Following the Thread: Elite Iconography on Weaving Objects at Poggio Civitate (Murlo)

Evidence for industrial scale production of numerous manufacturing processes has been attested in all phases of occupation at Poggio Civitate (Murlo). A subset of these, tools for the production of textiles and fibers, indicates that textile crafts were manufactured on a large scale as a part of a centralized and organized industry. These industrialized practices occurred within and around the monumental seventh and sixth century BCE complexes which displayed architectural decoration bearing iconographic themes that served to secure the positions of the aristocratic elites. This paper investigates the stamped decoration present on rocchetti and its relationship to the architectural decoration present on the monumental structures of the site. As small moveable objects used by members of the community, rocchetti present an opportunity to investigate the movement of elite images through the non-elite population of a community and their reception of aristocratic ideology presented in monumental structures.

Two Step Cost-Saving Process for Industrial Scale Production of 1-Amino-3,5-dimethyladamantane Hydrochloride (An Anti-Alzheimer’s Drug)

This work presents a predominant cost-saving process for the industrial scale synthesis of memantine hydrochloride (1) from 1,3-dimethyladamantane (2) by a two-step method in which both steps were carried out in one-pot. The conversion of 2 to N-(3,5-dimethyladamantane-1-yl)acetylamide (3) was synthesized first in mixture of sulfuric acid 96-98% and nitric acid 64-65% at 25 ºC for 2.5 h and then in acetonitrile at 40 ºC for 3.5 h as a key step. The yield of this process reached to 98%. Next, the deacetylation of 3 with potasium hydroxide in mixture of water-ethylene glycol at 140 ºC for 15 h to get memantine base which was then converted into memantine hydrochloride (1) by aq. HCl 14% (yield of 93%). Overall yield of this synthetic route was 91.65%. The advantages of this process is saving time, solvents, reagents and giving a higher yield as compared to reported procedures.

Techno-Economic Analysis on an Industrial-Scale Production System of Polyhydroxyalkanoates (PHA) from Cheese By-Products by Halophiles

Polyhydroxyalkanoates (PHA) are a family of biodegradable plastics used as an ecofriendly alternative for conventional plastics in various applications. In this study, an industrial-scale PHA production system was designed and analyzed for the material flows and economics with the use of SuperPro Designer. Haloferax mediterranei was utilized to produce poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). Byproduct streams from a local cheese plant, with an input of 168.7 metric ton/day (MT/day) lactose, were used as the feedstock. Three scenarios with different processes for the treatments of used enzyme and spent medium were investigated and the major factors that influence the overall economics were identified. The simulated system produces 9700 MT/year PHBV with a yield of 0.2 g PHBV/g lactose and an overall process efficiency of 87%. The breakeven price was found to be more sensitive to the lactose price than enzyme price. The scenario with enzyme reuse and spent medium recycling achieved the lowest breakeven price among others, which can be less than 4 $/kg PHA based on the delactosed permeate (DLP) unit price. The study suggests utilizing dairy derived feedstocks has the potential to make PHA competitive in the bioplastic market, which could be beneficial to both dairy and bioplastic industries.

Rapid Enabling of Gluconobacter oxydans Resistance to High D-Sorbitol Concentration and High Temperature by Microdroplet-Aided Adaptive Evolution

Gluconobacter oxydans is important in the conversion of D-sorbitol into l-sorbose, which is an essential intermediate for industrial-scale production of vitamin C. In a previous study, the strain G. oxydans WSH-004 could directly produce 2-keto-l-gulonic acid (2-KLG). However, its D-sorbitol tolerance was poor compared with that of other common industrial G. oxydans strains, which grew well in the presence of more than 200 g/L of D-sorbitol. This study aimed to use the microbial microdroplet culture (MMC) system for the adaptive evolution of G. oxydans WSH-004 so as to improve its tolerance to high substrate concentration and high temperature. A series of adaptively evolved strains, G. oxydans MMC1-MMC10, were obtained within 90 days. The results showed that the best strain MMC10 grew in a 300 g/L of D-sorbitol medium at 40°C. The comparative genomic analysis revealed that genetic changes related to increased tolerance were mainly in protein translation genes. Compared with the traditional adaptive evolution method, the application of microdroplet-aided adaptive evolution could improve the efficiency in terms of reducing time and simplifying the procedure for strain evolution. This research indicated that the microdroplet-aided adaptive evolution was an effective tool for improving the phenotypes with undemonstrated genotypes in a short time.

Dispersion of Micro Fibrillated Cellulose (MFC) in Poly(lactic acid) (PLA) from Lab-Scale to Semi-Industrial Processing Using Biobased Plasticizers as Dispersing Aids

In the present study, two commercial typologies of microfibrillated cellulose (MFC) (Exilva and Celish) with 2% wt % were firstly melt-compounded at the laboratory scale into polylactic acid (PLA) by a microcompounder. To reach an MFC proper dispersion and avoid the well-known aglomeration problems, the use of two kinds of biobased plasticisers (poly(ethylene glycol) (PEG) and lactic acid oligomer (OLA)) were investigated. The plasticizers had the dual effect of dispersing the MFC, and at the same time, they counterbalanced the excessive stiffness caused by the addition of MFC to the PLA matrix. Several preliminaries dilution tests, with different aqueous cellulose suspension/plasticizer weight ratios were carried out. These tests were accompanied by SEM observations and IR and mechanical tests on compression-molded films in order to select the best plasticizer content. The best formulation was then scaled up in a semi-industrial twin-screw extruder, feeding the solution by a peristaltic pump, to optimize the industrial-scale production of commercial MFC-based composites with a solvent-free method. From this study, it can be seen that the use of plasticisers as dispersing aids is a biobased and green solution that can be easily used in conventional extrusion techniques.

Autochthonous Natural Starter Cultures: A Chance to Preserve Biodiversity and Quality of Pecorino Romano PDO Cheese

During Pecorino Romano PDO cheese production, scotta (residual whey from ricotta cheese manufacturing) or siero (whey) can be integrated with autochthonous starters, natural and composed of an indefinite number of species and strains, or commercial selected starters to obtain scotta/siero-innesto. In this study, three biodiverse autochthonous natural starter cultures (SR30, SR56, and SR63) belonging to the Agris Sardegna BNSS microbial collection, composed of different strains belonging to the species Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Enterococcus faecium, and Limosilactobacillus reuteri were lyophilized and combined into two starter mixes (A and B). The scotta/siero-innesto and the Pecorino Romano obtained using natural starters were compared with those obtained using commercial selected starters during three seasons of the cheesemaking campaign. Different pH and microbial compositions for the scotta/siero-innesto obtained using natural or commercial starters were found, attributable to their different biodiversity. The six-month-ripened cheese microbiota was influenced mostly by the season of cheesemaking, whereas physico-chemical and sensory analyses did not highlight differences among the products obtained. In general, no effect attributable to the type of scotta/siero-innesto used was observed, allowing the conclusion that natural starter cultures can be used also in industrial-scale production, ensuring high stability in the technological performances and preserving the microbial, chemical, and sensory characteristics of Pecorino Romano PDO cheese.

Increased Protein Encapsulation in Polymersomes with Hydrophobic Membrane Anchoring Peptides in a Scalable Process

Hollow vesicles made from a single or double layer of block-copolymer molecules, called polymersomes, represent an important technological platform for new developments in nano-medicine and nano-biotechnology. A central aspect in creating functional polymersomes is their combination with proteins, especially through encapsulation in the inner cavity of the vesicles. When producing polymersomes by techniques such as film rehydration, significant proportions of the proteins used are trapped in the vesicle lumen, resulting in high encapsulation efficiencies. However, because of the difficulty of scaling up, such methods are limited to laboratory experiments and are not suitable for industrial scale production. Recently, we developed a scalable polymersome production process in stirred-tank reactors, but the statistical encapsulation of proteins resulted in fairly low encapsulation efficiencies of around 0.5%. To increase encapsulation in this process, proteins were genetically fused with hydrophobic membrane anchoring peptides. This resulted in encapsulation efficiencies of up to 25.68%. Since proteins are deposited on the outside and inside of the polymer membrane in this process, two methods for the targeted removal of protein domains by proteolysis with tobacco etch virus protease and intein splicing were evaluated. This study demonstrates the proof-of-principle for production of protein-functionalized polymersomes in a scalable process.

Immunogenicity and efficacy of the COVID-19 candidate vector vaccine MVA-SARS-2-S in preclinical vaccination

Severe acute respiratory syndrome (SARS) coronavirus 2 (SARS-CoV-2) has emerged as the infectious agent causing the pandemic coronavirus disease 2019 (COVID-19) with dramatic consequences for global human health and economics. Previously, we reached clinical evaluation with our vector vaccine based on modified vaccinia virus Ankara (MVA) against the Middle East respiratory syndrome coronavirus (MERS-CoV), which causes an infection in humans similar to SARS and COVID-19. Here, we describe the construction and preclinical characterization of a recombinant MVA expressing full-length SARS-CoV-2 spike (S) protein (MVA-SARS-2-S). Genetic stability and growth characteristics of MVA-SARS-2-S, plus its robust expression of S protein as antigen, make it a suitable candidate vaccine for industrial-scale production. Vaccinated mice produced S-specific CD8+ T cells and serum antibodies binding to S protein that neutralized SARS-CoV-2. Prime-boost vaccination with MVA-SARS-2-S protected mice sensitized with a human ACE2-expressing adenovirus from SARS-CoV-2 infection. MVA-SARS-2-S is currently being investigated in a phase I clinical trial as aspirant for developing a safe and efficacious vaccine against COVID-19.