Celecoxib-Induced Cytotoxic Effect Is Potentiated by Inhibition of Autophagy in Human Urothelial Carcinoma Cells

Celecoxib, a cyclooxygenase-2 (COX-2) inhibitor, can elicit anti-tumor effects in various malignancies. Here, we sought to clarify the role of autophagy in celecoxib-induced cytotoxicity in human urothelial carcinoma (UC) cells. The results shows celecoxib induced cellular stress response such as endoplasmic reticulum (ER) stress, phosopho-SAPK/JNK, and phosopho-c-Jun as well as autophagosome formation in UC cells. Inhibition of autophagy by 3-methyladenine (3-MA), bafilomycin A1 or ATG7 knockdown potentiated celecoxib-induced apoptosis. Up-regulation of autophagy by rapamycin or GFP-LC3B-transfection alleviated celecoxib-induced cytotoxicity in UC cells. Taken together, the inhibition of autophagy enhances therapeutic efficacy of celecoxib in UC cells, suggesting a novel therapeutic strategy against UC.     

Severe acute respiratory syndrome coronavirus 3a protein is a viral structural protein

The present study showed the association of a severe acute respiratory syndrome coronavirus (SCoV) accessory protein, 3a, with plasma membrane and intracellular SCoV particles in infected cells. 3a protein appeared to undergo posttranslational modifications in infected cells and was incorporated into SCoV particles, establishing that 3a protein was a SCoV structural protein.     

Effects of environmental factors on genetic diversity of Caragana microphylla in Horqin Sandy Land,northeast China

Caragana microphylla (Leguminosae) is a dominant climax semishrub species in northern China. We evaluated genetic variation within and among populations sampled from three different environmental gradients in Horqin Sandy Land in northern China using intersimple sequence repeats markers and investigated the possible existence of relationships between genetic diversity and environmental factors. The results showed that C. microphylla have high genetic diversity, and environmental gradients affected genetic diversity of C. microphylla populations. Genetic diversity of all populations was affected by many environmental factors and as well correlated with warm index and soil Olsen phosphorus (SOP) concentration. These results have important implications for restoration and management of these degraded ecosystems in arid and semi‐arid areas.     

Revised treatment of <Emphasis Type="Italic">Memecylon</Emphasis> sect. <Emphasis Type="Italic">Afzeliana</Emphasis> (<Emphasis Type="Italic">Melastomataceae: Olisbeoideae</Emphasis>), including three new species from Cameroon

Summary  Seven new names at species rank are proposed in Memecylon sect. Afzeliana Jacq.-Fél., a group of forest shrubs and small trees confined to Guineo-Congolian Africa. The group is centred in Cameroon, where 17 of the 20 species occur. A new flower type, the “star-flower” in Memecylon is revealed, and its taxonomic and ecological importance discussed. Three new, locally endemic species from the South West Province of Cameroon are described, mapped and illustrated: M. kupeanum R. D. Stone, Ghogue & Cheek, M. bakossiense R. D. Stone, Ghogue & Cheek, and M. rheophyticum R. D. Stone, Ghogue & Cheek. Two new names, M. accedens R. D. Stone, Ghogue & Cheek and M. hyleastrum R. D. Stone & Ghogue and one new combination, M. mamfeanum (Jacq.-Fél.) R. D. Stone, Ghogue & Cheek are provided at species level for three taxa originally proposed as varieties of M. afzelii G. Don. The taxon M. arcuatomarginatum var. simulans Jacq.-Fél. is also elevated to species status, as M. simulans (Jacq.-Fél.) R. D. Stone & Ghogue. Conservation assessments are provided for all the newly named taxa. A key is provided to the species of Memecylon sect. Afzeliana.     

NmeCas9 is an intrinsically high-fidelity genome-editing platform

Background

The development of CRISPR genome editing has transformed biomedical research. Most applications reported thus far rely upon the Cas9 protein from Streptococcus pyogenes SF370 (SpyCas9). With many RNA guides, wildtype SpyCas9 can induce significant levels of unintended mutations at near-cognate sites, necessitating substantial efforts toward the development of strategies to minimize off-target activity. Although the genome-editing potential of thousands of other Cas9 orthologs remains largely untapped, it is not known how many will require similarly extensive engineering to achieve single-site accuracy within large genomes. In addition to its off-targeting propensity, SpyCas9 is encoded by a relatively large open reading frame, limiting its utility in applications that require size-restricted delivery strategies such as adeno-associated virus vectors. In contrast, some genome-editing-validated Cas9 orthologs are considerably smaller and therefore better suited for viral delivery.

Results

Here we show that wildtype NmeCas9, when programmed with guide sequences of the natural length of 24 nucleotides, exhibits a nearly complete absence of unintended editing in human cells, even when targeting sites that are prone to off-target activity with wildtype SpyCas9. We also validate at least six variant protospacer adjacent motifs (PAMs), in addition to the preferred consensus PAM (5′-N₄GATT-3′), for NmeCas9 genome editing in human cells.

Conclusions

Our results show that NmeCas9 is a naturally high-fidelity genome-editing enzyme and suggest that additional Cas9 orthologs may prove to exhibit similarly high accuracy, even without extensive engineering.

     

Localized Surface Plasmon Resonance and Refractive Index Sensitivity of Metal–Dielectric–Metal Multilayered Nanostructures

The localized surface plasmon resonances of multilayered nanostructures are studied using finite difference time domain simulations and plasmon hybridization method. Concentric metal–dielectric–metal (MDM) structure with metal core and nanoshell separated by a thin dielectric layer exhibits a strong coupling between the core and nanoshell plasmon resonance modes. The coupled resonance mode wavelengths show dependence on the dielectric layer thickness and composition of core and outer layer metal. The aluminum-based MDM structures show lower plasmon wavelength compared with Ag- and Au-based MDM nanostructures. The calculated refractive index sensitivity (RIS) factor is in the order Ag–Air–Ag>Au–Air–Au>Al–Air–Al for monometallic multilayered nanostructures. Bimetallic multilayered nanostructures support strong and tunable plasmon resonance wavelengths as well as high RIS factor of 510 nm/refractive index unit (RIU) and 470 nm/RIU for Al–Air–Au and Ag-Air-Au, respectively. The MDM structures not only exhibit higher index sensitivity but also cover a wide ultraviolet–near-infrared wavelengths, making these structures very promising for index sensing, biomolecule sensing, and surface-enhanced Raman spectroscopy.     

Disrupted Topological Organization in Whole-Brain Functional Networks of Heroin-Dependent Individuals: A Resting-State fMRI Study

Neuroimaging studies have shown that heroin addiction is related to abnormalities in widespread local regions and in the functional connectivity of the brain. However, little is known about whether heroin addiction changes the topological organization of whole-brain functional networks. Seventeen heroin-dependent individuals (HDIs) and 15 age-, gender-matched normal controls (NCs) were enrolled, and the resting-state functional magnetic resonance images (RS-fMRI) were acquired from these subjects. We constructed the brain functional networks of HDIs and NCs, and compared the between-group differences in network topological properties using graph theory method. We found that the HDIs showed decreases in the normalized clustering coefficient and in small-worldness compared to the NCs. Furthermore, the HDIs exhibited significantly decreased nodal centralities primarily in regions of cognitive control network, including the bilateral middle cingulate gyrus, left middle frontal gyrus, and right precuneus, but significantly increased nodal centralities primarily in the left hippocampus. The between-group differences in nodal centralities were not corrected by multiple comparisons suggesting these should be considered as an exploratory analysis. Moreover, nodal centralities in the left hippocampus were positively correlated with the duration of heroin addiction. Overall, our results indicated that disruptions occur in the whole-brain functional networks of HDIs, findings which may be helpful in further understanding the mechanisms underlying heroin addiction.     

兔内毒素性急性肺损伤肺内NOS阳性细胞数及超微结构的变化

目的　从病理学和组织化学角度观察内毒素性肺损伤时肺组织超微结构和 NOS阳性染色细胞数的变化。方法　采用电镜、还原型辅酶 - -黄递酶 (NADPH- d)组织化学技术观察兔内毒素性急性肺损伤时肺组织超微结构的改变和肺内小血管、细小支气管、肺泡的 NOS阳性染色细胞数目的变化。结果 :内毒素注射后观察 30分钟组 (I3 0′组 )与内毒素注射后观察 1周组 (Iw 组 ) ,肺内小血管、 NOS阳性染色细胞数明显增多 ,细小支气管和肺泡则减少。 I3 0′组与 Iw 组分别与 C组比较 :(小动脉 :I3 0 vs C,P<0 .0 1;Iwvs C,P<0 .0 0 1;小静脉 :I3 0 vs C,P<0 .0 1;Iw vs C,P<0 .0 5 ;细小支气管 :I3 0vs C与 Iwvs C均为 P<0 .0 2 ;肺泡 :I3 0 vs C与 Iwvs C均为 P<0 .0 1)。电镜结果显示 ,I3 0′组部分肺毛细血管内见有大量红细胞集聚 ,毛细血管内皮变薄 ,微细结构不甚清楚 ,Iw组部分毛细血管内皮细胞损伤状况较 I3 0′组更显严重 ,除较多见红细胞渗入肺泡腔外 ,尚见部分肺泡上皮结构不甚完整而呈泡状改变。结论　内毒素性肺损伤 ,肺内 NOS阳性染色细胞数在小血管明显增加 ,在支气管内 ,肺泡内则明显减少。     

Pitcher plant facilitates prey capture in a sympatric congener

Carnivorous plants avoid below-ground competition for nitrogen by utilizing an alternative nitrogen resource—invertebrate prey, but it remains unclear if sympatric carnivorous plants compete for prey resources. The aim of this study was to investigate if exploitative prey-resource competition occurs between the two sympatric pitcher plant species, Nepenthes rafflesiana and N. gracilis in Singapore. We first investigated if prey-resource partitioning occurs between these two species, and then investigated niche shift in N. gracilis by examining its pitcher contents along an in situ gradient of N. rafflesiana interspecific competition. Our results showed clear evidence of resource partitioning between the two species, but contrary to the expectation of competition, proximity to N. rafflesiana pitchers correlated with higher total prey numbers in N. gracilis pitchers. Our multivariate model of prey assemblages further suggested that N. rafflesiana facilitates N. gracilis prey capture, especially in several ant taxa that are trapped by both species. Concurrently, we found strong evidence for intraspecific competition between N. gracilis pitchers, suggesting that prey resources are exhaustible by pitcher-predation. Our results show that resource partitioning can be associated with facilitative interactions, instead of competition as is usually assumed. Facilitation is more typically expected between phylogenetically distant species, but divergences in resource acquisition strategies can permit facilitation between congeners.