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1.  Mitochondrial dysfunction-associated OPA1 cleavage contributes to muscle degeneration: preventative effect of hydroxytyrosol acetate 
Wang, X | Li, H | Zheng, A | Yang, L | Liu, J | Chen, C | Tang, Y | Zou, X | Li, Y | Long, J | Liu, J | Zhang, Y | Feng, Z
Cell Death & Disease  2014;5(11):e1521-.
Mitochondrial dysfunction contributes to the development of muscle disorders, including muscle wasting, muscle atrophy and degeneration. Despite the knowledge that oxidative stress closely interacts with mitochondrial dysfunction, the detailed mechanisms remain obscure. In this study, tert-butylhydroperoxide (t-BHP) was used to induce oxidative stress on differentiated C2C12 myotubes. t-BHP induced significant mitochondrial dysfunction in a time-dependent manner, accompanied by decreased myosin heavy chain (MyHC) expression at both the mRNA and protein levels. Consistently, endogenous reactive oxygen species (ROS) overproduction triggered by carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), a mitochondrial oxidative phosphorylation inhibitor, was accompanied by decreased membrane potential and decreased MyHC protein content. However, the free radical scavenger N-acetyl-L-cysteine (NAC) efficiently reduced the ROS level and restored MyHC content, suggesting a close association between ROS and MyHC expression. Meanwhile, we found that both t-BHP and FCCP promoted the cleavage of optic atrophy 1 (OPA1) from the long form into short form during the early stages. In addition, the ATPase family gene 3-like 2, a mitochondrial inner membrane protease, was also markedly increased. Moreover, OPA1 knockdown in myotubes was accompanied by decreased MyHC content, whereas NAC failed to prevent FCCP-induced MyHC decrease with OPA1 knockdown, suggesting that ROS might affect MyHC content by modulating OPA1 cleavage. In addition, hydroxytyrosol acetate (HT-AC), an important compound in virgin olive oil, could significantly prevent t-BHP-induced mitochondrial membrane potential and cell viability loss in myotubes. Specifically, HT-AC inhibited t-BHP-induced OPA1 cleavage and mitochondrial morphology changes, accompanied by improvement on mitochondrial oxygen consumption capacity, ATP productive potential and activities of mitochondrial complex I, II and V. Moreover, both t-BHP- and FCCP-induced MyHC decrease was sufficiently inhibited by HT-AC. Taken together, our data provide evidence indicating that mitochondrial dysfunction-associated OPA1 cleavage may contribute to muscle degeneration, and olive oil compounds could be effective nutrients for preventing the development of muscle disorders.
doi:10.1038/cddis.2014.473
PMCID: PMC4260731  PMID: 25393477
2.  Cloning, expression and location of RNase9 in human epididymis 
Liu, J | Li, JY | Wang, HY | Zhang, CL | Li, N | Lin, YQ | Liu, J | Wang, WT
BMC Research Notes  2008;1:111.
Background
Mammalian spermatozoa become fully motile and fertile during transit through the luminal fluid of the epididymis. At least 200 proteins are present in the epididymal lumen, but the potential roles of these luminal proteins in male fertility are unknown. Investigation of the function of these proteins will elucidate the mechanism of sperm maturation, and also provide new drug targets for male contraception. We cloned RNase9 from a human epididymis cDNA library for characterization and analysis of its functions.
Findings
It was predicted that human RNase9 gene was located on chromosome 14q11.2 and encoded a 205 amino acids protein with a signal peptide of 26 amino acids at the N-terminus. The protein had eight conserved cysteine residues characteristic of the RNase A family members and several potential post-translational modification sites.
At the transcriptional level, RNase9 was expressed in a wide variety of tissues, and the expression was higher in men than in boys. RNase9 was localized to the post-equatorial region of the sperms' head. Immunofluorescence staining showed that RNase9 protein was present mostly in the epithelium of the epididymal tubule. Recombinant RNase9 had no ribonuclease activity. In addition, RNase9 had no detectable effect on sperm motility and fertilization as demonstrated by blocking spermatozoa with anti-RNase9 polyclonal serum.
Conclusion
RNase9 is expressed in a wide variety of tissues. It is located on the post-equatorial region of the sperm head and the epithelium of epididymal tubule. Although RNase9 belongs to the RNase A family, it has no ribonuclease activity.
doi:10.1186/1756-0500-1-111
PMCID: PMC2669477  PMID: 18992174
3.  Point and deletion mutations eliminate one or both methyl group transfers catalysed by the yeast TRM1 encoded tRNA (m22G26)dimethyltransferase. 
Nucleic Acids Research  1998;26(22):5102-5108.
Guanosine at position 26 in eukaryotic tRNAs is usually modified to N2 , N2 -dimethylguanosine (m22G26). In Saccharomyces cerevisiae , this reaction is catalysed by the TRM1 encoded tRNA (m22G26)dimethyltransferase. As a prerequisite for future studies, the yeast TRM1 gene was expressed in Escherichia coli and the His-tagged Trm1 protein (rTrm1p) was extensively purified. rTrm1p catalysed both the mono- and dimethylation of G26 in vivo in Escherichia coli tRNA and in vitro in yeast trm1 mutant tRNA. The TRM1 gene from two independent wild-type yeast strains differed at 14 base positions causing two amino acid exchanges . Exchange of the original Ser467 for Leu caused a complete loss of enzyme activity in vitro against trm1 yeast tRNA. Comparatively short N- or C-terminal deletions from the 570 amino acid long Trm1 polypeptide decreased or eliminated the enzyme activity, as did some point mutations within these regions. This indicated that the protein is not a two domain peptide with the enzyme activity localised to one of the domains, but rather that both ends of the polypeptide seem to interact to influence the conformation of those parts that make up the RNA-binding site and/or the active site of the enzyme.
PMCID: PMC147968  PMID: 9801306
4.  Adaptive Optical Scanning Holography 
Scientific Reports  2016;6:21636.
Optical Scanning Holography (OSH) is a powerful technique that employs a single-pixel sensor and a row-by-row scanning mechanism to capture the hologram of a wide-view, three-dimensional object. However, the time required to acquire a hologram with OSH is rather lengthy. In this paper, we propose an enhanced framework, which is referred to as Adaptive OSH (AOSH), to shorten the holographic recording process. We have demonstrated that the AOSH method is capable of decreasing the acquisition time by up to an order of magnitude, while preserving the content of the hologram favorably.
doi:10.1038/srep21636
PMCID: PMC4768316  PMID: 26916866
5.  VEGF promotes osteogenic differentiation of ASCs on ordered fluorapatite surfaces 
Vascular endothelial growth factor (VEGF) has been reported to mediate both osteogenesis and angiogenesis in bone regeneration. We previously found an up regulation of VEGF in adipose-derived stem cells (ASC) when obvious mineralization occurred on a novel fluorapatite (FA) coated surfaces. This study investigated the effect of FA and VEGF on the growth, differentiation and mineralization of (ASC) grown on ordered FA surfaces. Cells grown on FA and treated with VEGF demonstrated osteogenic differentiation as measured with ALP staining, and obvious mineralization as measured by Alizarin Red staining. A combined stimulating effect of FA and VEGF was seen using both indicators. VEGF signaling pathway perturbation using a specific VEGF receptor inhibitor showed the lowest levels of ALP and Alizarin Red staining, which was partially rescued when the cells were grown on FA and/or treated with the addition of VEGF. The osteogenic differentiation of ASCs stimulated by these FA surfaces as well as VEGF has been shown to be mediated through, but probably not only, the VEGF signaling pathway. The enhancement of osteogenic differentiation and mineralization supports the potential use of therapeutic VEGF and FA coatings in bone regeneration.
doi:10.1002/jbm.a.35215
PMCID: PMC4221573  PMID: 24797761
Fluorapatite; vascular endothelial growth factor; differentiation; mineralization; signaling pathway
6.  Renormalization-group approach to quantum Fisher information in an XY model with staggered Dzyaloshinskii-Moriya interaction 
Scientific Reports  2016;6:19359.
We investigate the quantum Fisher information and quantum phase transitions of an XY spin chain with staggered Dzyaloshinskii-Moriya interaction using the quantum renormalization-group method. The quantum Fisher information, its first-derivatives, and the finite-size scaling behaviors are rigorously calculated respectively. The singularity of the derivatives at the phase transition point as a function of lattice size is carefully discussed and it is revealed that the scaling exponent for quantum Fisher information at the critical point can be used to describe the correlation length of this model, addressing the substantial role of staggered Dzyaloshinskii-Moriya interaction in modulating quantum phase transitions.
doi:10.1038/srep19359
PMCID: PMC4726107  PMID: 26780973
7.  Magnetic phase separation in double layer ruthenates Ca3(Ru1−xTix)2O7 
Scientific Reports  2016;6:19462.
A phase transition from metallic AFM-b antiferromagnetic state to Mott insulating G-type antiferromagnetic (G-AFM) state was found in Ca3(Ru1−xTix)2O7 at about x = 0.03 in our previous work. In the present, we focused on the study of the magnetic transition near the critical composition through detailed magnetization measurements. There is no intermediate magnetic phases between the AFM-b and G-AFM states, which is in contrasted to manganites where a similar magnetic phase transition takes place through the presence of several intermediate magnetic phases. The AFM-b-to-G-AFM transition in Ca3(Ru1−xTix)2O7 happens through a phase separation process in the 2–5% Ti range, whereas similar magnetic transitions in manganites are tuned by 50–70% chemical substitutions. We discussed the possible origin of such an unusual magnetic transition and compared with that in manganites.
doi:10.1038/srep19462
PMCID: PMC4725874  PMID: 26771083
8.  Assessing the imaging performance of light sheet microscopies in highly scattering tissues 
Biomedical Optics Express  2016;7(2):454-466.
Light sheet microscopy (LSM) has emerged as an optical-imaging method for high spatiotemporal volumetric imaging of relatively transparent samples. While this capability has allowed the technique to be highly impactful in fields such as developmental biology, applications involving highly scattering thick tissues have been largely unexplored. Herein, we employ Monte Carlo simulations to explore the use of LSM for imaging turbid media. In particular, due to its similarity to dual-axis confocal (DAC) microscopy, we compare LSM performance to point-scanned (PS-DAC) and line-scanned (LS-DAC) dual-axis confocal microscopy techniques that have been previously shown to produce high-quality images at round-trip optical lengths of ~9 – 10 and ~3 – 4 respectively. The results of this study indicate that LSM using widefield collection (WF-LSM) provides comparable performance to LS-DAC in thick tissues, due to the fact that they both utilize an illumination beam focused in one dimension (i.e. a line or sheet). On the other hand, LSM using confocal line detection (CL-LSM) is more analogous to PS-DAC microscopy, in which the illumination beam is focused in two dimensions to a point. The imaging depth of LSM is only slightly inferior to DAC (~2 – 3 and ~6 – 7 optical lengths for WF-LSM and CL-LSM respectively) due to the use of a lower numerical aperture (NA) illumination beam for extended imaging along the illumination axis. Therefore, we conclude that the ability to image deeply is dictated most by the confocality of the microscope technique. In addition, we find that imaging resolution is mostly dependent on the collection NA, and is relatively invariant to imaging depth in a homogeneous scattering medium. Our results indicate that superficial imaging of highly scattering tissues using light sheet microscopy is possible.
doi:10.1364/BOE.7.000454
PMCID: PMC4771464  PMID: 26977355
(170.1790) Confocal microscopy; (170.2520) Fluorescence microscopy; (170.5810) Scanning microscopy; (110.0113) Imaging through turbid media; (170.3880) Medical and biological imaging
9.  Miniature in vivo MEMS-based line-scanned dual-axis confocal microscope for point-of-care pathology 
Biomedical Optics Express  2016;7(2):251-263.
There is a need for miniature optical-sectioning microscopes to enable in vivo interrogation of tissues as a real-time and noninvasive alternative to gold-standard histopathology. Such devices could have a transformative impact for the early detection of cancer as well as for guiding tumor-resection procedures. Miniature confocal microscopes have been developed by various researchers and corporations to enable optical sectioning of highly scattering tissues, all of which have necessitated various trade-offs in size, speed, depth selectivity, field of view, resolution, image contrast, and sensitivity. In this study, a miniature line-scanned (LS) dual-axis confocal (DAC) microscope, with a 12-mm diameter distal tip, has been developed for clinical point-of-care pathology. The dual-axis architecture has demonstrated an advantage over the conventional single-axis confocal configuration for reducing background noise from out-of-focus and multiply scattered light. The use of line scanning enables fast frame rates (16 frames/sec is demonstrated here, but faster rates are possible), which mitigates motion artifacts of a hand-held device during clinical use. We have developed a method to actively align the illumination and collection beams in a DAC microscope through the use of a pair of rotatable alignment mirrors. Incorporation of a custom objective lens, with a small form factor for in vivo clinical use, enables our device to achieve an optical-sectioning thickness and lateral resolution of 2.0 and 1.1 microns respectively. Validation measurements with reflective targets, as well as in vivo and ex vivo images of tissues, demonstrate the clinical potential of this high-speed optical-sectioning microscopy device.
doi:10.1364/BOE.7.000251
PMCID: PMC4771446  PMID: 26977337
(170.1790) Confocal microscopy; (170.2520) Fluorescence microscopy; (170.5810) Scanning microscopy; (170.3880) Medical and biological imaging; (230.4685) Optical microelectromechanical devices
10.  π Berry phase and Zeeman splitting of Weyl semimetal TaP 
Scientific Reports  2016;6:18674.
The recent breakthrough in the discovery of Weyl fermions in monopnictide semimetals provides opportunities to explore the exotic properties of relativistic fermions in condensed matter. The chiral anomaly-induced negative magnetoresistance and π Berry phase are two fundamental transport properties associated with the topological characteristics of Weyl semimetals. Since monopnictide semimetals are multiple-band systems, resolving clear Berry phase for each Fermi pocket remains a challenge. Here we report the determination of Berry phases of multiple Fermi pockets of Weyl semimetal TaP through high field quantum transport measurements. We show our TaP single crystal has the signatures of a Weyl state, including light effective quasiparticle masses, ultrahigh carrier mobility, as well as negative longitudinal magnetoresistance. Furthermore, we have generalized the Lifshitz-Kosevich formula for multiple-band Shubnikov-de Haas (SdH) oscillations and extracted the Berry phases of π for multiple Fermi pockets in TaP through the direct fits of the modified LK formula to the SdH oscillations. In high fields, we also probed signatures of Zeeman splitting, from which the Landé g-factor is extracted.
doi:10.1038/srep18674
PMCID: PMC4698660  PMID: 26726050
11.  Fluorapatite-modified Scaffold on Dental Pulp Stem Cell Mineralization 
Journal of Dental Research  2014;93(12):1290-1295.
In previous studies, fluorapatite (FA) crystal-coated surfaces have been shown to stimulate the differentiation and mineralization of human dental pulp stem cells (DPSCs) in two-dimensional cell culture. However, whether the FA surface can recapitulate these properties in three-dimensional culture is still unknown. This study examined the differences in behavior of human DPSCs cultured on electrospun polycaprolactone (PCL) NanoECM nanofibers with or without the FA crystals. Under near-physiologic conditions, the FA crystals were synthesized on the PCL nanofiber scaffolds. The FA crystals were evenly distributed on the scaffolds. DPSCs were cultured on the PCL+FA or the PCL scaffolds for up to 28 days. Scanning electron microscope images showed that DPSCs attached well to both scaffolds after the initial seeding. However, it appeared that more multicellular aggregates formed on the PCL+FA scaffolds. After 14 days, the cell proliferation on the PCL+FA was slower than that on the PCL-only scaffolds. Interestingly, even without any induction of mineralization, from day 7, the upregulation of several pro-osteogenic molecules (dmp1, dspp, runx2, ocn, spp1, col1a1) was detected in cells seeded on the PCL+FA scaffolds. A significant increase in alkaline phosphatase activity was also seen on FA-coated scaffolds compared with the PCL-only scaffolds at days 14 and 21. At the protein level, osteocalcin expression was induced only in the DPSCs on the PCL+FA surfaces at day 21 and then significantly enhanced at day 28. A similar pattern was observed in those specimens stained with Alizarin red and Von Kossa after 21 and 28 days. These data suggest that the incorporation of FA crystals within the three-dimensional PCL nanofiber scaffolds provided a favorable extracellular matrix microenvironment for the growth, differentiation, and mineralization of human DPSCs. This FA-modified PCL nanofiber scaffold shows promising potential for future bone, dental, and orthopedic regenerative applications.
doi:10.1177/0022034514547914
PMCID: PMC4462802  PMID: 25139361
odontogenesis; osteogenesis; biomaterial(s); cell differentiation; nanofibers; tissue engineering
12.  Treatment patterns among Canadian men diagnosed with localized low-risk prostate cancer 
Current Oncology  2015;22(6):427-429.
In general, guideline-recommended treatment options for men with low-risk prostate cancer (pca) include active surveillance, radical prostatectomy, and external-beam radiation therapy or brachytherapy. Because of the concern about overdiagnosis and consequent overtreatment of pca, patients with low-risk disease are increasingly being managed with active surveillance. Using data from six provincial cancer registries, we examined treatment patterns within a year of a diagnosis of localized low-risk pca, and we assessed differences by age.
Of patients diagnosed in 2010 in four of the six reporting provinces, most received surgery or radiation therapy within 1 year of diagnosis. Depending on the province, either surgery or radiation therapy was the most commonly used primary treatment. In the other two provinces, most patients had no record of treatment within a year of diagnosis. Examining treatment patterns by age demonstrated a lesser likelihood of receiving surgery or radiation therapy within 1 year of diagnosis among men more than 75 years of age than among men 75 years of age or younger (no record of treatment in 69.1% and 46.3% respectively).
In conclusion, we observed interprovincial and age-specific variations in the patterns of care for men with low-risk pca. The findings presented in this report are intended to identify opportunities for improvement in clinical practice that could lead to improved care and experience.
doi:10.3747/co.22.2895
PMCID: PMC4687664  PMID: 26715876
Low-risk prostate cancer; treatment; older men; active surveillance; observation
13.  Effects of PI3K inhibitor NVP-BKM120 on overcoming drug resistance and eliminating cancer stem cells in human breast cancer cells 
Hu, Y | Guo, R | Wei, J | Zhou, Y | Ji, W | Liu, J | Zhi, X | Zhang, J
Cell Death & Disease  2015;6(12):e2020-.
The multidrug resistance (MDR) phenotype often accompanies activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway, which renders a survival signal to withstand cytotoxic anticancer drugs and enhances cancer stem cell (CSC) characteristics. As a result, PI3K/AKT-blocking approaches have been proposed as antineoplastic strategies, and inhibitors of PI3K/AKT are currently being trailed clinically in breast cancer patients. However, the effects of PI3K inhibitors on MDR breast cancers have not yet been elucidated. In the present study, the tumorigenic properties of three MDR breast cancer cell lines to a selective inhibitor of PI3K, NVP-BKM120 (BKM120), were assessed. We found that BKM120 showed a significant cytotoxic activity on MDR breast cancer cells both in vitro and in vivo. When doxorubicin (DOX) was combined with BKM120, strong synergistic antiproliferative effect was observed. BKM120 activity induced the blockage of PI3K/AKT signaling and NF-κB expression, which in turn led to activate caspase-3/7 and caspase-9 and changed the expression of several apoptosis-related gene expression. Furthermore, BKM120 effectively eliminated CSC subpopulation and reduced sphere formation of these drug-resistant cells. Our findings indicate that BKM120 partially overcomes the MDR phenotype in chemoresistant breast cancer through cell apoptosis induction and CSC abolishing, which appears to be mediated by the inhibition of the PI3K/AKT/NF-κB axis. This offers a strong rationale to explore the therapeutic strategy of using BKM120 alone or in combination for chemotherapy-nonresponsive breast cancer patients.
doi:10.1038/cddis.2015.363
PMCID: PMC4720896  PMID: 26673665
14.  Porous VOxNy nanoribbons supported on CNTs as efficient and stable non-noble electrocatalysts for the oxygen reduction reaction 
Scientific Reports  2015;5:17385.
Novel nanocomposites of carbon nanotubes supported porous VOxNy nonoribbons (VOxNy-CNTs) have been synthesized by the annealing of the sol-gel mixture of CNTs and V2O5 under NH3 atmosphere as well as the ageing process in air. Besides the morphological and structural characterizations revealed by TEM, SEAD, EDS, XRD and XPS measurements, typical electrochemical tests including cyclic voltammetry (CV), rotating disk electrode (RDE) and chronoamperometry have been employed to determine the oxygen reduction reaction (ORR) performance of VOxNy-CNTs. Inspiringly, the results indicate that VOxNy-CNTs catalyst exhibits a 0.4 mA/cm2 larger diffusion-limited current density, a 0.10  V smaller onset potential value, a 10.73% less of ORR current decay and an excellent methanol-tolerance compared with commercial Pt/C catalyst. Therefore, we have reasonable grounds to believe that this new VOxNy-CNTs nanocomposites can be regarded as a promising non-precious methanol-tolerant ORR catalyst candidate for alkaline fuel cells.
doi:10.1038/srep17385
PMCID: PMC4663630  PMID: 26616719
15.  Atomistic Design of Favored Compositions for Synthesizing the Al-Ni-Y Metallic Glasses 
Scientific Reports  2015;5:16218.
For a ternary alloy system promising for obtaining the so-called bulk metallic glasses (BMGs), the first priority issue is to predict the favored compositions, which could then serve as guidance for the appropriate alloy design. Taking the Al-Ni-Y system as an example, here we show an atomistic approach, which is developed based on a recently constructed and proven realistic interatomic potential of the system. Applying the Al-Ni-Y potential, series simulations not only clarify the glass formation mechanism, but also predict in the composition triangle, a hexagonal region, in which a disordered state, i.e., the glassy phase, is favored energetically. The predicted region is defined as glass formation region (GFR) for the ternary alloy system. Moreover, the approach is able to calculate an amorphization driving force (ADF) for each possible glassy alloy located within the GFR. The calculations predict an optimized sub-region nearby a stoichiometry of Al80Ni5Y15, implying that the Al-Ni-Y metallic glasses designed in the sub-region could be the most stable. Interestingly, the atomistic predictions are supported by experimental results observed in the Al-Ni-Y system. In addition, structural origin underlying the stability of the Al-Ni-Y metallic glasses is also discussed in terms of a hybrid packing mode in the medium-range scale.
doi:10.1038/srep16218
PMCID: PMC4655373  PMID: 26592568
16.  Magnetic-charge ordering and phase transitions in monopole-conserved square spin ice 
Scientific Reports  2015;5:15875.
Magnetic-charge ordering and corresponding magnetic/monopole phase transitions in spin ices are the emergent topics of condensed matter physics. In this work, we investigate a series of magnetic-charge (monopole) phase transitions in artificial square spin ice model using the conserved monopole density algorithm. It is revealed that the dynamics of low monopole density lattices is controlled by the effective Coulomb interaction and the Dirac string tension, leading to the monopole dimerization which is quite different from the dynamics of three-dimensional pyrochlore spin ice. The condensation of the monopole dimers into monopole crystals with staggered magnetic-charge order can be predicted clearly. For the high monopole density cases, the lattice undergoes two consecutive phase transitions from high-temperature paramagnetic/charge-disordered phase into staggered charge-ordered phase before eventually toward the long-range magnetically-ordered phase as the ground state which is of staggered charge order too. A phase diagram over the whole temperature-monopole density space, which exhibits a series of emergent spin and monopole ordered states, is presented.
doi:10.1038/srep15875
PMCID: PMC4625371  PMID: 26511870
17.  Effects of Replacement of Concentrate Mixture by Broccoli Byproducts on Lactating Performance in Dairy Cows 
The objective of the present study was to determine the effects of feeding pelletized broccoli byproducts (PBB) on milk yield and milk composition in dairy cows. In Trial 1, an in vitro gas test determined the optimal replacement level of PBB in a concentrate mixture in a mixed substrate with Chinese wild ryegrass hay (50:50, w/w) at levels of 0, 10%, 20%, 30%, or 40% (dry matter basis). When the concentrate was replaced by PBB at a level of 20%, no adverse effects were found on the gas volume or its rate constant during ruminal fermentation. In trial 2, 24 lactating cows (days in milk = 170.4±35; milk yield = 30±3 kg/d; body weight = 580 ±13 kg) were divided into 12 blocks based on day in milk and milk yield and randomly allocated to two dietary treatments: a basic diet with or without PBB replacing 20% of the concentrate mixture. The feeding trial lasted for 56 days; the first week allowed for adaptation to the diet. The milk composition was analyzed once a week. No significant difference in milk yield was observed between the two groups (23.5 vs 24.2 kg). A significant increase was found in milk fat content in the PBB group (p<0.05). Inclusion of PBB did not affect milk protein, lactose, total solids or solids-not-fat (p>0.05). These results indicated that PBB could be included in dairy cattle diets at a suitable level to replace concentrate mixture without any adverse effects on dairy performance.
doi:10.5713/ajas.15.0016
PMCID: PMC4554852  PMID: 26323401
Broccoli Byproducts; Rumen Fermentation; Milk Yield; Milk Composition
18.  Measurement of the Intestinal Digestibility of Rumen Undegraded Protein Using Different Methods and Correlation Analysis 
Four methods were adopted, including the mobile nylon bag (MNB) method, modified three-step in vitro (MTS) method, original three-step in vitro (OTS) method, and acid detergent insoluble nitrogen (ADIN) estimating method, to evaluate the intestinal digestibility of rumen undegradable protein (DRUP) of 10 types of concentrates and 7 types of roughages. After correlation analysis to determine the DRUP values using the MNB, MTS, OTS, and ADIN methods, the study aimed to find out appropriate methods to replace the MNB method due to its disadvantages such as high price, long time period, and use of a duodenal T-fistula. Three dairy cows with a permanent ruminal fistula and duodenal T-fistula were used in a single-factor experimental design. The results showed that the determined DRUP values using the MNB method for soybean meal, cottonseed meal, rapeseed meal, sunflower meal, corn germ meal, corn, rice bran, barley, wheat bran, corn fiber feed, Alfalfa (Zhao dong), Alfalfa (Long mu 801), Alfalfa (Long mu 803), grass (North), Grass (Inner Mongolia), corn silage and corn straw were 98.13%, 87.37%, 88.47%, 82.60%, 75.40%, 93.23%, 69.27%, 91.27%, 72.37%, 79.03%, 66.72%, 68.64%, 73.57%, 50.47%, 51.52%, 54.05%, and 43.84%, respectively. The coefficient of determination (R2 = 0.964) of the results between the MTS method and the MNB method was higher than that (R2 = 0.942) between the OTS method and the MNB method. The coefficient of determination of the DRUP values of the concentrates among the in vitro method (including the MTS and OTS methods) and the MNB method was higher than that of the roughage. There was a weak correlation between the determined DRUP values in concentrates obtained from the ADIN method and those from the MNB method, and there was a significant correlation (p<0.01) between the determined DRUP values of the roughage obtained from the MNB method and those obtained from ADIN method. The DRUP values were significantly correlated with the nutritional ingredients of the feeds. The regression equation was DRUP =100.5566+0.4169CP − 0.4344SP − 0.7102NDF − 0.7950EE (R2 = 0.8668, p<0.01; CP, crude protein; SP, soluble protein; NDF, neutral detergent fiber; EE, ether extract). It was concluded that both the MTS method and the OTS may suitable to replace the MNB method for determining the DRUP values and the former method was more effective. Only the ADIN method could be used to predict the values of the roughages but conventional nutritional ingredients were available for all of the samples’ DRUP.
doi:10.5713/ajas.15.0085
PMCID: PMC4554853  PMID: 26323402
Protein Supplement; Energy Feed; Rumen Undegraded Protein; Correlation Analysis; Intestinal Digestibility
19.  Wait times for prostate cancer treatment and patient perceptions of care in Canada: a mixed-methods report 
Current Oncology  2015;22(5):361-364.
Background
Access to cancer care is a significant concern for Canadians. Prolonged delays between cancer diagnosis and treatment have been associated with anxiety, stress, and perceived powerlessness for patients and their family members. Longer wait times can also be associated with poorer prognosis, although the evidence is inconclusive. Here, we report national wait times for radiation therapy and surgery for localized prostate cancer (pca) and the effect of wait time on patient perceptions of their care.
Results
Treatment wait times showed substantial interprovincial variation. The longest 90th percentile wait times for radiation therapy and surgery were, respectively, 40 days and 105 days. In all provinces, waits for radiation therapy were longer for pca patients than for patients with breast, colorectal, or lung cancer.
In the focus groups and interviews conducted with 47 men treated for pca, many participants did not perceive that wait times for treatment were prolonged. Those who experienced delays between diagnosis and treatment voiced issues with a lack of communication about when they would receive treatment and a lack of support or information to make an informed decision about treatment. Minimizing treatment delays was an aspect of the cancer journey that participants would like to change because of the stress it caused.
Conclusions
Although wait time statistics are useful, a review of cancer control in Canada cannot be considered complete unless an effort is made to give voice to the experiences of individuals with cancer. The findings presented here are intended to provide a snapshot of national care delivery for localized pca and to identify opportunities for improvement in clinical practice.
doi:10.3747/co.22.2795
PMCID: PMC4608402  PMID: 26628869
Prostate cancer; treatment; surgery; radiation therapy; access to health care; qualitative research
20.  Graphite Carbon-Supported Mo2C Nanocomposites by a Single-Step Solid State Reaction for Electrochemical Oxygen Reduction 
PLoS ONE  2015;10(9):e0138330.
Novel graphite-molybdenum carbide nanocomposites (G-Mo2C) are synthesized by a typical solid state reaction with melamine and MoO3 as precursors under inert atmosphere. The characterization results indicate that G-Mo2C composites are composed of high crystallization and purity of Mo2C and few layers of graphite carbon. Mo2C nanoparticles with sizes ranging from 5 to 50 nm are uniformly supported by surrounding graphite layers. It is believed that Mo atom resulting from the reduction of MoO3 is beneficial to the immobilization of graphite carbon. Moreover, the electrocatalytic performances of G-Mo2C for ORR in alkaline medium are investigated by cyclic voltammetry (CV), rotating disk electrode (RDE) and chronoamperometry test with 3M methanol. The results show that G-Mo2C has a considerable catalytic activity and superior methanol tolerance performance for the oxygen reduction reaction (ORR) benefiting from the chemical interaction between the carbide nanoparticles and graphite carbon.
doi:10.1371/journal.pone.0138330
PMCID: PMC4575164  PMID: 26381266
21.  Bone marrow-derived mesenchymal stem cells ameliorate chronic high glucose-induced β-cell injury through modulation of autophagy 
Zhao, K | Hao, H | Liu, J | Tong, C | Cheng, Y | Xie, Z | Zang, L | Mu, Y | Han, W
Cell Death & Disease  2015;6(9):e1885-.
Chronic hyperglycemia causes a progressive decrease of β-cell function and mass in type 2 diabetic patients. Growing evidence suggests that augment of autophagy may be an effective approach to protect β cells against various extra-/intracellular stimuli. In this study, we thus investigated whether bone marrow-derived mesenchymal stem cells (BM-MSCs) could ameliorate chronic high glucose (HG)-induced β-cell injury through modulation of autophagy. Prolonged exposure to HG decreased cell viability, increased cell apoptosis and impaired basal insulin secretion and glucose-stimulated insulin secretion of INS-1 cells, but BM-MSC treatment significantly alleviated these glucotoxic alternations. In addition, western blotting displayed upregulated expression of Beclin1 and LC3-II in INS-1 cells co-cultured with BM-MSCs. Results from immunofluorescence staining and transmission electronic microscope analysis also revealed that BM-MSCs promoted autophagosomes and autolysosomes formation in HG-treated INS-1 cells. However, it should be noted that inhibition of autophagy significantly diminished the protective effects of BM-MSCs on HG-treated INS-1 cells, suggesting that the improvement of β-cell function and survival induced by BM-MSCs was mediated through autophagy. Furthermore, our results showed that BM-MSCs improved mitochondrial function and reduced reactive oxygen species production in HG-treated INS-1 cells, largely owing to autophagic clearance of impaired mitochondria. In vivo study was performed in rats with type 2 diabetes (T2D). BM-MSC infusion not only ameliorated hyperglycemia, but also promoted restoration of pancreatic β cells in T2D rats. Meanwhile, BM-MSC infusion upregulated LAMP2 expression and enhanced formation of autophagosomes and autolysosomes, combined with reduced β-cell apoptosis and increased number of insulin granules. These findings together indicated that BM-MSCs could protect β cells against chronic HG-induced injury through modulation of autophagy in vitro and in vivo. This study unveiled novel evidence of BM-MSCs as an ideal strategy to enhance autophagy for treatment of T2D mellitus.
doi:10.1038/cddis.2015.230
PMCID: PMC4650435  PMID: 26379190
22.  Subharmonic aided pressure estimation for monitoring interstitial fluid pressure in tumours -in vitro and in vivo proof of concept 
Ultrasonics  2014;54(7):1938-1944.
The feasibility of using subharmonic aided pressure estimation (SHAPE) to noninvasively estimate interstitial fluid pressure (IFP) was studied. In vitro, radiofrequency signals, from 0.2 ml/l of Definity (Lantheus Medical Imaging, N Billerica, MA) were acquired within a water-tank with a Sonix RP ultrasound scanner (Ultrasonix, Richmond, BC, Canada; fT/R=6.7/3.35 MHz and fT/R =10/5 MHz) and the subharmonic amplitudes of the signals were compared over 0–50 mmHg. In vivo, five swine with naturally occurring melanomas were studied. Subharmonic signals were acquired from tumours and surrounding tissue during infusion of Definity and compared to needle-based pressure measurements. Both in vitro and in vivo, an inverse linear relationship between hydrostatic pressure and subharmonic amplitude was observed with r2=0.63–0.95; p<0.05, maximum amplitude drop 11.36 dB at 10 MHz and −8 dB, and r2 as high as 0.97; p<0.02 (10 MHz and −4/−8 dB most promising), respectively, indicating that SHAPE may be useful in monitoring IFP.
doi:10.1016/j.ultras.2014.04.022
PMCID: PMC4120866  PMID: 24856899
Pressure estimation; Contrast agents; Subharmonic signals; Breast cancer; Ultrasound imaging
23.  Efficient and accurate treatment of electron correlations with Correlation Matrix Renormalization theory 
Scientific Reports  2015;5:13478.
We present an efficient method for calculating the electronic structure and total energy of strongly correlated electron systems. The method extends the traditional Gutzwiller approximation for one-particle operators to the evaluation of the expectation values of two particle operators in the many-electron Hamiltonian. The method is free of adjustable Coulomb parameters, and has no double counting issues in the calculation of total energy, and has the correct atomic limit. We demonstrate that the method describes well the bonding and dissociation behaviors of the hydrogen and nitrogen clusters, as well as the ammonia composed of hydrogen and nitrogen atoms. We also show that the method can satisfactorily tackle great challenging problems faced by the density functional theory recently discussed in the literature. The computational workload of our method is similar to the Hartree-Fock approach while the results are comparable to high-level quantum chemistry calculations.
doi:10.1038/srep13478
PMCID: PMC4551991  PMID: 26315767
24.  Prognostic significance of p62/SQSTM1 subcellular localization and LC3B in oral squamous cell carcinoma 
British Journal of Cancer  2014;111(5):944-954.
Background:
Autophagy is a programmed cell survival mechanism that has a key role in both physiologic and pathologic conditions. The relationship between autophagy and cancer is complex because autophagy can act as either a tumour suppressor or as a tumour promoter. The role of autophagy in oral squamous cell carcinoma (OSCC) is controversial. Several studies have claimed that either a high or low expression of autophagy-related proteins was associated with poor prognosis of OSCCs. The aims of the study were to compare autophagy in OSCCs, verrucous hyperplasias, and normal oral mucosas, and to inspect the prognostic role of autophagy in OSCCs.
Methods:
We used the autophagosome marker, LC3B, and autophagy flux marker, p62/SQSTM1 (p62), by using immunohistochemistry, and examined p62 mRNA by RNA in situ hybridization, to evaluate autophagy in 195 OSCCs, 47 verrucous hyperplasias, and 37 normal oral mucosas. The prognostic roles of LC3B and p62 protein expressions in OSCCs were investigated.
Results:
We discovered that the normal oral mucosa exhibited limited LC3B punctae and weak cytoplasmic p62 staining, whereas the OSCCs exhibited a marked increase in LC3B punctae and cytoplasmic p62 expression. The expression pattern of LC3B and cytoplasmic p62 of the verrucous hyperplasias were between normal oral mucosas and OSCCs. The normal oral mucosas, verrucous hyperplasias, and OSCCs presented no differences in nuclear p62 expression and the p62 mRNA level. p62 mRNA expression was elevated in a minority of cases. High p62 mRNA expression was associated with high p62 protein expression in the cytoplasm. Increased LC3B punctae, high cytoplasmic p62, and low nuclear p62 expressions in OSCCs were associated with aggressive clinicopathologic features and unfavourable prognosis. In addition, low nuclear p62 expression was an independent prognostic factor for overall and disease-specific survival rates. Furthermore, we disclosed that high cytoplasmic p62 expression accompanied with either a low or high LC3B expression, which indicated autophagy impairment under basal or activated autophagic activity, was associated with aggressive behaviour in advanced OSCCs.
Conclusions:
We suggested that autophagy was altered during cancer initiation and progression. Autophagy impairment contributed to cancer progression in advanced OSCCs.
doi:10.1038/bjc.2014.355
PMCID: PMC4150268  PMID: 24983366
autophagy; immunohistochemistry; in situ hybridization; LC3; p62/SQSTM1; oral; squamous cell carcinoma
25.  Survival of Recipients of Livers from Donation after Circulatory Death 
Use of donation after circulatory death (DCD) as a strategy to increase the pool of transplantable livers has been limited due to poorer recipient outcomes compared with donation after brain death (DBD). We examined outcomes of failed DCD grafts regarding wait-list mortality after relisting and patient and graft survival after retransplant. From the Scientific Registry of Transplant Recipients database, we identified 1820 adults who underwent first deceased donor liver transplant January 1, 2004–June 30, 2011, and were relisted due to graft failure; 12.7% were DCD recipients. Compared with DBD recipients, DCD recipients had better wait-list survival (90-day mortality: 8%, DCD recipients; 14%–21%, DBD recipients). Of 950 retransplant patients, 14.5% were prior DCD recipients. Graft survival after second liver transplant was similar for prior DCD (28% graft failure within 1 year) and DBD recipients (30%). Patient survival was slightly better for prior DCD (25% death within 1 year) than DBD recipients (28%). Despite higher graft failure and morbidity rates, survival of prior DCD recipients who were selected for relisting and retransplant was not worse than survival of DBD recipients.
doi:10.1111/ajt.12700
PMCID: PMC4546823  PMID: 24731165
Graft survival; liver transplantation; outcomes; patient survival; wait-list mortality

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