Increased yield of total RNA from fine-needle aspirates for use in expression microarray analysis

Fine-needle aspirate samples hold the potential for gaining valuable insight into the molecular details and prognostic indicators for certain types of cancer in a limited volume of relatively pure tumor cells. Although limited, such clinical samples can be used with high efficiency when analyzed in conjunction with gene-dense expression microarrays. For this reason, it is essential to retrieve as much high-quality genetic material as possible from each fine-needle aspirate sample. We have conducted a study to improve the efficiency of extracting high quality total RNA to use in microarray analysis from single ex vivo fine-needle aspirate samples of 11 breast cancers added to RNAlater RNA Stabilization Reagent immediately upon collection. Approximately half the total RNA from fine-needle aspirate samples of breast cancers was isolated from the supernatant, and that RNA had similar quality and gene expression profile to the RNA that was isolated from the corresponding cell pellet. We recommend that the supernatant not be discarded when extracting RNA from fine-needle aspirate samples stored in RNAlater.     

Antibody Detection and Molecular Characterization of Toxoplasma gondii from Bobcats (Lynx rufus), Domestic Cats (Felis catus), and Wildlife from Minnesota,USA

Little is known of the epidemiology of toxoplasmosis in Minnesota. Here, we evaluated Toxoplasma gondii infection in 50 wild bobcats (Lynx rufus) and 75 other animals on/near 10 cattle farms. Antibodies to T. gondii were assayed in serum samples or tissue fluids by the modified agglutination test (MAT, cut‐off 1:25). Twenty nine of 50 bobcats and 15 of 41 wildlife trapped on the vicinity of 10 farms and nine of 16 adult domestic cats (Felis catus) and six of 14 domestic dogs resident on farms were seropositive. Toxoplasma gondii oocysts were not found in feces of any felid. Tissues of all seropositive wild animals trapped on the farm were bioassayed in mice and viable T. gondii was isolated from two badgers (Taxidea taxus), two raccoons (Procyon lotor), one coyote (Canis latrans), and one opossum (Didelphis virginiana). All six T. gondii isolates were further propagated in cell culture. Multi‐locus PCR‐RFLP genotyping using 10 markers (SAG1, SAG2 (5′‐3′SAG2, and alt.SAG2), SAG3, BTUB, GRA6, c22‐8, c29‐2, L358, PK1, and Apico), and DNA from cell culture derived tachyzoites revealed three genotypes; #5 ToxoDataBase (1 coyote, 1 raccoon), #1 (1 badger, 1 raccoon, 1 opossum), and #2 (1 badger). This is the first report of T. gondii prevalence in domestic cats and in bobcats from Minnesota, and the first isolation of viable T. gondii from badger.     

An Upregulation of SENP3 After Spinal Cord Injury: Implications for Neuronal Apoptosis

SENP3 (SUMO-specific proteases 3), a member of the small ubiquitin-like modifier specific protease family, was identified as a molecule that deconjugates SUMOylation of modified protein substrates and functions as an isopeptidase by disrupting SUMO homeostasis to facilitate cancer development and progression. However, its expression and function in nervous system injury and repair are still unclear. In this study, we employed an acute spinal cord injury (SCI) model in adult rats and investigated the dynamic changes of SENP3 expression in the spinal cord. Western blot analysis indicated a gradual increase in SENP3 expression, which peaked 3?days after SCI, and then declined over the following days. Immunohistochemistry results further confirmed that SENP3 was expressed at low levels in the gray and white matter in the non-injured condition and increased after SCI. Moreover, immunofluorescence double-labeling showed that SENP3 was co-expressed with the neuronal marker, NeuN. Furthermore, the SENP3-positive cells that were co-expressed with NeuN had also expressed active caspase-3 after injury. To investigate whether SENP3 plays a role in neuronal apoptosis, we applied H₂O₂ to induce neuronal apoptosis in vitro. Western blot analysis showed a significant upregulation of SENP3 and active caspase-3 following H₂O₂ stimulation. Taken together, these results suggest that SENP3 may play important roles in the pathophysiology of SCI.     

BMP2/7 heterodimer enhances osteogenic differentiation of rat BMSCs via ERK signaling compared with respective homodimers

Bone morphogenetic protein (BMP)2/7 heterodimer shows greater efficacy in enhancing bone regeneration. However, the precise mechanism and the role of mitogen-activated protein kinase (MAPK) signaling network in BMP2/7-driven osteogenesis remain ambiguous. In this study, we evaluated the effects of BMP2/7 heterodimers on osteoblastic differentiation in rat bone marrow mesenchymal stem cells (BMSCs), with the aim to elaborate how MAPKs might be involved in this cellular process by treatment of rat BMSCs with BMP2/-7 with a special signal-pathway inhibitor. We found that BMP2/7 heterodimer induced a much stronger osteogenic response in rat BMSCs compared with either homodimer. Most interestingly, extracellular signal-regulated kinase (ERK) demonstrated a highly sustained phosphorylation and activation in the BMP2/7 heterodimer treatment groups, and inhibition of ERK cascades using U0126 special inhibitor that significantly reduced the activity of ALP and calcium mineralization to a substantial degree in rat BMSCs treated with BMP2/7 heterodimers. Collectively, we demonstrate that BMP2/7 heterodimer shows a potent ability to stimulate osteogenesis in rat BMSCs. The activated ERK signaling pathway involved in this process may contribute partially to an increased osteogenic potency of heterodimeric BMP2/7 growth factors.     

BioGPS and GXD: mouse gene expression data—the benefits and challenges of data integration

Mouse gene expression data are complex and voluminous. To maximize the utility of these data, they must be made readily accessible through databases, and those resources need to place the expression data in the larger biological context. Here we describe two community resources that approach these problems in different but complementary ways: BioGPS and the Mouse Gene Expression Database (GXD). BioGPS connects its large and homogeneous microarray gene expression reference data sets via plugins with a heterogeneous collection of external gene centric resources, thus casting a wide but loose net. GXD acquires different types of expression data from many sources and integrates these data tightly with other types of data in the Mouse Genome Informatics (MGI) resource, with a strong emphasis on consistency checks and manual curation. We describe and contrast the “loose” and “tight” data integration strategies employed by BioGPS and GXD, respectively, and discuss the challenges and benefits of data integration. BioGPS is freely available at http://biogps.org. GXD is freely available through the MGI web site (www.informatics.jax.org) or directly at www.informatics.jax.org/expression.shtml.     

Serum Macroelement and Microelement Concentrations in Patients with Polycystic Ovary Syndrome: a Cross-Sectional Study

Polycystic ovarian syndrome (PCOS) is one of the most common endocrine diseases. However, its pathogenesis is unclear. We aim to explore the potential relationships between serum macroelements/microelements and PCOS. A total of 1137 women were involved in the current study. PCOS was defined according to ESHRE/ASRM, and complete blood samples were collected. Serum macroelements (calcium and magnesium) and microelements (copper, zinc, and iron) were assayed through atomic absorption spectrophotometry. PCOS patients had significantly higher copper concentrations than patients without PCOS (P < 0.001). By contrast, PCOS patients had lower serum calcium levels than patients without PCOS (P < 0.001). No significant differences were observed in the levels of serum zinc, magnesium, and iron between PCOS and non-PCOS patients. PCOS patients with acne had higher magnesium levels than those without acne (P = 0.020), and PCOS patients with hirsutism had lower magnesium levels than those without hirsutism (P = 0.037). High serum copper and low calcium levels may be correlated with PCOS. Magnesium concentrations are correlated with acne and hirsutism in PCOS patients. These results provide clues to explore the mechanism of PCOS and guidance for element treatments in PCOS patients.     

Facile Patterning of Laser‐Induced Graphene with Tailored Li Nucleation Kinetics for Stable Lithium‐Metal Batteries

A facile and scalable approach is reported to stabilize the lithium‐metal anode by regulating the Li nucleation and deposition kinetics with laser‐induced graphene (LIG). By processing polyimide (PI) films on copper foils with a laser, a 3D‐hierarchical composite material is constructed, consisting of a highly conductive copper substrate, a pillared array of flexible PI, and most importantly, porous LIG on the walls of the PI pillars. The high number of defects and heteroatoms present in LIG significantly lowers the Li nucleation barrier compared to the copper foil. An overpotential‐free Li nucleation process is identified at current densities lower than 0.2 mA cm^?2. Theoretical computations reveal that the defects serve as nucleation centers during the heterogeneous nucleation of lithium. By adopting such composites, ultrastable lithium‐metal anodes are obtained with high Coulombic efficiencies of ≈99%. Full lithium‐metal cells based on LiFePO₄ cathodes with a material loading of ≈15 mg cm^?2 and a negative/positive ratio of 5/1 could be cycled over 250 times with a capacity loss of less than 10%. The current work highlights the importance of nucleation kinetics on the stability of metallic anodes and demonstrates a practical method toward long lasting Li‐metal batteries.     

街区尺度屋顶绿化热效应及其与城市形态结构之间的关系

研究探讨大面积屋顶绿化如何改善城市冠层内部三维空间的热环境,改善效果与城市形态结构之间存在的关系。基于三维小气候模型ENVI-met 4.2,开展街区尺度屋顶绿化热效应研究。在南京市选择具有不同形态结构特征的8个典型样区,每个样区设置2种朝向(街道与主导风向平行或相交)、3种屋顶绿化情景(传统光屋顶、简易型绿化、复合型绿化),共对48种情景进行了微气候模拟与分析。结果表明,街区尺度屋顶绿化产生的"冷岛"可从屋面扩散到地面空间,缓解城市冠层热岛效应。白天14:00时,8个样区屋顶1.5 m高处的降温强度最高可达0.64℃,样区平均降温强度最大值为0.44℃;地面1.5 m高处的降温最大值为0.55℃,样区平均降温强度最大值为0.25℃。建筑高度、高宽比、容积率与屋面、地面降温强度之间均存在显著负相关关系;建筑密度与屋面降温强度显著正相关,但与地面降温强度之间无显著相关性。总的来说,紧凑低层建筑区和开敞低层建筑区实施屋顶绿化后产生的降温效应明显高于紧凑和开敞高层建筑区;建筑高度和密度相同时,街道走向与主导风向呈45°夹角比与主导风向平行具有更高的降温强度。研究结果能够促进对亚热带城市地区建筑-植被-大气相互关系的深入认识,并为屋顶绿化实践提供指导。