Development of Anti-LRRC15 Small Fragments for Imaging Purposes Using a Phage-Display ScFv Approach

The human leucine-rich repeat-containing protein 15 (LRRC15) is a membrane protein identified as a marker of CAF (cancer-associated fibroblast) cells whose overexpression is positively correlated with cancer grade and outcome. Nuclear molecular imaging (i.e., SPECT and PET) to track LRRC15 expression could be very useful in guiding further therapeutic strategies. In this study, we developed an ScFv mouse phage-display library to obtain small fragment antibodies against human LRRC15 for molecular imaging purposes. Mice were immunized with recombinant human LRRC15 (hLRRC15), and lymph node cells were harvested for ScFv (single-chain variable fragment) phage-display analysis. The built library was used for panning on cell lines with constitutive or induced expression after transfection. The choice of best candidates was performed by screening various other cell lines, using flow cytometry. The selected candidates were reformatted into Cys-ScFv or Cys-diabody by addition of cysteine, and cloned in mammalian expression vectors to obtain batches of small fragments that were further used in site-specific radiolabeling tests. The obtained library was 1.2 × 10⁷ cfu/µg with an insertion rate >95%. The two panning rounds performed on cells permittedenrichment of 2 × 10⁻³. Screening with flow cytometry allowed us to identify 28 specific hLRRC15 candidates. Among these, two also recognized murine LRCC15 and were reformatted into Cys-ScFv and Cys-diabody. They were expressed transiently in a mammalian system to obtain 1.0 to 4.5 mg of Cys fragments ready for bioconjugation and radiolabeling. Thus, in this paper, we demonstrate the relevance of the phage-display ScFv library approach for the fast-track development of small antibodies for imaging and/or immunotherapy purposes.     

STEM nanoanalysis of Au/Pt/Ti-Si3N4 interfacial defects and reactions during local stress of SiGe HBTs

Nanoscaled materials are attractive building blocks for hierarchical assembly of functional nanodevices, which exhibit diverse performances and simultaneous functions. We innovatively fabricated semiconductor nano-probes of tapered ZnS nanowires through melting and solidifying by electro-thermal process; and then, as-prepared nano-probes can manipulate nanomaterials including semiconductor/metal nanowires and nanoparticles through sufficiently electrostatic force to the desired location without structurally and functionally damage. With some advantages of high precision and large domain, we can move and position and interconnect individual nanowires for contracting nanodevices. Interestingly, by the manipulating technique, the nanodevice made of three vertically interconnecting nanowires, i.e., diode, was realized and showed an excellent electrical property. This technique may be useful to fabricate electronic devices based on the nanowires' moving, positioning, and interconnecting and may overcome fundamental limitations of conventional mechanical fabrication.     

Properties of Injectable Apatite-Forming Premixed Cements

Previous studies reported premixed calcium phosphate cements (CPCs) that were stable in the package and form hydroxyapatite (HA) as the product after exposure to an aqueous environment. These cements had setting times of greater than 60 min, which are too long to be useful for some clinical applications. The present study investigated properties of fast-setting HA-forming premixed CPCs that initially consisted of two separate premixed pastes: (1) finely ground (1.0 μm in median size) dicalcium phosphate anhydrous (DCPA) mixed with an aqueous NaH(2)PO(4) solution, 1.5 mol/L or 3.0 mol/L in concentration, and (2) tetracalcium phosphate consisting of combinations of particles of two different size distributions, 5 μm (TTCP5) and 17 μm (TTCP17) in median size, mixed with glycerin. Equal volume of Pastes 1 and 2 were injected with the use of atwo-barrel syringe fitted with a static mixer into sample molds. The molar Ca/P ratio of combined paste was approximately 1.5. Cements were characterized in terms of setting time (Gilmore needle), diametral tensile strength (DTS), and phase composition (powder x-ray diffraction, XRD). Setting times were found to range from (4.3 ± 0.6 to 68 ± 3) min (mean ± sd; n = 3), and 1-d and 7-d DTS values were from (0.89 ± 0.08 to 2.44 ± 0.16) MPa (mean ± sd; n = 5). Both the NaH(2)PO(4) concentration and TTCP particle size distribution had significant (p < 0.01) effects on setting time and DTS. Powder XRD analysis showed that low crystallinity HA and unreacted DCPA were present in the 1-day specimens, and the extent of HA formation increased with increasing amount of TTCP5 in the TTCP paste. CONCLUSION: Injectable HA-forming premixed CPCs with setting times from 4 to 70 min can be prepared by using DCPA and TTCP as the ingredients. Compared to the conventional powder liquid cements, these premixed CPCs have the advantages of being easy to use and having a range of hardening times.     

西方学者看中国城市——社会科学及历史学的文献回顾

西方的社会科学工作者及历史学家,对中国城市的研究起步较早、牵涉面较广、视野较高、学科间的观念互用较多,并有较丰硕的成果。本文就西方学者在历史学、地理学、人类学、社会学、城市规划、政治学、经济学、及跨学科中,对中国城市研究的成果,特别是1980年之后的专著,作一简短的文献评介,以收他山之石之效。     

Implications of sequential use of UV and ozone for drinking water quality

The formation of bromate levels exceeding the drinking water standard of 10 microg L-1 may impose the reduction of ozone doses used in the treatment of drinking water. This paper illustrates the procedure of evaluating the use of reduced ozone doses while implementing an additional UV disinfection step for an actual drinking water treatment plant. Ozonation was performed at low ozone doses in bench-scale experiments with a pretreated river water from the Paris area (France). At the low ozone dose of 0.5 mg L-1, bromate formation could be kept below 0.4 microg L-1, while inactivation of vegetative bacteria and UV-resistant viruses was calculated to exceed 5 log units, and a substantial decoloration (31% of the absorption at lambda=254 nm) was achieved. Based on the measured transient ozone and OH radical concentrations, the oxidation of micropollutants was calculated. Fast reacting micropollutants containing phenol, amine or double bond moieties, such as sulfamethoxazole, diclofenac and 17-alpha-ethinylestradiol, were completely oxidized. Slow-reacting synthetic micropollutants, e.g., atrazine, iopromide and methyl tertiary butyl ether (MTBE), were oxidized by only 20%, 20% and 10%, respectively, and the taste and odor compounds 2-methylisoborneol (MIB) and geosmin by 40% and 50%, respectively. The addition of an UV treatment step to the existing treatment train, which should guarantee disinfection of ozone-resistant pathogenic microorganisms, including Cryptosporidium parvum oocysts, has negligible effects on water matrix components but may induce significant transformation of micropollutants. Overall, the combination of ozonation at reduced doses and UV treatment leads to an improved water quality with regard to disinfection, oxidation of micropollutants and minimization of bromate.     

Survey of 1012 moldy dwellings by culture fungal analysis: Threshold proposal for asthmatic patient management

Different countries have tried to define guidelines to quantify what levels of fungi are considered as inappropriate for housing. This retrospective study analyzes indoor fungi by cultures of airborne samples from 1012 dwellings. Altogether, 908 patients suffering from rhinitis, conjunctivitis, and asthma were compared to 104 controls free of allergies. Portuguese decree law no 118/2013 (PDL118), ANSES (a French environmental and health agency) recommendations, and health regulations of Besançon University Hospital were applied to determine the rates of non‐conforming dwellings, which were respectively 55.2%, 5.2%, and 19%. Environmental microbiological results and medical data were compared. The whole number of colonies per cubic meter of air was correlated with asthma (P < 0.001) and rhinitis (P = 0.002). Sixty‐seven genera and species were detected in bedrooms. Asthma was correlated to Aspergillus versicolor (P = 0.004) and Cladosporium spp. (P = 0.02). Thresholds of 300 cfu/m³ for A. versicolor or 495 cfu/m³ for Cladosporium spp. are able to discriminate 90% of the asthmatic dwellings. We propose a new protocol to obtain an optimal cost for indoor fungi surveys, excluding surface analyses, and a new guideline to interpret the results based on >1000 cfu/m³ of whole colonies and/or above threshold levels for A. versicolor or Cladosporium spp.     

Co-creation of value and the project context: Towards application on the case of Hinkley Point C Nuclear Power Station

The research analyses decisions as evaluative outcomes regarding project value. The UK-French Hinkley Point C Nuclear Power Station provides the case study. Value is traditionally assessed as inputs and outputs. Here, value is conceptualized as a co-created value proposition at the front-end with implications for realization post-completion. Service-dominant logic (SDL) provides the theoretical lens and contributes to a methodological approach for examining projects. Decision-making outcomes provide the evidence from a range of data sources, including reports and commentaries. The methods are interpretative. The findings show that decision-making extends beyond the time-cost-quality/scope dimensions. The long-term issues regarding value realized are often overlooked. Stakeholders and individual actors have mainly focused upon managing political and financial risks, especially time and cost. The research poses challenges to project management analysis, SDL and research design in assessing evidence. Addressing these issues facilitates a knowledge contribution to SDL theorization and the field of project management.