First report of Puccinia xanthii causing rust disease on Xanthium orientale in Korea.

Puccinia xanthii Schw. is a microcyclic rust fungus, first found on Xanthium strumarium Lour in North Carolina, the United States. This rust fungus is native to the continental United States, Hawaii, Mexico, and the West Indies (Arthur 1934). It has become notoriously invasive and is now distributed in the Europe (Bulgaria, France, Hungary, Italy, Romania, Spain, and the former Yugoslavia), India, Indonesia, Australia, and South Africa (Parmelee 1969; Alcorn 1976; Wahyuno 2012). In East Asia, the fungus has been reported in Japan (Hiratsuka et al. 1992) and China (Zhao et al. 2014) but not in Korea. It has been reported mainly on the invasive weeds Xanthium and Ambrosia species. In addition, it rarely occurs on sunflowers (Helianthus spp.) in Australia (Alcorn 1976), South Africa (Pretorius et al. 2000), and North America (Gulya and Charlet 2002). In Korea, rust disease symptoms caused by a Puccinia fungus were first found on X. orientale L. at the roadside of Okcheon-gun, Chungcheongbuk-do (36 27'95.428"N 127 66'26.378"E) in October 2021 and were repeatedly observed in the same site in 2022. The similar symptom was additionally found on X. orientale in Yesan-gun, Oct. 2022. The symptoms were brown spots on round chlorotic haloes on the adaxial leaf surface and dark brown pustules on the abaxial leaf surface. Telia were brown to dark brown, round, mostly grouped, 0.28-0.61 mm in diameter, and mainly formed on the abaxial leaf surface but sometimes on the adaxial leaf surface. Teliospores were two-celled, pedicellate, and measured 37.6-110 × 12.4-21.5 µm in size; the wall was yellowish or almost colorless, smooth, 1.2-2.6 µm thick at the sides, and up to 7.4 µm thick at the apex. The morphological characteristics of the teliospores were identical to those of P. xanthii described by Arthur (1934) and Parmelee (1969). Based on phylogenetic analyses (e-Xtra 2) of the internal transcribed spacer (ITS) and partial large subunit (LSU) rDNA extracted from the teliospores, they were identified as P. xanthii. BLAST analysis showed that the sequences had high homologies (over 99.82%) with the reference strains of P. xanthii (EF635903 and KX999896). The representative specimens were preserved at the Animal and Plant Quarantine Agency Herbarium (PQK211005 for Okcheon-gun isolate and PQK220913 for Yesan-gun) and the sequences were deposited in GenBank (OR958716 and OR958692). A pathogenicity test was performed by dropping a suspension of germinating teliospores and basidiospores onto the adaxial leaf surfaces of apparently healthy X. orientale plants in Oct. 2022, using the isolate PQK220913 (OR958692). The three inoculated plants were placed together with three controls treated with only distilled water, in the dark at saturated humidity for 24 hours in an isolated greenhouse. After two weeks, typical rust symptoms were observed in the three infected plants, whereas no symptoms appeared in the control plants (e-Xtra 1). The causal fungus was identified as P. xanthii based on host relationships, successful experimental inoculation, morphological characteristics, and sequence similarity of partial DNA fragments. To our knowledge, this is the first report of P. xanthii on X. orientale in Korea. P. xanthii was additionally confirmed on X. orientale in Gumi-si, Boeun-gun, Seongju-gun, Naju-si, and Gunsan-si in 2023, indicating its wide distribution in Korea. It is expected that P. xanthii could be a candidate as a biological agent for controlling the invasive weed, X. orientale.

First report of a rust disease of Lycium chinense caused by Puccinia tumidipes in Korea.

Lycium chinense Mill is a deciduous broad-leaved shrub belonging to the Solanaceae family and, is widely distributed throughout Korea. This plant is native to, or cultivated for various oriental medicinal purposes in, multiple regions of Asia, including Korea, China, and Japan (Lee 1982; Kim et al. 1994). Eleven Puccinia species have been reported to infect Lycium species (Otálora et al. 2018). In May and October 2022, symptoms of rust disease caused by Puccinia sp. were observed on almost all the leaves of about 60 sprawling stems of L. chinense plants on the seashore of Jeju island, Korea (33°14'15.0835″N, 126°30'53.40E). Brownish red (uredinium) or blackish brown (telium) pustules were observed on upper and lower surfaces of infected leaves. These symptoms were observed on about 40 L. chinense plants, barely growing between rocks on the seashore of Ulsan Metropolitan City, and on the about 20 L. chinense plants on a small home garden of Jindo-gun, Korea, in June and October 2023, respectively. Uredinia were amphigenous, individually scattered, but sometimes formed groups of two or three on leaves and sepals, ferruginous, pulverulent, and surrounded by a ruptured epidermis, often developing into blackish telia. Urediniospores were either ellipsoid or ovoid, approximately 29.3-34.9 × 17-24.3 µm, with yellowish walls, 1-2 µm thick. The germ pores were bizonate, and each band contained four pores covered by low papillae. Blackish-brown telia were observed on both leaf surfaces. Teliospores were broadly ellipsoidal, and rounded at the apex and towards the base. They were measured approximately 37.1-53.4 × 25-34.5 µm. The walls were light chestnut-brown and 2-3.7 µm thick, apically up to 5-9 µm thick. The swollen pedicel was persistent, basal, hyaline, smooth, and similar in length to the spores (Fig. 1). These morphological characteristics were similar to those of P. tumidipes, as described by Otálora et al. (2018). The representative specimens were preserved at the Animal and Plant Quarantine Agency Herbarium (PQK220531, -230605, and -231026). The fungal internal transcribed spacer (ITS2) and cytochrome oxidase subunit 3(CO3) regions were amplified from the total DNA of the isolates, using the primer pairs ITS5, ITS4, CO3F1, and CO3R1 for phylogenetic analysis (White et al. 1990; Vialle et al. 2009). PCR products were sequenced (Celemics, Seoul, Korea), and deposited in GenBank (Accession numbers are shown in Fig. 2.). The combined ITS2 and CO3 sequences were grouped with those of other isolates of P. tumidipes in the phylogenetic tree (Fig. 2). In November 2022, three pathogenicity tests were conducted using a urediniospore suspension made with the PQK220531 isolate in sterile distilled water. The suspension was smeared onto the upper surface of healthy L. chinense leaves. The three inoculated plants were kept in the dark at saturated moisture levels for 24 hours and placed in an isolated glasshouse together with the three control plants. After two weeks, uredinia of P. tumidipes were observed on the leaves of the inoculated plants, but not on the control plants. Although no spermogonial or aecial stage has been observed in Korea, our study has proven that P. tumidipes is the causal fungus of rust disease in L. chinense. This result is also the first discovery of the New-World P. tumidipes in Asia, showing this fungus is not limitedly distributed in America and suggesting further surveys be done on its exact geographical distribution.

Malva sylvestris, a new host of Puccinia modiolae in Korea.

Malva sylvestris (Malvaceae), known as common mallow, is native to Europe, western Asia, and northern Africa. It was intentionally introduced to Korea as an ornamental plant in the early 20th century, and has become partly naturalized in several areas including the woods (Jung et al. 2017). Among nine microcylic Puccinia species attacking the Malvaceae plants, three species of P. heterospora, P. malvacearum, and P. modiolae have been reported on M. sylvestris (Classen et al. 2000, Colenso 1885, McKenzie 1998 and Melo et al. 2012). Only P. modiolae has been found on Alcea rosea and M. verticillata, and not M. sylvestris in Korea (Lee et al. 2022; Ryu et al. 2022). In August 2022, rust disease symptoms of a Puccinia fungus were observed on some overgrown seedlings of M. sylvestris, which were neglected in containers after sales at a wholesale nursery (36°50'19.8″N, 128°55'28.7″E) in Bonghwa, Korea. Typical rust spots were observed around 60% (on 111 seedlings of the 186 seedlings of M. sylvestris). The brown spots were produced on round chlorotic haloes on the adaxial leaf surface, and brown to dark brown pustules on the abaxial. Subepidermal spermogonia on the adaxial, were obovoid, and 112.1-160.0 × 88.7-149.3 µm in size. Telia were golden-brown to dark brown, round, mostly grouped, and 0.30-0.72 mm in diameter, and mainly hypophyllus. Fusoid teliospores were two-celled, rarely one- or three-celled, 36.2-92.3 × 10.6-19.3 µm in size, with many anomalies appearing notched at apex; wall was yellowish or almost colorless, smooth, 1.0-2.6 µm thick at the sides, and up to 6.8 µm thick at the apex; pedicel was hyaline, thick wall, persistent, and (39.3-)60.4-154.6(-189.9) µm long. Based on these morphological features together with the results of the phylogenetic analyses (e-Xtra 2) using internal transcribed spacer (ITS) and partial large subunit (LSU) sequences according to the method described by Ryu et al. (2022), the fungus was identified as an autoecious P. modiolae, recently reported on M. verticillate and A. rosea in Korea (Lee et al. 2022; Ryu et al. 2022). A representative sample was deposited in the Animal and Plant Quarantine Agency Herbarium (PQK220818). Pathogenicity tests were done using three host plants: M. sylvestris, M. verticillate and A. rosea. Three to four leaf discs with basidiospore-bearing telia were placed on the upper surfaces of healthy young leaves of the seedlings. Three replicates of each host plant set including an untreated control were tested. The plants were kept in an isolated glass house. At ten to twelve days after inoculation, typical telial spots of P. modiolae were recovered, but not in the control plants, showing all three tested species were highly susceptible (e-Xtra 1). The ITS and LSU sequences obtained from the genomic DNAs of each newly recovered rust spot were consistent with that of the inoculum (accession no. OQ542745). The previous A. rosea isolate (OP369290 by Ryu et al. 2022) also showed the pathogenesis on M. sylvestris and M. verticillata by the same tests, mentioned above (e-Xtra 1). To date, only one collection of P. modiolae on M. sylvestris has been reported in Louisiana, the United States (Aime and Abbasi 2018). The results of this study show that P. modiolae is firstly confirmed as the causal rust fungus of M. sylvestris and the same causal agent of M. verticillate and A. rosea rust disease, recently reported in Korea.

First Report of Rust Disease on Alcea rosea Caused by Puccinia modiolae in Korea.

Alcea rosea, in the family Malvaceae, is a biennial plant native to China and is grown typically for gardening in Korea (Lee 2003). Seven microcyclic Puccinia species have been reported on A. rosea: P. heterogenea, P. heterospora, P. lobata, P. malvacearum, P. platyspora, P. sherardiana, and P. modiolae (Demers et al. 2015; Aime and Abbasi 2018). In early May 2022, characteristic symptoms of rust were observed on four of ten seedlings of A. rosea purchased at a wholesale nursery (36°50'19.8″N, 128°55'28.7″E) in Bonghwa, Korea. Rust spots were present on almost 90% of the 1,000 seedlings of A. rosea in that nursery during our survey in late May. Through a distribution survey from June to July 2022, similar symptomatic leaves were additionally collected from the leaves of A. rosea grown in gardens at five sites in Gimcheon (two sites), Gumi (one), Seongju (one), and Busan (one). Spots were yellow-orange the center surrounded by chlorotic haloes on the adaxial leaf surface, and reddish-brown or dark brown pustules on the abaxial leaf surface. Over time, the spots enlarged and coalesced, causing the decay of large sections of the leaves, and heavily infected leaves fell early. Spermogonia, produced at the center of the chlorotic spot on the adaxial leaf surface, were subepidermal, obovoid, and 113.2-164.5 × 97.6-153.3 µm in size. Telia were reddish-brown to dark brown, round, mostly grouped, 0.28-0.61 mm in diameter, and mainly formed on the abaxial leaf surface but sometimes on the adaxial leaf surface also. Teliospores were two-celled, but rarely one- or three-celled, and were fusoid and 37. 6-110 × 12.4-21.5 µm in size; the wall was yellowish or almost colorless, smooth, 1.2-2.6 µm thick at the sides, and up to 7.4 µm thick at the apex. The morphological characteristics were similar to those of P. modiolae, although the teliospores in our study were longer than those observed by Aime and Abbasi (2018). For phylogenetic analysis, genomic DNA was extracted from the teliospores of each regional specimen. Partial 18S, internal transcribed spacer (ITS), and partial 28S sequences were amplified using primers NS1, ITS4, ITS5, and LR11. The PCR products were sequenced (Celemics, Seoul, Korea) and deposited in GenBank. The ITS-partial large subunit (LSU) sequence and 28S sequences had 100% homology with other P. modiolae sequences deposited in GenBank (accession numbers are shown in Fig. 2). In the phylogenetic trees of the ITS and LSU sequences, the isolates collected in this study were grouped with the reference sequences of P. modiolae, including the Korean isolate (ON631218) recently reported on Malva verticillata by Lee et al. (2022). For the pathogenicity test, the teliospores with germinating basidiospores were suspended in sterile distilled water and smeared on the upper surface of asymptomatic A. rosea leaves in August. The inoculated plants were sprayed with distilled water and kept in the dark with saturated moisture for 24 h in an isolated glass house of the Animal & Plant Quarantine Agent. After 2 weeks, typical rust spots and telia of P. modiolae were observed on the leaves of the inoculated plants, but not in the control plants, which were only sprayed with distilled, sterilized water and otherwise treated similarly to the inoculated plants. The results of this study show that the casual fungus is P. modiolae, which has been commonly found in A. rosea in Korea. To the best of our knowledge, this is the first report of P. modiolae in A. rosea in Korea.

Effects of community-based rehabilitation program on activities of daily living and cognition in elderly chronic stroke survivors.

[Purpose] The aim of this study was to examine the effects of community-based rehabilitation program in chronic stroke patients. [Subjects and Methods] Eleven subjects received community-based rehabilitation program ten times for ten months. The main outcome measures were the Modified Barthel Index score for activities of daily living and the Korean Mini-Mental State Examination score for cognition. [Results] The results of the study demonstrated that the community-based rehabilitation program improved activities of daily living performance and cognition significantly. [Conclusion] Based on the study results, the community-based rehabilitation program is an effective method for improving activities of daily living performance and cognitive function in elderly patients with chronic stroke.

MicroRNA844-Guided Downregulation of Cytidinephosphate Diacylglycerol Synthase3 (CDS3) mRNA Affects the Response of Arabidopsis thaliana to Bacteria and Fungi.

Despite the fact that a large number of miRNA sequences have been determined in diverse plant species, reports demonstrating the functional roles of miRNAs in the plant response to pathogens are severely limited. Here, Arabidopsis thaliana miRNA844 (miR844) was investigated for its functional role in the defense response to diverse pathogens. Transgenic Arabidopsis plants overexpressing miR844 (35S::miR844) displayed much more severe disease symptoms than the wild-type plants when challenged with the bacterium Pseudomonas syringae pv. tomato DC3000 or the fungus Botrytis cinerea. By contrast, a loss-of-function mir844 mutant showed an enhanced resistance against the pathogens. Although no cleavage was observed at the predicted cleavage site of the putative target mRNA, cytidinephosphate diacylglycerol synthase3 (CDS3), cleavage was observed at 6, 12, 21, or 52 bases upstream of the predicted cleavage site of CDS3 mRNA, and the level of CDS3 mRNA was downregulated by the overexpression of miR844, implying that miR844 influences CDS3 transcript level. To further confirm that the miR844-mediated defense response was due to the decrease in CDS3 mRNA level, the disease response of a CDS3 loss-of-function mutant was analyzed upon pathogen challenge. Increased susceptibility of both cds3 mutant and 35S::miR844 plants to pathogens confirmed that miR844 affected the defense response by downregulating CDS3 mRNA. The expression of miR844 was decreased, and the CDS3 transcript level increased upon pathogen challenge. Taken together, these results provide evidence that downregulation of miR844 and a concomitant increase in CDS3 expression is a defensive response of Arabidopsis to bacteria and fungi.

MicroRNA400-guided cleavage of Pentatricopeptide repeat protein mRNAs Renders Arabidopsis thaliana more susceptible to pathogenic bacteria and fungi.

Although a large number of microRNAs (miRNAs) have been identified in different plant species, the functional roles and targets of the majority of miRNAs have not yet been determined. Here, Arabidopsis thaliana miRNA400 (miR400) was investigated for its functional role in the defense response to diverse pathogens. Transgenic Arabidopsis plants that overexpress MIR400 (35S::MIR400) displayed much more severe disease symptoms than the wild-type plants when infected with the bacterium Pseudomonas syringae pv. tomato DC3000 or the fungus Botrytis cinerea. MiR400 guided the cleavage of two genes (At1g06580 and At1g62720) encoding pentatricopeptide repeat (PPR) proteins. To confirm further that the miR400-mediated defense response was due to the cleavage of PPR mRNAs, loss-of-function mutant and artificial miRNA-mediated knockdown mutants of PPR were generated, and their disease responses were analyzed upon pathogen challenge. Similar to the 35S::MIR400 plants, the ppr mutants displayed much more severe disease symptoms than the wild-type plants when challenged with the pathogens, indicating that miR400 affects the defense response by cleaving PPR mRNAs. Expression of miR400 was down-regulated, whereas the PPR1 and PPR2 transcripts increased upon pathogen challenge. Collectively, the present study reveals that miR400-mediated dysfunction of PPR proteins renders Arabidopsis more susceptible to pathogenic bacteria and fungi, which emphasizes the importance of PPR proteins in plant defense against diverse pathogens.

Decrease in gastrointestinal absorption of roxithromycin in bile duct cannulated rats due to depletion of bile salts.

The purpose of this study was to investigate the effect of bile salts on gastrointestinal absorption and the plasma second peak phenomenon of roxithromycin in rats. The pharmacokinetic parameters of roxithromycin were calculated after single oral administration at a dose of 20 mg/kg in sham-operated (control), bile duct cannulated (BDC) and bile salt co-administered bile duct cannulated (BSBDC) rats. In BDC rats, the total area under the plasma concentration-time curve from time zero to time infinity (AUC0-∞) and the peak plasma concentration (Cmax) were significantly smaller (0.572-fold) and lower (0.412-fold), respectively, than those in the control rats. These values were recovered by co-administration of bile salt (0.831- and 0.828-fold for AUC0-∞ and Cmax compared with the control, respectively). Thus, the decreased absorption of roxithromycin in BDC rats could be due to a depletion of bile. The solubility of roxithromycin was 3.09-fold increased at 30 mm of sodium taurocholate. The oral dosage regimen of roxithromycin could be changed in patients with bile deficiency or when the drug is administered to individuals on a high-fat diet, if the present rat data can be extrapolated to humans.

New bone formation between bare titanium surface and hydroxyapatite coating by the aerosol deposition technique in the nasal mucosal penetration model.

OBJECTIVE: The aim of this study was to compare new bone formation with titanium (Ti) surface and hydroxyapatite (HA)-coated titanium surface in mucosal perforation model. MATERIALS AND METHODS: HA coating to the Ti disc and implant were done by aerosol deposition technique. Alkaline phosphatase assay and cell migration assay were done in Ti and HA surface disc with MG63 cells. For the in vivo test, 5 New Zealand white rabbits were used. Two penetration defects were prepared in the nasal bone. Subsequently, 2 types of implants were installed into the defect (diameter: 3.0 mm, length: 6.0 mm). Approximately 5.0 mm of the fixture's surface penetrated into the nasal cavity. In the experimental group, HA-coated implants were used. The same design of implants without coating was used in the control group. The animals were sacrificed 8 weeks postoperatively. Subsequently, a histomorphometric analysis was done. RESULTS: Alkaline phosphatase activity was significantly higher in HA-coated surface than in titanium surface (P < 0.05). In addition, more cells were migrated into the HA-coated surface when compared to Ti surface. In the animal experiments, mean new bone formation was 30.68 ± 14.16% in the experimental group and 6.92 ± 5.12% in the control group (P = 0.001). Mean bone-to-implant contact was 31.71 ± 8.41% in the experimental group and 7.98 ± 5.58% in the control group (P < 0.001). Mean height of the bone regeneration was 3.70 ± 0.76 mm in the experimental group and 1.04 ± 0.67 mm in the control group. The difference between the 2 groups was statistically significant (P < 0.001). CONCLUSIONS: HA-coated implants exhibited more bone regeneration in the mucosal penetration model than the uncoated implants.

Transcriptomic evaluation on methyl bromide-induced phytotoxicity in Arabidopsis thaliana and its mode of phytotoxic action via the occurrence of reactive oxygen species and uneven distribution of auxin hormones.

The increase in worldwide trade has caused the quality maintenance of commercialized agriproducts to be crucial in keeping its economic value. In recent years, methyl bromide (MB) has been used dominantly during quarantine and pre-shipment, despite it being an environmental hazard with global repercussions. Through this study, it was shown that Arabidopsis thaliana's 2 h exposure to the MB treatment displayed no signs of phytotoxicity, whereas its 4 h exposure significantly interfered with growth. The transcriptomic analysis found the molecular modifications in A. thaliana after the MB fumigation with the up-regulation of genes specifically relative to the abiotic and oxidative stress, and the down-regulation of auxin transporter genes. Some important gene expressions were verified by RT-qPCR and their expression patterns were similar. Oxidative stresses via the reactive oxygen species (ROS) in relation to MB phytotoxicity were confirmed with the increased malondialdehyde in MB-4h-treated A. thaliana. Uneven distribution of auxins via lower expression of auxin transporter genes was also determined using UPLC-ESI-QqQ MS. Application of two ROS scavengers such as N-acetyl-cysteine and L-glutathione minimized MB phytotoxic effect in A. thaliana. Therefore, MB caused severe oxidative stress, and alternatives regarding the use of MB should be considered.

Effects of installation depth on survival of an hydroxyapatite-coated Bicon implant for single-tooth restoration.

PURPOSE: The present retrospective study evaluated various implant surface factors associated with Bicon implant survival for single-tooth restoration in the healthy individual. PATIENTS AND METHODS: A retrospective cohort study design was used. A total of 613 Bicon (Bicon System, Boston, MA) implants (272 patients) were included. Because the use of hydroxyapatite (HA) coating has been controversial, the surface type was chosen according to the patient's preference. A total of 308 HA-coated implants and 305 titanium plasma-sprayed (TPS) implants were used. The macroanatomy of both implant types was identical. Patients who had systemic disease, poor quality bone, or removable prosthetics were excluded from the present study. A chart review was conducted to record age, gender, implant diameter, implant length, installation depth, installation location, and the use of bone grafting. Implant failure was recorded, and the data were analyzed using the chi(2) test and logistic regression analysis. RESULTS: The installation depth was an important prognostic factor in the HA-coated implants. The failure rate for the HA-coated implants installed at margin level and 2 mm below level was 10.29% and 3.01%, respectively (chi(2) = 6.035, P = .014). The implant length was an important prognostic factor for the TPS-treated implants. The failure rate recorded for the TPS-treated implants installed with a length of less than 10 mm and 10 mm or longer was 15.46% and 2.40%, respectively (chi(2) = 18.414, P < .001). CONCLUSION: Many factors can influence the failure rate of TPS and HA-coated Bicon implants. Among these, installation depth played an especially significant role in the success of HA-coated Bicon implants in the present study.

Susceptibility of the Cigarette Beetle Lasioderma serricorne (Fabricius) to Phosphine, Ethyl Formate and Their Combination, and the Sorption and Desorption of Fumigants on Cured Tobacco Leaves.

The susceptibility of Lasioderma serricorne to phosphine (PH3), ethyl formate (EF) and their combination (PH3 + EF) was evaluated in this study. Eggs, larvae, pupae and adults were subjected to treatment with fumigants to determine the 90% lethal concentration time (LCt90) values. Treatment with PH3 for 20 h resulted in LCt90 values of 1.15, 1.39, 14.97 and 1.78 mg h/L while treatment with EF resulted in values of 157.96, 187.75, 126.06 and 83.10 mg h/L, respectively. By contrast, the combination of PH3 + EF resulted in LCt90 values of 36.05, 44.41, 187.17 and 35.12 mg h/L after 4 h. These results show that, through treatment with PH3 + EF, control can be achieved at lower concentrations than for treatment with EF alone and at lower exposure times than for treatment with PH3 alone. The sorption rates of the fumigants on cured tobacco leaves were determined for filling ratios of 2.5%, 5.0% and 10.0% (w/v). Cured tobacco leaves were treated with either 2 mg/L PH3, 114 mg/L EF or 0.5 mg/L PH3 + 109 mg/L EF. Treatment with PH3 showed sorption rates of 0.0%, 7.1% and 14.3%. EF, however, showed higher sorption rates of 64.9%, 68.5% and 75.5%, respectively, for the indicated filling ratios. When PH3 and EF were combined, the sorption rate of PH3 was 0.0%, while the sorption rates of EF were lower (9.1%, 12.0% and 23.2%) than treatment with only EF. EF required a ventilation time of longer than 22 h to desorb from cured tobacco leaves. Therefore, PH3 + EF can effectively control L. serricorne in cured tobacco leaves, with sufficient ventilation time required after treatment for the safety of workers.

Novel therapeutic roles of MC-4 in combination with everolimus against advanced renal cell carcinoma by dual targeting of Akt/pyruvate kinase muscle isozyme M2 and mechanistic target of rapamycin complex 1 pathways.

Current clinical trials of new anticancer therapies against metastatic renal cell carcinoma (RCC), including molecular-targeted therapies, have not shown promise. The purpose of this study was to preclinically assess the antitumor effects of MC-4, a partially purified material of Artemisia annua L., as a monotherapy or in combination with the known mechanistic target of rapamycin complex 1 (mTORC1) inhibitor, everolimus, against Caki-1 (Von Hippel-Lindau (VHL)+/+) and 786-O (VHL-/-) human RCC cells. MC-4 monotherapy significantly increased tumor growth inhibition and autophagic cell death in RCC cells in vitro and in vivo. Everolimus led to compensatory Akt activation by inhibiting only mTORC1 signaling pathway. In contrast to everolimus, MC-4 enhanced phosphatase and tensin homolog expression and reduced its downstream effector, Akt/pyruvate kinase muscle isozyme M2 (PKM2), leading to decreased expression of glucose transporter 1, which is associated with cancer cell metabolism. The synergistic antitumor and anti-metastatic effects induced by co-administration of MC-4 and everolimus involve cell growth inhibition and autophagic cell death via dual targeting of phosphatidylinositol 3-kinase (PI3K)/Akt/PKM2 and mTORC1. These findings suggest that MC-4 is a novel Akt/PKM2 inhibitor that can overcome the limitation of existing mTOR inhibitors and can be considered a novel strategy to treat patients with rapidly progressing advanced RCC.

Liquid chromatographic resolution of secondary amino alcohols on a chiral stationary phase based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid Dependence of temperature effect on analyte structure.

Two types of secondary amino alcohols were successfully resolved on a liquid chromatographic chiral stationary phase based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid. The secondary amino alcohols containing a secondary amino group connected directly to the stereogenic center were resolved much better than those containing a hydroxyl group attached directly to the stereogenic center. In addition, the dependence of the separation factor (alpha) on column temperature was exactly opposite for the two different types of secondary amino alcohols. From the thermodynamic calculations based on Van't Hoff plots, we conclude that the enantioselectivity for the resolution of the secondary amino alcohols containing a secondary amino group connected directly to the stereogenic center is enthalpically controlled while that for the resolution of the secondary amino alcohols containing a hydroxyl group attached directly to the stereogenic center is entropically controlled.

Effects of Photoperiod Manipulation on Gonadal Activity of the Damselfish, Chromis notata.

This study investigated the effect of different photoperiods (14L: 10D, 12L:12D and 10L:14D) on the gonadal development and GtH mRNA expression in the pituitary of damselfish. The results showed that gonadosomatic index (GSI) was significantly lower in shot photoperiod (10L:14D), in comparison with other photoperiodic group during the spawning season. After 60 days treatment, histological analysis of gonad tissue showed that the gonad of 10L:14D and 12L: 12D treatment groups were resting phase with spermatogonia and perinucleolus stage oocytes but the gonad of 14L:10D treatment group was still ripe phase with spermatozoa and mature stage oocyte. The FSHß and LHß mRNA expression in pituitary drastically decreased shot photoperiod treatment from July (spawning period). These results suggest that photoperiod is considered to be the most effective environmental factor in controlling the reproductive cycle of damselfish.

Large scale pattern graphene electrode for high performance in transparent organic single crystal field-effect transistors.

High quality, large grain size graphene on polycrystalline nickel film on two inch silicon wafers was successfully synthesized by the chemical vapor deposition (CVD) method. The polydimethylsiloxane (PDMS) stamping method was used for graphene transferring in this experiment. The graphene transferred onto Al2O3/ITO substrates was patterned into macroscopic dimension electrodes using conventional lithography followed by oxygen plasma etching. Experimental results show that this graphene can serve as transparent source and drain electrodes in high performance organic semiconductor nanoribbon organic field-effect transistors (OFETs), facilitating high hole injection efficiency due to the preferred work function match with the channel material: single crystalline copper phthalocyanine (CuPc) nanoribbons. The nanoribbons were grown on top of the patterned graphene via evaporate-deposition to form the FET device. The carrier mobility and on/off current ratio of such devices were measured to be as high as 0.36 cm2/(V s) and 10(4).

Isolation and characterization of DMRT1 and its putative regulatory region in the protogynous wrasse, Halichoeres tenuispinis.

A full-length cDNA of doublesex and mab-3-related transcription factor 1 gene (DMRT1) from wrasse testis was isolated by cDNA library screening. Wrasse DMRT1 was 3116 bp in size and contained the DM domain, with a zinc finger DNA-binding motif, and the male-specific motif. Northern blot analysis identified a 3.2-kb transcript approximately equal in size to the DMRT1 nucleotide sequence detected in the testis, but not in the ovary, confirming that this sequence is male-specific in protogynous wrasse. Southern blot analysis suggested that the wrasse genome contains two copies of the DMRT1 gene. The ORF consisted of five exons and four introns with conserved donor-acceptor splice sites at all exon-intron junctions. The 5'-flanking region of the wrasse DMRT1 gene was isolated by DNA walking, and putative regulatory sites were identified by searching data bases. The 5'-flanking region was divided into 9 elements, then 17 DMRT-luciferase chimeric plasmids were constructed. By transient transfection into Cos-1 and TM4 cells, distal element I which contains GATA-binding sites and proximal element B containing the sex-determining region on Y chromosome gene (SRY) binding site were revealed to have an important role in transcriptional regulation of the wrasse DMRT1 when an enhancer sequence was provided.

The CYP3A4*18 allele, the most frequent coding variant in asian populations, does not significantly affect the midazolam disposition in heterozygous individuals.

The objective of this study was to identify CYP3A4 variants in Koreans and to characterize their functional consequences in vitro and in vivo. Four single nucleotide polymorphisms were identified in 50 Koreans by direct DNA sequencing. In an additional genotyping using 248 subjects, CYP3A4(*)18 was confirmed as the most frequent coding variant in Koreans at 1.7%, and its frequency was similar to that of Asians, suggesting that CYP3A4(*)18 would be the highest coding variant in Asians. The recombinant CYP3A4.18 protein prepared in baculovirus expression system showed 67.4% lower Vmax and 1.8-fold higher K(m) for midazolam 1'-hydroxylation compared with the wild type. The mean values of Cmax and area under the concentration curve (AUC) in the CYP3A4(*)1/(*)18 and CYP3A5(*)1/(*)3 subjects (n = 8) were 63% and 32% higher than in CYP3A4(*)1/(*)1 and CYP3A5(*)1/(*)3 carriers (n = 8), respectively. Although the in vitro assay exhibited a significant reduction of the enzyme activity for midazolam, the in vivo differences associated with the CYP3A4(*)1/(*)18 tend to be low (P < 0.07 in Cmax and P < 0.09 in AUC). In summary, the heterozygous CYP3A4(*)1/(*)18 does not appear to cause a significant change of midazolam disposition in vivo; however, the clinical relevance of CYP3A4(*)18/(*)18 remains to be evaluated.