Transport of an anionic substrate by the H+/monosaccharide symport inRhodotorula gracilis: Only the protonated form of the carrier is catalytically active

Summary The yeastRhodotorula gracilis accumulated glucuronate by an H⁺/symport. The transport was electroneutral, driven by the chemical gradient of protons pH. The observed stoichiometry amounted to 1 proton per molecule glucuronate. At pH 4, the half-saturation constantK _T was at its lowest value (K _T =8mm), whereas the maximal velocityV _T reached a maximum (V _T =15 nmol/min×mg dry wt). Monosaccharides competitively inhibited the uptake of glucuronate and vice versa. Hence, the two substrates share the same transport system. The steady-state accumulation of glucuronate reflected the course of the pH gradient. It is concluded that glucuronate is transported as an anionic substrate by the protonated carrier, the driving force being the chemical gradient of the H⁺ (pH). The ternary carrier/H⁺/glc-COOO-complex is electroneutral and independent of the membrane potential. Simultaneous uptake of organic acids (acetic or propionic acid) which is also energized by the pH gradient led to a noncompetitive inhibition of glucuronate transport. Thus, manipulation of the driving force, pH, reducedV _T without affectingK _T . Kinetic and energetic arguments are presented which stronly suggest that only the protonated carrier is catalytically active inR. gracilis.     

Pseudomonas aeruginosa acid phosphatase contains an anionic site with a trimethyl subsite

Summary In this work the action of the following compounds upon Ps. aeruginosa acid phosphatase has been studied: 1) alkylammonium compounds; 2) aminoalcohols and aminoacids with different substituents (–H, –CH₃OH and –CH₃) attached to the nitrogen atom; 3) alcohols analogous to some compounds of the above series, but without the amino group.It was found that the enzyme inhibition was more effective with N-trimethylated compounds than with the triethylated ones. The degree of inhibition depended on the number of methyl groups bound to the nitrogen atom. Taking into account the choline and betaine series the hydroxyl derivatives showed more affinity for the enzyme than the carboxylated ones. In each series the Ki values increased with the decrease of methyl groups bound to the nitrogen atom. The presence of a positively charged nitrogen atom in the molecule of the effector was essential. These results enable us to confirm that in the molecule of Ps. aeruginosa acid phosphatase there exists an anionic site with one subsite with affinity for methyl groups.     

Complexation of a Ligand with a Surfactant Micelle for Soil Heavy Metal Desorption

The ligands iodide (I^?) and thiocyanate (SCN^?), alone or in admixture, in combination with a non-ionic surfactant, Triton X-100, were evaluated as washing agents for heavy metal desorption from a contaminated soil. After seven successive washings, selective sequential extraction (SSE) was performed to determine the heavy metal content that remained associated with each geo-chemical fraction of the soil. The surfactant with 0.336 mol L^?1 of ligand I^? removed 75% Cd and 23% Cu, whereas the mobilization of Zn and Pb were not significant after 7 washings. At a concentration of 0.286 mol L^?1, the ligand SCN^? in the presence of surfactant removed 36% Cd, 44% Cu and 77% Zn. Among the washing agents, the combination of I^? and SCN^? produced the highest desorption efficiencies 95% Cd, 48% Cu, and 3.1% Pb, but not for Zn. The SCN^? ligand extracted the most Zn (77%). The SSE procedure indicated that the I^? removed metals from the exchangeable, carbonate and oxide fractions whereas SCN^? removed metals only from the exchangeable fraction. Both ligands, in the presence of surfactant, removed Cu from all fractions except the exchangeable sites, whereas only SCN^? plus surfactant removed Zn from all fractions. The ligand mixture plus surfactant mobilized only limited quantities of Pb from the oxide and residual fractions.     

改良微创肺泡表面活性物质技术对极早早产儿呼吸窘迫综合征患儿Th1/Th2细胞因子含量及心肌损伤指标的影响

摘要 目的：观察应用改良微创肺泡表面活性物质技术对极早早产儿呼吸窘迫综合征患儿Th1/Th2细胞因子含量及心肌损伤指标的影响。方法：以自2018年1月-2020年3月于本院院收治的104例极早早产儿呼吸窘迫综合征患儿为研究对象,按随机数字表法分为研究组（52例,给予改良微创肺泡表面活性物质技术治疗）和对照组（52例,给予气管插管-肺表面活性物质-拔管（INSURE）技术）。观察两组患儿治疗前、治疗7 d后的Th1/Th2细胞因子含量及心肌损伤指标,统计治疗后不良反应及并发症的发生率。结果：（1）干预前,两组患儿干扰素-γ（IFN-γ）、白介素-4（IL-4）及IFN-γ/IL-4含量比较,差异不显著（P＞0.05）;治疗后,两组IFN-γ及IFN-γ/IL-4含量均较治疗前降低,而IL-4含量明显上升;其中研究组IFN-γ及IFN-γ/IL-4含量均低于对照组,IL-4含量高于对照组,差异显著（P<0.05）。（2）干预前,两组患儿肌酸激酶同工酶（CK-MB）、肌酐蛋白（cTnI）及乳酸脱氢酶（LDH）含量比较,差异不显著（P＞0.05）;治疗后,两组CK-MB、cTnI及LDH含量均较治疗前降低,其中研究组CK-MB、cTnI及LDH含量均明显低于对照组（P<0.05）。（3）在治疗过程中,两组不良反应中心动过缓、经皮血氧饱和度（SpO2）下降、72 h内机械通气率及再次使用肺泡表面活性物质率比较,差异不显著（P＞0.05）。（4）在治疗过程中,两组并发症中早产儿视网膜病、坏死性小肠结肠炎、支气管肺发育不良、气胸、脑室内出血及死亡率比较,差异不显著（P＞0.05）。结论：应用改良微创肺泡表面活性物质技术可有效调节极早早产儿呼吸窘迫综合征患儿免疫功能,保护心肌细胞,且安全性高,值得临床推广使用。     

Novel coumarin derivatives as potent acetylcholinesterase inhibitors: insight into efficacy,mode and site of inhibition

The inhibitory efficacy of two substituted coumarin derivatives on the activity of neurodegenerative enzyme acetylcholinesterase (AChE) was assessed in aqueous buffer as well as in the presence of human serum albumin (HSA) and compared against standard cholinergic AD drug, Donepezil (DON). The experimental data revealed the inhibition to be of non-competitive type with both the systems showing substantial inhibitory activity on AChE. In fact, one of the tested compounds Chromenyl Coumarate (CC) was found to be better inhibitor (IC₅₀ = 48.49 ± 5.6 nM) than the reference drug DON (IC₅₀ = 74.13 ± 8.3 nM), unequivocally amplifying its importance. The structure of the compound was found to play a vital role in the inhibitory efficiency, validating previous Structure Activity Relationship (SAR) reviews for coumarin. The mechanism of inhibition remained impervious when the experimental medium was switched from aqueous buffer to HSA, albeit noticeable change in the inhibition potency of the compound 3, 3′- Methylene-bis (4-hydroxy coumarin) (MHC) (38%) and CC (35%). Both the coumarin derivatives were observed to bind to the peripheral anionic site (PAS) of AChE and also found to displace the fluorescence marker thioflavinT (ThT) from AChE binding pocket. All experimental observations were seconded by molecular docking and MD simulation results. The inferences drawn in this study form a foundation for further investigation on these compounds; magnifying the probability of their usage as AD drugs and re-emphasizes the significance of drug delivery media while considering the inhibition potencies of targeted drugs.     

Molecular simulation and experimental studies on the interfacial properties of a mixed surfactant SDS/C4mimBr

Mixed surfactants have potential applications in various fields. The understanding and prediction of their macro- and microscopic properties are of great importance in the designing of these materials. We used molecular dynamics (MD) and experiments to study the interfacial tension and the microscopic structures of the sodium dodecyl sulfate (SDS)/C₄mimBr mixed surfactant at the water/hexane interface. The interfacial tension, density profile, radial distribution function (RDF), orientation distribution of the tails and order parameters have been examined. It seems that the addition of C₄mimBr decreased the interfacial tension; a higher C₄mimBr concentration resulted in a thicker interface, a smaller droplet, and more disordered SDS tails. The competition between free volume and electrostatic shielding seems to be the primary mechanism behind these phenomena.     

Comparative Evaluation of α-Amylase Refolding Through Two Different Artificial Chaperone Systems

Two different artificial chaperone systems were evaluated in this work using either detergents or CDs as the stripping agents. Upon dilution of urea-denatured α-amylase to a non-denaturing urea concentration in the presence of the capturing agent, complexes of the detergent and non-native protein molecules are formed and thereby the formation of protein aggregates is prevented. The so-called captured protein is unable to refold from the detergent-protein complex states unless a stripping agent is used to remove the detergent molecules. Our results by fluorescence, UV, turbidity measurement, circular dichroism, surface tension and activity assay indicated that the extent of refolding assistance was different due to different inter- and intra- molecular interactions in the two different systems. However, the high activity recovery in the presence of detergents, as the stripping agent, suggests that they can constitute suitable replacement for the more expensive and common stripping agent of cyclodextrins.     

Biological Degradation of Crude Oil Refinery Sludge with Commercial Surfactant and Sewage Sludge by Co-Composting

This study determined the potential of surfactant and sewage sludge in enhancing degradation of oil sludge. A mixture of oil sludge, surfactant, and sewage sludge was co-composted for 24 weeks in the laboratory. Physical and chemical parameters in the compost were measured every four weeks. Isolated microorganisms were characterized by molecular techniques. The pH in all experiments remained between 8 and 6.4. CO₂ evolution reached 5503 µg/dwt/day by the twenty-fourth week. The dominant bacterial species were Acinectobacter, Rodococcus, mycobacterium, Pseudomonas, Bacillus, Arthrobacter, and Staphylococcus species and fungi were Pleurotus, Penicillium, and Aspergillus sp. TPH was reduced by 92% in the sewage sludge and surfactant treatment, 75 and 81% in other treatments, and 44.2% in the control. PAH concentrations were reduced by between 75 and 100%. The results indicate that a careful application of surfactant and sewage sludge could enhance oil sludge degradation in a compost system.     

Physiological aspects. Part 1 in a series of papers devoted to surfactants in microbiology and biotechnology

Surfactants, both chemical and biological, are amphiphilic compounds which can reduce surface and interfacial tensions by accumulating at the interface of immiscible fluids and increase the solubility, mobility, bioavailability and subsequent biodegradation of hydrophobic or insoluble organic compounds. Investigations on their impacts on microbial activity have generally been limited in scope to the most common and best characterized surfactants. Recently a number of new biosurfactants have been described and accelerated advances in molecular and cellular biology are expected to expand our insights into the diversity of structures and applications of biosurfactants. Biosurfactants play an essential natural role in the swarming motility of microorganisms and participate in cellular physiological processes of signaling and differentiation as well as in biofilm formation. Biosurfactants also exhibit natural physiological roles in increasing bioavailability of hydrophobic molecules and can complex with heavy metals, and some also possess antimicrobial activity. Chemical- and indeed bio-surfactants may also be added exogenously to microbial systems to influence behaviour and/or activity, mimicking the latter effects of biosurfactants. They have been exploited in this way, for example as antimicrobial agents in disease control and to improve degradation of chemical contaminants. Chemical surfactants can interact with microbial proteins and can be manipulated to modify enzyme conformation in a manner that alters enzyme activity, stability and/or specificity. Both chemical- and bio-surfactants are potentially toxic to specific microbes and may be exploited as antimicrobial agents against plant, animal and human microbial pathogens. Because of the widespread use of chemical surfactants, their potential impacts on microbial communities in the environment are receiving considerable attention.