効果が不十分であり他剤に変更した．上室性頻拍の1例では心拍数の安定を得た．心室性では4例全てで頻拍症（VT）が消失した．血行動態：オノアクト投与により脈拍は140±42/分から95±21/分へと有意に（p＜0.05）低下した．収縮期血圧は，投与前の118±24 mmHgから106±21 mmHgへと軽度低下したが有意差はなかった．心係数（CI）は投与前の2.4±0.5 l/min/m2 より2.7±0.6 min/m2 へと有意に（p＜0.05）上昇した．拍出係数（SI）は17±7 ml/回/m2 より27±4 ml/回/m2 へと有意に（p＜0.05）増加した．副作用：血圧低下，徐脈，気管支喘息発作はなかった．血液生化学検査では異常を認めなかった．短時間作用型β遮断薬（オノアクト®）は心大血管手術後の頻脈性不整脈に対して有用であった．少量持続投与法は心血行動態には大きな影響が無く，開心術後でも安全に用い得る．

症例は60歳男性．2年前に大動脈炎症候群による大動脈弁閉鎖不全症，上行大動脈拡大に対して，大動脈弁置換術，上行大動脈置換術を受けた．以後ステロイドの内服を行いながら外来通院していたが，小脳梗塞で入院した．人工弁感染による弁輪部膿瘍と診断され，準緊急手術を施行した．手術では破壊された弁輪部を郭清・修復後，Ross手術を行い救命した．Ross手術は大動脈弁位人工弁感染に対する選択肢の一つと考えられた．

SummaryA 65-year-old female with catecholamine-dependent ischemic cardiomyopathy was admitted to our hospital. Preoperative examination revealed severe triple vessel coronary artery disease, severe mitral valve regurgitation and left ventricular (LV) dilatation associated with low ejection fraction. Coronary artery bypass grafting with four distal anastomoses, mitral valve plasty using original papillary muscle application method, LV volume reduction using overlapping method and biventricular pacing were performed. Postoperative course was uneventful and quality of life at 2 years after surgery was good. Active combined surgery has the possibility of improving the outcome of patients with severe ischemic cardiomyopathy.

Immature vascular smooth muscle cells (VSMCs) proliferate responding to extrinsic mitogens and accumulate in neointima after arterial injuries. Cell proliferation is positively regulated by cyclin/cyclin-dependent kinase (CDK) complex and negatively controlled by CDK inhibitors; CKIs such as p27 kip1 and p57 kip2 . In this study, embryonic rat thoracic aorta VSMCs; A10 were G0/G1 arrested by serum starvation, re-stimulated with serum, and harvested every four hours. Both CKIs co-expressed in quiescent VSMCs and rapidly diminished by stimulation. Protein level of p27 kip1 was regulated by both transcription and post-transcription, but that of p57 kip2 was mainly by post-transcription. Supplemental overexpression of p57 kip2 inhibited the activations of G1 cyclin/CDKs and subsequent hyperphosphorylations of all three retinoblastoma pocket proteins as well as G1/S transition of cell cycle. Our findings suggest that the downregulations of not only p27 kip1 , but also p57 kip2 responding to mitogenic stimulation, play key roles in the cell cycle progression of VSMCs.

We developed a new apparatus for long-term heart preservation that combines simple immersion with coronary perfusion. In a previous study, we reported that suppression of pro-inflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta), improved results after transplantation. In this study, we evaluated whether long-term preservation using our apparatus for continuous coronary perfusion, combined with suppression of pro-inflammatory cytokines, improves donor heart function after transplantation in a canine model.We used adult mongrel dogs in this study. Coronary vascular beds were washed with University of Wisconsin (UW) solution after arresting hearts with glucose-insulin-potassium solution. The heart was then excised and preserved for 12 hours with a combination of immersion and coronary perfusion using a preservation apparatus. Adult mongrel dogs were divided into 2 groups: the coronary perfusion (CP) group (n = 7) and the FR167653 (FR-CP) group (n = 6). In the CP group, we used a 4 degrees C UW solution for immersion and coronary perfusion. In the FR-CP group, we used a 4 degrees C UW solution supplemented with 20 mg/liter of the anti-inflammatory agent FR167653 for immersion and coronary perfusion. At 2 and at 3 hours after orthotopic transplantation, we compared hemodynamic parameters with pre-operative values in donor animals, with right atrial pressure at 10 mm Hg and with 5 microg/kg/min dopamine infusion. We compared serum concentrations of TNF-alpha from the coronary sinus and compared electron microscopic studies between the 2 groups.Three hours after transplantation, cardiac output (CO), left ventricular pressure (LVP), and -LVdp/dt were significantly greater (p0.05) in the FR-CP group than in the CP group (CO, 178% +/- 65% vs 93% +/- 40%; LVP, 115% +/- 22% vs 73% +/-26%; -LVdp/dt, 168% +/- 13% vs 61% +/- 17%, respectively). Electron microscopic studies showed that glycogen was well preserved in the FR-CP group compared with the CP group. Serum concentrations of TNF-alpha were decreased significantly in the FR-CP group compared with the CP group at 3 hours after reperfusion (161 +/- 54 pg/dl vs 642 +/- 636 pg/dl, respectively).Hemodynamics after transplantation were significantly better in the FR-CP group than in the CP group. The combined preservation method of continuous perfusion and immersion using our apparatus in conjunction with suppression of pro-inflammatory cytokines improves donor heart function after transplantation.

Summary Background Mast cells (MCs) arise from haematopoietic stem cells. We have recently reported that CD34+ progenitors derived from human bone marrow (BM) develop into tryptase+, chymase+ MCs when cultured in the presence of recombinant human stem cell factor (rhSCF) and recombinant human IL-6 (rhIL-6). In an experiment for the expression of chymase during differentiation, chymase+ cells were detected in human BM, but tryptase+ cells were not found. Objective The purpose of this study was to show the appearance of chymase+ cells in CD34+ cells with an origin different from MC differentiation. Methods CD34+ cells from human BM were sorted with anti-CD117 monoclonal antibody (mAb), and cytospins of CD34+, CD34+CD117+, or CD34+CD117− were prepared. These cells were cultured with rhSCF+rhIL-6 for 12 weeks. Some of the cells were subjected to either histological stain with Wright–Giemsa or immunocytochemistry with anti-chymase mAb. Real-time RT-PCR was also performed to compare the transcriptional level of chymase from each cell preparation. Results Chymase was expressed in CD34+ cells as well as human MCs by immunocytochemistry. Substantial CD34+CD117− cells, but not CD34+CD117+ cells, were stained immunocytochemically with anti-chymase mAb. For 1 week culture with rhSCF+rhIL-6, no cells expressed chymase in any preparation. Real-time RT-PCR revealed positivity for chymase mRNA in CD34+ cells, but it reduced at 1 week of culture, and increased as cells developed into MCs. Chymase mRNA in CD34+CD117+ cells was negligible compared with that in CD34+CD117−. Tryptase mRNA was below the detectable level in CD34+ cells, and increased along with MC differentiation. After 12 weeks of culture, CD34+CD117+ developed predominantly into MCs, whereas CD34+CD117− developed into monocytes/macrophages. Conclusion Our findings suggested that chymase is present not only in MCs but also in CD34+CD117− BM progenitors, but that its origin is different from the MC lineage.

A new member of the Y-box protein family of the silkworm Bombyx mori (BYB) was co-purified with the fibroin gene enhancer-binding protein FMBP-1, and stimulated the binding of FMBP-1 to its cognate DNA element. However, the stimulatory effect was not specific to FMBP-1, BYB also enhancing the binding of mammalian transcription factors OTF2, SP1 and AP2 to their specific binding elements. Besides the above transcription regulatory factors, BYB facilitated the binding of basal transcription factor TBP, and enhanced transcription from the adenovirus 2 major late promoter in a reconstituted transcription system. Moreover, BYB stimulated the reactions of some restriction endonucleases under cold conditions. The C-terminal region of BYB was sufficient for these stimulatory effects, and the highly conserved cold shock domain (CSD) in the N-terminal region was dispensable. GST-pull down experiments showed that the C-terminal region could interact with DNA independently of the CSD. The above results suggest that the C-terminal region of BYB causes the active interaction of various DNA binding proteins with their targets. Such a function of the C-terminal region of BYB may partly explain the functional diversity of Y-box proteins.

The CagA protein of Helicobacter pylori, which is injected from the bacteria into bacteria-attached gastric epithelial cells, is associated with gastric carcinoma. CagA is tyrosine-phosphorylated by Src family kinases, binds the SH2 domain-containing SHP-2 phosphatase in a tyrosine phosphorylation-dependent manner, and deregulates its enzymatic activity. We established AGS human gastric epithelial cells that inducibly express wild-type or a phosphorylation-resistant CagA, in which tyrosine residues constituting the EPIYA motifs were substituted with alanines. Upon induction, wild-type CagA, but not the mutant CagA, elicited strong elongation of cell shape, termed the "hummingbird" phenotype. Time-lapse video microscopic analysis revealed that the CagA-expressing cells exhibited a marked increase in cell motility with successive rounds of elongation-contraction processes. Inhibition of CagA phosphorylation by an Src kinase inhibitor, PP2, or knockdown of SHP-2 expression by small interference RNA (siRNA) abolished the CagA-mediated hummingbird phenotype. The morphogenetic activity of CagA also required Erk MAPK but was independent of Ras or Grb2. In AGS cells, CagA prolonged duration of Erk activation in response to serum stimulation. Conversely, inhibition of SHP-2 expression by siRNA abolished the sustained Erk activation. Thus, SHP-2 acts as a positive regulator of Erk activity in AGS cells. These results indicate that SHP-2 is involved in the Ras-independent modification of Erk signals that is necessary for the morphogenetic activity of CagA. Our work therefore suggests a key role of SHP-2 in the pathological activity of H. pylori virulence factor CagA.

Aims : The hemodynamic and metabolic effects of hypertonic saline (HTS) infusion were studied in normovolemic dogs. Methods : Seven adult mongrel dogs, weighing 10 to 15kg, were used in this study. Under general anesthesia, the pericardium was opened via a right thoracotomy. High-dosage of HTS (10% NaCl, 5ml/kg) was infused intravenously within a five minute period. Results : Systemic blood pressure significantly increased from 110± 10mmHg to 125 ± 10mmHg. Central venous pressure and left atrial pressure also significantly increased (4.6 ± 1.7 → 9.3±1.9mmHg, 3.5 ± 1.4 → 5.7 ± 2.4mmHg, respectively). Cardiac output increased significantly from 2.0±0.2L/min to 3.0±0.2L/min 10 minutes after HTS infusion. Arterial blood gas analysis revealed that pH, base excess and HCO3- had decreased significantly 10 minutes after HTS infusion. Hemoglobin and hematocrit levels significantly decreased after HTS infusion. Although O2 content decreased significantly, the myocardial O2 extraction rate did not change. Serum sodium and chloride values increased significantly and sustained those levels until 60 minutes after HTS infusion. Conclusions : In conclusion, HTS infusion remains a simple and valuable clinical procedure to use in restoring adequate hemodynamic conditions after hypovolemic shock.

慢性肺動脈血栓塞栓症に対して, 2種類の湾曲鉗子を使用し超低体温循環停止法を用いず血栓内膜摘除を行い得たので報告する.患者は44歳の男性で, 呼吸困難, 左下肢浮腫のため平成14年6月当院に紹介された.胸部CT上, 両側肺動脈中枢側に血栓を認め抗凝固療法が無効で, 主肺動脈まで血栓塞栓が増悪したため緊急手術を施行した.完全体外循環, 中等度低体温下に主肺動脈を横切開し, 心耳鉗子および心房鉗子を各々右および左肺動脈末梢側に挿入して21gの血栓内膜摘除を行った.循環停止とはせず肺動脈末梢からの血液back flowによる視野障害を予測し, back flowによるwash out効果と最小限の肺動脈切開による鈍的摘除を組み合わせて低侵襲手術を試みた.大動脈遮断時間43分, 体外循環時間98分であった.術後CTで左肺動脈下葉枝の血栓残存を認めたが, 術前のNYHA IV度からII度まで改善し日常生活に復帰している.

[Purpose] In this report, the clinical features and surgical treatment highlighting areas of concern in treatment of inflammatory abdominal aortic aneurysm (IAAA) are discussed together with a review of literature. [Patients and Methods] Among 287 patients undergoing repair for abdominal aortic aneurysm (AAA) between 1992 and 2003, IAAA was observed in six (2.1%). All six patients were male, and the mean age at the time of surgery was 66 ± 8 years, ranging from 52 to 72. Four patients (67%) had been previously treated for hypertension, and no genitourinary dysfunction occurred in any patients. Two patients (33%) complained of lumbago prior to surgery. The preoperative diagnosis of IAAA was made in only two patients (33%) because the mantle sign and other typical appearances of IAAA were identified in only these patients. [Results] Five patients underwent elective surgery and there was one case of urgent surgery. There was massive adhesion in five patients developed between the aneurysm and the mesentery and/or intestine. Postoperative ileus due to duodenal stenosis occurred in one patient. [Conclusion] The preoperative diagnosis of IAAA is sometimes difficult, and the operation for IAAA should be modified to avoid excessive dissection.

A 62-year-old female with Osler's disease was admitted to our hospital because of fever and cardiac failure. The patient had undergone a mitral valve replacement (MVR) using a Carpentier-Edwards prosthetic valve 14 years earlier. A bacterial examination of arterial blood identified Streptococcus mitis. No arteriovenous malformations were detected in visceral organs. The patient underwent MVR using the same prosthetic xenograft after conservative treatment and management of repetitive epistaxis and decayed teeth. Intra- and postoperative bleeding were typical of a mitral valve reoperation. This is the first reported experience, to the best of our knowledge, of a second MVR in a patient with Osler's disease.