Human Cellular Retinol Binding Protein II Forms a Domain-Swapped Trimer Representing a Novel Fold and a New Template for Protein Engineering

Domain-swapping is a mechanism for evolving new protein structure from extant scaffolds, and has been an efficient protein-engineering strategy for tailoring functional diversity. However, domain swapping can only be exploited if it can be controlled, especially in cases where various folds can coexist. Herein, we describe the structure of a domain-swapped trimer of the iLBP family member hCRBPII, and suggest a mechanism for domain-swapped trimerization. It is further shown that domain-swapped trimerization can be favored by strategic installation of a disulfide bond, thus demonstrating a strategy for fold control. We further show the domain-swapped trimer to be a useful protein design template by installing a high-affinity metal binding site through the introduction of a single mutation, taking advantage of its threefold symmetry. Together, these studies show how nature can promote oligomerization, stabilize a specific oligomer, and generate new function with minimal changes to the protein sequence.     

Defining Dry Eye from a Clinical Perspective

Over the past decades, the number of patients with dry eye disease (DED) has increased dramatically. The incidence of DED is higher in Asia than in Europe and North America, suggesting the involvement of cultural or racial factors in DED etiology. Although many definitions of DED have been used, discrepancies exist between the various definitions of dry eye disease (DED) used across the globe. This article presents a clinical consensus on the definition of DED, as formulated in four meetings with global DED experts. The proposed new definition is as follows: “Dry eye is a multifactorial disease characterized by a persistently unstable and/or deficient tear film (TF) causing discomfort and/or visual impairment, accompanied by variable degrees of ocular surface epitheliopathy, inflammation and neurosensory abnormalities.” The key criteria for the diagnosis of DED are unstable TF, inflammation, ocular discomfort and visual impairment. This definition also recommends the assessment of ocular surface epitheliopathy and neurosensory abnormalities in each patient with suspected DED. It is easily applicable in clinical practice and should help practitioners diagnose DED consistently. This consensus definition of DED should also help to guide research and clinical trials that, to date, have been hampered by the lack of an established surrogate endpoint.     

Lactic fermentation of cooked navy beans by Lactobacillus paracasei CBA L74 aimed at a potential production of functional legume-based foods

In recent years, scientific interest in the development of non-dairy-based functional foods is increasing progressively and the fermentation of cereals, legumes, fruits and vegetable-based foods is becoming an important scientific research topic for the production of new probiotic products. In particular, legumes represent a possible alternative to protein foods from animal origins and an adequate fermentation substrate as they contain high amount of nutrients, such as proteins, carbohydrates, fibres, vitamins, and minerals, which are all useful to the growth and metabolic activity of certain microorganisms. This work focuses on the feasibility of developing a dry legume-based functional product using a fermentation process carried out on a 10% w/v navy bean suspension, in a lab-scale stirred batch reactor. After soaking and cooking dried navy beans, the fermentation tests performed on the resulting medium using Lactobacillus paracasei CBA L74 showed a maximum bacterial count of 10⁹ CFU/mL after 20 hours and a maximum lactic acid concentration of 1.9 g/L after 16 hours of process time. A freeze-drying process was performed on the fermented bean suspension, showing a 2-log microbial reduction and a bacterial viability in the resulting probiotic powder of 3.7 × 10⁸ CFU/g.     

Effect of Alumina Reactivity on the Densification and Properties of Al2O3–Cr2O3 Refractories

Alumina‐chrome (Al₂O₃–Cr₂O₃) refractories with Al₂O₃:Cr₂O₃ molar ratio 1:1 were synthesized in the temperature range of 1400–1700°C by conventional solid–oxide reaction route. The effect of different aluminas (viz., hydrated and calcined) on the densification, microstructure, and properties of Al₂O₃–Cr₂O₃ refractories was investigated without changing the Cr₂O₃ source. The starting materials were analyzed to determine the chemical composition, mineralogy, density, surface area, and particle size. Sintered materials were characterized in terms of densification, phase assemblage, and mechanical strength at room temperature and at higher temperatures. Microstructural evolution at different sintering temperature was correlated with sintering characteristics. It can be concluded that the Al₂O₃–Cr₂O₃ refractories prepared with hydrated alumina as Al₂O₃ source show better densification and hot mechanical strength than corresponding calcined variety.     

Miscible blends of biobased poly(lactide) with poly(methyl methacrylate): Effects of chopped glass fiber incorporation

Poly(lactide) (PLA) and poly(methyl methacrylate) (PMMA) are melt compounded with chopped glass fiber using laboratory scale twin‐screw extrusion. Physical properties are examined using differential scanning calorimetry (DSC), dynamic mechanical thermal analysis (DMTA), thermogravimetric analysis (TGA), tensile testing, impact testing, X‐ray computed tomography (CT) scanning, and field emission scanning electron microscopy (FE‐SEM). Molecular weight is determined using gel permeation chromatography (GPC). Miscibility of the blends is implied by the presence of a single glass transition temperature and homogeneous morphology. PLA/PMMA blends tend to show positive deviations from a simple linear mixing rule in their mechanical properties (e.g., tensile toughness, modulus, and stress at break). The addition of 40 wt % glass fiber to the system dramatically increases physical properties. Across all blend compositions, the tensile modulus increases from roughly 3 GPa to roughly 10 GPa. Estimated heat distortion temperatures (HDTs) are also greatly enhanced; the pure PLA sample HDT increases from 75 °C to 135 °C. Fiber filled polymer blends represent a sustainable class of earth abundant materials which should prove useful across a range of applications. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 44868.     

Improvement in volume resistivity and morphology of a blend of polyolefin elastomer with linear low-density polyethylene

A silane moisture-cured polyolefin elastomer/linear low-density polyethylene (LLDPE) blend was prepared through a two-step silane-grafting method (Sioplas Process) in an industrial scale twin-screw extruder. The silane-grafted compound was used to make wire and cable coatings. In this work, the effect of some interactive parameters on quality of the products prepared by the above method has been studied, while so far, there have been less experimental investigations. The volume resistivity of cross-linked compound was changed from 2.96 × 10¹⁴ to 7.41 × 10¹⁴ Ω cm with increasing LLDPE component by maximum 10 wt%. Surface morphology of the product was corrected with reduction in benzoyl peroxide (BPO) concentration from 0.2 wt% to 0.13 wt%. BPO at this level acted as an initiator in grafting reaction of vinyl trimethoxysilane. The curing condition and specimen preparation method by injection molding and/or extrusion were factors which influenced the hot-set test results at 200 °C. The results of tensile and elongation studies showed a maximum value of 9 MPa and 397% for the tests, after 6 h curing. With increases in curing time at a specified temperature, the gel content of the cross-linked compound was increased and reached its maximum value. The maximum gel content values were found to be approximately 60%, 80%, and 82% at temperatures of 25, 60, and 85 °C, respectively. The hardness, density, and tear strength of the samples did not vary significantly with the curing temperature.

     

高产刺糖菌素发酵菌株的选育

采用紫外线结合氯化锂以及硫酸二乙酯结合氯化锂2种常规诱变方法对刺糖多孢菌进行复合诱变,分别筛选鼠李糖抗性突变株、2-脱氧-D-葡萄糖抗性突变株和刺糖菌素抗性突变株,并结合自然分离纯化,最终获得高产菌株D-S-23,其产量达N89.57 mg/L,较出发菌株提高了104.3%.     

Evaluation of alkane sulfonic acid catalysts and derivatives for curing activity in a melamine/acrylic polyol coating formulation using fourier transform infrared spectroscopy

FTIR measurements may be able to replace more laborious physical property determinations and thus serve as an efficient screening methodology in catalysis driven processes. This possibility was considered in evaluating alkane sulfonic acids catalyst performance to promote the cure of a melamine resin with acrylic polyols. The example chosen here is an evaluation of alkane sulfonic acid derivatives to measure catalytic efficiency of the cure reaction between hexamethoxymethylmelamine (HMMM) and an acrylicpolyol. The IR measurements provided information on the extent of reaction, via monitoring the disappearance of the OH stretching mode at 3472 cm⁻¹ associated with the acrylic polyol. This method may also be used to provide detailed information on reaction kinetics and insight into the cure mechanism. As the reaction proceeds, the coating’s properties change and IR measurements can serve as a probe for these changes. 900 First Ave., King of Prussia, PA 19406.     

A Michael-type reaction between acrylato ions and ethylenediamine coordinated to Ni(II). Synthesis,crystal structure and magnetic properties of [Ni2(EDDP)2(H2O)2] · 2H2O (H2EDDP = ethylenediamine-N,N-dipropionic acid)

The reaction between NiCO₃ · Ni(OH)₂, acrylic acid and ethylenediamine in a 2:4:1 molar ratio affords the binuclear complex, [Ni₂(EDDP)₂(H₂O)₂] · 2H₂O 1. The organic ligand, EDDP^2? (the dianion of the ethylenediamine-N,N-dipropionic acid ligand), results from the addition of one amine group to the carbon–carbon double bonds of two acrylato ions. The crystal structure of 1 consists of neutral centrosymmetric entities, with the nickel ions connected by two carboxylato groups, each one acting as a monoatomic bridge. The intramolecular Ni?Ni distance is 3.212 Å. The metal ions exhibit an octahedral geometry. The cryomagnetic investigation of 1 reveals an antiferromagnetic coupling of the nickel(II) ions (J = ?21.8 cm^?1, H = ?JS_Ni1S_Ni2).     

Isolation,Culture and Comprehensive Characterization of Biological Properties of Human Urine-Derived Stem Cells

Mesenchymal stem cells (MSCs) represent an attractive source within the field of tissue engineering. However, their harvesting often requires invasive medical procedures. Urine-derived stem cells (UDSCs) display similar properties to MSCs, and their obtention and further processing is non-invasive for the donors as well as low cost. Here, we offer a comprehensive analysis of their biological properties. The goal of this study was to analyze their morphology, stemness, differentiation potential and cytokine profile. We have successfully isolated UDSCs from 25 urine samples. First colonies emerged up to 9 days after the initial seeding. Cell doubling time was 45 ± 0.24 SD, and when seeded at the density of 100 cells/cm², they formed 42 ± 6.5 SD colonies within 10 days. Morphological analyzes revealed that two different types of the cell populations have been present. The first type had a rice-grain shape and the second one was characterized by a polyhedral shape. In several cell cultures, dome-shaped cells were observed as well. All examined UDSCs expressed typical MSC-like surface markers, CD73, CD90 and CD105. Moreover, conditioned media from UDSCs were harvested, and cytokine profile has been evaluated showing a significantly higher secretory rate of IL-8, IL-6 and chemokines MCP-1 and GM-CSF. We have also successfully induced human UDSCs into chondrogenic, osteogenic and myogenic cell lineages. Our findings indicate that UDSCs might have immense potential in the regeneration of the damaged tissues.