Pergolide Presynaptically Inhibits Calcium-Stimulated Release of γ-Aminobutyric Acid

The release of gamma-aminobutyric acid (GABA) in rat dorsolateral striatum was studied using in vivo microdialysis. Dialysis was conducted 2 days after probe implantation in awake, freely moving rats using a modified Ringer solution. Calcium induced a reversible increase in GABA release that was abolished by tetrodotoxin but was only slightly attenuated by a maximally effective dose of pergolide, a D2 receptor agonist. It was thus concluded that pergolide inhibits calcium-stimulated release of GABA presynaptically by a mechanism distinct from that of tetrodotoxin.     

High sequence coverage by in-capillary proteolysis of native proteins and simultaneous analysis of the resulting peptides by nanoelectrospray ionization-mass spectrometry and tandem mass spectrometry

Losses of proteolytic peptides during extraction and/or purification procedures succeeding in-gel or in-solution digests of proteins frequently occur in the course of protein identification investigations. In order to overcome this disadvantage, the method of in-capillary digest was developed: native proteins were incubated in the presence of endoproteases in the electrospray capillary and the resulting peptides were analyzed by nanoelectrospray-mass spectrometry during the ongoing proteolysis. In-capillary digest of apomyglobin by use of trypsin in a molar ratio of 25:1 yielded complete degradation already after 15 min. The sequence coverage based on formation of molecular ions was 100% and peptide ions could be fragmented by collision-induced dissociation and sequenced. When myoglobin was incubated in the electrospray capillary with trypsin in a molar ratio of 500:1, a clear shift from molecular ions and miscleaved peptide ions to the expected final tryptic peptide ions was observed over a 2 h period. The peptide spectra obtained from tryptic in-capillary proteolysis of bovine serum albumin and apotransferrin, respectively, gave rise to sequence coverages of more than 40% for both proteins. The data obtained from the peptide maps as well as from collision-induced dissociation (CID) of selected peptides were more than sufficient for protein identification by database searches. An elephant milk protein preparation was used to demonstrate the application of in-capillary proteolysis on protein mixtures. Tryptic digest, simultaneous analysis of the proteolytic peptides by use of CID, and subsequent sequencing allowed the identification of lactoferrin, alphas1-casein, beta-casein, delta-casein, and kappa-casein by homology search.     

Metal elements associate with in vitro fertilization (IVF) outcomes in 195 couples

PurposeEnvironmental factors may affecting reproductive function reduction and embryonic development. Couples who are exposed to heavy metals for a long time may affect the outcome of in vitro fertilization (IVF). To evaluate the effect of elements on IVF outcomes, a total of 195 couples undergoing IVF were included in this study.MethodsElements including V, Cr, Mn, Ni, Cu, Zn, As, Mo, Cd, Ba, Hg, Tl, Pb were measured in serum and follicular fluid (FF) samples of female and semen samples of male by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Multiple linear regression were applied to evaluate the association between metal elements and semen quality parameters and the number of oocytes in MII stage. Poisson regression and the robust variance estimation of the generalized estimation equation were used to evaluate the association between elements and IVF outcomes.ResultsThe statistical results showed that Cr had a significant negative correlation with total sperm concentration (TSC) and total motile sperm count (TMC), the correlation coefficients were -0.52 (-0.27∼1.43) and -0.4(-1.24, 0.45), respectively. At the same time, Ba was significantly correlated with TSC and TMC, the correlation coefficients were 0.1(-0.15∼0.34) and 0.12(-0.13, 0.36), respectively. Cr, Ba and Pb in follicular fluid (FF) had a significant positive correlation with the number of oocytes in MII stage. The correlation coefficients were 3.15 (0.79, 5.52), 1.54 (-0.27, 3.36), 12.27 (7.49, 17.04). The Tl level of FF was significantly associated with the high probability of blastocyst formation and high-quality blastocysts (RR: 2.83, 95 % CI: 0.92∼7.95; RR: 3.12, 95 % CI: 0.64, 12.84). The Hg level (RR: 3.98, 95 % CI: 0.78∼14.77) and the Ba level in serum (RR: 12.75 95 % CI: 1.31∼89.71) were significantly correlated with high-quality blastocysts. The levels of Ni, Cu, Mo in seminal plasma of men were significantly correlated with blastocyst formation and high-quality blastocysts (RR values were all greater than 1.5). In addition, the level of Ba was significantly correlated with the high probability of blastocyst formation (RR: 1.7, 95 % CI: 1.14∼2.52).ConclusionOur results reveal that Cr, Ba and Pb may affect TSC, TMC and MII oocytes. Moreover, Ba, Cr, As, Hg and Tl in serum and Mo in seminal plasma were related to fertilization results, good embryos, blastocyst formation, high-quality embryos, and pregnancy and live birth rates. Tl in FF may related to the quality of embryonic development, Ba was an important risk factor which closely related to the outcomes of IVF in both male and female. Through our detection and statistical analysis of clinical samples, it is shown that although not all elements will affect the outcome of IVF the key elements we have selected need to arouse our attention, which benifit to the diagnosis and prevention of clinical infertility.     

Efficiency Boost in All-Small-Molecule Organic Solar Cells: Insights from the Re-Ordering Kinetics

Achieving high-performance in all-small-molecule organic solar cells (ASM-OSCs) significantly relies on precise nanoscale phase separation through domain size manipulation in the active layer. Nonetheless, for ASM-OSC systems, forging a clear connection between the tuning of domain size and the intricacies of phase separation proves to be a formidable challenge. This study investigates the intricate interplay between domain size adjustment and the creation of optimal phase separation morphology, crucial for ASM-OSCs’ performance. It is demonstrated that exceptional phase separation in ASM-OSCs’ active layer is achieved by meticulously controlling the continuity and uniformity of domains via re-packing process. A series of halogen-substituted solvents (Fluorobenzene, Chlorobenzene, Bromobenzene, and Iodobenzene) is adopted to tune the re-packing kinetics, the ASM-OSCs treated with CB exhibited an impressive 16.2% power conversion efficiency (PCE). The PCE enhancement can be attributed to the gradual crystallization process, promoting a smoothly interconnected and uniformly distributed domain size. This, in turn, leads to a favorable phase separation morphology, enhanced charge transfer, extended carrier lifetime, and consequently, reduced recombination of free charges. The findings emphasize the pivotal role of re-packing kinetics in achieving optimal phase separation in ASM-OSCs, offering valuable insights for designing high-performance ASM-OSCs fabrication strategies.     

钙库调控的钙通道Orai1体外研究方法的建立

目的:建立钙通道Orai1的体外研究方法。方法:利用脂质体重组技术,将体外纯化的Orai1蛋白重组到脂质体膜上,利用蔗糖密度梯度离心来检测其重组效率及Orai1蛋白在脂质体膜上的结构,并利用钙染料Fura-2检测脂质体内钙离子的释放。结果:成功制备了脂质体及体外纯化了GST-Orai1融合蛋白,蔗糖密度梯度离心结果证明GST-Orai1蛋白成功重组到脂质体上,以及Orai1蛋白以多聚体的形式定位在脂质体膜上。钙离子释放实验证明脂质体内钙离子包装完好,可用于后续Orai1钙通道的功能研究。结论:利用脂质体重组技术建立了一种新的Orai1的研究方法,能够更直接有效地研究其功能及其活化机制。