The cell growth is inhibited by GNPs in cells transfected with GCLC siRNA.

<p>The A549 cells were transfected with negative control or GCLC siRNA-1 for 24 hours, followed by cultured for an additional 3 days in the absence of GNPs, subsequently harvested and counted. Each bar represents the mean (±SD n = 4) of triplicate determinations. *p<0.05 versus negative control.</p

Knockdown of Glutamate Cysteine Ligase Catalytic Subunit by siRNA Causes the Gold Nanoparticles-Induced Cytotoxicity in Lung Cancer Cells

<div><p>Gold nanoparticles (GNPs) have shown promising medical applications in cancer treatment involved in the regulation of intracellular redox balance. Previously, we have reported that GNPs can trigger apoptosis and necrosis in human lung cancer cells (A549) when L-buthionine-sulfoximine (BSO) was used to decrease the expression of intracellular glutathione (GSH). Herein, we investigated the cytotoxicity of GNPs toward lung cancer cells under the glutamate cysteine ligase catalytic subunit (GCLC) was silenced by siRNA. Our results showed that GNPs cause apoptosis and necrosis in cells transfected with GCLC siRNA by elevating intracellular reactive oxygen species (ROS). These findings demonstrated that the regulation of glutathione synthesis by GCLC siRNA in A549 cells can initiate the gold nanoparticles-induced cytotoxicity.</p></div

Effect of GCLC siRNA on intracellular GSH levels.

<p>Cytosol was isolated from cells transfected with negative control siRNA, GCLC siRNA-1, GCLC siRNA-2 and GCLC siRNA-3. The intracellular GSH levels were determined at 412 nm absorbance with a multiwell plate reader. Data represent the mean percentage of negative control (n = 3) ±SD for 4 independent experiments. *p<0.05, ** P<0.01, compared to negative control.</p

Effects of dissolved oxygen, starvation, temperature, and salinity on the locomotive ability of juvenile Chinese shrimp <i>Fenneropenaeus chinensis</i>

Locomotion is considered a fundamental property defining the fitness of animals. Shrimp locomotion includes walking, swimming, and tail-flipping. Little is known about the locomotive ability of juvenile shrimp. We studied the acute effects of key environmental factors on the ability of swimming and tail-flipping of juvenile Chinese shrimp Fenneropenaeus chinensis (1.10 ± 0.07 cm, body length), which was indicated by critical swimming speed (Ucrit) and tail-flipping speed (Utail), respectively. The key environmental factors include dissolved oxygen [DO; 2.2, 3.2, 4.2, 5.2, and 6.2 (control) mg L−1], starvation [1 (control), 3, 5, 7, and 9 days], temperature [9, 13, 17, 21 (control), and 25 °C (degrees Celsius)], and salinity [10, 15, 20, 25, and 30 (control) ppt]. The effect comparisons between these environmental factors were also obtained. Results showed that the changes of DO, starvation, temperature and salinity can affect not only the swimming, but also the tail-flipping of juvenile shrimp. The tail-flipping may be more important than swimming for survival in most shrimps, and therefore, Utail was more stable than Ucrit under hypoxia and starvation exposures. Ucrit and Utail showed similar sensibility to the changes of temperature and salinity. As in the effect comparison, Ucrit of juvenile F. chinensis was most affected by temperature, followed by DO and starvation, and the effect of salinity was the lowermost. These findings can be of value in understanding the physiological ecology of juvenile shrimp, and improving the stock enhancement of F. chinensis.</p

Additional file 2 of Integrative transcriptomic and metabolomic analysis of D-leaf of seven pineapple varieties differing in N-P-K% contents

Additional file 2

GNPs induce apoptosis and necrosis in cells transfected with GCLC siRNA.

<p>Cells were transfected with either non-target control siRNA or GCLC-specific siRNA-1. One day later, cells were treated with GNPs (20μM), GNPs (20μM)﹢GSH (1mM) and GNPs (20μM)+NAC(1mM) for additional 72 hours. AnnexinV-FITC and PI cells were measured with flow cytometry. (A) The fluorescence pattern of AnnexinV-FITC and PI-stained A549 cells after treatment. (B) Percentages of Annexin V positive or PI positive cells for different treatments. Each bar represents the mean (±SD n = 3). **p<0.01, ***P<0.001, versus control.</p

Table_1_Stable Isotope Analysis Revealed Ontogenetic Changes in Trophic Ecology and Migration Patterns of Sepia esculenta in the Northern Coastal Waters of China.DOCX

Golden cuttlefish (Sepia esculenta), the most economically important cephalopod in the northern coastal seas of China, had experienced greatly reduced population biomass due to continuous fishing pressure in recent decades. Understanding the trophic ecology and clarifying the spatial distribution of wintering ground can help develop management plans for this species. In this study, the ontogenetic changes in the trophic ecology of S. esculenta in the Qingdao coastal water were determined, and the migration patterns were studied using stable isotope analysis. Cluster analysis based on isotopic values divided S. esculenta with different lengths into groups: 11–20, 21–100, and 121–200 mm. A significant difference in the δ13C values between the groups 11–20 mm (−17.10‰) and 21–100 mm (−15.89‰) illustrates an ontogenetic change in the feeding habits. Due to the migratory habits of S. esculenta, the δ13C value of the group 121–200 mm (−16.39‰) was lower than that of the group 21–100 mm. The δ15N values of S. esculenta were found to increase in length, and there was a clear linear relationship between different S. esculenta groups, suggesting that the wintering ground may locate in the same latitude as the spawning ground (i.e., the middle Yellow Sea). Furthermore, the trophic relationship between S. esculenta and coexisting species was assessed, revealing that the group 11–20 mm of S. esculenta has some overlap of carbon isotope space with other species, suggesting that these species may feed on it as prey. Thus, slightly increasing the length to more than 20 mm may reduce the pressure of being the prey of post-release juveniles of S. esculenta and improve the effect of release.</p

Additional file 1 of Integrative transcriptomic and metabolomic analysis of D-leaf of seven pineapple varieties differing in N-P-K% contents

Additional file 1

Flow cytometry analysis of mitochondrial membrane potential in GNPs-treated cells.

<p>After transfected with negative control siRNA or GCLC siRNA-1, cells were treated with GNPs (20μM) for additional 72 hours then collected and stained with JC-1 in darkness at 37°C, rinsed by PBS. The fluorescence shift (red to green) of samples was measured by flow cytometry. Each bar represents the mean (±SD n = 4) of triplicate determinations. *<i>p</i><0.05, compared with negative control group.</p

GNPs induce caspase activation in cells transfected with GCLC siRNA.

<p>Activation of caspase-3 was measured using specific antibodies by flow cytometry. Intracellular GSH was depleted by GCLC siRNA-1, after approximately 72 hours of GNPs treatment, the cells were collected, treated with 0.1% Triton X-100 and blocked with 1% BSA, then incubated with cleaved caspase-3 (Asp175) antibody (Alexa fluor 488 conjugate) for 30 minutes. The fluorescence intensity was measured by flow cytometry. Each bar represents the mean (±SD n = 4) of triplicate determinations. *<i>p</i><0.05, compared with negative control group.</p