Complexity of projected images of convex subdivisions

AbstractLet S be a subdivision of Rd into n convex regions. We consider the combinatorial complexity of the image of the (k - 1)-skeleton of S orthogonally projected into a k-dimensional subspace. We give an upper bound of the complexity of the projected image by reducing it to the complexity of an arrangement of polytopes. If k = d − 1, we construct a subdivision whose projected image has Ω(n⌊(3d−2)/2⌋) complexity, which is tight when d ⩽ 4. We also investigate the number of topological changes of the projected image when a three-dimensional subdivision is rotated about a line parallel to the projection plane

One-Step Generation of Multiple Gene-Edited Pigs by Electroporation of the CRISPR/Cas9 System into Zygotes to Reduce Xenoantigen Biosynthesis

Xenoantigens cause hyperacute rejection and limit the success of interspecific xenografts. Therefore, genes involved in xenoantigen biosynthesis, such as GGTA1, CMAH, and B4GALNT2, are key targets to improve the outcomes of xenotransplantation. In this study, we introduced a CRISPR/Cas9 system simultaneously targeting GGTA1, CMAH, and B4GALNT2 into in vitro-fertilized zygotes using electroporation for the one-step generation of multiple gene-edited pigs without xenoantigens. First, we optimized the combination of guide RNAs (gRNAs) targeting GGTA1 and CMAH with respect to gene editing efficiency in zygotes, and transferred electroporated embryos with the optimized gRNAs and Cas9 into recipient gilts. Next, we optimized the Cas9 protein concentration with respect to the gene editing efficiency when GGTA1, CMAH, and B4GALNT2 were targeted simultaneously, and generated gene-edited pigs using the optimized conditions. We achieved the one-step generation of GGTA1/CMAH double-edited pigs and GGTA1/CMAH/B4GALNT2 triple-edited pigs. Immunohistological analyses demonstrated the downregulation of xenoantigens; however, these multiple gene-edited pigs were genetic mosaics that failed to knock out some xenoantigens. Although mosaicism should be resolved, the electroporation technique could become a primary method for the one-step generation of multiple gene modifications in pigs aimed at improving pig-to-human xenotransplantation

Anti-angiogenic effect of siphonaxanthin from green alga, Codium fragile.

Since anti-angiogenic therapy has becoming a promising approach in the prevention of cancer and related diseases, the present study was aimed to examine the anti-angiogenic effect of siphonaxanthin from green alga (Codium fragile) in cell culture model systems and ex vivo approaches using human umbilical vein endothelial cells (HUVECs) and rat aortic ring, respectively. Siphonaxanthin significantly suppressed HUVEC proliferation (p<0.05) at the concentration of 2.5 μM (50% as compared with control) and above, while the effect on chemotaxis was not significant. Siphonaxanthin exhibited strong inhibitory effect on HUVEC tube formation. It suppressed the formation of tube length by 44% at the concentration of 10 μM, while no tube formation was observed at 25 μM, suggesting that it could be due to the suppression of angiogenic mediators. The ex vivo angiogenesis assay exhibited reduced microvessel outgrowth in a dose dependent manner and the reduction was significant at more than 2.5 μM. Our results imply a new insight on the novel function of siphonaxanthin in preventing angiogenesis related diseases

Efficient generation of GGTA1-deficient pigs by electroporation of the CRISPR/Cas9 system into in vitro-fertilized zygotes

Background: Xenoantigens are a major source of concern with regard to the success of interspecific xenografts. GGTA1 encodes α1,3-galactosyltransferase, which is essential for the biosynthesis of galactosyl-alpha 1,3-galactose, the major xenoantigen causing hyperacute rejection. GGTA1-modified pigs, therefore, are promising donors for pig-to-human xenotransplantation. In this study, we developed a method for the introduction of the CRISPR/Cas9 system into in vitro-fertilized porcine zygotes via electroporation to generate GGTA1-modified pigs. Results: We designed five guide RNAs (gRNAs) targeting distinct sites in GGTA1. After the introduction of the Cas9 protein with each gRNA via electroporation, the gene editing efficiency in blastocysts developed from zygotes was evaluated. The gRNA with the highest gene editing efficiency was used to generate GGTA1-edited pigs. Six piglets were delivered from two recipient gilts after the transfer of electroporated zygotes with the Cas9/gRNA complex. Deep sequencing analysis revealed that five out of six piglets carried a biallelic mutation in the targeted region of GGTA1, with no off-target events. Furthermore, staining with isolectin B4 confirmed deficient GGTA1 function in GGTA1 biallelic mutant piglets. Conclusions: We established GGTA1-modified pigs with high efficiency by introducing a CRISPR/Cas9 system into zygotes via electroporation. Multiple gene modifications, including knock-ins of human genes, in porcine zygotes via electroporation may further improve the application of the technique in pig-to-human xenotransplantation

A Rare Complication:Misdirection of an Indwelling Urethral Catheter into the Ureter

We report 3 patients with the rare complication of an indwelling urethral catheter misdirected into the ureter. This is the largest series to date. Patients were referred to us for a variety of reasons following exchange of their chronic indwelling urinary catheters. CT in all cases demonstrated the urinary catheters residing in the left ureter. The ages of the patients were 37, 67 and 81 years old. All patients suffered from neurogenic bladder. Two patients were female, one was male, and 2 of the 3 had a sensory disorder inhibiting their pain response. The catheters were replaced with open-end Foley catheters. Extensive follow-up CT scans were obtained in one case, demonstrating improvement of hydronephrosis and no evidence of ureteral stenosis. Cystoscopy in this patient demonstrated normally positioned and functioning ureteral orifices. Although the placement of an indwelling urethral catheter is a comparatively safe procedure, one must keep in mind that this complication can occur, particularly in female patients with neurogenic bladder. CT without contrast is a noninvasive, definitive diagnostic tool

The Combination of Gemcitabine, Cisplatin, and Paclitaxel as Salvage Chemotherapy for Advanced Urothelial Carcinoma

There is no standard second-line or salvage treatment for advanced urothelial carcinoma (UC). Here we investigated the efficacy and safety of gemcitabine, cisplatin, and paclitaxel (GCP) combination chemotherapy as salvage chemotherapy for advanced UC. We retrospectively analyzed the cases of 23 patients with advanced UC who showed progression or recurrence after cisplatin-based chemotherapy. Gemcitabine (1000 mg/m2), and paclitaxel (80 mg/m2) were administered on days 1 and 8. Cisplatin (70 mg/m2) was administered on day 1. The 3-week cycle regimen was repeated until disease progression if it had no intolerable toxicity. The overall response rate was 61% (95%CI, 41-78%). The median overall survival and progression-free survival times were 14 months and 5.5 months, respectively. Of the already known risk factors of chemotherapy for advanced UC, only the performance status was a prognostic factor for OS. Overall, 16 of the 23 patients (70%) experienced grade 3/4 toxicities, and no fatal adverse events were observed. GCP therapy was a promising option as second-line or salvage therapy for advanced UC

Salmonella Contamination of Rivers and Sewages in Fukuyama City

地方都市におけるサルモネラによる環境汚染の実態を知る目的で，昭和41・42年に広島県東部地区福山市における河水，下水などを対象としてサルモネラの検索を行なった.その結果，以下のような成績が得られた. 1. サルモネラ陽性率は河水が23.6% (13/55)，下水が22.2% (8/36)，と場汚水が36.8% (7/19)の成績であった. 2. これらの陽性率は，東京，大阪など大都市における河川および下水のサルモネラ陽性率に比べると，まだ低い状況にある. 3. 28例の陽性検体から分離された33株の血清型は，S. derby，S. typhimurium，S. montevideo，S. thompson，S. mikawashima，S. narashino，S. enteritidis，S. meleagridis，S. give，S. senftenberg，S. westerstedeの11種類に分類された. 4. S. westerstedなど，広島県で今までにみられたことが無い新しい菌型が多数検出されたことから，当地方におけるサルモネラによる環境汚染の進みつつあることが推定できる. 5. このようなサルモネラによる環境汚染の実態から，今後の食品衛生上に及ぼす影響などについて指摘した.One hundred and ten water samples collected from the rivers, sewages and slaughterhouse in Fukuyama city in the eastern district of Hiroshima prefecture were examined for Salmonella organisms，from October 1967 to December 1968. It was found that 23.6% (13/55) of the rivers, 22.2% (8/36) of the sewages, and 36.8% (7/19) of the sewages of the slaughterhouse were contaminated by this organism. These results show that the percentage of contamination is smaller in Fukuyama city than that in a large cities，such as Tokyo or Osaka. Thirty three strains of Salmonella isolated from water samples were divided into the following 11 serotypes: S. derby，S. typhimurium，S. montevideo，S. thompson，S. mikawashima，S. narashino，S. enteritidis，S. meleagridis，S. give，S. senftenberg and S. westerstede. It might be presumed that the environmental contamination of the area by the organisms is increasing gradually，since new types of Salmonella，e.g. S. westerstede etc. never found in Hiroshima prefecture before，were isolated in the area

Prostaglandin D2 Reinforces Th2 Type Inflammatory Responses of Airways to Low-dose Antigen through Bronchial Expression of Macrophage-derived Chemokine

PGD2, a lipid mediator released from mast cells, is known to participate in allergic reactions. However, the mechanism by which PGD2 contributes to such reactions remains unclear. We established a novel experimental model of asthma that permitted direct assessment of the role of PGD2 in airway inflammation. Antigen-sensitized mice were exposed to aerosolized prostaglandin D2 (PGD2) 1 d before challenge with low-dose aerosolized antigen. Not only the numbers of eosinophils, lymphocytes, and macrophages but also the levels of IL-4 and IL-5 in bronchoalveolar lavage fluid were higher in PGD2-pretreated mice than in control mice. The expression of macrophage-derived chemokine (MDC), a chemoattractant for Th2 cells, was greater in PGD2-pretreated mice than in control. Injection of anti-MDC antibody into PGD2-pretreated mice markedly inhibited inflammatory cell infiltration as well as Th2 cyto-kine production after antigen challenge. These results indicate that PGD2 accelerates Th2 type inflammation by induction of MDC. Our results suggest that this mechanism may play a key role in the development of human asthma and that MDC might be a target molecule for therapeutic intervention