Stard 3 (Steroidogenic Acute Regulatory-Related Lipid Transfer Domain-Containing Protein 3) Play Important Role in Preadipocyte Differentiation

Aim: To evaluate Stard 3’s effects and relative mechanisms in preadipocyto differentiation by vitro study. Materials and Methods: The 3T3-L1 cell were divided into 5 groups as NC, si-Stard 3, ROS agonist, ROS inhibitor and si-Stard 3+ROS agonist groups. The cell of different groups were evaluated by Oil red O staining and Triglyceride. Evaluating ROS production by DHE and NBT assay. Using RT-qPCR and WB methods to evaluate gene and protein expressions. Results: Compared with NC group, Triglyceride, DHE fluorescence intensity and NBT positive rate were significantly down-regulation in si-Stard 3 and ROS inhibitor groups (P < 0.001, respectively), and were significantly up-regulation in ROS agonist group (P < 0.001, respectively); However, with si-Stard 3 transfection and ROS agonist treatment, compared with si-Stard 3 group, Triglyceride, DHE fluorescence intensity and NBT positive rate were significantly increased in si-Stard 3+ROS agonist group (P < 0.001, respectively). With RT-qPCR and WB assay, Compared with NC group, Stard 3 gene and protein expressions of si-Stard 3 and si-Stard 3+ROS agonist group were significantly depressed (P < 0.001, respectively), AMPK, PPARγ, CEBPα and FABP4 gene expressions were significantly differences in si-Stard 3, ROS agonist and ROS inhibitor groups (P < 0.001, respectively) and p-AMPK, PPARγ, CEBPα and FABP4 protein expressions were significantly differences in si-Stard 3, ROS agonist and ROS inhibitor groups (P < 0.001, respectively), with si-Stard 3 transfection and ROS agonist the relative gene and protein expressions were significantly resumed compared with si-Stard 3 group (P < 0.001, respectively). Conclusion: Stard 3 knockdown had effects to suppress 3T3-L1 cells transformation into adipocytes in vitro study.

Effect of Particle Concentration and Pipe Materials on the Formation of Biofilms in Drinking Water Distribution Systems

Microorganism rebreeding and biofilm shedding enter the water body in the process of a drinking water distribution system (DWDS), which poses a threat to public health. Particles in water can gather pollutants as well as providing favorable growth conditions for bacteria. To date, there are a few studies which focus on the relationship between particles and biofilm formation. Therefore, the microbial diversity of biofilms in the different pipe materials and the effect on particle concentration on biofilm formation were investigated in this study. Experiments were carried out under a simulative DWDS (including iron (DI) and polyvinyl chloride (PVC) pipe). The results showed that the microbial diversity in biofilms followed this order: DI pipe > PVC pipe > DI pipe (upper). Moreover, the microbial biomass of biofilm and the fluorescence intensity of extracellular polymeric substances (EPS, produced by microorganisms) were the largest in the absence of particles. The amount of biofilm bacterial and the fluorescence intensity of EPS both showed first an increasing and then decreasing trend with particle concentration increasing. When particle concentration was relatively low, the absorption of particles and bacteria played a major role, however, with the increasing particle concentration, more stable particle–particle were formed and thus, EPS was easily extracted, resulting in the increase of fluorescence intensity of EPS.

CD4 and CD8 co-receptors modulate functional avidity of CD1b-restricted T cells

T cells recognize mycobacterial glycolipid (mycolipid) antigens presented by CD1b molecules, but the role of CD4 and CD8 co-receptors in mycolipid recognition is unknown. Here we show CD1b-mycolipid tetramers reveal a hierarchy in which circulating T cells expressing CD4 or CD8 co-receptor stain with a higher tetramer mean fluorescence intensity than CD4-CD8- T cells. CD4+ primary T cells transduced with mycolipid-specific T cell receptors bind CD1b-mycolipid tetramer with a higher fluorescence intensity than CD8+ primary T cells. The presence of either CD4 or CD8 also decreases the threshold for interferon-γ secretion. Co-receptor expression increases surface expression of CD3ε, suggesting a mechanism for increased tetramer binding and activation. Targeted transcriptional profiling of mycolipid-specific T cells from individuals with active tuberculosis reveals canonical markers associated with cytotoxicity among CD8+ compared to CD4+ T cells. Thus, expression of co-receptors modulates T cell receptor avidity for mycobacterial lipids, leading to in vivo functional diversity during tuberculosis disease.

Synthesis and Properties of Nitrogen-Doped Carbon Quantum Dots Using Lactic Acid as Carbon Source

Nitrogen-doped carbon quantum dots (N-CQDs) were synthesized in a one-step hydrothermal technique utilizing L-lactic acid as that of the source of carbon and ethylenediamine as that of the source of nitrogen, and were characterized using dynamic light scattering, X-ray photoelectron spectroscopy ultraviolet-visible spectrum, Fourier-transformed infrared spectrum, high-resolution transmission electron microscopy, and fluorescence spectrum. The generated N-CQDs have a spherical structure and overall diameters ranging from 1–4 nm, and their surface comprises specific functional groups such as amino, carboxyl, and hydroxyl, resulting in greater water solubility and fluorescence. The quantum yield of N-CQDs (being 46%) is significantly higher than that of the CQDs synthesized from other biomass in literatures. Its fluorescence intensity is dependent on the excitation wavelength, and N-CQDs release blue light at 365 nm under ultraviolet light. The pH values may impact the protonation of N-CQDs surface functional groups and lead to significant fluorescence quenching of N-CQDs. Therefore, the fluorescence intensity of N-CQDs is the highest at pH 7.0, but it decreases with pH as pH values being either more than or less than pH 7.0. The N-CQDs exhibit high sensitivity to Fe3+ ions, for Fe3+ ions would decrease the fluorescence intensity of N-CQDs by 99.6%, and the influence of Fe3+ ions on N-CQDs fluorescence quenching is slightly affected by other metal ions. Moreover, the fluorescence quenching efficiency of Fe3+ ions displays an obvious linear relationship to Fe3+ concentrations in a wide range of concentrations (up to 200 µM) and with a detection limit of 1.89 µM. Therefore, the generated N-CQDs may be utilized as a robust fluorescence sensor for detecting pH and Fe3+ ions.

Continuous Monitoring of IgG using Immobilized Fluorescent Reporters

In the manufacture of therapeutic monoclonal antibodies (mAbs), the clarified cell culture fluid is typically loaded onto an initial protein A affinity capture column. Imperfect mass transfer and loading to maximum capacity can risk antibody breakthrough and loss of valuable product, but conservative underloading wastes expensive protein A resin. In addition, the effects of column fouling and ligand degradation require the frequent optimization of IgG loading to avoid wastage. Therefore, continuous real-time monitoring of IgG flowthrough is of great interest. We previously developed a fluorescence-based monitoring technology that allows mix-and-read mAb detection in cell culture fluid. Here we report the use of reporters immobilized on CNBr-activated Sepharose 4B resin for continuous detection of IgG in column breakthrough. The column effluent is continuously contacted with immobilized fluorescein-labeled Fc-binding ligands to produce an immediately detectable change in fluorescence intensity. The technology allows rapid and reliable monitoring of IgG in a flowing stream of clarified cell culture fluid emerging from a Protein A column, without prior sample preparation. We observed a significant change in fluorescence intensity at 0.5 g/L human IgG, sufficient to detect a 5% breakthrough of a 10 g/L load, within 2 minutes at a flow rate of 0.5 mL/min.

Флуктуирующая флуоресценция одиночных центров окраски в кристаллах фторида лития

Single F2 and F3+- color centers in the LiF crystal were studied by confocal fluorescence microscopy. The time dependences of their fluorescence intensity were analyzed and statistically processed. Our studies show that, the F3+- color center, being photoexcited, is able enter the triplet state, while in ground (singlet) state it changes orientation with a frequency of 1.5 – 2 Hz at room temperature, due to reorientational diffusion, unlike the F2- center, which is reoriented only being in the triplet state. This subtype of rotational diffusion of the center does not lead to its translational diffusion.

“Turn on” Fluorescence Sensor of Glutathione Based on Inner Filter Effect of Co-Doped Carbon Dot/Gold Nanoparticle Composites

Glutathione (GSH) is a thiol that plays a significant role in nutrient metabolism, antioxidant defense and the regulation of cellular events. GSH deficiency is related to variety of diseases, so it is useful to develop novel approaches for GSH evaluation and detection. In this study we used nitrogen and phosphorus co-doped carbon dot-gold nanoparticle (NPCD–AuNP) composites to fabricate a simple and selective fluorescence sensor for GSH detection. We employed the reductant potential of the nitrogen and phosphorus co-doped carbon dots (NPCDs) themselves to form AuNPs, and subsequently NPCD–AuNP composites from Au3+. The composites were characterized by using a range of spectroscopic and electron microscopic techniques, including electrophoretic light scattering and X-ray diffraction. The overlap of the fluorescence emission spectrum of NPCDs and the absorption spectrum of AuNPs resulted in an effective inner filter effect (IFE) in the composite material, leading to a quenching of the fluorescence intensity. In the presence of GSH, the fluorescence intensity of the composite was recovered, which increased proportionally to increasing the GSH concentration. In addition, our GSH sensing method showed good selectivity and sensing potential in human serum with a limit of detection of 0.1 µM and acceptable results.

Spectrofluorimetric Determination of Some N. Containing Medicines Using Rhodamine 6G as a Chromogenic Reagent

A sensitive spectrofluorimetric method has been developed for the analysis of some medicines containing primary, secondary, and tertiary amino groups, namely Diclofenac (DIC), Domperidone (DOM), Famotidine (FAM), and Propranolol (PRO), in their pure and medicinal forms. The method is based on the quenching of the fluorescence intensity of rhodamine 6G (R6G) through the formation of ion-pair complexes between the above medicines and the R-6G reagent, which is measured at 552 nm after excitation at 402 nm. The calibration graphs were rectilinear in the concentration ranges of 0.10- 9.00, 0.05-15.00, 0.10-14.0 and 0.05-5.00 µg mL-1 for above medicines respectively. The recovery (%) values were ranged between 99.45%- 100.97%. The detection limits ranged in the concentration of 0.243-0.754 µg/mL, and the limits of quantitation were 0.806- 2.420 µgmL-1 for all drugs. The method was successfully applied for the determination of these drugs in their pharmaceutical preparations.

Self-confocal NIR-II fluorescence microscopy for in vivo imaging

Benefiting from low scatter of NIR-II light in biological tissues and high spatial resolution of confocal microscopy, NIR-II fluorescence confocal microscopy has been developed recently and achieve deep imaging in vivo. However, independence of excitation point and detection point makes this system difficult to be adjusted. New, improved, self-confocal NIR-II fluorescence confocal systems are created in this work. Based on a shared pinhole for excitation light and fluorescence, the system is easy and controlled to be adjusted. The fiber-pinhole confocal system is constructed for cerebrovascular and hepatocellular NIR-II fluorescence intensity imaging. The air-pinhole confocal system is constructed for cerebrovascular NIR-II fluorescence intensity imaging, hepatic NIR-II fluorescence lifetime imaging, and hepatic multiphoton imaging.