Hydrogen gas decreased the •OH levels in PC12 cells during HBO therapy.

<p>PC12 cells in logarithmic growth phase were exposed to the experimental gases for 3 h at 1 ATA or 5 ATA. The •OH levels (A and B) were determined using HPF staining. Fluorescence images of PC12 cells were obtained by Laser-Scanning Confocal Microscopy. Fluorescence was quantified from 100 cells of each independent experiment. *<i>p</i><0.05, compared with the Air group. ^#<i>p</i><0.05, compared with the HBO group. ^&<i>p</i><0.05, compared with the Air-H₂ group. Representative results were shown from at least three repeats.</p

Hydrogen gas reduced the levels of lipid peroxidation and DNA oxidation in PC12 cells during HBO therapy.

<p>PC12 cells in logarithmic growth phase were exposed to the experimental gases for 3 h at 1 ATA or 5 ATA. (A) The MDA levels in the culture medium as an indicator for lipid peroxidation were determined using a TBARS assay kit. (B) The 8-OH-dG levels in the culture medium as an indicator for DNA oxidation were determined using a kit. The data were analyzed using ANOVA followed by Fisher’s LSD test. *<i>p</i><0.05 (n = 6), compared with the Air group, ^#<i>p</i><0.05 (n = 6), compared with the HBO group, and ^&<i>p</i><0.05 (n = 6), compared with the Air-H₂ group. Representative results were shown from at least three repeats.</p

Hydrogen gas promoted the viability and inhibited the damage in the cell membrane of PC12 cells during HBO therapy.

<p>PC12 cells in logarithmic growth phase were exposed to the experimental gases for 3 h at 1 ATA or 5 ATA. (A) The viability of PC12 cells was determined using WST-1 assay. (B) The integrity of cell membrane in PC12 cells was determined using LDH assay. The data were analyzed by ANOVA followed by Fisher’s LSD test. *<i>p</i><0.05 (n = 6), compared with the Air group, ^#<i>p</i><0.05 (n = 6⁾, compared with the HBO group, and ^&<i>p</i><0.05 (n = 6), compared with the Air-H₂ group. Representative results were shown from at least three repeats.</p

Hydrogen gas inhibited decrease in MMP levels of PC12 cells during HBO therapy.

<p>PC12 cells in logarithmic growth phase were exposed to the experimental gases for 3 h at 5 ATA and then stained using TMRM and MTGreen dyes. Fluorescence was detected using a Laser-Scanning Confocal Microscope. After HBO treatment, H₂ prevented the decline of the mitochondrial membrane potential, as detected by fluorescence of TMRM, which depends upon the mitochondrial membrane potential, whereas fluorescence levels of MTGreen, which are independent of the membrane potential, were unchanged. Representative results were shown from at least three repeats.</p

Shear Effects on Stability of DNA Complexes in the Presence of Serum

The behavior of nanocarriers, even though they are well-defined at equilibrium conditions, is unpredictable in living system. Using the complexes formed by plasmid DNA (pDNA) and K₂₀ (K: lysine), protamine, or polylysine (PLL) as models, we studied the dynamic behavior of gene carriers in the presence of fetal bovine serum (FBS) and under different shear rates, a condition mimicking the internal physical environment of blood vessels. Without shear, all the positively charged complexes bind to the negatively charged proteins in FBS, leading to the formation of large aggregates and even precipitates. The behaviors are quite different under shear. The shear generates two effects: a mechanical force to break down the complex into smaller size particles above a critical shear rate and a stirring effect leading to secondary aggregation of complexes below the critical shear rate. In the studied shear rate from 100 to 3000 s^–1, the mechanical force plays a key role in K₂₀/pDNA and protamine/pDNA, while the stirring effect is dominant in PLL/pDNA. A model study shows that the interfacial tension, the chain density, and the elasticity of the complexes determine their responsiveness to shear force. This study is helpful to understand the fate of drug/gene carriers under physiological conditions. It also gains insight in designing drug/gene carriers with desirable properties for in vivo applications

Inward Budding and Endocytosis of Membranes Regulated by de Novo Designed Peptides

Protein-mediated endocytosis of membrane is a key event in biological system. The mechanism, however, is still not clear. Using a de novo designed bola-type peptide KKKLLLLLLLLKKK (K₃L₈K₃) as a protein mimic, we studied how it induced giant unilamellar vesicle (GUV) to form inward buds or endocytosis at varying conditions. Results show that the inward budding is initiated as the charged lipids are neutralized by K₃L₈K₃, which results in a negative spontaneous curvature. If the charged lipids have unsaturated tails, the buddings are slim fibrils, which can further wrap into a spherical structure. In the case of saturated charged lipids, the buddings are rigid tubules, stable in the studied time period. The unsaturated lipid to saturated lipid ratio in the mother membrane is another key parameter governing the shape and dynamics of the buds. A complete endocytosis is observed when K₃L₈K₃ is attached with a hydrophobic moiety, suggesting that hydrophobic interaction helps the buds to detach from the mother membrane. The molecules in the surrounding medium, such as negatively charged oligonucleotides, are engulfed into the GUV via endocytosis pathway induced by K₃L₈K₃. Our study provides a novel strategy for illustrating the endocytosis mechanism by using peptides of simple sequence

Inward Budding and Endocytosis of Membranes Regulated by de Novo Designed Peptides

Protein-mediated endocytosis of membrane is a key event in biological system. The mechanism, however, is still not clear. Using a de novo designed bola-type peptide KKKLLLLLLLLKKK (K₃L₈K₃) as a protein mimic, we studied how it induced giant unilamellar vesicle (GUV) to form inward buds or endocytosis at varying conditions. Results show that the inward budding is initiated as the charged lipids are neutralized by K₃L₈K₃, which results in a negative spontaneous curvature. If the charged lipids have unsaturated tails, the buddings are slim fibrils, which can further wrap into a spherical structure. In the case of saturated charged lipids, the buddings are rigid tubules, stable in the studied time period. The unsaturated lipid to saturated lipid ratio in the mother membrane is another key parameter governing the shape and dynamics of the buds. A complete endocytosis is observed when K₃L₈K₃ is attached with a hydrophobic moiety, suggesting that hydrophobic interaction helps the buds to detach from the mother membrane. The molecules in the surrounding medium, such as negatively charged oligonucleotides, are engulfed into the GUV via endocytosis pathway induced by K₃L₈K₃. Our study provides a novel strategy for illustrating the endocytosis mechanism by using peptides of simple sequence

Inward Budding and Endocytosis of Membranes Regulated by de Novo Designed Peptides

Protein-mediated endocytosis of membrane is a key event in biological system. The mechanism, however, is still not clear. Using a de novo designed bola-type peptide KKKLLLLLLLLKKK (K₃L₈K₃) as a protein mimic, we studied how it induced giant unilamellar vesicle (GUV) to form inward buds or endocytosis at varying conditions. Results show that the inward budding is initiated as the charged lipids are neutralized by K₃L₈K₃, which results in a negative spontaneous curvature. If the charged lipids have unsaturated tails, the buddings are slim fibrils, which can further wrap into a spherical structure. In the case of saturated charged lipids, the buddings are rigid tubules, stable in the studied time period. The unsaturated lipid to saturated lipid ratio in the mother membrane is another key parameter governing the shape and dynamics of the buds. A complete endocytosis is observed when K₃L₈K₃ is attached with a hydrophobic moiety, suggesting that hydrophobic interaction helps the buds to detach from the mother membrane. The molecules in the surrounding medium, such as negatively charged oligonucleotides, are engulfed into the GUV via endocytosis pathway induced by K₃L₈K₃. Our study provides a novel strategy for illustrating the endocytosis mechanism by using peptides of simple sequence

Inward Budding and Endocytosis of Membranes Regulated by de Novo Designed Peptides

Protein-mediated endocytosis of membrane is a key event in biological system. The mechanism, however, is still not clear. Using a de novo designed bola-type peptide KKKLLLLLLLLKKK (K₃L₈K₃) as a protein mimic, we studied how it induced giant unilamellar vesicle (GUV) to form inward buds or endocytosis at varying conditions. Results show that the inward budding is initiated as the charged lipids are neutralized by K₃L₈K₃, which results in a negative spontaneous curvature. If the charged lipids have unsaturated tails, the buddings are slim fibrils, which can further wrap into a spherical structure. In the case of saturated charged lipids, the buddings are rigid tubules, stable in the studied time period. The unsaturated lipid to saturated lipid ratio in the mother membrane is another key parameter governing the shape and dynamics of the buds. A complete endocytosis is observed when K₃L₈K₃ is attached with a hydrophobic moiety, suggesting that hydrophobic interaction helps the buds to detach from the mother membrane. The molecules in the surrounding medium, such as negatively charged oligonucleotides, are engulfed into the GUV via endocytosis pathway induced by K₃L₈K₃. Our study provides a novel strategy for illustrating the endocytosis mechanism by using peptides of simple sequence

Inward Budding and Endocytosis of Membranes Regulated by de Novo Designed Peptides

Protein-mediated endocytosis of membrane is a key event in biological system. The mechanism, however, is still not clear. Using a de novo designed bola-type peptide KKKLLLLLLLLKKK (K₃L₈K₃) as a protein mimic, we studied how it induced giant unilamellar vesicle (GUV) to form inward buds or endocytosis at varying conditions. Results show that the inward budding is initiated as the charged lipids are neutralized by K₃L₈K₃, which results in a negative spontaneous curvature. If the charged lipids have unsaturated tails, the buddings are slim fibrils, which can further wrap into a spherical structure. In the case of saturated charged lipids, the buddings are rigid tubules, stable in the studied time period. The unsaturated lipid to saturated lipid ratio in the mother membrane is another key parameter governing the shape and dynamics of the buds. A complete endocytosis is observed when K₃L₈K₃ is attached with a hydrophobic moiety, suggesting that hydrophobic interaction helps the buds to detach from the mother membrane. The molecules in the surrounding medium, such as negatively charged oligonucleotides, are engulfed into the GUV via endocytosis pathway induced by K₃L₈K₃. Our study provides a novel strategy for illustrating the endocytosis mechanism by using peptides of simple sequence