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SCON—a Short Conditional intrON for conditional knockout with one-step zygote injection
Wu Szu-Hsien Sam,Lee Heetak,Szép-Bakonyi Réka,Colozza Gabriele,Boese Ayse,Gert Krista R.,Hallay Natalia,Lee Ji-Hyun,Kim Jihoon,Zhu Yi,Linssen Margot M.,Pilat-Carotta Sandra,Hohenstein Peter,Theussl Ha 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-
The generation of conditional alleles using CRISPR technology is still challenging. Here, we introduce a Short Conditional intrON (SCON, 189 bp) that enables the rapid generation of conditional alleles via one-step zygote injection. In this study, a total of 13 SCON mouse lines were successfully generated by 2 different laboratories. SCON has conditional intronic functions in various vertebrate species, and its target insertion is as simple as CRISPR/Cas9-mediated gene tagging.
True Digestibility of Phosphorus in Different Resources of Feed Ingredients in Growing Pigs
Wu, X.,Ruan, Z.,Zhang, Y.G.,Hou, Y.Q.,Yin, Y.L.,Li, T.J.,Huang, R.L.,Chu, W.Y.,Kong, X.F.,Gao, B.,Chen, L.X. Asian Australasian Association of Animal Productio 2008 Animal Bioscience Vol.21 No.1
To determine the true digestible phosphorus (TDP) requirement of growing pigs, two experiments were designed with the experimental diets containing five true digestible P levels (0.16%, 0.20%, 0.23%, 0.26% and 0.39%) and the ratio of total calcium to true digestible P (TDP) kept at 2:1. In Experiment 1, five barrows (Duroc${\times}$Landrace${\times}$Yorkshire) with an average initial body weight of 27.9 kg were used in a $5{\times}5$ Latin-square design to evaluate the effect of different dietary P levels on the digestibility and output of P and nitrogen. In Experiment 2, sixty healthy growing pigs (Duroc${\times}$Landrace${\times}$Yorkshire) with an average body weight (BW) of 21.4 kg were assigned randomly to one of the five dietary treatments (12 pigs/diet), and were used to determine the true digestible phosphorus (TDP) requirement of growing pigs on the basis of growth performance and serum biochemical indices. The results indicated that the true digestibility of P increased (p<0.05) linearly with increasing dietary TDP level below 0.26%. The true P digestibility was highest (56.6%) when dietary TDP was 0.34%. Expressed as g/kg dry matter intake (DMI), fecal P output increased (p<0.05) linearly with increasing P input. On the basis of g/kg fecal dry matter (DM), fecal P output was lowest for Diet 4 and highest (p<0.05) for Diet 5. The apparent digestibility of crude protein (CP) did not differ (p>0.05) among the five diets, with the average nitrogen output of 12.14 g/d and nitrogen retention of 66% to 74% (p>0.05), which suggested that there was no interaction between dietary P and CP protein levels. During the 28-d experimental period of Experiment 2, the average daily gain (ADG) of pigs was affected by dietary TDP levels as described by Eq. (1): $y=-809,532x^4+788,079x^3-276,250x^2+42,114x-1,759$; ($R^2=0.99$; p<0.01; y = ADG, g/d; x = dietary TDP, %), F/G for pigs by Eq. (2): $y=3,651.1x^4-3,480.4x^3+1,183.8x^2-172.5x+10.9$ ($R^2=0.99$; p<0.01; y = F/G; x = dietary TDP, %), and Total P concentrations in serum by Eq. (3): $y=-3,311.7x^4+3,342.7x^3-1,224.6x^2+195.6x-8.7$ (R2 = 0.99; p<0.01; y = total serum P concentration and x = dietary TDP, %). The highest ADG (782 g/d), the lowest F/G (1.07) and the highest total serum P concentration (3.1 mmol/L) were obtained when dietary TDP level was 0.34%. Collectively, these results indicate that the optimal TDP requirement of growing pigs is 0.34% of the diet at a total Ca to TDP ratio of 2:1.
Wang, Yong-Hao,Wang, Songhu,Liu, Hui-Gen,Hinse, Tobias C.,Laughlin, Gregory,Wu, Dong-Hong,Zhang, Xiaojia,Zhou, Xu,Wu, Zhenyu,Zhou, Ji-Lin,Wittenmyer, R. A.,Eastman, Jason,Zhang, Hui,Hori, Yasunori,Nar American Astronomical Society 2017 The Astronomical journal Vol.154 No.2
<P>We present 10. R-band photometric observations of eight different transits of the hot Jupiter HAT-P-33b, which has been targeted by our Transiting Exoplanet Monitoring Project. The data were obtained by two telescopes at the Xinglong Station of National Astronomical Observatories of China (NAOC) from 2013 December through 2016 January, and exhibit photometric scatter of 1.6-3.0 mmag. After jointly analyzing the previously published photometric data, radial-velocity (RV) measurements, and our new light curves, we revisit the system parameters and orbital ephemeris for the HAT-P-33b system. Our results are consistent with the published values except for the planet to. star radius ratio (RP/R-*), the ingress/egress duration (tau) and the total duration (T-14), which together indicate a slightly shallower and shorter transit shape. Our results are based on more complete light curves, whereas the previously published work had only one complete transit light curve. No significant anomalies in Transit Timing Variations (TTVs) are found, and we place upper mass limits on potential perturbers, largely supplanting the loose constraints provided by the extant RV data. The TTV limits are stronger near mean-motion resonances, especially for the low-order commensurabilities. We can exclude the existence of a perturber with mass larger than 0.6, 0.3, 0.5, 0.5, and 0.3 M-circle plus near the 1: 3, 1: 2, 2: 3, 3: 2, and 2: 1 resonances, respectively.</P>
Zheng, W.,Shen, R. F.,Sakamoto, T.,Beardmore, A. P.,De Pasquale, M.,Wu, X. F.,Gorosabel, J.,Urata, Y.,Sugita, S.,Zhang, B.,Pozanenko, A.,Nissinen, M.,Sahu, D. K.,Im, M.,Ukwatta, T. N.,Andreev, M.,Klun IOP Publishing 2012 The Astrophysical journal Vol.751 No.2
<P>We present a comprehensive analysis of a bright, long-duration (T-90 similar to 257 s) GRB 110205A at redshift z = 2.22. The optical prompt emission was detected by Swift/UVOT, ROTSE-IIIb, and BOOTES telescopes when the gamma-ray burst (GRB) was still radiating in the gamma-ray band, with optical light curve showing correlation with gamma-ray data. Nearly 200 s of observations were obtained simultaneously from optical, X-ray, to gamma-ray (1 eV to 5 MeV), which makes it one of the exceptional cases to study the broadband spectral energy distribution during the prompt emission phase. In particular, we clearly identify, for the first time, an interesting two-break energy spectrum, roughly consistent with the standard synchrotron emission model in the fast cooling regime. Shortly after prompt emission (similar to 1100 s), a bright (R = 14.0) optical emission hump with very steep rise (alpha similar to 5.5) was observed, which we interpret as the reverse shock (RS) emission. It is the first time that the rising phase of an RS component has been closely observed. The full optical and X-ray afterglow light curves can be interpreted within the standard reverse shock (RS) + forward shock (FS) model. In general, the high-quality prompt and afterglow data allow us to apply the standard fireball model to extract valuable information, including the radiation mechanism (synchrotron), radius of prompt emission (R-GRB similar to 3 x 10(13) cm), initial Lorentz factor of the outflow (Gamma(0) similar to 250), the composition of the ejecta (mildly magnetized), the collimation angle, and the total energy budget.</P>
Pancreatic regulation of glucose homeostasis
Pia V Röder,Bingbing Wu,Yixian Liu,Weiping Han 생화학분자생물학회 2016 Experimental and molecular medicine Vol.48 No.-
In order to ensure normal body function, the human body is dependent on a tight control of its blood glucose levels. This is accomplished by a highly sophisticated network of various hormones and neuropeptides released mainly from the brain, pancreas, liver, intestine as well as adipose and muscle tissue. Within this network, the pancreas represents a key player by secreting the blood sugar-lowering hormone insulin and its opponent glucagon. However, disturbances in the interplay of the hormones and peptides involved may lead to metabolic disorders such as type 2 diabetes mellitus (T2DM) whose prevalence, comorbidities and medical costs take on a dramatic scale. Therefore, it is of utmost importance to uncover and understand the mechanisms underlying the various interactions to improve existing anti-diabetic therapies and drugs on the one hand and to develop new therapeutic approaches on the other. This review summarizes the interplay of the pancreas with various other organs and tissues that maintain glucose homeostasis. Furthermore, anti-diabetic drugs and their impact on signaling pathways underlying the network will be discussed.
Jia, Lina,Lee, Hun Seok,Wu, Chun Fu,Kundu, Juthika,Park, Sang Gyu,Kim, Ryong Nam,Wang, Li-Hui,Erkin, Ö,zgü,r Cem,Choi, Jong-Sun,Chae, Seoung Wan,Yang, Ho Bin,Choi, Yoon-La,Shin, Young Kee American Association for Cancer Research 2014 Molecular Cancer Research Vol.12 No.12
<P>SMAD4 has been suggested to inhibit the activity of the WNT/β-catenin signaling pathway in cancer. However, the mechanism by which SMAD4 antagonizes WNT/β-catenin signaling in cancer remains largely unknown. Aurora A kinase (AURKA), which is frequently overexpressed in cancer, increases the transcriptional activity of β-catenin/T-cell factor (TCF) complex by stabilizing β-catenin through the inhibition of GSK-3β. Here, SMAD4 modulated AURKA in a TGFβ-independent manner. Overexpression of SMAD4 significantly suppressed AURKA function, including colony formation, migration, and invasion of cell lines. In addition, SMAD4 bound to AURKA induced degradation of AURKA by the proteasome. A luciferase activity assay revealed that the transcriptional activity of the β-catenin/TCF complex was elevated by AURKA, but decreased by SMAD4 overexpression. Moreover, target gene analysis showed that SMAD4 abrogated the AURKA-mediated increase of β-catenin target genes. However, this inhibitory effect of SMAD4 was abolished by overexpression of AURKA or silencing of AURKA in SMAD4-overexpressed cells. Meanwhile, the SMAD4-mediated repression of AURKA and β-catenin was independent of TGFβ signaling because blockage of TGFβR1 or restoration of TGFβ signaling did not prevent suppression of AURKA and β-catenin signaling by SMAD4. These results indicate that the tumor-suppressive function of SMAD4 is mediated by downregulation of β-catenin transcriptional activity via AURKA degradation in a TGFβ-independent manner.</P><P><B>Implications:</B> SMAD4 interacts with AURKA and antagonizes its tumor-promoting potential, thus demonstrating a novel mechanism of tumor suppression. <I>Mol Cancer Res; 12(12); 1779–95. ©2014 AACR</I>.</P>
Scheffler, T. L.,Scheffler, J. M.,Park, S.,Kasten, S. C.,Wu, Y.,McMillan, R. P.,Hulver, M. W.,Frisard, M. I.,Gerrard, D. E. American Physiological Society 2014 American journal of physiology. Cell physiology Vol.306 No.4
<P>An inverse relationship between skeletal muscle fiber cross-sectional area (CSA) and oxidative capacity suggests that muscle fibers hypertrophy at the expense of oxidative capacity. Therefore, our objective was to utilize pigs possessing mutations associated with increased oxidative capacity [AMP-activated protein kinase (AMPKγ<SUB>3</SUB><SUP>R200Q</SUP>)] or fiber hypertrophy [ryanodine receptor 1 (RyR1<SUP>R615C</SUP>)] to determine if these events occur in parallel. Longissimus muscle was collected from wild-type (control), AMPKγ<SUB>3</SUB><SUP>R200Q</SUP>, RyR1<SUP>R615C</SUP>, and AMPKγ<SUB>3</SUB><SUP>R200Q</SUP>-RyR1<SUP>R615C</SUP> pigs. Regardless of AMPK genotype, RyR<SUP>R615C</SUP> increased fiber CSA by 35%. In contrast, AMPKγ<SUB>3</SUB><SUP>R200Q</SUP> pig muscle exhibited greater citrate synthase and β-hydroxyacyl CoA dehydrogenase activity. Isolated mitochondria from AMPKγ<SUB>3</SUB><SUP>R200Q</SUP> muscle had greater maximal, ADP-stimulated oxygen consumption rate. Additionally, AMPKγ<SUB>3</SUB><SUP>R200Q</SUP> muscle contained more (∼50%) of the mitochondrial proteins succinate dehydrogenase and cytochrome <I>c</I> oxidase and more mitochondrial DNA. Surprisingly, RyR1<SUP>R615C</SUP> increased mitochondrial proteins and DNA, but this was not associated with improved oxidative capacity, suggesting that altered energy metabolism in RyR1<SUP>R615C</SUP> muscle influences mitochondrial proliferation and protein turnover. Thus pigs that possess both AMPKγ3<SUP>R200Q</SUP> and RyR<SUP>R615C</SUP> exhibit increased muscle fiber CSA as well as greater oxidative capacity. Together, our findings support the notion that hypertrophy and enhanced oxidative capacity can occur simultaneously in skeletal muscle and suggest that the signaling mechanisms controlling these events are independently regulated.</P>
ORBITAL-PHASE-DEPENDENT γ-RAY EMISSIONS FROM THE BLACK WIDOW PULSAR
Wu, E. M. H.,Takata, J.,Cheng, K. S.,Huang, R. H. H.,Hui, C. Y.,Kong, A. K. H.,Tam, P. H. T.,Wu, J. H. K. IOP Publishing 2012 The Astrophysical journal Vol.761 No.2
<P>We report on evidence for orbital phase dependence of the gamma-ray emission from the PSR B1957+20 black widow system using data from the Fermi Large Area Telescope. We divide an orbital cycle into two regions: one containing the inferior conjunction and the other containing the rest of the orbital cycle. We show that the observed spectra for the different orbital regions are fitted by different functional forms. The spectrum of the orbital region containing the inferior conjunction can be described by a power law with an exponential cutoff (PLE) model, which also gives the best-fit model for the orbital phase without the inferior conjunction, plus an extra component above similar to 2.7 GeV. The emission above 3 GeV in this region is detected with a similar to 7 sigma confidence level. The gamma-ray data above similar to 2.7 GeV are observed to be modulated at the orbital period at the similar to 2.3 sigma level. We anticipate that the PLE component dominant below similar to 2.7 GeV originates from the pulsar magnetosphere. We also show that inverse Compton scattering of the thermal radiation of the companion star off a 'cold' ultrarelativistic pulsar wind can explain the extra component above similar to 2.7 GeV. The black widow pulsar PSR B1957+20 may be a member of a new class of object, in the sense that the system is showing gamma-ray emission with both magnetospheric and pulsar wind origins.</P>