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1
Moro, L. N., M. I. Hiriart, C. Buemo, J. Jarazo, A. Sestelo, D. Veraguas, L. Rodriguez-Alvarez, and D. F. Salamone. "Cheetah interspecific SCNT followed by embryo aggregation improves in vitro development but not pluripotent gene expression." REPRODUCTION 150, no. 1 (July 2015): 1–10. http://dx.doi.org/10.1530/rep-15-0048.
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The aim of this study was to evaluate the capacity of domestic cat (Dc,Felis silvestris) oocytes to reprogram the nucleus of cheetah (Ch,Acinonyx jubatus) cells by interspecies SCNT (iSCNT), by using embryo aggregation. Dc oocytes werein vitromatured and subjected to zona pellucida free (ZP-free) SCNT or iSCNT, depending on whether the nucleus donor cell was of Dc or Ch respectively. ZP-free reconstructed embryos were then cultured in microwells individually (Dc1X and Ch1X groups) or in couples (Dc2X and Ch2X groups). Embryo aggregation improvedin vitrodevelopment obtaining 27.4, 47.7, 16.7 and 28.3% of blastocyst rates in the Dc1X, Dc2X, Ch1X and Ch2X groups, respectively (P<0.05). Moreover, aggregation improved the morphological quality of blastocysts from the Dc2X over the Dc1X group. Gene expression analysis revealed that Ch1X and Ch2X blastocysts had significantly lower relative expression of OCT4, CDX2 and NANOG than the Dc1X, Dc2X and IVF control groups. The OCT4, NANOG, SOX2 and CDX2 genes were overexpressed in Dc1X blastocysts, but the relative expression of these four genes decreased in the Dc2X, reaching similar relative levels to those of Dc IVF blastocysts. In conclusion, Ch blastocysts were produced using Dc oocytes, but with lower relative expression of pluripotent and trophoblastic genes, indicating that nuclear reprogramming could be still incomplete. Despite this, embryo aggregation improved the development of Ch and Dc embryos, and normalized Dc gene expression, which suggests that this strategy could improve full-term developmental efficiency of cat and feline iSCNT embryos.
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Moro, L. N., M. I. Hiriart, J. Jarazo, C. Buemo, A. Sestelo, and D. F. Salamone. "41 EFFICIENT STRATEGY FOR INTERSPECIFIC CLONING IN FELIDS." Reproduction, Fertility and Development 26, no. 1 (2014): 134. http://dx.doi.org/10.1071/rdv26n1ab41.
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Most of the 36 species of wild felids are at a level of threat, and interspecific SCNT (iSCNT) comes as a strategy to contribute to these species conservation. The aim of this study was to evaluate the effect of embryo aggregation in cheetah (Ch, Acinonyx jubatus), bengal (Ben, a hybrid between Felis silvestris and Prionailurus bengalensis), and domestic cat (DC, Felis silvestris) embryos generated by cloning. DC oocytes were in vitro matured and zona-free SCNT (with DC fibroblasts) or iSCNT (with Ch or Ben fibroblasts) was performed. The reconstructed embryos were activated with 5 μM ionomycin and 1.9 mM 6-DMAP, and cultured in SOF using microwells. Cloned embryos were cultured individually or as 2-embryo aggregates. The experimental groups were Ch1X, Ch2X, Ben1X, Ben2X, and the control groups were DC1X and DC2X. Embryo development was compared by Fisher's exact test (P ≤ 0.05). Embryo aggregation improved cleavage (Day 2) and blastocyst (Day 7) rates per well in all the groups (87.2% v. 96.7%, 83.8% v. 93.3% and 87.6% v. 98.2% for cleavage; and 13.7% v. 28.6%, 33.3% v. 43.8% and 27.4% v. 47.7% for blastocyst, for Ch1X (n = 102), Ch2X (n = 91), Ben1X (n = 154), Ben2X (n = 105), DC1X (n = 113), and DC2X (n = 109), respectively. Moreover, the Ch2X blastocyst rate was statistically similar as the control group DC1X. The mean total cell numbers of the blastocysts obtained were 264 ± 211 and 400.8 ± 97 for Ch1X and Ch2X, 278 ± 62 and 517 ± 104 for Ben1X and Ben2X, 385 ± 127 and 625 ± 183 for DC1X and DC2X, respectively. Although no statistical differences were obtained between the 1X and 2X groups, the 2X groups nearly doubled the average number of cells compared with the 1X groups. Blastocysts were also classified as grade 1 (expanded blastocysts with a well-defined ICM), grade 2 (expanded blastocysts without a well-defined ICM), and grade 3 (not expanded blastocysts). This classification showed an increase in grade 1 DC2X blastocyst compared with DC1X blastocysts (36.7% v. 16.1%), but no differences were observed in the other species. Expression of OCT-4 was assessed by inmunocytochemistry. The cheetah blastocysts markedly over-expressed this protein: the percentage of cells that expressed OCT-4 in Ch1X, Ch2X, Ben1X, Ben2X, DC1X, and DC2X was 88.2, 80.2, 46.3, 45.4, 51, and 47.4%, respectively, with statistical differences among all the groups except Ben1X and Ben2X. The proportion of OCT-4 expressing cells over total cell numbers was analysed by the difference of proportions test (P ≤ 0.05). In conclusion, iSCNT resulted in high rates of blastocyst formation, especially when embryo aggregation was applied. This strategy has not been previously evaluated in felids or iSCNT procedures, and has been demonstrated to improve blastocyst formation, the number of cells in the 3 groups, and the blastocyst quality in the DC. Other pluripotent genes besides OCT-4 should be studied to determine whether the overexpression of this gene in cheetah embryos is the consequence of an inefficient nuclear reprogramming that prevents a correct regulation. Finally, the iSCNT and embryo aggregation could contribute to species conservation in felids.
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Moro, L. N., D. Veraguas, L. Rodriguez-Alvarez, M. I. Hiriart, C. Buemo, A. Sestelo, and D. Salamone. "212 INTERSPECIFIC CLONING AND EMBRYO AGGREGATION INFLUENCE THE EXPRESSION OF oct4, nanog, sox2, AND cdx2 IN CHEETAH AND DOMESTIC CAT BLASTOCYSTS." Reproduction, Fertility and Development 27, no. 1 (2015): 196. http://dx.doi.org/10.1071/rdv27n1ab212.
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The cheetah (Ch, Acinonyx jubatus) is a species considered globally endangered and cloning is one of the assisted reproductive techniques that can help to preserve it and to study early embryo development. However, the production of cloned felid embryos remains inefficient, probably because of the difficulty to control the process of nuclear reprogramming and obtain adequate gene expression. Embryo aggregation has been demonstrated to improve the cloning efficiency in several species and to normalise cdx2 in the mouse by lowering its expression (Balbach et al. 2010), but it has not been evaluated in felids before. To better understand the effect of interspecific somatic-cell nuclear transfer (iSCNT) and embryo aggregation in nuclear reprogramming, we analysed the expression of oct4, sox2, nanog, and cdx2 in cheetah blastocysts generated by iSCNT, domestic cat blastocysts (Dc) generated by SCNT, and IVF blastocysts as control. To achieve this, domestic cat oocytes were in vitro matured and zona-free SCNT or iSCNT was performed, as previously described (Moro et al. 2014, Reprod. Fertil. Dev.). Zona-free reconstructed embryos were then cultured individually (1X) or two embryo were cultured together (2X) in microwells, in synthetic oviductal fluid (SOF) medium. The experimental groups were Dc1X, Dc2X, Ch1X, Ch2X, and IVF. After 8 days of in vitro culture the blastocysts obtained were stored in RNA-later at –20°C. For gene expression analysis, blastocysts were pooled as follows: Dc1X, 4 replicates of 3 blastocysts each; Dc2X, 4 replicates of 3 blastocysts each; Ch1X, 2 replicates of 2 blastocysts and 1 replicate of 1 blastocyst; Ch2X, 4 replicates of 3 blastocysts each; IVF 3 replicates of 3 blastocysts each. Embryos were treated with a Cells-to-cDNA TM II kit (Life Technologies, Carlsbad, CA, USA) lyses buffer and treated with DNase I (0.04 U μL–1) for genomic DNA digestion. Gene expression analysis was performed by real-time qPCR using the standard curve method. In all qPCRs, GAPDH was used as an internal control. The statistical analysis was performed using a non-parametric Kruskal–Wallis test (P < 0.05). We observed that Dc1X blastocysts overexpressed the 4 genes evaluated respect to the IVF control. However, the gene expression of the aggregated group (Dc2X) was lower for all the genes, achieving the same levels of nanog and sox2 as the IVF blastocysts. The expression of oct4 and cdx2 were also closer to the expression levels of the control in the Dc2X group than in the Dc1X group. With respect to interspecific embryos, the amount of oct4 and cdx2 was also significantly reduced in the Ch2X blastocysts respect to Ch1X blastocysts. Both cheetah groups showed significantly lower expression of oct4, cdx2, and nanog than the IVF control. In conclusion, transcription of pluripotent and early differentiation factors in cheetah embryos was not as efficient as in the domestic cat embryos, probably caused by interspecific transfer. Our study demonstrated for the first time that defects in gene expression of domestic cat embryos can be corrected by embryo aggregation, providing a simple strategy to improve felid cloning.
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Rubin, Daniel J., Naveen Maliakkal, Huaqing Zhao, and Eli E. Miller. "Hospital Readmission Risk and Risk Factors of People with a Primary or Secondary Discharge Diagnosis of Diabetes." Journal of Clinical Medicine 12, no. 4 (February 6, 2023): 1274. http://dx.doi.org/10.3390/jcm12041274.
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Hospital readmission among people with diabetes is common and costly. A better understanding of the differences between people requiring hospitalization primarily for diabetes (primary discharge diagnosis, 1°DCDx) or another condition (secondary discharge diagnosis, 2°DCDx) may translate into more effective ways to prevent readmissions. This retrospective cohort study compared readmission risk and risk factors between 8054 hospitalized adults with a 1°DCDx or 2°DCDx. The primary outcome was all-cause hospital readmission within 30 days of discharge. The readmission rate was higher in patients with a 1°DCDx than in patients with a 2°DCDx (22.2% vs. 16.2%, p < 0.01). Several independent risk factors for readmission were common to both groups including outpatient follow up, length of stay, employment status, anemia, and lack of insurance. C-statistics for the multivariable models of readmission were not significantly different (0.837 vs. 0.822, p = 0.15). Readmission risk of people with a 1°DCDx was higher than that of people with a 2°DCDx of diabetes. Some risk factors were shared between the two groups, while others were unique. Inpatient diabetes consultation may be more effective at lowering readmission risk among people with a 1°DCDx. These models may perform well to predict readmission risk.
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Wei, Yuqi. "LLC and CLLC Resonant Converters Based DC Transformers (DCXs): Characteristics, Issues, and Solutions." CPSS Transactions on Power Electronics and Applications 6, no. 4 (December 2021): 332–48. http://dx.doi.org/10.24295/cpsstpea.2021.00031.
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Conventional line frequency transformers have the disadvantages of large volume and low efficiency. The medium or high frequency transformers based on power converters can achieve high power conversion with small footprint have drawn popularity in numerous industrial applications. Unregulated resonant converters, LLC and CLLC resonant converters, with fixed voltage conversion ratio operating at resonant frequency, which are also known as DC transformers (DCXs), are attractive owning to their high efficiency characteristic. Nevertheless, there are issues associated with DCXs in real applications. Regulation capability and automatic resonant frequency tracking capability are the two most important issues for DCXs. The main work of this paper is to characterize the resonant converters based DCXs, and overview the issues and solutions associated with DCXs. Guidelines can be provided for researchers and engineers when designing the resonant converters based DCXs.
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Ma, Shiyu, Lin Zheng, Xiao Lin, Yi Feng, Ming Yang, and Lan Shen. "Network Pharmacology and Metabolomics Studies on Antimigraine Mechanisms of Da Chuan Xiong Fang (DCXF)." Evidence-Based Complementary and Alternative Medicine 2021 (April 20, 2021): 1–16. http://dx.doi.org/10.1155/2021/6665137.
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Background. Da Chuan Xiong Fang (DCXF) is a traditional Chinese medicine (TCM) formula used to treat migraines. Previously, we uncovered partial mechanisms involved in the therapeutic actions of DCXF on migraines. Methods. In this study, we further elucidated its antimigraine mechanisms in vivo by using an integrated strategy coupling with network pharmacology and metabolomics techniques. Results. Network pharmacology identified 33 genes linked with both migraine and DCXF, most of which were 5-hydroxytryptamine receptors, dopamine, and peptide receptors. The results of GO and KEGG enrichment analysis showed that DCXF significantly regulated tyrosine metabolism, tryptophan metabolism, dopamine metabolic process, glucose transmembrane transport, lipid metabolism, and fatty acid transport. The results of metabolomics analysis found that the metabolism of tryptophan and tyrosine in the brain tissue and energy and lipid metabolism of rats tended towards normal and reached normal levels after administering DCXF. The metabolomics and network pharmacology approaches demonstrated similar antimigraine effects of DCXF on endogenous neurotransmitters and overall trends in serum and brain tissue. Using both approaches, 62 hub genes were identified from the protein-protein interaction (PPI) network of DCXF and gene-metabolite interaction network, with hub genes and different metabolites in serum and brain tissue. The hub genes of DCXF, which were mostly linked with inflammation, might affect mainly neurotransmitters in serum and brain tissue metabolisms. Conclusion. Network pharmacology and metabolomics study may help identify hub genes, metabolites, and possible pathways of disease and treatment. Additionally, two parts of the results were integrated to confirm each other. Their combination may help elucidate the relationship between hub genes and metabolites and provide the further understanding of TCM mechanisms.
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Tho Son, Le, Nguyen Hai Dang, Lai Thi Phuong, Nguyen Thi Thu, and Nguyen Huy Hoang. "New inhibitors for dicarbonyl/L-xylulose reductase in Caenorhabditis elegans." Vietnam Journal of Biotechnology 20, no. 3 (September 30, 2022): 517–26. http://dx.doi.org/10.15625/1811-4989/16673.
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Dicarbonyl/L-xylulose reductase (DCXR) is an enzyme reducing dicarbonyl (-CO-) groups in single sugar molecules and participates in the sugar metabolism of several sugar metabolism cycles in different living organisms (humans, other animals, fungi, and microorganisms). Caenorhabditis elegans, a model organism, has a unique DCXR (referred to as Ce DCXR) which plays a biochemical function similar to its homologs in the other organisms. However, the catalytic regulation of the enzyme has not been fully elucidated yet. Therefore, in this study, we investigated the chemicals which could inhibit the enzyme and found three inhibitor compounds including hexanoic acid, phosphoenolpyruvic acid, and DL-α-aminobutyrate hydrochloride for the enzyme. Because these inhibitors and their derivatives suppressed DCXRs in humans, mice, rabbits, and rats, they possibly inhibit the DCXR homologs at least in other animals. If the inhibitors and their derivatives are components in pharmaceutical products, foods, and drinks, they may come into cells, interact with DCXRs and inactivate them, causing toxicity for the host organisms.
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Faisalina, A. F., Fabio Sonvico, Paolo Colombo, A. A. Amirul, H. A. Wahab, and Mohamed Isa Abdul Majid. "Docetaxel-Loaded Poly(3HB-co-4HB) Biodegradable Nanoparticles: Impact of Copolymer Composition." Nanomaterials 10, no. 11 (October 26, 2020): 2123. http://dx.doi.org/10.3390/nano10112123.
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Polyhydroxyalkanoate (PHA) copolymers show a relatively higher in vivo degradation rate compared to other PHAs, thus, they receive a great deal of attention for a wide range of medical applications. Nanoparticles (NPs) loaded with poorly water-soluble anticancer drug docetaxel (DCX) were produced using poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB), copolymers biosynthesised from Cupriavidus malaysiensis USMAA1020 isolated from the Malaysian environment. Three copolymers with different molar proportions of 4-hydroxybutirate (4HB) were used: 16% (PHB16), 30% (PHB30) and 70% (PHB70) 4HB-containing P(3HB-co-4HB). Blank and DCX-loaded nanoparticles were then characterized for their size and size distribution, surface charge, encapsulation efficiency and drug release. Preformulation studies showed that an optimised formulation could be achieved through the emulsification/solvent evaporation method using PHB70 with the addition of 1.0% PVA, as stabilizer and 0.03% VitE-TPGS, as surfactant. DCX-loaded PHB70 nanoparticles (DCX-PHB70) gave the desired particle size distribution in terms of average particle size around 150 nm and narrow particle size distribution (polydispersity index (PDI) below 0.100). The encapsulation efficiency result showed that at 30% w/w drug-to-polymer ratio: DCX- PHB16 NPs were able to encapsulate up to 42% of DCX; DCX-PHB30 NPs encapsulated up to 46% of DCX and DCX-PHB70 NPs encapsulated up to 50% of DCX within the nanoparticle system. Approximately 60% of DCX was released from the DCX-PHB70 NPs within 7 days for 5%, 10% and 20% of drug-to-polymer ratio while for the 30% and 40% drug-to-polymer ratios, an almost complete drug release (98%) after 7 days of incubation was observed.
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Wang, Lu, Jinming Zhang, Yanlong Hong, Yi Feng, Meiwan Chen, and Yitao Wang. "Phytochemical and Pharmacological Review of Da Chuanxiong Formula: A Famous Herb Pair Composed ofChuanxiong RhizomaandGastrodiae Rhizomafor Headache." Evidence-Based Complementary and Alternative Medicine 2013 (2013): 1–16. http://dx.doi.org/10.1155/2013/425369.
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Chronic headache such as migraine and nervous headache has become one of the most common locations of pain and one of the most difficult diseases to recover due to its numerous causes and inconvenience to keep acesodyne administration for a long time. However, there are a series of treatment theories and herbal formulas for this disease in traditional Chinese medicine (TCM), in which Da Chuanxiong formula (DCXF), a herb pair composed ofChuanxiong Rhizoma(CR), Chuanxiong in Chinese, andGastrodiae Rhizoma(GR) called as Tianma in China, is a greatly classic representative. This formula has been used for headaches via dispelling wind pathogen and dissipating blood stasis for many years in TCM. In recent years, the efficiency and representativeness of DCXF have garnered many researchers’ attention. To reveal the compatibility mechanism and develop innovative Chinese herb, herein ethnopharmacological relevance, chemical characters, and pharmacological actions of DCXF are detailed. It is expected to give a comprehensive interpretation of DCXF, namely, Chuanxiong Tianma herb pair (CTHP), to inherit the essence of herb pair and innovate drug delivery system of this prescription.
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Luty, Marcin, Katarzyna Piwowarczyk, Anna Łabędź-Masłowska, Tomasz Wróbel, Małgorzata Szczygieł, Jessica Catapano, Grażyna Drabik, et al. "Fenofibrate Augments the Sensitivity of Drug-Resistant Prostate Cancer Cells to Docetaxel." Cancers 11, no. 1 (January 11, 2019): 77. http://dx.doi.org/10.3390/cancers11010077.
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Metronomic agents reduce the effective doses and adverse effects of cytostatics in cancer chemotherapy. Therefore, they can enhance the treatment efficiency of drug-resistant cancers. Cytostatic and anti-angiogenic effects of fenofibrate (FF) suggest that it can be used for the metronomic chemotherapy of drug-resistant prostate tumors. To estimate the effect of FF on the drug-resistance of prostate cancer cells, we compared the reactions of naïve and drug-resistant cells to the combined treatment with docetaxel (DCX)/mitoxantrone (MTX) and FF. FF sensitized drug-resistant DU145 and PC3 cells to DCX and MTX, as illustrated by their reduced viability and invasive potential observed in the presence of DCX/MTX and FF. The synergy of the cytostatic activities of both agents was accompanied by the inactivation of P-gp-dependent efflux, dysfunction of the microtubular system, and induction of polyploidy in DCX-resistant cells. Chemical inhibition of PPARα- and reactive oxygen species (ROS)-dependent pathways by GW6471 and N-acetyl-L-cysteine, respectively, had no effect on cell sensitivity to combined DCX/FF treatment. Instead, we observed the signs of adenosine triphosphate (ATP) deficit and autophagy in DCX/FF-treated drug-resistant cells. Furthermore, the cells that had been permanently propagated under DCX- and DCX/FF-induced stress did not acquire DCX/FF-resistance. Instead, relatively slow proliferation of DCX-resistant cells was efficiently inhibited by FF. Collectively, our observations show that FF reduces the effective doses of DCX by interfering with the drug resistance and energy metabolism of prostate cancer cells. Concomitantly, it impairs the chemotherapy-induced microevolution and expansion of DCX/FF-resistant cells. Therefore, FF can be applied as a metronomic agent to enhance the efficiency of palliative chemotherapy of prostate cancer.
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Tanaka, Teruyuki, Finley F. Serneo, Christine Higgins, Michael J. Gambello, Anthony Wynshaw-Boris, and Joseph G. Gleeson. "Lis1 and doublecortin function with dynein to mediate coupling of the nucleus to the centrosome in neuronal migration." Journal of Cell Biology 165, no. 5 (June 1, 2004): 709–21. http://dx.doi.org/10.1083/jcb.200309025.
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Humans with mutations in either DCX or LIS1 display nearly identical neuronal migration defects, known as lissencephaly. To define subcellular mechanisms, we have combined in vitro neuronal migration assays with retroviral transduction. Overexpression of wild-type Dcx or Lis1, but not patient-related mutant versions, increased migration rates. Dcx overexpression rescued the migration defect in Lis1+/− neurons. Lis1 localized predominantly to the centrosome, and after disruption of microtubules, redistributed to the perinuclear region. Dcx outlined microtubules extending from the perinuclear “cage” to the centrosome. Lis1+/− neurons displayed increased and more variable separation between the nucleus and the preceding centrosome during migration. Dynein inhibition resulted in similar defects in both nucleus–centrosome (N-C) coupling and neuronal migration. These N-C coupling defects were rescued by Dcx overexpression, and Dcx was found to complex with dynein. These data indicate Lis1 and Dcx function with dynein to mediate N-C coupling during migration, and suggest defects in this coupling may contribute to migration defects in lissencephaly.
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Odrzywolski, Adrian, Bożena Jarosz, Michał Kiełbus, Ilona Telejko, Dominik Ziemianek, Sebastian Knaga, and Radosław Rola. "Profiling Glioblastoma Cases with an Expression of DCX, OLIG2 and NES." International Journal of Molecular Sciences 22, no. 24 (December 8, 2021): 13217. http://dx.doi.org/10.3390/ijms222413217.
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Glioblastoma (GBM) remains the leading cause of cancer-related deaths with the lowest five-year survival rates among all of the human cancers. Multiple factors contribute to its poor outcome, including intratumor heterogeneity, along with migratory and invasive capacities of tumour cells. Over the last several years Doublecortin (DCX) has been one of the debatable factors influencing GBM cells’ migration. To resolve DCX’s ambiguous role in GBM cells’ migration, we set to analyse the expression patterns of DCX along with Nestin (NES) and Oligodendrocyte lineage transcription factor 2 (OLIG2) in 17 cases of GBM, using immunohistochemistry, followed by an analysis of single-cell RNA-seq data. Our results showed that only a small subset of DCX positive (DCX+) cells was present in the tumour. Moreover, no particular pattern emerged when analysing DCX+ cells relative position to the tumour margin. By looking into single-cell RNA-seq data, the majority of DCX+ cells were classified as non-cancerous, with a small subset of cells that could be regarded as glioma stem cells. In conclusion, our findings support the notion that glioma cells express DCX; however, there is no clear evidence to prove that DCX participates in GBM cell migration.
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Tabrizi, Shervin, Mohammed Alshalalfa, Brandon Arvin Virgil Mahal, Elai Davicioni, Yang Liu, Kent William Mouw, Felix Y. Feng, Paul L. Nguyen, and Vinayak Muralidhar. "Doublecortin expression in prostate adenocarcinoma and neuroendocrine tumors." Journal of Clinical Oncology 38, no. 6_suppl (February 20, 2020): 161. http://dx.doi.org/10.1200/jco.2020.38.6_suppl.161.
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161 Background: Recent work using prostate cancer mouse models implicated doublecortin (DCX)-expressing neural progenitor cells in prostate adenocarcinoma, reporting a strong association between DCX expression and disease progression and outcome. We sought to evaluate the relationship between DCX expression and these outcomes in human prostate cancer. Methods: DCX expression was measured in transcriptome-wide microarray data from 18,501 patients with localized prostate cancer and 290 patients with metastatic castration-resistant prostate cancer (mCRPC). Pairwise comparisons were performed using the Mann–Whitney U test. Metastasis-free survival (MFS) and overall survival (OS) were analyzed using Cox-proportional hazards. Results: DCX expression was not significantly different between normal prostate (n=29), primary prostate cancer (n=131), or metastases (n=19) (p > 0.5), and did not differ across Gleason score in a large cohort of RP samples (n=17,967, p=0.21). The lack of difference persisted after adjusting for stromal contribution using a 141-gene stromal signature. Those with DCX expression above and below the median did not have significant differences in MFS (HR 1.2 [0.84-1.7], p=0.3) or OS (HR 1.15 [0.7-1.84], p =0.56). In a cohort of untreated prostate cancer, DCX expression was higher in neuroendocrine tumors (n=10) compared to Gleason 9-10 prostate adenocarcinoma (n=110) (p=0.007). Similarly, in two cohorts with mCRPC (n=290), DCX expression was higher in lesions with neuroendocrine features than adenocarcinoma (p<0.001). Consistently, in a patient-derived xenograft model subjected to host castration, DCX expression was initially low, but increased significantly once tumors underwent neuroendocrine differentiation and treatment escape. Conclusions: Contrary to recent data using mouse models, DCX expression did not differ by disease state, grade, or outcome in a dataset of human patients with prostate adenocarcinoma. However, DCX expression appeared to correlate with neuroendocrine prostate cancers, a subgroup that can arise de novo or in the castrate-resistant setting. Further work is needed to define the role of DCX expression and its prognostic significance in prostate cancer.
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Roy, Avik, Madhuchhanda Kundu, Sudipta Chakrabarti, Dhruv R. Patel, and Kalipada Pahan. "Oleamide, a Sleep-Inducing Supplement, Upregulates Doublecortin in Hippocampal Progenitor Cells via PPARα." Journal of Alzheimer's Disease 84, no. 4 (December 7, 2021): 1747–62. http://dx.doi.org/10.3233/jad-215124.
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Background: Doublecortin (DCX), a microtubule associated protein, has emerged as a central biomarker of hippocampal neurogenesis. However, molecular mechanisms by which DCX is regulated are poorly understood. Objective: Since sleep is involved with the acquisition of memory and oleamide or 9-Octadecenamide (OCT) is a sleep-inducing supplement in human, we examined whether OCT could upregulate DCX in hippocampal progenitor cells (HPCs). Methods: We employed real-time PCR, western blot, immunostaining, chromatin immunoprecipitation, lentiviral transduction in HPCs, and the calcium influx assay. Results: OCT directly upregulated the transcription of Dcx in HPCs via activation of peroxisome proliferator-activated receptor α (PPARα), a lipid-lowering transcription factor. We observed that, HPCs of Ppara-null mice displayed significant impairment in DCX expression and neuronal differentiation as compared to that of wild-type mice. Interestingly, treatment with OCT stimulated the differentiation process of HPCs in wild-type, but not Ppara-null mice. Reconstruction of PPARα in mouse Ppara-null HPCs restored the expression of DCX, which was further stimulated with OCT treatment. In contrast, a dominant-negative mutant of PPARα significantly attenuated the stimulatory effect of OCT on DCX expression and suppressed neuronal differentiation of human neural progenitor cells. Furthermore, RNA microarray, STRING, chromatin immunoprecipitation, site-directed mutagenesis, and promoter reporter assay have identified DCX as a new target of PPARα. Conclusion: These results indicate that OCT, a sleep supplement, directly controls the expression of DCX and suggest that OCT may be repurposed for stimulating the hippocampal neurogenesis.
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Ni, Ni, Qingqing Wang, Xiao Lin, Yanlong Hong, Yi Feng, and Lan Shen. "Studies on the Mechanism of Glutamate Metabolism in NTG-Induced Migraine Rats Treated with DCXF." Evidence-Based Complementary and Alternative Medicine 2019 (December 31, 2019): 1–11. http://dx.doi.org/10.1155/2019/1324797.
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Objective. To explore the mechanism of the antimigraine effect by active components extracted from the Dachuanxiong prescription (DCXF), nitroglycerin- (NTG-) induced migraine rats were used to detect the change of glutamate metabolism and the overall metabolic profile at different time points in the serum and Trigeminocervical complex(TCC) samples. Method. The biological samples that were obtained at 30 minutes, 60 minutes, and 90 minutes after model establishment or drug administration were tested by GC-TOF-MS. Then, real-time PCR and western blot were applied to detect changes in the expression of some substances involved in glutamate metabolism. Result. DCXF could improve the metabolic profile of serum and TCC in migraine rats and showed the time trend of treatment, mainly involved by amino acid metabolism (glutamate, aspartic acid, and alanine metabolism). In addition, DCXF could increase the expressions of GS at 60 min and 90 min and EAAT1 at 90 min. The results of GS protein were similar to that of mRNA. Conclusion. The antimigraine effect of DCXF could be achieved by improving the metabolic profile and increasing the expressions of GS and EAAT1 to promote the glutamate cycle of TCC and serum samples in NTG-induced migraine rats to a certain extent.
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DeOliveira-Mello, Laura, Isabel Vicente, Veronica Gonzalez-Nunez, Adrian Santos-Ledo, Almudena Velasco, Rosario Arévalo, Juan M. Lara, and Andreas F. Mack. "Doublecortin in the Fish Visual System, a Specific Protein of Maturing Neurons." Biology 11, no. 2 (February 6, 2022): 248. http://dx.doi.org/10.3390/biology11020248.
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Doublecortin (DCX) is a microtubule associated protein, essential for correct central nervous system development and lamination in the mammalian cortex. It has been demonstrated to be expressed in developing—but not in mature—neurons. The teleost visual system is an ideal model to study mechanisms of adult neurogenesis due to its continuous life-long growth. Here, we report immunohistochemical, in silico, and western blot analysis to detect the DCX protein in the visual system of teleost fish. We clearly determined the expression of DCX in newly generated cells in the retina of the cichlid fish Astatotilapia burtoni, but not in the cyprinid fish Danio rerio. Here, we show that DCX is not associated with migrating cells but could be related to axonal growth. This work brings to light the high conservation of DCX sequences between different evolutionary groups, which make it an ideal marker for maturing neurons in various species. The results from different techniques corroborate the absence of DCX expression in zebrafish. In A. burtoni, DCX is very useful for identifying new neurons in the transition zone of the retina. In addition, this marker can be applied to follow axons from maturing neurons through the neural fiber layer, optic nerve head, and optic nerve.
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Brégère, Catherine, Urs Fisch, Florian Samuel Halbeisen, Christian Schneider, Tanja Dittmar, Sarah Stricker, Soheila Aghlmandi, and Raphael Guzman. "Doublecortin and Glypican-2 concentrations in the cerebrospinal fluid from infants are developmentally downregulated." PLOS ONE 18, no. 2 (February 17, 2023): e0279343. http://dx.doi.org/10.1371/journal.pone.0279343.
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Objective Doublecortin (DCX) and glypican-2 (GPC2) are neurodevelopmental proteins involved in the differentiation of neural stem/progenitor cells (NSPCs) to neurons, and are developmentally downregulated in neurons after birth. In this study, we investigated whether the concentrations of DCX and GPC2 in the cerebrospinal fluid (CSF) from human pediatric patients reflect this developmental process or are associated with cerebral damage or inflammatory markers. Methods CSF was collected from pediatric patients requiring neurosurgical treatment. The concentrations of DCX, GPC2, neuron-specific enolase (NSE), S100 calcium-binding protein B (S100B), and cytokines (IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-13, IFN-γ, and TNF-⍺) were measured using immunoassays. Results From March 2013 until October 2018, 63 CSF samples were collected from 38 pediatric patients (20 females; 17 patients with repeated measurements); the median term born-adjusted age was 3.27 years [Q1: 0.31, Q3: 7.72]. The median concentration of DCX was 329 pg/ml [Q1: 192.5, Q3: 1179.6] and that of GPC2 was 26 pg/ml [Q1: 13.25, Q3: 149.25]. DCX and GPC2 concentrations independently significantly associated with age, and their concentration declined with advancing age, reaching undetectable levels at 0.3 years for DCX, and plateauing at 1.5 years for GPC2. Both DCX and GPC2 associated with hydrocephalus, NSE, IL-1β, IL-2, IL-8, IL-13. No relationship was found between sex, acute infection, S100B, IL-4, IL-6, IL-10, IFN-γ, TNF-α and DCX or GPC2, respectively. Conclusions Concentrations of DCX and GPC2 in the CSF from pediatric patients are developmentally downregulated, with the highest concentrations measured at the earliest adjusted age, and reflect a neurodevelopmental stage rather than a particular disease state.
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Cho, Hee Kyung. "A Study on the Digital Customer Experience Design in Virtual Reality AdverGame Contents." JOURNAL OF THE KOREAN SOCIETY DESIGN CULTURE 27, no. 1 (March 31, 2021): 375–87. http://dx.doi.org/10.18208/ksdc.2020.27.1.375.
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Won, Jongwook. "A Study on the Design of Digital Customer Experience(DCX) for the Activation of Public Services." Journal of Communication Design 71 (April 30, 2020): 7–17. http://dx.doi.org/10.25111/jcd.2020.71.01.
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Ünal, Sedat, Osman Doğan, and Yeşim Aktaş. "Orally administered docetaxel-loaded chitosan-decorated cationic PLGA nanoparticles for intestinal tumors: formulation, comprehensive in vitro characterization, and release kinetics." Beilstein Journal of Nanotechnology 13 (November 23, 2022): 1393–407. http://dx.doi.org/10.3762/bjnano.13.115.
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Intestinal cancers are the third most lethal cancers globally, beginning as polyps in the intestine and spreading with a severe metastatic tendency. Chemotherapeutic drugs used in the treatment of intestinal tumors are usually formulated for parenteral administration due to poor solubility and bioavailability problems. Pharmaceutically, clinical failure due to a drug’s wide biodistribution and non-selective toxicity is one of the major challenges of chemotherapy. In addition, parenteral drug administration in chronic diseases that require long-term drug use, such as intestinal tumors, is challenging in terms of patient compliance and poses a burden in terms of health economy. Especially in the field of chemotherapy research, oral chemotherapy is a subject that has been intensively researched in recent years, and developments in this field will provide serious breakthroughs both scientifically and socially. Development of orally applicable nanodrug formulations that can act against diseases seen in the distant region of the gastrointestinal tract (GIT), such as intestinal tumor, brings with it a series of difficulties depending on the drug and/or GIT physiology. The aim of this study is to develop an oral nanoparticle drug delivery system loaded with docetaxel (DCX) as an anticancer drug, using poly(lactic-co-glycolic acid) (PLGA) as nanoparticle material, and modified with chitosan (CS) to gain mucoadhesive properties. In this context, an innovative nanoparticle formulation that can protect orally administered DCX from GIT conditions and deliver the drug to the intestinal tumoral region by accumulating in mucus has been designed. For this purpose, DCX-PLGA nanoparticles (NPs) and CS/DCX-PLGA NPs were prepared, and their in vitro characteristics were elucidated. Nanoparticles around 250–300 nm were obtained. DCX-PLGA NPs had positive surface charge with CS coating. The formulations have the potential to deliver the encapsulated drug to the bowel according to the in vitro release studies in three different simulated GIT fluids for approximately 72 h. Mucin interaction and penetration into the artificial mucus layer were also investigated in detail, and the mucoadhesive and mucus-penetration characteristics of the formulations were examined. Furthermore, in vitro release kinetic studies of the NPs were elucidated. DCX-PLGA NPs were found to be compatible with the Weibull model, and CS/DCX-PLGA NPs were found to be compatible with the Peppas–Sahlin model. Within the scope of in vitro cytotoxicity studies, the drug-loaded NPs showed significantly higher cytotoxicity than a DCX solution on the HT-29 colon cell line, and CS/DCX-PLGA showed the highest cytotoxicity (p < 0.05). According to the permeability studies on the Caco-2 cell line, the CS/DCX-PLGA formulation increased permeability by 383% compared to free DCX (p < 0.05). In the light of all results, CS/DCX-PLGA NPs can offer a promising and innovative approach as an oral anticancer drug-loaded nanoformulation for intestinal tumors.
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Burke, Rachael M., Saulos Nyirenda, Hussein H. Twabi, Marriott Nliwasa, Elizabeth Joekes, Naomi Walker, Rose Nyirenda, et al. "Design and protocol for a cluster randomised trial of enhanced diagnostics for tuberculosis screening among people living with HIV in hospital in Malawi (CASTLE study)." PLOS ONE 17, no. 1 (January 10, 2022): e0261877. http://dx.doi.org/10.1371/journal.pone.0261877.
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Background People living with HIV (PLHIV) have a high risk of death if hospitalised in low-income countries. Tuberculosis has long been the leading cause of admission and death, in part due to suboptimal diagnostics. Two promising new diagnostic tools are digital chest Xray with computer-aided diagnosis (DCXR-CAD) and urine testing with Fujifilm SILVAMP LAM (FujiLAM). Neither test has been rigorously evaluated among inpatients. Test characteristics may be complementary, with FujiLAM especially sensitive for disseminated tuberculosis and DCXR-CAD especially sensitive for pulmonary tuberculosis, making combined interventions of interest. Design and methods An exploratory unblinded, single site, two-arm cluster randomised controlled trial, with day of admission as the unit of randomisation. A third, smaller, integrated cohort arm (4:4:1 random allocation) contributes to understanding case-mix, but not trial outcomes. Participants are adults living with HIV not currently on TB treatment. The intervention (DCXR-CAD plus urine FujiLAM plus usual care) is compared to usual care alone. The primary outcome is proportion of participants started on tuberculosis treatment by day 56, with secondary outcomes of mortality (time to event) measured to to 56 days from enrolment, proportions with undiagnosed tuberculosis at death or hospital discharge and comparing proportions with enrolment-day tuberculosis treatment initiation. Discussion Both DCXR-CAD and FujiLAM have potential clinical utility and may have complementary diagnostic performance. To our knowledge, this is the first randomised trial to evaluate these tests among hospitalised PLHIV.
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A. Razak, S. Aishah, Amirah Mohd Gazzali, Faisalina Ahmad Fisol, Ibrahim M. Abdulbaqi, Thaigarajan Parumasivam, Noratiqah Mohtar, and Habibah A. Wahab. "Advances in Nanocarriers for Effective Delivery of Docetaxel in the Treatment of Lung Cancer: An Overview." Cancers 13, no. 3 (January 22, 2021): 400. http://dx.doi.org/10.3390/cancers13030400.
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Docetaxel (DCX) is a highly effective chemotherapeutic drug used in the treatment of different types of cancer, including non-small cell lung cancer (NSCLC). The drug is known to have low oral bioavailability due to its low aqueous solubility, poor membrane permeability and susceptibility to hepatic first-pass metabolism. To mitigate these problems, DCX is administered via the intravenous route. Currently, DCX is commercially available as a single vial that contains polysorbate 80 and ethanol to solubilize the poorly soluble drug. However, this formulation causes short- and long-term side effects, including hypersensitivity, febrile neutropenia, fatigue, fluid retention, and peripheral neuropathy. DCX is also a substrate to the drug efflux pump P-glycoprotein (P-gp) that would reduce its concentration within the vicinity of the cells and lead to the development of drug resistance. Hence, the incorporation of DCX into various nanocarrier systems has garnered a significant amount of attention in recent years to overcome these drawbacks. The surfaces of these drug-delivery systems indeed can be functionalized by modification with different ligands for smart targeting towards cancerous cells. This article provides an overview of the latest nanotechnological approaches and the delivery systems that were developed for passive and active delivery of DCX via different routes of administration for the treatment of lung cancer.
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Huang, Fei, Yunyi Lan, Liyue Qin, Huaihuai Dong, Hailian Shi, Hui Wu, Qinrui Zou, Zhibi Hu, and Xiaojun Wu. "Astragaloside IV Promotes Adult Neurogenesis in Hippocampal Dentate Gyrus of Mouse through CXCL1/CXCR2 Signaling." Molecules 23, no. 9 (August 29, 2018): 2178. http://dx.doi.org/10.3390/molecules23092178.
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Astragaloside IV (ASI) has been reported to promote neural stem cells proliferation in vitro and CXCR2 expression on neutrophils. The present study was aimed to investigate the influence of ASI on adult neurogenesis in hippocampal dentate gyrus (DGs) of mouse and to discuss the possible underlying mechanisms. Total number of proliferative cells (BrdU+), pre-mature neurons (DCX+), early proliferative cells (BrdU+/DCX+), proliferative radial gila-like cells (BrdU+/GFAP+) and newly generated neurons (BrdU+/NeuN+) after ASI or vehicle administration for two weeks were counted, respectively. The results showed that BrdU+ cells and DCX+ cells were significantly increased in DGs of mice administered with ASI. The numbers of BrdU+/DCX+, BrdU+/GFAP+ cells and BrdU+/NeuN+ cells were also elevated in the ASI group. Correspondingly, ASI increased the protein expression of hippocampal DCX, GFAP and NeuN. Further study disclosed that ASI remarkably up-regulated the mRNA and protein expressions of CXCL1 as well as that of CXCR2 in the hippocampus. The promotive effect of ASI on DCX, GFAP and NeuN protein expression was abolished by SB225002, the inhibitor of CXCR2. Our results indicated that ASI modulated the homeostasis of the CXCL1/CXCR2 signaling pathway, which might be responsible for the increased neurogenesis within the hippocampal DGs of mice.
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Liu, Xian Shuang, Michael Chopp, Xue Guo Zhang, Rui Lan Zhang, Ben Buller, Ann Hozeska-Solgot, Sara R. Gregg, and Zheng Gang Zhang. "Gene Profiles and Electrophysiology of Doublecortin-Expressing Cells in the Subventricular Zone after Ischemic Stroke." Journal of Cerebral Blood Flow & Metabolism 29, no. 2 (October 15, 2008): 297–307. http://dx.doi.org/10.1038/jcbfm.2008.119.
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Stroke increases neuroblasts in the subventricular zone (SVZ) of the lateral ventricle and these neuroblasts migrate toward the ischemic boundary to replace damaged neurons. Using brain slices from the nonischemic adult rat and transgenic mice that expressed enhanced green fluorescent protein (EGFP) concomitantly with doublecortin (DCX), a marker for migrating neuroblasts, we recorded electrophysiological characteristics while simultaneously analyzing the gene expression in single SVZ cells. We found that SVZ cells expressing the DCX gene from the nonischemic rat had a mean resting membrane potential (RMP) of −30 mV. DCX—EGFP-positive cells in the nonischemic SVZ of the transgenic mouse had a mean RMP of −25 ± 7 mV and did not exhibit Na+ currents, characteristic of immature neurons. However, DCX—EGFP-positive cells in the ischemic SVZ exhibited a hyperpolarized mean RMP of −54 ± 18 mV and displayed Na+ currents, indicative of more mature neurons. Single-cell multiplex RT-PCR analysis revealed that DCX—EGFP-positive cells in the nonischemic SVZ of the transgenic mouse expressed high neural progenitor marker genes, Sox2 and nestin, but not mature neuronal marker genes. In contrast, DCX—EGFP-positive cells in the ischemic SVZ expressed tyrosine hydroxylase, a mature neuronal marker gene. Together, these data indicate that stroke changes gene profiles and the electrophysiology of migrating neuroblasts.
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Sơn, Lê Thọ, Joohong Ahnn, Jeong Hoon Cho, and Nguyễn Huy Hoàng. "Promoters of the dhs-21 gene encoding dicarbonyl/l-xylulose reductase in Caenorhabditis elegans." Vietnam Journal of Biotechnology 15, no. 2 (April 20, 2018): 359–65. http://dx.doi.org/10.15625/1811-4989/15/2/12353.
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Dicarbonyl/L-xylulose (DCXR) was identified as a dehydrogenase. This type of enzyme was presented in
various forms of lives including bacteria, fungi, plants and animals. Generally, it converts L-xylulose to xylitol in the presence of either cofactor NADH or NADPH in vitro. Previous studies reported the biochemistry properties and crystal structure but largely uncovered biological roles of DCXRs. It was impossible to dissect the functions in mice or human cells that had many DCXR homologs in their genomes. Interestingly, the wild-type Caenorhabditis elegans, a well-known model organism in biological research, has only nuclear genomic dhs-21 that encodes a unique homologous DCXR. Thus Ce.dhs-21 and the host C. elegans were relevant for investigation of the physiologically-vital functions of the DCXR. This research aimed to the expression of dhs-21 in vivo. We defined three promoters , manipulated three relative reporter-constructs that conjugated the dhs-21 gene and Green Flouresent Protein (known as GFP) one. The construct vectors were transferred into wild-type C. elegans N2 and as well as the hermaphroditic loss of function dhs-21(jh129) by microinjection. In the results, we found that the expression pattern of dhs-21 under the only p2-promoter construct was stable and similar to immunogold Electric Microscopy (EM) images. The dhs-21 gene was expressed in both sexes of at all larval stages till the deaths of worms. DHS-21 was expressed in the cytosol of the intestinal, gonad sheath and uterous seam cell (utse).
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Venkatesh, G., M. Govindaraju, C. Kamal, P. Vennila, and S. Kaya. "Structural, electronic and optical properties of 2,5-dichloro-p-xylene: experimental and theoretical calculations using DFT method." RSC Advances 7, no. 3 (2017): 1401–12. http://dx.doi.org/10.1039/c6ra25535c.
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GRAHAM, Mark E., Patricia RUMA-HAYNES, Amanda G. CAPES-DAVIS, Joanne M. DUNN, Timothy C. TAN, Valentina A. VALOVA, Phillip J. ROBINSON, and Peter L. JEFFREY. "Multisite phosphorylation of doublecortin by cyclin-dependent kinase 5." Biochemical Journal 381, no. 2 (July 6, 2004): 471–81. http://dx.doi.org/10.1042/bj20040324.
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Doublecortin (DCX) is a 40 kDa microtubule-associated protein required for normal neural migration and cortical layering during development. Mutations in the human DCX gene cause a disruption of cortical neuronal migration. Defects in cdk5 (cyclin-dependent kinase 5) also cause defects in neural migration and cortical layering. DCX is a substrate for cdk5 in vitro and in vivo and the major site of in vitro phosphorylation is Ser-297. We used a highly developed MS strategy to identify the cdk5 phosphorylation sites and determine the major and minor sites. Several phosphopeptides were identified from a tryptic digest of 32P-labelled, cdk5-phosphorylated DCX using a combination of off-line HPLC and matrix-assisted laser-desorption ionization-MS with alkaline phosphatase treatment. Tandem MS/MS enabled the identification of seven phosphorylation sites for cdk5. Monitoring of 32P label indicated that there was one major site, Ser-28, at the N-terminus, and a major site, Ser-339, in the serine/proline-rich domain at the C-terminus. Five other sites, Ser-287, Thr-289, Ser-297, Thr-326 and Ser-332, were also found in the tail. Site-directed mutagenesis largely supported these findings. Single mutation of Ser-28 reduced but did not abolish phosphorylation. Double, rather than single, mutation for Ser-332 and Ser-339 was required to reduce overall phosphorylation, suggesting an interaction between these sites. Truncations of the tail produced a significant reduction in cdk5 phosphorylation of DCX. These results do not support Ser-297 as the major cdk5 phosphorylation site in DCX, but indicate that DCX is subject to complex multisite phosphorylation. This illustrates the importance of a well-developed MS strategy to identify phosphorylation sites.
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Osacka, Jana, Alexander Kiss, Zuzana Bacova, and Andrej Tillinger. "Effect of Haloperidol and Olanzapine on Hippocampal Cells’ Proliferation in Animal Model of Schizophrenia." International Journal of Molecular Sciences 23, no. 14 (July 12, 2022): 7711. http://dx.doi.org/10.3390/ijms23147711.
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Aberrant neurogenesis in the subventricular zone (SVZ) and hippocampus (HIP) contributes to schizophrenia pathogenesis. Haloperidol (HAL) and olanzapine (OLA), commonly prescribed antipsychotics for schizophrenia treatment, affect neurogenesis too. The effect of HAL and OLA on an mHippoE-2 cell line was studied in vitro where we measured the cell number and projection length. In vivo, we studied the gene expression of DCX, Sox2, BDNF, and NeuN in the SVZ and HIP in an MK-801-induced animal schizophrenia model. Cells were incubated with HAL, OLA, and MK-801 for 24, 48, and 72 h. Animals were injected for 6 days with saline or MK801 (0.5 mg/kg), and from the 7th day with either vehicle HAL (1 mg/kg) or OLA (2 mg/kg), for the next 7 days. In vitro, HAL and OLA dose/time-dependently suppressed cells’ proliferation and shortened their projection length. HAL/OLA co-treatment with MK-801 for 24 h reversed HAL’s/OLA’s inhibitory effect. In vivo, HAL and OLA suppressed DCX and NeuN genes’ expression in the HIP and SVZ. MK-801 decreased DCX and NeuN genes’ expression in the HIP and OLA prevented this effect. The data suggest that subchronic HAL/OLA treatment can inhibit DCX and NeuN expression. In an MK-801 schizophrenia model, OLA reversed the MK-801 inhibitory effect on DCX and NeuN and HAL reversed the effect on DCX expression; however, only in the HIP.
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Werner, Lars, Helge Müller-Fielitz, Manuela Ritzal, Tim Werner, Moritz Rossner, and Markus Schwaninger. "Involvement of Doublecortin-Expressing Cells in the Arcuate Nucleus in Body Weight Regulation." Endocrinology 153, no. 6 (April 4, 2012): 2655–64. http://dx.doi.org/10.1210/en.2011-1760.
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Hypothalamic functions, including feeding behavior, show a high degree of plasticity throughout life. Doublecortin (DCX) is a marker of plasticity and neuronal migration expressed in the hypothalamus. Therefore, we wanted to map the fate of DCX+ cells in the arcuate nucleus (ARC) of the hypothalamus. For this purpose, we generated a BAC transgenic mouse line that expresses the inducible recombinase CreERT2 under control of the DCX locus. Crossing this line with the Rosa26 or Ai14 reporter mouse lines, we found reporter+ cells in the ARC upon tamoxifen treatment. They were born prenatally and expressed both DCX and the plasticity marker TUC-4. Immediately after labeling, reporter+ cells had an enlarged soma that normalized over time, suggesting morphological remodeling. Reporter+ cells expressed β-endorphin and BSX, neuronal markers of the feeding circuit. Furthermore, leptin treatment led to phosphorylation of STAT3 in reporter+ cells in accordance with the concept that they are part of the feeding circuits. Indeed, we found a negative correlation between the number of reporter+ cells and body weight and epididymal fat pads. Our data suggest that DCX+ cells in the ARC represent a cellular correlate of plasticity that is involved in controlling energy balance in adult mice.
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Fohlen-Walter, Anne, Christine Jacob, Thomas Lecompte, and Jean-François Lesesve. "Laboratory Identification of Cryoglobulinemia From Automated Blood Cell Counts, Fresh Blood Samples, and Blood Films." American Journal of Clinical Pathology 117, no. 4 (April 2002): 606–14. http://dx.doi.org/10.1309/qxpp-dc4x-n3q8-kw62.
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FINDLEY, HENRY M., EARL INGRAM, and SEBRENA MOTEN. "THE WARN ACT AND ITS LEGAL HISTORY." Journal of Individual Employment Rights 10, no. 4 (April 1, 2002): 309–23. http://dx.doi.org/10.2190/cfgy-dcbx-kwch-x3hr.
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Meester, Ludolf E., and J. George Shanthikumar. "Regularity of Stochastic Processes: A Theory Based on Directional Convexity." Probability in the Engineering and Informational Sciences 7, no. 3 (July 1993): 343–60. http://dx.doi.org/10.1017/s0269964800002965.
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We define a notion of regularity ordering among stochastic processes called directionally convex (dcx) ordering and give examples of doubly stochastic Poisson and Markov renewal processes where such ordering is prevalent. Further-more, we show that the class of segmented processes introduced by Chang, Chao, and Pinedo [3] provides a rich set of stochastic processes where the dcx ordering can be commonly encountered. When the input processes to a large class of queueing systems (single stage as well as networks) are dcx ordered, so are the processes associated with these queueing systems. For example, if the input processes to two tandem /M/c1→/M/c2→…→/M/cm queueing systems are dcx ordered, so are the numbers of customers in the systems. The concept of directionally convex functions (Shaked and Shanthikumar [15]) and the notion of multivariate stochastic convexity (Chang, Chao, Pinedo, and Shanthikumar [4]) are employed in our analysis.
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Reiner, Orly, Amos Gdalyahu, Indraneel Ghosh, Talia Levy, Sivan Sapoznik, Ronit Nir, and Tamar Sapir. "DCXs Phosphorylation by Not Just aNother Kinase (JNK)." Cell Cycle 3, no. 6 (June 2004): 745–49. http://dx.doi.org/10.4161/cc.3.6.909.
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Sandberg, Anne, Klaus Skovbo Jensen, Pierre Baudoux, Françoise Van Bambeke, Paul M. Tulkens, and Niels Frimodt-Møller. "Intra- and Extracellular Activities of Dicloxacillin against Staphylococcus aureus In Vivo and In Vitro." Antimicrobial Agents and Chemotherapy 54, no. 6 (March 22, 2010): 2391–400. http://dx.doi.org/10.1128/aac.01400-09.
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ABSTRACT Antibiotic treatment of Staphylococcus aureus infections is often problematic due to the slow response and recurrences. The intracellular persistence of the staphylococci offers a plausible explanation for the treatment difficulties because of the impaired intracellular efficacies of the antibiotics. The intra- and extracellular time- and concentration-kill relationships were examined in vitro with THP-1 cells and in vivo by use of a mouse peritonitis model. The in vivo model was further used to estimate the most predictive pharmacokinetic/pharmacodynamic (PK/PD) indices (the ratio of the maximum concentration of drug in plasma/MIC, the ratio of the area under the concentration-time curve/MIC, or the cumulative percentage of a 24-h period that the free [f] drug concentration exceeded the MIC under steady-state pharmacokinetic conditions [fT MIC]) for dicloxacillin (DCX) intra- and extracellularly. In general, DCX was found to have similar intracellular activities, regardless of the model used. Both models showed (i) the relative maximal efficacy (1-log-unit reduction in the numbers of CFU) of DCX intracellularly and (ii) the equal relative potency of DCX intra- and extracellularly, with the MIC being a good indicator of the overall response in both situations. Discordant results, based on data obtained different times after dosing, were obtained from the two models when the extracellular activity of DCX was measured, in which the in vitro model showed a considerable reduction in the number of CFU from that in the original inoculum (3-log-unit decrease in the number of CFU after 24 h), whereas the extracellular CFU reduction achieved in vivo after 4 h did not exceed 1 log unit. Multiple dosing of DCX in vivo revealed increased extra- and intracellular efficacies (2.5 log and 2 log units of reduction in the numbers of CFU after 24 h, respectively), confirming that DCX is a highly active antistaphylococcal antibiotic. PK/PD analysis revealed that fT MIC is the index that is the most predictive of the outcome of infection both intra- and extracellularly.
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Moreno, Javier, Angeles Vicente, Alberto Varas, and Agustín G. Zapata. "T-Cell Development in Early Partially Decapitated Chicken Embryos." Developmental Immunology 4, no. 3 (1995): 211–26. http://dx.doi.org/10.1155/1995/81462.
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We have evaluated the immunohistological and cytofluorometric changes that occur in the thymus of chicken embryos partially decapitated at 33-38 hr of incubation (DCx embryos) in an attempt to analyze possible neuroendocrinological influences on T-cell differentiation and, indirectly, the ontogeny of the so-called neuroendocrine-immune network. The thymus of DCx embryos shows important variations that profoundly and selectively affect different T-cell subsets, but not the nonlymphoid cell components of thymic stroma. These modifications include the accumulation of cell precursors, mainly DN (CD4-CD8-) cells and immature CD8highCD4-cells, which expand but do not differentiate, resulting in an extreme decline of both DP (CD4+CD8+) cells and TcR c-expressing cells. Accordingly, both subcapsulary and outer cortex increase in size, whereas the deep cortex and principally the thymic medulla almost disappear in DCx embryos. In contrast, other T-cell subsets of DCx embryos, largely CDgglowCD4-cells and TcRγδ-expressing cells do not undergo significant variations throughout thymic ontogeny.
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Fourniol, Franck J., Charles V. Sindelar, Béatrice Amigues, Daniel K. Clare, Geraint Thomas, Mylène Perderiset, Fiona Francis, Anne Houdusse, and Carolyn A. Moores. "Template-free 13-protofilament microtubule–MAP assembly visualized at 8 Å resolution." Journal of Cell Biology 191, no. 3 (October 25, 2010): 463–70. http://dx.doi.org/10.1083/jcb.201007081.
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Microtubule-associated proteins (MAPs) are essential for regulating and organizing cellular microtubules (MTs). However, our mechanistic understanding of MAP function is limited by a lack of detailed structural information. Using cryo-electron microscopy and single particle algorithms, we solved the 8 Å structure of doublecortin (DCX)-stabilized MTs. Because of DCX’s unusual ability to specifically nucleate and stabilize 13-protofilament MTs, our reconstruction provides unprecedented insight into the structure of MTs with an in vivo architecture, and in the absence of a stabilizing drug. DCX specifically recognizes the corner of four tubulin dimers, a binding mode ideally suited to stabilizing both lateral and longitudinal lattice contacts. A striking consequence of this is that DCX does not bind the MT seam. DCX binding on the MT surface indirectly stabilizes conserved tubulin–tubulin lateral contacts in the MT lumen, operating independently of the nucleotide bound to tubulin. DCX’s exquisite binding selectivity uncovers important insights into regulation of cellular MTs.
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Höder, Steffen. "Phonological elements and Diasystematic Construction Grammar." Reflections on Constructions across Grammars 6, no. 2 (December 31, 2014): 202–31. http://dx.doi.org/10.1075/cf.6.2.04hod.
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Usage-based CxG approaches share the central assumption that any grammar has to be acquired and organised through input-based abstraction and categorisation. Diasystematic Construction Grammar (DCxG) is based on the idea that these processes are not sensitive to language boundaries. Multilingual input thus results in multilingual grammars which are conceived of as constructicons containing language-specific as well as language-unspecific constructions. Within such systems, phonological structures play an important part in the identification of schematic constructions. However, the status of phonology in DCxG, as in CxG in general, yet remains unclear. This paper presents some arguments for including phonological elements systematically in the construction-based analysis of (multilingual) constructional systems.
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Roque, João Vitor Oribka, Lilian Cristiane Baeza, and Eduardo Alexandre Loth. "Efeito do extrato da própolis e do digluconato de clorexidina sobre a formação de biofilme por Candida albicans em resina acrílica." Revista da Faculdade de Odontologia - UPF 25, no. 1 (December 16, 2020): 74–80. http://dx.doi.org/10.5335/rfo.v25i1.10998.
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Objetivo: avaliar in vitro a ação antimicrobiana do extrato da própolis (EP) e do digluconato de clorexidina (DCHX) na formação de biofilme por C. albicans em resina acrílica termopolimerizada. Métodos: o efeito do EP e DCHX em biofilmes de C. albicans foram avaliados através da quantificação de Unidades Formadoras de Colônias (UFCs) e pela quantificação da biomassa por cristal violeta e polissacarídeos por safranina. Para tanto, C. albicans foram pré-aderidas em corpos-de-prova e somente em microplacas de poliestireno e posteriormente foi realizado o tratamento com diferentes concentrações de EP (221 e 443 μg/mL) e DCHX (0,25% e 0,5%). Resultados: foi demostrado uma redução significativa na formação de biofilme por C. albicans, com ambas as substâncias testadas e em todas as concentrações. Conclusão: tendo em vista as dificuldades e os vários fatores que interferem no tratamento da candidose oral com antifúngicos e ao fato de essa ter sido relatada como problema de saúde pública, com alta incidência em usuários de próteses removíveis, faz-se necessários lançar-se mão de outras alternativas que possam ser mais eficazes no tratamento desta infecção, levando-se em consideração os efeitos colaterais para o paciente, sendo bem indicado o EP, por ser produto de uso tópico e, portanto, de fácil aplicação, possuindo custo menos elevado.
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Sandermann, Heinrich, Werner Heller, Norbert Hertkorn, Enamul Hoque, Dietmar Pieper, and Reinhard Winkler. "A New Intermediate in the Mineralization of 3,4-Dichloroaniline by the White Rot FungusPhanerochaete chrysosporium." Applied and Environmental Microbiology 64, no. 9 (September 1, 1998): 3305–12. http://dx.doi.org/10.1128/aem.64.9.3305-3312.1998.
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ABSTRACT Phanerochaete chrysosporium ATCC 34541 has been reported to be unable to mineralize 3,4-dichloroaniline (DCA). However, high mineralization is now shown to occur when a fermentation temperature of 37° and gassing with oxygen are used. Mineralization did not correlate with lignin peroxidase activity. The latter was high under C limitation and low under N limitation, whereas the reverse was true for mineralization. The kinetics of DCA metabolism was studied in low-N and low-C and C- and N-rich culture media by metabolite analysis and 14CO2determination. In all cases, DCA disappeared within 2 days, and a novel highly polar conjugate termed DCAX accumulated in the growth medium. This metabolite was a dead-end product under C and N enrichment. In oxygenated low-C medium and in much higher yield in oxygenated low-N medium, DCAX was converted to DCA-succinimide and then mineralized. DCAX was purified by high-performance liquid chromatography and identified asN-(3,4-dichlorophenyl)-α-ketoglutaryl-δ-amide by high-performance liquid chromatography and mass spectroscopy, gas chromatography and mass spectroscopy, and nuclear magnetic resonance spectroscopy. The formation of conjugate intermediates is proposed to facilitate mineralization because the sensitive amino group of DCA needs protection so that ring cleavage rather than oligomerization can occur.
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Coquelle, Frédéric M., Talia Levy, Sven Bergmann, Sharon Grayer Wolf, Daniela Bar-El, Tamar Sapir, Yehuda Brody, et al. "The DCX Superfamily 1: Common and Divergent Roles for Members of the Mouse DCX Superfamily." Cell Cycle 5, no. 9 (April 7, 2006): 976–83. http://dx.doi.org/10.4161/cc.5.9.2715.
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Zare, Isabelle, Dustin Paul, and Shade Moody. "Doublecortin Mutation in an Adolescent Male." Child Neurology Open 6 (January 1, 2019): 2329048X1983658. http://dx.doi.org/10.1177/2329048x19836589.
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Doublecortin (DCX) mutations cause abnormal development of the DCX protein that normally aids in neuronal migration during fetal development. These mutations lead to lissencephaly, or the appearance of a “smooth brain,” which is varying levels of pachygyria or agyria in severe cases. Many genetic variants of the mutation have been identified, and an even greater range of phenotypic presentations have been described in the literature. The X-linked lissencephaly (DCX) mutation leads to an X-linked gender-dependent condition that causes subcortical heterotopia in females and lissencephaly in males. The authors report the case of a 13-year-old male who presented to our clinic for new-onset seizure disorder. He had a past medical history of developmental delay and features of autism spectrum disorder which was diagnosed at age 5 years at an outside clinic. Magnetic resonance imaging (MRI) brain at age 5 years showed pachygyria of the frontal and temporal lobes. After extensive genetic testing over the course of over a decade, the patient was found to have a de novo mutation in the DCX gene diagnosed via whole-exome sequencing. Specifically, he was found to have a mosaic mutation of the DCX gene as a c.30-31 deletion. His previous MRI findings were consistent with a diagnosis of X-linked sporadic lissencephaly sequence and included mainly a diffuse bilateral pachygyria (isolated lissencephaly sequence X chromosome). Thickening of the cortex and pachygyria or agyria are classic findings of lissencephaly, but do not help specify any mutation in the gene, of which there are over 70 possibilities. Our patient is unique in that most individuals with DCX mutation have infantile seizures, severe intellectual disability, orthopedic complications, and postnatal microcephaly, which our patient does not have.
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Cho-Vega, Jeong Hee, Francisco Vega, Mary R. Schwartz, and Victor G. Prieto. "Expression of dicarbonyl/l-xylulose reductase (DCXR) in human skin and melanocytic lesions: morphological studies supporting cell adhesion function of DCXR." Journal of Cutaneous Pathology 34, no. 7 (July 2007): 535–42. http://dx.doi.org/10.1111/j.1600-0560.2006.00661.x.
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Rosen, Ilana. "Jelinek, Yeshayahu A.: "The Carpathian Diaspora: The Jews of Subcarpathian Rus' and Mukachevo"." Hungarian Cultural Studies 4 (January 1, 2011): 295–97. http://dx.doi.org/10.5195/ahea.2011.64.
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Çilingir Kaya, Özlem Tuğçe, Sercan Doğukan Yıldız, Nisva Hilal Levent, Esra Bihter Gürler, Ümit Süleyman Şehirli, and Serap Sirvancı. "Gender differences in doublecortin expression in the dentate gyrus of the Wistar rat during development." Anatomy 14, no. 2 (August 31, 2020): 111–16. http://dx.doi.org/10.2399/ana.20.058.
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Objectives: Neurogenesis is the formation process of functional neurons from progenitor cells which continues during lifetime. Alterations in neurogenesis is associated with neurodegenerative disorders (ND). Different mechanisms underlie the ND in males and females which may be related to neurogenesis. In this study, we aimed to investigate the developmental process of neurogenesis in the hippocampus of male and female rats at different ages and shed light on the effect of gender difference on ND. Methods: Brains were obtained from 7, 14, 21 days and 3-month-old male and female Wistar rats following intracardiac perfusion and processed for immunohistochemical and immunoflorescence staining. Doublecortin protein (DCX) was used as a marker of newly-born neuroblasts to determine neurogenesis. Results: DCX immunoreactive (-ir) cells were dispersed throughout the granular and subgranular layers of DG in 7-days-old group in both genders. However, in the 14 and 21 days old groups, DCX-ir cells were observed only in the subgranular zone in the sections labelled with both immunohistochemistry (IHC) and immunoflourescent (IF) methods. In all age groups, female rats had a tendency to increase in DCX immunoreactivity when compared to that of male Wistar rats. Conclusion: DCX-ir cells may be localized in different parts of DG during development. The number of newly born neurons showed a tendency to increase in female rats in all groups. Further studies are needed to understand the reason for differences in the normal developmental neurogenesis process between two genders.
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Benedetti, Bruno, Dominik Dannehl, Richard König, Simona Coviello, Christina Kreutzer, Pia Zaunmair, Dominika Jakubecova, et al. "Functional Integration of Neuronal Precursors in the Adult Murine Piriform Cortex." Cerebral Cortex 30, no. 3 (October 22, 2019): 1499–515. http://dx.doi.org/10.1093/cercor/bhz181.
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Abstract The extent of functional maturation and integration of nonproliferative neuronal precursors, becoming neurons in the adult murine piriform cortex, is largely unexplored. We thus questioned whether precursors eventually become equivalent to neighboring principal neurons or whether they represent a novel functional network element. Adult brain neuronal precursors and immature neurons (complex cells) were labeled in transgenic mice (DCX-DsRed and DCX-CreERT2 /flox-EGFP), and their cell fate was characterized with patch clamp experiments and morphometric analysis of axon initial segments. Young (DCX+) complex cells in the piriform cortex of 2- to 4-month-old mice received sparse synaptic input and fired action potentials at low maximal frequency, resembling neonatal principal neurons. Following maturation, the synaptic input detected on older (DCX−) complex cells was larger, but predominantly GABAergic, despite evidence of glutamatergic synaptic contacts. Furthermore, the rheobase current of old complex cells was larger and the maximal firing frequency was lower than those measured in neighboring age-matched principal neurons. The striking differences between principal neurons and complex cells suggest that the latter are a novel type of neuron and new coding element in the adult brain rather than simple addition or replacement for preexisting network components.
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Lee, Sun-Kyung, Le Tho Son, Hee-Jung Choi, and Joohong Ahnn. "Dicarbonyl/l-xylulose reductase (DCXR): The multifunctional pentosuria enzyme." International Journal of Biochemistry & Cell Biology 45, no. 11 (November 2013): 2563–67. http://dx.doi.org/10.1016/j.biocel.2013.08.010.
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Papuc, Sorina Mihaela, Magdalena Budisteanu, Alina Erbescu, Virgil Ionescu, Catrinel Iliescu, Carmen Sandu, and Aurora Arghir. "Novel DCX pathogenic variant in a girl with subcortical band heterotopia." Revista Romana de Medicina de Laborator 30, no. 3 (July 1, 2022): 345–52. http://dx.doi.org/10.2478/rrlm-2022-0031.
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Abstract Subcortical band heterotopia (SBH), is a brain malformation defined by symmetrical and bilateral heterotopic gray matter bands localized deep within the white matter, between the cortex and lateral ventricles. SBH is the result of abnormal neuronal migration, with improper positioning of the cortical neurons. DCX gene (doublecortin), a microtubule-associated protein with essential roles in neuronal migration and differentiation during brain development, is one of the main contributors to the X-linked Lissencephaly spectrum pathogenesis (OMIM #300067). DCX variants are responsible for SBH in females and isolated lissencephaly in males. Herein, we present a 7-year-old girl with a de novo frameshift variant in DCX gene, unreported by date. The patient has focal complex seizures with onset at 23 months of age, fully controlled with medication, mild tremor and coordination impairment of fine movements and some learning difficulties, otherwise with normal development. The brain magnetic resonance imaging revealed the presence of thick SBH. Direct sequencing of DCX gene revealed a pathogenic heterozygous cytosine duplication in exon 3; this frameshift variant leads to a premature stop codon in position 164 (p.Gln160Profs*5). The variant type and its predicted consequence at protein level correlates with the severity of radiological findings. The clinical presentation of our patient is, however, milder than expected. Our research expands the mutational spectrum of DCX gene in SBH females and provides a detailed clinical and imagistic description of the patient. This paper highlights the utility of single gene sequencing as a first-tier diagnostic test of patients with gene-specific phenotypic features.
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Taylor, Sabrina R., Colin Smith, Brent T. Harris, Beth A. Costine, and Ann-Christine Duhaime. "Maturation-dependent response of neurogenesis after traumatic brain injury in children." Journal of Neurosurgery: Pediatrics 12, no. 6 (December 2013): 545–54. http://dx.doi.org/10.3171/2013.8.peds13154.
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Object Traumatic brain injury (TBI) is the leading cause of acquired disability in children, yet innate repair mechanisms are incompletely understood. Given data from animal studies documenting neurogenesis in response to trauma and other insults, the authors investigated whether similar responses could be found in children of different ages after TBI. Methods Immunohistochemistry was used to label doublecortin (DCX), a protein expressed by immature migrating neuroblasts (newborn neurons), in specimens from patients ranging in age from 3 weeks to 10 years who had died either after TBI or from other causes. Doublecortin-positive (DCX+) cells were examined in the subventricular zone (SVZ) and periventricular white matter (PWM) and were quantified within the granule cell layer (GCL) and subgranular zone (SGZ) of the dentate gyrus to determine if age and/or injury affect the number of DCX+ cells in these regions. Results The DCX+ cells decreased in the SVZ as patient age increased and were found in abundance around a focal subacute infarct in a 1-month-old non-TBI patient, but were scarce in all other patients regardless of age or history of trauma. The DCX+ cells in the PWM and dentate gyrus demonstrated a migratory morphology and did not co-localize with markers for astrocytes, microglia, or macrophages. In addition, there were significantly more DCX+ cells in the GCL and SGZ of the dentate gyrus in children younger than 1 year old than in older children. The density of immature migrating neuroblasts in infants (under 1 year of age) was significantly greater than in young children (2–6 years of age, p = 0.006) and older children (7–10 years of age, p = 0.007). Conclusions The main variable influencing the number of migrating neuroblasts observed in the SVZ, PWM, and hippocampus was patient age. Trauma had no discernible effect on the number of migrating neuroblasts in this cohort of patients in whom death typically occurred within hours to days after TBI.
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Sakaguchi, A., S. Ajimura, H. Bhang, L. Busso, M. Endo, D. Faso, T. Fukuda, et al. "PRODUCTION OF NEUTRON-RICH LAMBDA HYPERNUCLEI AT J-PARC." International Journal of Modern Physics E 19, no. 12 (December 2010): 2632–37. http://dx.doi.org/10.1142/s0218301310017198.
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We discuss the usefulness of the double charge-exchange reactions (DCX) for the production of the neutron-rich Λ-hypernuclei. We believe the (π-, K+) reaction is one of the most promising DCX reactions, and propose to produce the neutron-rich Λ-hypernuclei, [Formula: see text] and [Formula: see text], at the J-PARC 50 GeV PS by the reaction (J-PARC E10 experiment). The design of the experiment is presented.
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Hubscher, Charles H., and Richard D. Johnson. "Effects of Chronic Dorsal Column Lesions on Pelvic Viscerosomatic Convergent Medullary Reticular Formation Neurons." Journal of Neurophysiology 92, no. 6 (December 2004): 3596–600. http://dx.doi.org/10.1152/jn.00310.2004.
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Single medullary reticular formation (MRF) neurons receive multiple somatovisceral convergent inputs originating from many different spinal and cranial nerves, including the pelvic nerve (PN), dorsal nerve of the penis (DNP), and the abdominal branches of the vagus. In a previous study, the input to MRF from the male genitalia was shown to be eliminated with chronic 30-day dorsal hemisection at the T8 spinal level. In this study, the effect of a smaller chronic lesion [dorsal column lesion (DCx)] on MRF neuronal responses was examined. Responses to bilateral electrical stimulation of the DNP remained. MRF neuronal responses to non-noxious (touch/stroke) levels of penile stimulation, however, were eliminated; only responses to noxious pinch remained. No differences were found for the number of neurons responding to noxious distention of the colon between the DCx and control groups. Although no differences were found across these groups for the percent MRF responses to vagal stimulation, the mean response latency for the DCx group was twice the sham-DCx/intact control group. Taken together, these results indicate that the MRF receives at least some of its input from the male genitalia via pathways located within the dorsal columns at the mid-thoracic spinal level.