Bacteria form surface attached biofilm communities as one of the most important survival strategies in nature. Biofilms consist of water, bacterial cells and a wide range of self-generated extracellular polymeric substances (EPS). Biofilm formation is a dynamic self-assembly process and several distinguishable stages are observed during bacterial biofilm development. Biofilm formation is shown to be coordinated by EPS production, cell migration, subpopulation differentiation and interactions. However, the ways these different factors affect each other and contribute to community structural differentiation remain largely unknown. The distinct roles of different EPS have been addressed in the present report. Both Pel and Psl polysaccharides are required for type IV pilus-independent microcolony formation in the initial stages of biofilm formation by Pseudomonas aeruginosa PAO1. Both Pel and Psl polysaccharides are also essential for subpopulation interactions and macrocolony formation in the later stages of P. aeruginosa PAO1 biofilm formation. Pel and Psl polysaccharides have different impacts on Pseudomonas quinolone signal-mediated extracellular DNA release in P. aeruginosa PAO1 biofilms. Psl polysaccharide is more important than Pel polysaccharide in P. aeruginosa PAO1 biofilm formation and antibiotic resistance. Our study thus suggests that different EPS materials play distinct roles during bacterial biofilm formation.
Different genetic determinants were involved in controlling seed yield and yield-related traits in B. napus under normal and low P conditions. The QTLs detected under reduced P supply may provide useful information for improving the seed yield of B. napus in soils with low P availability in marker-assisted selection.
Mutations in ryanodine receptors (RyRs), intracellular Ca2+ channels, are associated with deadly disorders. Despite abundant functional studies, the molecular mechanism of RyR malfunction remains elusive. We studied two single-point mutations at an equivalent site in the skeletal (RyR1 R164C) and cardiac (RyR2 R176Q) isoforms using ryanodine binding, Ca2+ imaging, and cryo–electron microscopy (cryo-EM) of the full-length protein. Loss of the positive charge had greater effect on the skeletal isoform, mediated via distortion of a salt bridge network, a molecular latch inducing rotation of a cytoplasmic domain, and partial progression to open-state traits of the large cytoplasmic assembly accompanied by alteration of the Ca2+ binding site, which concur with the major “hyperactive” feature of the mutated channel. Our cryo-EM studies demonstrated the allosteric effect of a mutation situated ~85 Å away from the pore and identified an isoform-specific structural effect.
Honeybee caste development is nutritionally regulated by royal jelly (RJ). Major royal jelly protein 1 (MRJP1), the most abundant glycoprotein among soluble royal jelly proteins, plays pivotal roles in honeybee nutrition and larvae development, and exhibits broad pharmacological activities in humans. However, its structure has long remained unknown. Herein, we identify and report a 16-molecule architecture of native MRJP1 oligomer containing four MRJP1, four apisimin, and eight unanticipated 24-methylenecholesterol molecules at 2.65 Å resolution. MRJP1 has a unique six-bladed β-propeller fold with three disulfide bonds, and it interacts with apisimin mainly by hydrophobic interaction. Every four 24-methylenecholesterol molecules are packaged by two MRJP1 and two apisimin molecules. This assembly dimerizes to form an H-shaped MRJP14-apisimin4-24-methylenecholesterol8 complex via apisimin in a conserved and pH-dependent fashion. Our findings offer a structural basis for understanding the pharmacological effects of MRJPs and 24-methylenecholesterol, and provide insights into their unique physiological roles in bees.
The accumulation of mineral elements in seeds is controlled by multiple genes. Common physiological and molecular mechanisms could be involved in the accumulation of several mineral elements, and genes involved in these processes in B. napus are suggested. These results offer insights to the genetic basis of seed mineral accumulation across different P levels in B. napus.
2 Application of phosphorus (P) fertilizer is important in crop production because of the low bioavailability of phosphorus to plants in both acidic and calcareous soils. Although rapeseed (Brassica napus) is generally sensitive to P deficiency, different cultivars differ widely in this respect. Differences in P uptake and utilization between two rapeseed cultivars, one P-efficient ('97081') and one P-inefficient ('97009'), were evaluated in solution culture by studying the changes in root morphology and parameters of P uptake kinetics in response to low-P stress. The P-efficient cultivar had lower Km and Cmin values and higher Vmax and developed longer and denser lateral root hair with greater number of root tips and branches under low-P stress, which resulted in a better developed root system and more efficient uptake of P. That, in turn, led to higher concentration and accumulation of P in the plants, culminating in higher biomass production. However, P utilization efficiency (biomass production per unit P accumulated in plant) of the P-efficient '97081' was lower than that of '97009' when P was deficient. These results suggest that P efficiency in rapeseed is due to a better developed root system as well as efficient uptake of P.
Psoriasis is an immune-mediated polygenic inherited skin disease. Many biologic agents have been approved for the treatment of moderate-to-severe plaque psoriasis. The most commonly utilized biologics include TNF-α antagonists (etanercept, infliximab, and adalimumab), IL-12/23P40 antagonist (ustekinumab), IL-23P19 antagonist (guselkumab), IL-17A antagonist (secukinumab and ixekizumab), and IL-17RA antagonist (brodalumab). However, some patients may fail to respond well to their first biologic agent. Reasons for failure include primary failure (lack of initial efficacy), secondary failure (loss of efficacy over time) or the development of adverse effects. For patients desiring maximum skin clearance and better quality of life, switching to a second biologic agent might be a worthwhile option. This review discusses recent clinical studies on switching therapies in treating psoriasis, and found that switching biologic agents can significantly improve outcomes for patients. Some clinical guidelines are also discussed. This research provides some advice on establishing individualized treatment regimens based on clinical needs and pharmacologic characteristics.
We have studied Streptococcus mutans (S. mutans) biofilm growth and growth inhibition on Au(111)-surfaces using atomic force microscopy (AFM) and interfacial electrochemistry of a number of redox probe molecules. AFM of the biofilm growth and growth inhibition on both mica and Au(111)-surfaces was followed by sampling at given times, drying the samples naturally, and imaging. The electrochemical investigations were based on single-crystal Au(111)-electrode surfaces to exclude polycrystallinity as a cause of inhomogeneous voltammetric broadening on the biofilm covered electrode surfaces. The redox couples were chosen for their positive ([Ru(NH(3))(6)](3+/2+), [Co(terpy)(2)](3+/2+), terpy = 2,2',2''-terpyridine) or negative ([Fe(CN)(6)](3-/4-), [IrCl(6)](3-/4-)) electrostatic charge. [Co(NH(3))(6)](3+/2+) and [Co(phen)(3)](3+/2+) (phen = 1,10-phenanthroline) were other inhibition factors investigated. The positively and negatively charged redox probe couples displayed antagonistic inhibition and voltammetric patterns. [Ru(NH(3))(6)](3+/2+) and the homologous compound [Co(NH(3))(6)](3+/2+) were the only probe compounds to effect growth inhibition. On the other hand, cyclic voltammetry (CV) of both [Ru(NH(3))(6)](3+/2+) (positively charged, biofilm growth inhibitor) and [Co(terpy)(2)](3+/2+) (positively charged, no biofilm growth inhibition) displayed fully reversible CV on biofilm covered electrodes, almost indistinguishable from CV at bare Au(111)-electrode surfaces. In comparison, CVs of [Fe(CN)(6)](3-/4-) and [IrCl(6)](3-/4-) (both negatively charged and no growth inhibition) were distorted from planar diffusion behavior on bare Au(111)-electrode surfaces toward spherical diffusion behavior on S. mutans biofilm covered Au(111)-electrode surfaces. DNAase treatment of the biofilm covered Au(111)-electrode surface partly restores planar diffusion CV of [Fe(CN)(6)](3-/4-) and [IrCl(6)](3-/4-). This is reflected in a decrease of the growth rate and the appearance of molecular-scale structures near the bacterial edges as imaged by AFM after DNAase treatment. A rationale for the different voltammetric behavior of positively and negatively charged redox probe molecules based on electrostatic properties of the local surface environment is offered.
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