Industrial production of recycled cement: energy consumption and carbon dioxide emission estimation.

The urge to reduce greenhouse gas emissions, in particular carbon dioxide, is a global problem, not only in spatial terms but also in terms of the scope of activities and sectors involved. Nevertheless, some sectors/industries are more critical due to their overall contribution to the problem, which is the case of the Portland cement industry. The present research estimates the energy consumption and carbon emissions associated with a novel process for producing cement by recycling used concrete and mortars. The novel process assessed resorts to the magnetic separation of the cement paste from the aggregates, followed by the thermal reactivation of the cement paste. Comparing the recycled cement production with the clinker production, higher energy consumption (over 9000 MJ/t compared with roughly 4000 MJ/t for Portland cement) and lower carbon dioxide emissions (average 730 kg CO2/t compared with more than 800 kg CO2/t for Portland cement) were estimated. However, the potential benefits in an industrial application are potentially much higher with the optimization of the production process. In particular, improvements in the washing and drying of the material prior to the magnetic separation will be critical since most of the energy is consumed in the process of drying.

Life Cycle Assessment of Thermoactivated Recycled Cement Production.

The urgent need to tackle the effects of global warming has led to a worldwide compromise and ever-more demanding regulations. In this respect, as an important greenhouse gas emitter, the cement industry has to implement major changes in its production processes to achieve future goals. In this perspective, low-carbon eco-efficient cement, such as the thermoactivated recycled cement from concrete waste (RCC), seem to be a promising alternative to current carbon-intensive binders, such as ordinary Portland cement (OPC). This study aimed to demonstrate the potential contribution of RCC to the reduction in the environmental impacts of the cement industry, by means of a comparative life cycle assessment of three production methods of this binder (wet (WM), dry (DM) and air clean (ACM) methods) and OPC. Overall, RCC WM did not turn out to be a good alternative to OPC, essentially owing to the amount of fuel and electricity required for washing and drying the particles before the magnetic separation. On the other hand, RCC DM and RCC ACM proved to be promising alternatives to RCC WM and OPC, with a relevant reduction in all impact categories.

Influence of the Treatment Temperature on the Microstructure and Hydration Behavior of Thermoactivated Recycled Cement.

This paper intends to contribute to a better knowledge of the production and rehydration of thermoactivated recycled cement and its incorporation in cement-based materials. To this end, the influence of the treatment temperature on the properties of recycled cements and recycled cement pastes was assessed by means of a wide array of tests. Anhydrous recycled cement as well as the resulting pastes were characterized through density and particle size, water demand and setting time, thermogravimetry, X-ray diffraction, field emission gun scanning electron microscopy, isothermal calorimetry, 29Si nuclear magnetic resonance spectroscopy, flowability, mechanical strength, mercury intrusion porosimetry and scanning electron microscopy. The treatment temperature had a significant influence on the dehydration and hydration of recycled cement, essentially resulting in the formation of C2S polymorphs of varying reactivity, which led to pastes of different fresh and hardened behaviors. The high water demand and the pre-hydration of recycled cement resulted in high setting times and low compressive strengths. The highest mechanical strength was obtained for a treatment temperature of 650 °C.

Synthetic antibody discovery against native antigens by CRISPR/Cas9-library generation and endoplasmic reticulum screening.

Despite the significant advances of antibodies as therapeutic agents, there is still much room for improvement concerning the discovery of these macromolecules. Here, we present a new synthetic cell-based strategy that takes advantage of eukaryotic cell biology to produce highly diverse antibody libraries and, simultaneously, link them to a high-throughput selection mechanism, replicating B cell diversification mechanisms. The interference of site-specific recognition by CRISPR/Cas9 with error-prone DNA repair mechanisms was explored for the generation of diversity, in a cell population containing a gene for a light chain antibody fragment. We achieved up to 93% of cells containing a mutated antibody gene after diversification mechanisms, specifically inside one of the antigen-binding sites. This targeted variability strategy was then integrated into an intracellular selection mechanism. By fusing the antibody with a KDEL retention signal, the interaction of antibodies and native membrane antigens occurs inside the endoplasmic reticulum during the process of protein secretion, enabling the detection of high-quality leads for expression and affinity by flow cytometry. We successfully obtained antibody lead candidates against CD3 as proof of concept. In summary, we developed a novel antibody discovery platform against native antigens by endoplasmic synthetic library generation using CRISPR/Cas9, which will contribute to a faster discovery of new biotherapeutic molecules, reducing the time-to-market.

Author Correction: A zebrafish drug screening platform boosts the discovery of novel therapeutics for spinal cord injury in mammals.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

A Review on the Carbonation and Chloride Penetration Resistance of Structural Lightweight Aggregate Concrete.

This paper presents a comprehensive review on structural lightweight aggregate concrete (SLWAC) durability. The main transport properties and degradation mechanisms of reinforced concrete are addressed, namely, carbonation and chloride attack. The influence of the main composition parameters, such as type of aggregate, type of binder and water/binder ratio, as well as the influence of cracking, are also analysed. Finally, the current knowledge of SLWAC's service life prediction is assessed. Although the knowledge of SLWAC's durability behaviour is still limited, investigation works performed either in laboratory or in real environments indicate that SLWAC can have similar to better durability performance than normal weight concrete, especially when the same strength level is considered. The importance of the quality of the paste over the characteristics of the lightweight aggregates is highlighted. Durability standardization regarding SLWAC is still insufficient and is one of the main gaps of current knowledge. The objective of this review is to foster a better understanding on the durability and service life prediction of SLWAC, contributing to a greater confidence in using this type of concrete.

A zebrafish drug screening platform boosts the discovery of novel therapeutics for spinal cord injury in mammals.

Spinal cord injury (SCI) is a complex condition, with limited therapeutic options, that results in sensory and motor disabilities. To boost discovery of novel therapeutics, we designed a simple and efficient drug screening platform. This innovative approach allows to determine locomotor rescue properties of small molecules in a zebrafish (Danio rerio) larval spinal cord transection model. We validated our screening platform by showing that Riluzole and Minocycline, two molecules that are in clinical trials for SCI, promote rescue of the locomotor function of the transected larvae. Further validation of the platform was obtained through the blind identification of D-Cycloserine, a molecule scheduled to enter phase IV clinical trials for SCI. Importantly, we identified Tranexamic acid and further showed that this molecule maintains its locomotor recovery properties in a rodent female contusion model. Our screening platform, combined with drug repurposing, promises to propel the rapid translation of novel therapeutics to improve SCI recovery in humans.

Quality and safety requirements for sustainable phage therapy products.

The worldwide antibiotic crisis has led to a renewed interest in phage therapy. Since time immemorial phages control bacterial populations on Earth. Potent lytic phages against bacterial pathogens can be isolated from the environment or selected from a collection in a matter of days. In addition, phages have the capacity to rapidly overcome bacterial resistances, which will inevitably emerge. To maximally exploit these advantage phages have over conventional drugs such as antibiotics, it is important that sustainable phage products are not submitted to the conventional long medicinal product development and licensing pathway. There is a need for an adapted framework, including realistic production and quality and safety requirements, that allows a timely supplying of phage therapy products for 'personalized therapy' or for public health or medical emergencies. This paper enumerates all phage therapy product related quality and safety risks known to the authors, as well as the tests that can be performed to minimize these risks, only to the extent needed to protect the patients and to allow and advance responsible phage therapy and research.

Wound healing potential of topical bacteriophage therapy on diabetic cutaneous wounds.

Chronic wounds that fail to heal are a common complication of diabetes mellitus and the most common precipitating reason for nontraumatic lower limb amputation. Unfortunately, the bacterial species that cause these infections are becoming more resistant to antibiotics, making them increasingly difficult to treat. We assessed the feasibility of combating chronic bacterial infections with a topically delivered bacteriophage cocktail in two animal models of diabetes mellitus. Microbiological, planimetric, and histological parameters were compared in debrided infected wounds with or without topical bacteriophage treatment. We determined that bacteriophage treatment effectively decreased bacterial colony counts and improved wound healing, as indicated by smaller epithelial and dermal gaps, in Staphylococcus aureus and Pseudomonas aeruginosa infections but was not as effective against Acinetobacter baumannii. Although the improvements were more significant in the rodent model than in the porcine model, our results suggest that topically administered bacteriophage treatment may be effective in resolving chronic infections, especially when applied in conjunction with wound debridement. These findings have important implications for the feasibility of using topical antimicrobial therapies to safely treat chronic infections in diabetes mellitus patients.

Recombinant antibodies as therapeutic agents: pathways for modeling new biodrugs.

Hybridoma fusion technology, proposed by Köhler and Milstein in 1975, started major developments in the field of monoclonal antibodies (mAbs). During the following 2 decades, their high potential as laboratory tools was rapidly exploited for biotechnology and biomedical applications. Today, mAbs represent over 30% of all biological proteins undergoing clinical trials and are the second largest class of biodrugs after vaccines. With the help of antibody engineering, mAbs have been reduced in size, rebuilt into multivalent molecules, and conjugated with drugs, toxins, or radioisotopes for the treatment of cancer, autoimmune disorders, graft rejection, and infectious diseases. Additionally, in the past few years, important advances have been made in the design, selection, and production of these new types of engineered antibodies. The present review focuses on the structural and functional characteristics of mAbs and their fragments, and also provides a walk through the most important methods used in antibody selection. In addition, the recent trends in antibody engineering for improving antibody clinical efficacy are also reviewed.

Intrabodies targeting the Kaposi sarcoma-associated herpesvirus latency antigen inhibit viral persistence in lymphoma cells.

Kaposi sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen-1 (LANA1) is essential for the maintenance and segregation of viral episomes in KSHV latently infected B cells. We report development of intracellular, rabbit-derived antibodies generated by phage display technology, which bind to N-terminal LANA1 epitopes and neutralize the chromosome-binding activity of LANA1. Although these cloned single-chain variable fragments (scFvs) show relatively low binding affinities for the LANA1 viral antigen in in vitro assays, they nonetheless outcompete KSHV-seropositive human sera for LANA1 epitope binding. In heterologous cells, intracellular intrabody expression inhibits LANA1-dependent plasmid maintenance of both an artificial plasmid containing KSHV LANA1 binding sequences and a bacterial artificial chromosome containing the entire KSHV genome. In KSHV naturally infected primary effusion lymphoma cells, intracellular intrabody expression causes a reduction or loss of the typical LANA1 punctate, nuclear pattern. This morphologically apparent LANA1 dispersion correlates to loss of viral episome by molecular analysis. These data suggest a novel approach to antiherpes viral therapy and confirm LANA1 is critical target for neutralization of KSHV viral latency.

Cell type-specific targeting with sindbis pseudotyped lentiviral vectors displaying anti-CCR5 single-chain antibodies.

Lentiviral vectors are among the most efficient tools for gene delivery into mammalian cells. A major goal of lentiviral gene delivery systems is to develop vectors that can efficiently target specific cell types. In the present work, we attempt to generate viral particles for targeting gene delivery. We have used CCR5-positive cells as the target for our strategy. Therefore, we developed a novel Sindbis pseudotyped lentiviral vector where the Sindbis receptor binding envelope protein was modified to directly encode a single-chain antibody fragment (scFv) against the CCR5 chemokine receptor. We have generated two chimeric scFv-Sindbis envelopes, varying the length of the peptide linker that connects the heavy chain and light chain of anti-CCR5 scFv. The two chimeric scFv-Sindbis envelopes were successfully incorporated into lentiviral-derived vectors, and the resulting pseudotyped viral particles showed specific targeting to CCR5-expressing cells. However, our data demonstrate that the length of the peptide linker significantly affects the efficiency of infection. Pseudotyped viral particles, which display single-chain antibody fragments with longer peptide linkers, allowed higher titers of infection. The present study can be a model strategy for specific gene delivery mediated by lentiviral vectors pseudotyped with Sindbis envelope displaying scFv that recognizes specific cellular surface proteins. Furthermore, this strategy has the potential to become a powerful approach for targeting gene delivery in anti- HIV gene therapy due to the important role of CCR5 expression in disease progression.