Monoclonal antibody-based serological methods for detection of Cucumber green mottle mosaic virus

<p>Abstract</p> <p>Background</p> <p>Cucumber green mottle mosaic virus (CGMMV), a member of the genus <it>Tobamovirus</it>, can be transmitted by seeds and infects many cucurbit species, causing serious yield losses in cucumber and watermelon plants. In this paper, five serological methods including antigen-coated plate enzyme-linked immunosorbent assay (ACP-ELISA), triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA), Dot-immunobinding assay (DBIA), direct tissue blot immunoassay (DTBIA) and immunocapture reverse transcriptase polymerase chain reaction (IC-RT-PCR) were described for detection and diagnosis of CGMMV.</p> <p>Results</p> <p>Using the purified CGMMV particles as immunogens, six murine monoclonal antibodies (MAbs) were produced. Five serological methods were established using the MAb 4H1 and detection sensitivity was compared using purified preparations and infected-plant tissue extracts. The detection sensitivity of ACP-ELISA was 0.16 ng of purified CGMMV, whereas TAS-ELISA was more sensitive than ACP-ELISA with a minimum detection of 0.04 ng of purified CGMMV. The sensitivities of TAS-ELISA and DBIA were similar for detecting CGMMV in infected-plant tissue extracts, and were four times higher than ACP-ELISA. The IC-RT-PCR was the most sensitive method, which could detect as little as 0.1 pg of purified virus. The detection sensitivity of IC-RT-PCR for CGMMV-infected plant tissues was about 400 times higher than that of TAS-ELISA and DBIA.</p> <p>Conclusions</p> <p>The established ACP-ELISA, TAS-ELISA, DBIA and DTBIA are suitable for routine CGMMV detection of large-scale samples in the field survey, while IC-RT-PCR is more sensitive and suitable for acquiring information about the viral genome.</p

Quantity and clinical relevance of circulating endothelial progenitor cells in human ovarian cancer

<p>Abstract</p> <p>Background</p> <p>Circulating bone marrow-derived endothelial progenitor cells (EPCs) have been reported to participate in tumor angiogenesis and growth; however, the role of circulating EPCs in tumor progression is controversial. The role of circulating EPCs in ovarian cancer progression and angiogenesis has not yet been investigated.</p> <p>Methods</p> <p>The number of circulating EPCs in the peripheral blood in 25 healthy volunteers and 42 patients with ovarian cancer was determined by flow cytometry. EPCs were defined by co-expression of CD34 and vascular endothelial growth factor receptor 2 (VEGFR2). In addition, we determined CD34 and VEGFR2 mRNA levels by real-time reverse transcription-polymerase chain reaction. Plasma levels of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) were determined by enzyme-linked immunosorbent assay.</p> <p>Results</p> <p>Circulating levels of EPCs were significantly increased in ovarian cancer patients, correlating with tumor stage and residual tumor size. Higher levels of EPCs were detected in patients with stage III and IV ovarian cancer than in patients with stage I and II disease. After excision of the tumor, EPCs levels rapidly declined. Residual tumor size greater than 2 cm was associated with significantly higher levels of EPCs. In addition, high circulating EPCs correlated with poor overall survival. Pretreatment CD34 mRNA levels were not significantly increased in ovarian cancer patients compared with healthy controls; however, VEGFR2 expression was increased, and plasma levels of VEGF and MMP-9 were also elevated.</p> <p>Conclusions</p> <p>Our results demonstrate the clinical relevance of circulating EPCs in ovarian cancer. EPCs may be a potential biomarker to monitor ovarian cancer progression and angiogenesis and treatment response.</p

Equipo simulador de agua de lluvia para experimentos a campo en ecosistemas áridos y semiáridos

The predicted changes in precipitation patterns because of global change have profound effects on terrestrial ecosystems. In the present study, the principle and design details of a rainfall enrichment system (RAINES) for open field experiments in semi-arid and arid ecosystems are presented. The rainfall intensity, validity and uniformity of this experimental facility were also tested. During the period from 2008 to 2010, our data showed that the RAINES was able to simulate rainfall events with different rainfall sizes, frequencies and timing. The greatest advantage of the RAINES was its high uniformity in rainfall distribution over a relatively large experimental surface area (>65 m2), which was important for experimental studies of semi-arid and arid ecosystems where vegetation distribution is sparse. The rainfall validity of RAINES was steadily at 66% or higher as long as the hydraulic pressure exceeded 1.4 KPa and the wind speed was below 2.5 m s-1. Since the RAINES is light-weight, inexpensive and versatile enough to be used to simulate various rainfall events with needed properties in remote fields, it is able to provide reliable simulated rainfall in the field for studying possible responses of soil and vegetation processes to rainfall change in arid and semi-arid ecosystems. The application of the RAINES will improve our understanding on the relationship between water availability and ecosystem processes in arid and semi-arid ecosystems, which will provide useful knowledge for the protection, restoration and sustainable management of semi-arid and arid desert ecosystems worldwide.Los cambios predichos en los modelos de precipitación como resultado del cambio global tienen efectos profundos en los ecosistemas terrestres. Un equipo que simule la lluvia es una herramienta de investigación efectiva para explorar los efectos de los cambios en los modelos de lluvia sin varias restricciones naturales. En el presente estudio, se presentan los principios y detalles de diseño de un equipo simulador de lluvia (RAINES) para estudios a campo en ecosistemas áridos y semiaridos. También se determinaron la intensidad de lluvia, validez y uniformidad del RAINES. Durante el período 2008 al 2010, nuestros datos mostraron que el RAINES fue capaz de simular eventos de lluvia con diferentes cantidades, frecuencias y momentos de lluvia. La mayor ventaja del RAINES fue su gran uniformidad en la distribución de la lluvia sobre una superficie experimental relativamente grande (>65 m2 ). Esto es importante para estudios experimentales en ecosistemas áridos y semiáridos donde la distribución de la vegetación es dispersa. La validez de la lluvia provista por el RAINES fue al menos de 66% siempre y cuando la presión hidráulica excedió los 1.4 KPa y la velocidad del viento fue menor que 2.5 m s-1. El RAINES es de bajo peso, no costoso y lo suficientemente versátil como para ser usado para simular varios eventos de lluvia bajo condiciones de campo distantes. Es capaz de proveer lluvia simulada en forma confiable en el campo para estudiar la respuesta de procesos en el suelo y la vegetación a cambios en la cantidad de lluvia en ecosistemas áridos y semiáridos. El uso del RAINES mejorará nuestro entendimiento en la relación entre la disponibilidad de agua y los procesos en ecosistemas áridos y semiáridos. También proveerá conocimiento útil para la protección, restauración y menejo sustentable de ecosistemas de desierto áridos y semiáridos a escala mundial.Fil: Xin, Zhiming. Chinese Academy of Forestry; ChinaFil: Qian, Jianqiang. Henan Agricultural University; ChinaFil: Busso, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Centro de Recursos Naturales Renovables de la Zona Semiárida. Universidad Nacional del Sur. Centro de Recursos Naturales Renovables de la Zona Semiárida; Argentina. Universidad Nacional del Sur. Departamento de Agronomía; ArgentinaFil: Wu, Bo. Chinese Academy of Forestry; ChinaFil: Zhu, Yajuan. Chinese Academy of Forestry; ChinaFil: Zhang, Jinxin. Chinese Academy of Forestry; ChinaFil: Li, Yonghua. Chinese Academy of Forestry; China. State Forestry Administration; ChinaFil: Lu, Qi. Chinese Academy of Forestry; China. State Forestry Administration; Chin

Laser-Scribed Conductive, Photoactive Transition Metal Oxide on Soft Elastomers for Janus On-Skin Electronics and Soft Actuators

Laser-assisted fabrication of conductive materials on flexible substrates has attracted intense interests because of its simplicity, easy customization, and broad applications. However, it remains challenging to achieve laser scribing of conductive materials on tissue-like soft elastomers, which can serve as the basis to construct bioelectronics and soft actuators. Here, we report laser scribing of metallic conductive, photoactive transition metal oxide (molybdenum dioxide) on soft elastomers, coated with molybdenum chloride precursors, under ambient conditions. Laser-scribed molybdenum dioxide (LSM) exhibits high electrical conductivity, biocompatibility, chemical stability, and compatibility with magnetic resonance imaging. In addition, LSM can be made on various substrates (polyimide, glass, and hair), showing high generality. Furthermore, LSM-based Janus on-skin electronics are developed to record information from human skin, human breath, and environments. Taking advantage of its outstanding photothermal effect, LSM-based soft actuators are developed to build light-driven reconfigurable three-dimensional architectures, reshapable airflow sensors, and smart robotic worms with bioelectronic sensors

Laser-Scribed Conductive, Photoactive Transition Metal Oxide on Soft Elastomers for Janus On-Skin Electronics and Soft Actuators

Laser-assisted fabrication of conductive materials on flexible substrates has attracted intense interests because of its simplicity, easy customization, and broad applications. However, it remains challenging to achieve laser scribing of conductive materials on tissue-like soft elastomers, which can serve as the basis to construct bioelectronics and soft actuators. Here, we report laser scribing of metallic conductive, photoactive transition metal oxide (molybdenum dioxide) on soft elastomers, coated with molybdenum chloride precursors, under ambient conditions. Laser-scribed molybdenum dioxide (LSM) exhibits high electrical conductivity, biocompatibility, chemical stability, and compatibility with magnetic resonance imaging. In addition, LSM can be made on various substrates (polyimide, glass, and hair), showing high generality. Furthermore, LSM-based Janus on-skin electronics are developed to record information from human skin, human breath, and environments. Taking advantage of its outstanding photothermal effect, LSM-based soft actuators are developed to build light-driven reconfigurable three-dimensional architectures, reshapable airflow sensors, and smart robotic worms with bioelectronic sensors

The PI3K/Akt pathway upregulates Id1 and integrin α4 to enhance recruitment of human ovarian cancer endothelial progenitor cells

<p>Abstract</p> <p>Background</p> <p>Endothelial progenitor cells (EPCs) contribute to tumor angiogenesis and growth. We aimed to determine whether inhibitors of differentiation 1 (Id1) were expressed in circulating EPCs of patients with ovarian cancer, whether Id1 could mediate EPCs mobilization and recruitment, and, if so, what underlying signaling pathway it used.</p> <p>Methods</p> <p>Circulating EPCs cultures were from 25 patients with ovarian cancer and 20 healthy control subjects. Id1 and integrin α4 expression were analyzed by real-time reverse transcription-polymerase chain reaction and western blot. EPCs proliferation, migration, and adhesion were detected by MTT, transwell chamber, and EPCs-matrigel adhesion assays. Double-stranded DNA containing the interference sequences were synthesized according to the structure of a pGCSIL-GFP viral vector and then inserted into a linearized vector. Positive clones were identified as lentiviral vectors that expressed human Id1 short hairpin RNA (shRNA).</p> <p>Results</p> <p>Id1 and integrin α4 expression were increased in EPCs freshly isolated from ovarian cancer patients compared to those obtained from healthy subjects. siRNA-mediated Id1 downregulation substantially reduced EPCs function and integrin α4 expression. Importantly, Inhibition of PI3K/Akt inhibited Id1 and integrin α4 expression, resulting in the decreasing biological function of EPCs.</p> <p>Conclusions</p> <p>Id1 induced EPCs mobilization and recruitment is mediated chiefly by the PI3K/Akt signaling pathway and is associated with activation of integrin α4.</p

Correlation between the intestinal microflora and peripheral blood Th1/Th2 balance in hypothyroidism during the first half of pregnancy

ObjectiveThis study aimed to investigate the relationship between intestinal microflora characteristics and the peripheral blood T helper cell (Th)1/Th2 balance in patients with hypothyroidism during the first half of pregnancy.MethodsThe Th1/Th2 ratios in the peripheral blood of pregnant women in the hypothyroidism and control groups were determined using flow cytometry. The cytometric bead array assay was used to determine the serum levels of interleukin-2 (IL-2), IL-4, IL-6, IL-10, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ. Moreover, 16S rRNA amplicon sequencing was used to determine the intestinal microbial composition in the two groups. Finally, the relationships between intestinal microflora, Th1/Th2 cells, cytokines, and clinical indicators were analyzed.ResultsC-reactive protein levels were higher in the hypothyroidism group than in the control group. In contrast to the control group, the hypothyroidism group showed an increase in Th1 cells and the Th1/Th2 ratio, and a decrease in Th2 cells. The hypothyroidism group had higher serum IL-2, TNF-α, and IFN-γ levels, and lower IL-10 levels, than the control group. The richness of the intestinal microflora in the hypothyroidism group increased whereas the diversity decreased. The linear discriminant analysis effect size revealed that the hypothyroidism group had a higher abundance of Prevotella and Faecalibacterium, but a lower abundance of Bacteroides, compared to the control group. Prevotella was positively correlated with Th1 cells, the Th1/2 ratio, and TNF-α. Bacteroides was positively correlated with Th2 cells and IL-10, but negatively correlated with Th1 cells, the Th1/2 ratio, TNF-α, and IFN-γ. The thyroid peroxidase antibody level was directly proportional to TNF-α.ConclusionA Th1/Th2 imbalance occurs in patients with hypothyroidism during the first half of pregnancy. Disorders of the intestinal microflora may lead to hypothyroidism during pregnancy by affecting the Th1/Th2 balance