Search results for: polymeric chain friction

Authors: Thi Thi Zin, Pyke Tin, Ikuo Kobayashi, Yoichiro Horii

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Today dairy farm experts and farmers have well recognized the importance of dairy cow Body Condition Score (BCS) since these scores can be used to optimize milk production, managing feeding system and as an indicator for abnormality in health even can be utilized to manage for having healthy calving times and process. In tradition, BCS measures are done by animal experts or trained technicians based on visual observations focusing on pin bones, pin, thurl and hook area, tail heads shapes, hook angles and short and long ribs. Since the traditional technique is very manual and subjective, the results can lead to different scores as well as not cost effective. Thus this paper proposes an algebraic geometric imaging approach for an automatic dairy cow BCS system. The proposed system consists of three functional modules. In the first module, significant landmarks or anatomical points from the cow image region are automatically extracted by using image processing techniques. To be specific, there are 23 anatomical points in the regions of ribs, hook bones, pin bone, thurl and tail head. These points are extracted by using block region based vertical and horizontal histogram methods. According to animal experts, the body condition scores depend mainly on the shape structure these regions. Therefore the second module will investigate some algebraic and geometric properties of the extracted anatomical points. Specifically, the second order polynomial regression is employed to a subset of anatomical points to produce the regression coefficients which are to be utilized as a part of feature vector in scoring process. In addition, the angles at thurl, pin, tail head and hook bone area are computed to extend the feature vector. Finally, in the third module, the extracted feature vectors are trained by using Markov Classification process to assign BCS for individual cows. Then the assigned BCS are revised by using multiple regression method to produce the final BCS score for dairy cows. In order to confirm the validity of proposed method, a monitoring video camera is set up at the milk rotary parlor to take top view images of cows. The proposed method extracts the key anatomical points and the corresponding feature vectors for each individual cows. Then the multiple regression calculator and Markov Chain Classification process are utilized to produce the estimated body condition score for each cow. The experimental results tested on 100 dairy cows from self-collected dataset and public bench mark dataset show very promising with accuracy of 98%.

Keywords: algebraic geometric imaging approach, body condition score, Markov classification, polynomial regression

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Authors: Bagireanu Astrid, Bros-Williamson Julio, Duncheva Mila, Currie John

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The Architecture, Engineering, and Construction (AEC) sector plays a critical role in the global transition toward sustainable and net-zero built environments. However, the industry faces unique challenges in planning for net-zero while struggling with low productivity, cost overruns and overall resistance to change. Traditional practices fall short due to their inability to meet the requirements for systemic change, especially as governments increasingly demand transformative approaches. Working in silos and rigid hierarchies and a short-term, client-centric approach prioritising immediate gains over long-term benefit stands in stark contrast to the fundamental requirements for the realisation of net-zero objectives. These practices have limited capacity to effectively integrate AEC stakeholders and promote the essential knowledge sharing required to address the multifaceted challenges of achieving net-zero. In the context of built environment, procurement may be described as the method by which a project proceeds from inception to completion. Collaborative procurement methods under the Integrated Practices (IP) umbrella have the potential to align more closely with net-zero objectives. This paper explores the synergies between collaborative procurement principles and the pursuit of net zero in the AEC sector, drawing upon the shared values of cross-disciplinary collaboration, Early Supply Chain involvement (ESI), use of standards and frameworks, digital information management, strategic performance measurement, integrated decision-making principles and contractual alliancing. To investigate the role of collaborative procurement in advancing net-zero objectives, a structured research methodology was employed. First, the study focuses on a systematic review on the application of collaborative procurement principles in the AEC sphere. Next, a comprehensive analysis is conducted to identify common clusters of these principles across multiple procurement methods. An evaluative comparison between traditional procurement methods and collaborative procurement for achieving net-zero objectives is presented. Then, the study identifies the intersection between collaborative procurement principles and the net-zero requirements. Lastly, an exploration of key insights for AEC stakeholders focusing on the implications and practical applications of these findings is made. Directions for future development of this research are recommended. Adopting collaborative procurement principles can serve as a strategic framework for guiding the AEC sector towards realising net-zero. Synergising these approaches overcomes fragmentation, fosters knowledge sharing, and establishes a net-zero-centered ecosystem. In the context of the ongoing efforts to amplify project efficiency within the built environment, a critical realisation of their central role becomes imperative for AEC stakeholders. When effectively leveraged, collaborative procurement emerges as a powerful tool to surmount existing challenges in attaining net-zero objectives.

Keywords: collaborative procurement, net-zero, knowledge sharing, architecture, built environment

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Authors: Maria Jose Granero Paris, Ana Isabel Jimenez Zarco, Agustin Pablo Alvarez Herranz

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In the industrial sector collaboration with suppliers is key to the development of innovations in the field of processes. Access to resources and expertise that are not available in the business, obtaining a cost advantage, or the reduction of the time needed to carry out innovation are some of the benefits associated with the process. However, the success of this collaborative process is compromised, when from the beginning not clearly rules have been established that govern the relationship. Abundant studies developed in the field of innovation emphasize the strategic importance of the concept of “Goverance”. Despite this, there have been few papers that have analyzed how the governance process of the relationship must be designed and managed to ensure the success of the cooperation process. The lack of literature in this area responds to the wide diversity of contexts where collaborative processes to innovate take place. Thus, in sectors such as the car industry there is a strong collaborative tradition between manufacturers and suppliers being part of the value chain. In this case, it is common to establish mechanisms and procedures that fix formal and clear objectives to regulate the relationship, and establishes the rights and obligations of each of the parties involved. By contrast, in other sectors, collaborative relationships to innovate are not a common way of working, particularly when their aim is the development of process improvements. It is in this case, it is when the lack of mechanisms to establish and regulate the behavior of those involved, can give rise to conflicts, and the failure of the cooperative relationship. Because of this the present paper analyzes the similarities and differences in the processes of governance in collaboration with service providers in engineering R & D in the European aerospace industry. With these ideas in mind, we present research is twofold: - Understand the importance of governance as a key element of the success of the cooperation in the development of process innovations, - Establish the mechanisms and procedures to ensure the proper management of the processes of cooperation. Following the methodology of the case study, we analyze the way in which manufacturers and suppliers cooperate in the development of new processes in two industries with different levels of technological intensity and collaborative tradition: the automotive and aerospace. The identification of those elements playing a key role to establish a successful governance and relationship management and the compression of the mechanisms of regulation and control in place at the automotive sector can be use to propose solutions to some of the conflicts that currently arise in aerospace industry. The paper concludes by analyzing the strategic implications for the aerospace industry entails the adoption of some of the practices traditionally used in other industrial sectors. Finally, it is important to highlight that in this paper are presented the first results of a research project currently in progress describing a model of governance that explains the way to manage outsourced engineering services to suppliers in the European aerospace industry, through the analysis of companies in the sector located in Germany, France and Spain.

Keywords: innovation management, innovation governance, managing collaborative innovation, process innovation

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Authors: Yuriko Takayama, Norihiro Kato

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Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

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Authors: Thauane Silva, Gabrielle do Vale, Andre Ferreira, Marilda Siqueira, Thiago Moreno L. Souza, Milene D. Miranda

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The exposure of A(H1N1)pdm09-infected epithelial cells (HeLa) to HIV-1 viral particles, or its gp120, enhanced interferon-induced transmembrane protein (IFITM3) content, a viral restriction factor (RF), resulting in a decrease in influenza replication. The gp120 binds to CCR5 (R5) or CXCR4 (X4) cell receptors during HIV-1 infection. Then, it is possible that the endogenous ligands of these receptors also modulate the expression of IFITM3 and other cellular factors that restrict influenza virus replication. Thus, the aim of this study is to analyze the role of cellular receptors R5 and X4 in modulating RFs in order to inhibit the replication of the influenza virus. A549 cells were treated with 2x effective dose (ED50) of endogenous R5 or X4 receptor agonists, CCL3 (20 ng/ml), CCL4 (10 ng/ml), CCL5 (10 ng/ml) and CXCL12 (100 ng/mL) or exogenous agonists, gp120 Bal-R5, gp120 IIIB-X4 and its mutants (5 µg/mL). The interferon α (10 ng/mL) and oseltamivir (60 nM) were used as a control. After 24 h post agonists exposure, the cells were infected with virus influenza A(H3N2) at 2 MOI (multiplicity of infection) for 1 h. Then, 24 h post infection, the supernatant was harvested and, the viral titre was evaluated by qRT-PCR. To evaluate IFITM3 and SAM and HD domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (SAMHD1) protein levels, A549 were exposed to agonists for 24 h, and the monolayer was lysed with Laemmli buffer for western blot (WB) assay or fixed for indirect immunofluorescence (IFI) assay. In addition to this, we analyzed other RFs modulation in A549, after 24 h post agonists exposure by customized RT² Profiler Polymerase Chain Reaction Array. We also performed a functional assay in which SAMHD1-knocked-down, by single-stranded RNA (siRNA), A549 cells were infected with A(H3N2). In addition, the cells were treated with guanosine to assess the regulatory role of dNTPs by SAMHD1. We found that R5 and X4 agonists inhibited influenza replication in 54 ± 9%. We observed a four-fold increase in SAMHD1 transcripts by RFs mRNA quantification panel. After 24 h post agonists exposure, we did not observe an increase in IFITM3 protein levels through WB or IFI assays, but we observed an upregulation up to three-fold in the protein content of SAMHD1, in A549 exposed to agonists. Besides this, influenza replication enhanced in 20% in cell cultures that SAMDH1 was knockdown. Guanosine treatment in cells exposed to R5 ligands further inhibited influenza virus replication, suggesting that the inhibitory mechanism may involve the activation of the SAMHD1 deoxynucleotide triphosphohydrolase activity. Thus, our data show for the first time a direct relationship of SAMHD1 and inhibition of influenza replication, and provides perspectives for new studies on the signaling modulation, through cellular receptors, to induce proteins of great importance in the control of relevant infections for public health.

Keywords: chemokine receptors, gp120, influenza, virus restriction factors

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Authors: Taha Anjamrooz, Sareh Rajabi, Ayman Alzaatreh

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Smart cities are playing important roles in real life. Integration and automation between different features of modern cities and information technologies improve smart city efficiency, energy management, human and equipment resource management, life quality and better utilization of resources for the customers. One of difficulties in this path, is use, interface and link between software, hardware, and other IT technologies to develop and optimize processes in various business fields such as construction, supply chain management and transportation in parallel to cost-effective and resource reduction impacts. Also, Smart cities are certainly intended to demonstrate a vital role in offering a sustainable and efficient model for smart houses while mitigating environmental and ecological matters. Energy management is one of the most important matters within smart houses in the smart cities and communities, because of the sensitivity of energy systems, reduction in energy wastage and maximization in utilizing the required energy. Specially, the consumption of energy in the smart houses is important and considerable in the economic balance and energy management in smart city as it causes significant increment in energy-saving and energy-wastage reduction. This research paper develops features and concept of smart city in term of overall efficiency through various effective variables. The selected variables and observations are analyzed through data analysis processes to demonstrate the efficiency of smart city and compare the effectiveness of each variable. There are ten chosen variables in this study to improve overall efficiency of smart city through increasing effectiveness of smart houses using an automated solar photovoltaic system, RFID System, smart meter and other major elements by interfacing between software and hardware devices as well as IT technologies. Secondly to enhance aspect of energy management by energy-saving within smart house through efficient variables. The main objective of smart city and smart houses is to reproduce energy and increase its efficiency through selected variables with a comfortable and harmless atmosphere for the customers within a smart city in combination of control over the energy consumption in smart house using developed IT technologies. Initially the comparison between traditional housing and smart city samples is conducted to indicate more efficient system. Moreover, the main variables involved in measuring overall efficiency of system are analyzed through various processes to identify and prioritize the variables in accordance to their influence over the model. The result analysis of this model can be used as comparison and benchmarking with traditional life style to demonstrate the privileges of smart cities. Furthermore, due to expensive and expected shortage of natural resources in near future, insufficient and developed research study in the region, and available potential due to climate and governmental vision, the result and analysis of this study can be used as key indicator to select most effective variables or devices during construction phase and design

Keywords: smart city, traditional housing, RFID, photovoltaic system, energy efficiency, energy saving

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Authors: Lishuang Ma

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Charge transfer (CT) complex, also known as Electron donor-acceptor (EDA) complex, has received attentions increasingly in the field of synthetic chemistry community, due to the CT complex can absorb the visible light through the intermolecular charge transfer excited states, various of catalyst-free photochemical transformations under mild visible-light conditions. However, a number of fundamental questions are still ambiguous, such as the origin of visible light absorption, the photochemical and photophysical properties of the CT complex, as well as the detailed mechanism of the radical coupling pathways mediated by CT complex. Since these are critical factors for target-specific design and synthesis of more new-type CT complexes. To this end, theoretical investigations were performed in our group to answer these questions based on multiconfigurational perturbation theory. The photo-induced fluoroalkylation reactions are mediated by CT complexes, which are formed by the association of an acceptor of perfluoroalkyl halides RF−X (X = Br, I) and a suitable donor molecule such as β-naphtholate anion, were chosen as a paradigm example in this work. First, spectrum simulations were carried out by both CASPT2//CASSCF/PCM and TD-DFT/PCM methods. The computational results showed that the broadening spectra in visible light range (360-550nm) of the CT complexes originate from the 1(σπ*) excitation, accompanied by an intermolecular electron transfer, which was also found closely related to the aggregate states of the donor and acceptor. Moreover, from charge translocation analysis, the CT complex that showed larger charge transfer in the round state would exhibit smaller charge transfer in excited stated of 1(σπ*), causing blue shift relatively. Then, the excited-state potential energy surface (PES) was calculated at CASPT2//CASSCF(12,10)/ PCM level of theory to explore the photophysical properties of the CT complexes. The photo-induced C-X (X=I, Br) bond cleavage was found to occur in the triplet state, which is accessible through a fast intersystem crossing (ISC) process that is controlled by the strong spin-orbit coupling resulting from the heavy iodine and bromine atoms. Importantly, this rapid fragmentation process can compete and suppress the backward electron transfer (BET) event, facilitating the subsequent effective photochemical transformations. Finally, the reaction pathways of the radical coupling were also inspected, which showed that the radical chain propagation pathway could easy to accomplish with a small energy barrier no more than 3.0 kcal/mol, which is the key factor that promote the efficiency of the photochemical reactions induced by CT complexes. In conclusion, theoretical investigations were performed to explore the photophysical and photochemical properties of the CT complexes, as well as the mechanism of radical coupling reactions mediated by CT complex. The computational results and findings in this work can provide some critical insights into mechanism-based design for more new-type EDA complexes

Keywords: charge transfer complex, electron transfer, multiconfigurational perturbation theory, radical coupling

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Authors: Timothy Akinwande, Eddie Hui, Karien Dekker

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Housing the urban poor remains a persistent challenge, despite evident research attention over many years. It is, therefore, pertinent to investigate affordable housing provision challenges with novel approaches. For innovative solutions to affordable housing constraints, it is apposite to thoroughly examine housing solutions vis a vis the key elements of the housing supply value chain (HSVC), which are housing finance, housing construction and land acquisition. A pragmatic analysis will examine affordable housing solutions from demand and supply perspectives to arrive at consolidated solutions from bilateral viewpoints. This study thoroughly examined informal housing finance strategies of the urban poor and diligently investigated expert opinion on affordable housing finance solutions. The research questions were: (1) What mutual grounds exist between informal housing finance solutions of the urban poor and housing expert solutions to affordable housing finance constraints in developing economies? (2) What are effective approaches to affordable housing finance in developing economies from an integrated demand - supply perspective? Semi-structured interviews were conducted in the 5 largest slums of Lagos, Nigeria, with 40 informal settlers for demand-oriented solutions, while focus group discussion and in-depth interviews were conducted with 12 housing experts in Nigeria for supply-oriented solutions. Following a rigorous thematic, content and descriptive analyses of data using NVivo and Excel, findings ascertained mutual solutions from both demand and supply standpoints that can be consolidated into more effective affordable housing finance solutions in Nigeria. Deliberate finance models that recognise and include the finance realities of the urban poor was found to be the most significant supply-side housing finance solution, representing 25.4% of total expert responses. Findings also show that 100% of sampled urban poor engage in vocations where they earn little irregular income or zero income, limiting their housing finance capacities and creditworthiness. Survey revealed that the urban poor are involved in community savings and employ microfinance institutions within the informal settlements to tackle their housing finance predicaments. These are informal finance models of the urban poor, revealing common grounds between demand and supply solutions for affordable housing financing. Effective, affordable housing approach will be to modify, institutionalise and incorporate the informal finance strategies of the urban poor into deliberate government policies. This consolidation of solutions from demand and supply perspectives can eliminate the persistent misalliance between affordable housing demand and affordable housing supply. This study provides insights into mutual housing solutions from demand and supply perspectives, and findings are informative for effective, affordable housing provision approaches in developing countries. This study is novel in consolidating affordable housing solutions from demand and supply viewpoints, especially in relation to housing finance as a key component of HSVC. The framework for effective, affordable housing finance in developing economies from a consolidated viewpoint generated in this study is significant for the achievement of sustainable development goals, especially goal 11 for sustainable, resilient and inclusive cities. Findings are vital for future housing studies.

Keywords: affordable housing, affordable housing finance, developing economies, effective affordable housing, housing policy, urban poor, sustainable development goal, sustainable affordable housing

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Authors: Ravi Chaturvedi, Ghanshyam Beniwal

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Without going much into the detail of long history of Sufism in the world and the etymological definition of the word ‘Sufi’, it will be sufficient to underline that the concept of Sufism was to focus the mystic power on the spiritual dimension of Islam with a view-shielding the believers from the outwardly and unrealistic dogma of the faith. Sufis adopted rather a liberal view in propagating the religious order of Islam suitable to the cultural and social environment of the land. It is, in fact, a mission of higher religious order of any faith, which disdains strife and conflict in any form. The joy of self-realization being the essence of religion is experienced after a long spiritual practice. India had Sufi and Bhakti (devotion) traditions in Islam and Hinduism, respectively. Both Sufism and Bhakti traditions were based on respect for different religions. The poorer and lower caste Hindus and Muslims were greatly influenced by these traditions. Unlike Ulemas and Brahmans, the Sufi and Bhakti saints were highly tolerant and open to the truth in other faiths. They never adopted sectarian attitudes and were never involved in power struggles. They kept away from power structures. Sufism is integrated with the Indian cinema since its initial days. In the earliest Bollywood movies, Sufism was represented in the form of qawwali which made its way from dargahs (shrines). Mixing it with pop influences, Hindi movies began using Sufi music in a big way only in the current decade. However, of late, songs with Sufi influences have become de rigueur in almost every film being released these days, irrespective of the genre, whether it is a romantic Gangster or a cerebral Corporate. 'Sufi is in the DNA of the Indian sub-continent', according to several contemporary filmmakers, critics, and spectators.The inherent theatricality motivates the performer of the 'Sufi' rituals for a dramatic behavior. The theatrical force of these stages of Sufi practice is so powerful that even the spectator cannot resist himself from being moved. In a multi-cultural country like India, the mediating streams have acquired a multi-layered importance in recent history. The second half of Indian post-colonial era has witnessed a regular chain of some conflicting religio-political waves arising from various sectarian camps in the country, which have compelled the counter forces to activate for keeping the spirit of composite cultural ethos alive. The study has revealed that the Sufi practice methodology is also being adapted for inclusion of spirituality in life at par to Yoga practice. This paper, a part of research study, is an attempt to establish that the Sufism in Indian cinema is one such mediating voice which is very active and alive throughout the length and width of the country continuously bridging the gap between various religious and social factions, and have a significant role to play in future as well.

Keywords: Indian cinema, mediating voice, Sufi, yoga practice

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Authors: Rongrong Liu, Li Wang, Hui Xu, Jianpei Fang, Sixi Liu, Xiaolin Yin, Junbin Liang, Gaohui Yan, Yaoyun Li, Yali Zhou, Xinyu Li, Yue Li, Lei Shi, Yongrong Lai, Junjiu Huang, Xinhua Zhang

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Background: Beta-Thalassemia is caused by reduced (β+) or absent (β0) synthesis of the β-globin chains of hemoglobin. Transfusions and oral iron chelation therapy have improved the quality of life for patients with Transfusion-Dependent thalassemia (TDT). Recent advances in genome editing platforms of CRISPR-Cas9 have paved the way for induction of HbF by reactivating expression of γ-chain.Aims: We performed CRISPR-Cas9-mediated genome editing of hematopoietic stem cells to mutate HBG1/HBG2 promoter sequence, thereby representing a naturally occurring HPFH-liked mutation, producing RM-001. Here, we present an initial assessment of safety and efficacy of RM-001 in patients with TDT. Methods: Patients (6–35 y of age) with TDT receiving packed red blood cell (pRBC) transfusions of ≥100 mL/kg/y or ≥10 units/y in the previous 2 y were eligible. CD34+ cells were edited with CRISPR-Cas9 using a guide RNA specific for the binding site of BCL11A on the HBG1/2 promoter. Prior to RM-001 product infusion (day 0), patients received myeloablative conditioning with Busulfan from day-7 to day-4. Patients were monitored for AEs Hb expression.Results: Data cut as of 28 Feb 2024, 16 TDT patients have been treated with RM-001 and followed ≥3 months. 5 of these 16 patients had finished their 24 months follow up. Eleven patients have β0/β0 genotype and five patients have β0/β+ genotype. In addition to β-thalassemia, two patients had α- deletion with the genotype of --/αα. Efficacy:All patients received a single dose intravenous infusion of RM-001 cells. 5 of them had been followed 24 months or longer. All patients achieved transfusion-independent (TI, total Hb continued ≥ 9g/dL) (Figure1). Patients demonstrated sustained and clinically meaningful increases in HbF levels since 4 month post-RM-001 infusion (Figure.2). Total hemoglobin in all patients was stable at 10-12g/dL during the follow-up period. Safety:The adverse events observed after RM-001 infusion were consistent with those that are typical of Busulfan-based myeloablation. The allelic editing analysis at 6-month visit showed that the on-target allelic editing frequency in bone marrow cells was 73.44% (64.65% to 84.6%, n=13).Summary/Conclusion: This interim analysis, in which all the 19 patients age from 7.9 to 25yo met the success criteria for the trial with respect to transfusion independence, showed that autologous HBG1/2 promoter-modified CD34+ HSPCs gene therapy resulted in an adequate amount of HbF as early as 2 months after infusion led to near-normal hemoglobin levels, remained transfusion-free through the reported period without product related SAE. After RM-001 infusion, high levels of HbF proportion and on-target editing in bone marrow cells were maintained. Submitted on behalf of the RM-001 Investigators.

Keywords: thalassemian, genetherapy, CRISPR/Cas9, HbF

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Authors: Ben Otange, Wolfgang Parak, Florian Schulz, Michael Alexander Rubhausen

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The feasibility of extending the usage of molecular imprinting technique in complex biomolecules is demonstrated in this research. This technique is promising in diverse applications in areas such as drug delivery, diagnosis of diseases, catalysts, and impurities detection as well as treatment of various complications. While molecularly imprinted polymers MIP remain robust in the synthesis of molecules with remarkable binding sites that have high affinities to specific molecules of interest, extending the usage to complex biomolecules remains futile. This work reports on the successful synthesis of MIP from complex proteins: BSA, Transferrin, and MUC1. We show in this research that despite the heterogeneous binding sites and higher conformational flexibility of the chosen proteins, relying on their respective epitopes and motifs rather than the whole template produces highly sensitive and selective MIPs for specific molecular binding. Introduction: Proteins are vital in most biological processes, ranging from cell structure and structural integrity to complex functions such as transport and immunity in biological systems. Unlike other imprinting templates, proteins have heterogeneous binding sites in their complex long-chain structure, which makes their imprinting to be marred by challenges. In addressing this challenge, our attention is inclined toward the targeted delivery, which will use molecular imprinting on the particle surface so that these particles may recognize overexpressed proteins on the target cells. Our goal is thus to make surfaces of nanoparticles that specifically bind to the target cells. Results and Discussions: Using epitopes of BSA and MUC1 proteins and motifs with conserved receptors of transferrin as the respective templates for MIPs, significant improvement in the MIP sensitivity to the binding of complex protein templates was noted. Through the Fluorescence Correlation Spectroscopy FCS measurements on the size of protein corona after incubation of the synthesized nanoparticles with proteins, we noted a high affinity of MIPs to the binding of their respective complex proteins. In addition, quantitative analysis of hard corona using SDS-PAGE showed that only a specific protein was strongly bound on the respective MIPs when incubated with similar concentrations of the protein mixture. Conclusion: Our findings have shown that the merits of MIPs can be extended to complex molecules of higher biomolecular mass. As such, the unique merits of the technique, including high sensitivity and selectivity, relative ease of synthesis, production of materials with higher physical robustness, and higher stability, can be extended to more templates that were previously not suitable candidates despite their abundance and usage within the body.

Keywords: molecularly imprinted polymers, specific binding, drug delivery, high biomolecular mass-templates

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Authors: Pinar Erturk, Sevde Altuntas, Fatih Buyukserin

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In order to obtain an effective integration between an implant and a bone, implant surfaces should have similar properties to bone tissue surfaces. Especially mimicry of the chemical, mechanical and topographic properties of the implant to the bone is crucial for fast and effective osseointegration. Titanium-based biomaterials are more preferred in clinical use, and there are studies of coating these implants with oxide layers that have chemical/nanotopographic properties stimulating cell interactions for enhanced osseointegration. There are low success rates of current implantations, especially in craniofacial implant applications, which are large and vital zones, and the oxide layer coating increases bone-implant integration providing long-lasting implants without requiring revision surgery. Our aim in this study is to examine bone-cell behavior on titanium implants with an aluminum oxide layer (AAO) on effective osseointegration potential in the deformation of large zones with difficult spontaneous healing. In our study, aluminum layer coated titanium surfaces were anodized in sulfuric, phosphoric, and oxalic acid, which are the most common used AAO anodization electrolytes. After morphologic, chemical, and mechanical tests on AAO coated Ti substrates, viability, adhesion, and mineralization of adult bone cells on these substrates were analyzed. Besides with atomic layer deposition (ALD) as a sensitive and conformal technique, these surfaces were coated with pure alumina (5 nm); thus, cell studies were performed on ALD-coated nanoporous oxide layers with suppressed ionic content too. Lastly, in order to investigate the effect of the topography on the cell behavior, flat non-porous alumina layers on silicon wafers formed by ALD were compared with the porous ones. Cell viability ratio was similar between anodized surfaces, but pure alumina coated titanium and anodized surfaces showed a higher viability ratio compared to bare titanium and bare anodized ones. Alumina coated titanium surfaces, which anodized in phosphoric acid, showed significantly different mineralization ratios after 21 days over other bare titanium and titanium surfaces which anodized in other electrolytes. Bare titanium was the second surface that had the highest mineralization ratio. Otherwise, titanium, which is anodized in oxalic acid electrolyte, demonstrated the lowest mineralization. No significant difference was shown between bare titanium and anodized surfaces except AAO titanium surface anodized in phosphoric acid. Currently, osteogenic activities of these cells on the genetic level are investigated by quantitative real-time polymerase chain reaction (qRT-PCR) analysis results of RUNX-2, VEGF, OPG, and osteopontin genes. Also, as a result of the activities of the genes mentioned before, Western Blot will be used for protein detection. Acknowledgment: The project is supported by The Scientific and Technological Research Council of Turkey.

Keywords: alumina, craniofacial implant, MG-63 cell line, osseointegration, oxalic acid, phosphoric acid, sulphuric acid, titanium

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Authors: Álvaro Turriago-Hoyos

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Most research tends to see innovation as an explanatory factor in achieving high levels of competitiveness and productivity. More recent studies have begun to analyze the determinants of innovation in the services sector as opposed to the much-discussed industrial sector of a country’s economy. This research paper focuses on the services sector in Colombia, one of Latin America’s fastest growing and biggest economies. Over the past decade, much of Colombia’s economic expansion has relied on commodity exports (mainly oil and coffee) whilst the industrial sector has performed relatively poorly. Such developments highlight the potential of the innovative role played by the services sector of the Colombian economy and its future growth prospects. This research paper analyzes the relationship between inputs, which at the same time are internal sources of innovation (such as R&D activities), and external sources that are improved by technology acquisition. The outputs are basically the four kinds of innovation that the OECD Oslo Manual recognizes: product, process, marketing and organizational innovations. The instrument used to measure this input-output relationship is based on Knowledge Production Function approaches. We run Probit models in order to identify the existing relationships between the above inputs and outputs, but also to identify spill-overs derived from interactions of the components of the value chain of the services firms analyzed: customers, suppliers, competitors, and complementary firms. Data are obtained from the Colombian National Administrative Department of Statistics for the period 2008 to 2013 published in the II and III Colombian National Innovation Survey. A short summary of the results obtained lead to conclude that firm size and a firm’s level of technological development turn out to be important discriminating factors for the description of the innovative process at the firm level. The model’s outcomes show a positive impact on the probability of introducing any kind of innovation both on R&D and Technology Acquisition investment. Also, cooperation agreements with customers, research institutes, competitors, and the suppliers are significant. Belonging to a particular industrial group is an important determinant but only to product and organizational innovation. It is possible to establish that Health Services, Education, Computer, Wholesale trade, and Financial Intermediation are the ISIC sectors, which report the highest number of frequencies of the considered set of firms. Those five sectors of the sixteen considered, in all cases, explained more than half of the total of all kinds of innovations. Product Innovation, which is followed by Marketing Innovation, gets the highest results. Displaying the same set of firms distinguishing by size, and belonging to high and low tech services sector shows that the larger the firms the larger a number of innovations, but also that always high-tech firms show a better innovation performance.

Keywords: Colombia, determinants of innovation, innovation, services sector

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Authors: Leo Nnamdi Ozurumba-Dwight

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Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.

Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq

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Authors: Anita Bajpai, Sarah Duan, Ashlee Erskine, Shehzein Khan, Raymond Kramer

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Introduction: This research focuses on exploring the beneficial effects if any, of Vitamin-D in reducing the need for supplemental oxygen among hospitalized COVID-19 patients. Two questions are investigated – Q1)Doeshaving a healthy level of baselineVitamin-D 25-OH (≥ 30ng/ml) help,andQ2) does administering Vitamin-D therapy after-the-factduring inpatient hospitalization help? Methods/Study Design: This is a comprehensive, retrospective, observational study of all inpatients at RUHS from March through December 2020 who tested positive for COVID-19 based on real-time reverse transcriptase–polymerase chain reaction assay of nasal and pharyngeal swabs and rapid assay antigen test. To address Q1, we looked atall N1=182 patients whose baseline plasma Vitamin-D 25-OH was known and who needed supplemental oxygen. Of this, a total of 121 patients had a healthy Vitamin-D level of ≥30 ng/mlwhile the remaining 61 patients had low or borderline (≤ 29.9ng/ml)level. Similarly, for Q2, we looked at a total of N2=893 patients who were given supplemental oxygen, of which713 were not given Vitamin-D and 180 were given Vitamin-D therapy. The numerical value of the maximum amount of oxygen flow rate(dependent variable) administered was recorded for each patient. The mean values and associated standard deviations for each group were calculated. Thesetwo sets of independent data served as the basis for independent, two-sample t-Test statistical analysis. To be accommodative of any reasonable benefitof Vitamin-D, ap-value of 0.10(α< 10%) was set as the cutoff point for statistical significance. Results: Given the large sample sizes, the calculated statistical power for both our studies exceeded the customary norm of 80% or better (β< 0.2). For Q1, the mean value for maximumoxygen flow rate for the group with healthybaseline level of Vitamin-D was 8.6 L/min vs.12.6L/min for those with low or borderline levels, yielding a p-value of 0.07 (p < 0.10) with the conclusion that those with a healthy level of baseline Vitamin-D needed statistically significant lower levels of supplemental oxygen. ForQ2, the mean value for a maximum oxygen flow rate for those not administered Vitamin-Dwas 12.5 L/min vs.12.8L/min for those given Vitamin-D, yielding a p-valueof 0.87 (p > 0.10). We thereforeconcludedthat there was no statistically significant difference in the use of oxygen therapy between those who were or were not administered Vitamin-D after-the-fact in the hospital. Discussion/Conclusion: We found that patients who had healthy levels of Vitamin-D at baseline needed statistically significant lower levels of supplemental oxygen. Vitamin-D is well documented, including in a recent article in the Lancet, for its anti-inflammatory role as an adjuvant in the regulation of cytokines and immune cells. Interestingly, we found no statistically significant advantage for giving Vitamin-D to hospitalized patients. It may be a case of “too little too late”. A randomized clinical trial reported in JAMA also did not find any reduction in hospital stay of patients given Vitamin-D. Such conclusions come with a caveat that any delayed marginal benefits may not have materialized promptly in the presence of a significant inflammatory condition. Since Vitamin-D is a low-cost, low-risk option, it may still be useful on an inpatient basis until more definitive findings are established.

Keywords: COVID-19, vitamin-D, supplemental oxygen, vitamin-D in primary care

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Authors: Q. R. Cheah, Y. F. Tan

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In modern times, the push for a low carbon footprint entails achieving carbon neutrality as a goal for future generations. One possible step towards carbon footprint reduction is the use of more durable materials with longer lifespans, for example, silicone data cableswhich show at least double the lifespan of similar plastic products. By having greater durability and longer lifespans, silicone data cables can reduce the amount of trash produced as compared to plastics. Furthermore, silicone products don’t produce micro contamination harmful to the ocean. Every year the electronics industry produces an estimated 5 billion data cables for USB type C and lightning data cables for tablets and mobile phone devices. Material usage for outer jacketing is 6 to 12 grams per meter. Tests show that the product lifespan of a silicone data cable over plastic can be doubled due to greater durability. This can save at least 40,000 tonnes of material a year just on the outer jacketing of the data cable. The facility in this study specialises in compounding of liquid silicone rubber (LSR) material for the extrusion process in jacketing for the silicone data cable. This study analyses the carbon emissions from the facility, which is presently capable of producing more than 1,000 tonnes of LSR annually. This study uses guidelines from the World Business Council for Sustainable Development (WBCSD) and World Resources Institute (WRI) to define the boundaries of the scope. The scope of emissions is defined as 1. Emissions from operations owned or controlled by the reporting company, 2. Emissions from the generation of purchased or acquired energy such as electricity, steam, heating, or cooling consumed by the reporting company, and 3. All other indirect emissions occurring in the value chain of the reporting company, including both upstream and downstream emissions. As the study is limited to the compounding facility, the system boundaries definition according to GHG protocol is cradle-to-gate instead of cradle-to-grave exercises. Malaysia’s present electricity generation scenario was also used, where natural gas and coal constitute the bulk of emissions. Calculations show the LSR produced for the silicone data cable with high fire retardant capability has scope 1 emissions of 0.82kg CO2/kg, scope 2 emissions of 0.87kg CO2/kg, and scope 3 emissions of 2.76kg CO2/kg, with a total product carbon footprint of 4.45kg CO2/kg. This total product carbon footprint (Cradle-to-gate) is comparable to the industry and to plastic materials per tonne of material. Although per tonne emission is comparable to plastic material, due to greater durability and longer lifespan, there can be significantly reduced use of LSR material. Suggestions to reduce the calculated product carbon footprint in the scope of emissions involve 1. Incorporating the recycling of factory silicone waste into operations, 2. Using green renewable energy for external electricity sources and 3. Sourcing eco-friendly raw materials with low GHG emissions.

Keywords: carbon footprint, liquid silicone rubber, silicone data cable, Malaysia facility

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Authors: K. S. Vimal, D. Bodhini, K. Ramya, N. Lakshmipriya, R. M. Anjana, V. Sudha, J. A. Lovegrove, V. Mohan, V. Radha

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Gene-lifestyle interaction studies have been carried out in various populations. However, to date there are no studies in an Asian Indian population. Hence, we examined whether lifestyle factors such as diet and physical activity modify the association between fat mass and obesity–associated (FTO) gene variants and obesity and type 2 diabetes (T2D) in an Asian Indian population. We studied 734 unrelated T2D and 884 normal glucose-tolerant (NGT) participants randomly selected from the Chennai Urban Rural Epidemiology Study (CURES) in Southern India. Obesity was defined according to the World Health Organization Asia Pacific Guidelines (non-obese, BMI < 25 kg/m2; obese, BMI ≥ 25 kg/m2). Six single nucleotide polymorphisms (SNPs) in the FTO gene (rs9940128, rs7193144, rs8050136, rs918031, rs1588413 and rs11076023) identified from recent genome-wide association studies for T2D were genotyped by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing. Dietary assessment was carried out using a validated food frequency questionnaire and physical activity was based upon the self-report. Interaction analyses were performed by including the interaction terms in the model. A joint likelihood ratio test of the main SNP effects and the SNP-diet/physical activity interaction effects was used in the linear regression analyses to maximize statistical power. Statistical analyses were performed using STATA version 13. There was a significant interaction between FTO SNP rs8050136 and carbohydrate energy percentage (Pinteraction=0.04) on obesity, where the ‘A’ allele carriers of the SNP rs8050136 had 2.46 times higher risk of obesity than those with ‘CC’ genotype (P=3.0x10-5) among individuals in the highest tertile of carbohydrate energy percentage. Furthermore, among those who had lower levels of physical activity, the ‘A’ allele carriers of the SNP rs8050136 had 1.89 times higher risk of obesity than those with ‘CC’ genotype (P=4.0x10-5). We also found a borderline interaction between SNP rs11076023 and carbohydrate energy percentage (Pinteraction=0.08) on T2D, where the ‘A’ allele carriers in the highest tertile of carbohydrate energy percentage, had 1.57 times higher risk of T2D than those with ‘TT’ genotype (P=0.002). There was also a significant interaction between SNP rs11076023 and physical activity (Pinteraction=0.03) on T2D. No further significant interactions between SNPs and macronutrient intake or physical activity on obesity and T2D were observed. In conclusion, this is the first study to provide evidence for a gene-diet and gene-physical activity interaction on obesity and T2D in an Asian Indian population. These findings suggest that the association between FTO gene variants and obesity and T2D is influenced by carbohydrate intake and physical activity levels. Greater understanding of how FTO gene influences obesity and T2D through dietary and exercise interventions will advance the development of behavioral intervention and personalised lifestyle strategies predicted to reduce the development of metabolic diseases in ‘A’ allele carriers of both SNPs in this Asian Indian population.

Keywords: dietary intake, FTO, obesity, physical activity, type 2 diabetes, Asian Indian.

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Authors: Beth Jacobs

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The landmark movement of Western clinical psychology in the 20th century was the development of psychotherapy. The landmark movement of clinical psychology in the 21st century will be the absorption of meditation practices from Buddhist psychology. While millions of people explore meditation and related philosophy, very few people are exposed to the materials of original Buddhism on this topic, especially to the Theravadan Abhidharma. The Abhidharma is an intricate system of lists and matrixes that were used to understand and remember Buddha’s teaching. The Abhidharma delineates the first psychological system of Buddhism, how the mind works in the universe of reality and why meditation training strengthens and purifies the experience of life. Its lists outline the psychology of mental constructions, perception, emotion and cosmological causation. While the Abhidharma is technical, elaborate and complex, its essential purpose relates to the central purpose of clinical psychology: to relieve human suffering. Like Western depth psychology, the methodology rests on understanding underlying processes of consciousness and perception. What clinical psychologists might describe as therapeutic improvement, the Abhidharma delineates as a specific pathway of purified actions of consciousness. This paper discusses the concept of 'path' as presented in aspects of the Theravadan Abhidharma and relates this to current clinical psychological views of therapy outcomes and gains. The core path in Buddhism is the Eight-Fold Path, which is the fourth noble truth and the launching of activity toward liberation. The path is not composed of eight ordinal steps; it’s eight-fold and is described as opening the way, not funneling choices. The specific path in the Abhidharma is described in many steps of development of consciousness activities. The path is not something a human moves on, but something that moments of consciousness develop within. 'Cittas' are extensively described in the Abhidharma as the atomic-level unit of a raw action of consciousness touching upon an object in a field, and there are 121 types of cittas categorized. The cittas are embedded in the mental factors, which could be described as the psychological packaging elements of our experiences of consciousness. Based on these constellations of infinitesimal, linked occurrences of consciousness, citta are categorized by dimensions of purification. A path is a chain of citta developing through causes and conditions. There are no selves, no pronouns in the Abhidharma. Instead of me walking a path, this is about a person working with conditions to cultivate a stream of consciousness that is pure, immediate, direct and generous. The same effort, in very different terms, informs the work of most psychotherapies. Depth psychology seeks to release the bound, unconscious elements of mental process into the clarity of realization. Cognitive and behavioral psychologies work on breaking down automatic thought valuations and actions, changing schemas and interpersonal dynamics. Understanding how the original Buddhist concept of positive human development relates to the clinical psychological concept of therapy weaves together two brilliant systems of thought on the development of human well being.

Keywords: Abhidharma, Buddhist path, clinical psychology, psychotherapeutic outcome

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Authors: M. Kazou, A. Gavriil, O. Kalagkatsi, T. Paschos, E. Tsakalidou

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Kopanisti cheese is a Greek soft Protected Designation of Origin (PDO) cheese made of raw cow, sheep or goat milk, or mixtures of them, with similar organoleptic characteristics to that of Roquefort cheese. Traditional manufacturing of Kopanisti cheese is limited in small-scale dairies, without the addition of starter cultures. Instead, an amount of over-mature Kopanisti cheese, called Mana Kopanisti, is used to initiate ripening. Therefore, the selection of proper starter cultures and the understanding of the contribution of various microbial groups to its overall quality is crucial for the production of a high-quality final product with standardized organoleptic and physicochemical characteristics. Taking the above into account, the aim of the present study was the investigation of Kopanisti cheese microbiota and its role in cheese quality. For this purpose, four different types of Kopanisti were produced in triplicates, all with pasteurized cow milk, with the addition of (A) the typical mesophilic species Lactococcus lactis and Lactobacillus paracasei used as starters in the production of soft spread cheeses, (B) strains of Lactobacillus acidipiscis and Lactobacillus rennini previously isolated from Kopanisti and Mana Kopanisti, (C) all the species from (A) and (B) as inoculum, and finally (D) the species from (A) and Mana Kopanisti. Physicochemical and microbiological analysis was performed for milk and cheese samples during ripening. Enumeration was performed for major groups of lactic acid bacteria (LAB), total mesophilic bacteria, yeasts as well as hygiene indicator microorganisms. Bacterial isolates from all the different LAB groups, apart from enterococci, alongside yeasts isolates, were initially grouped using repetitive sequence-based polymerase chain reaction (rep-PCR) and then identified at the species level using 16S rRNA gene and internal transcribed spacer (ITS) DNA region sequencing, respectively. Sensory evaluation was also performed for final cheese samples at the end of the ripening period (35 days). Based on the results of the classical microbiological analysis, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, ranged between 7 and 10 log colony forming unit (CFU) g⁻¹, phychrotrophic bacteria, and yeast extract glucose chloramphenicol (YGC) isolates between 4 and 8 log CFU g⁻¹, while coliforms and enterococci up to 2 log CFU g⁻¹ throughout ripening in cheese samples A, C and D. In contrast, in cheese sample B, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, phychrotrophic bacteria, and YGC isolates ranged between 0 and 10 log CFU g⁻¹ and coliforms and enterococci up to 2 log CFU g⁻¹. Although the microbial counts were not that different among samples, identification of the bacterial and yeasts isolates revealed the complex microbial community structure present in each cheese sample. Differences in the physicochemical characteristics among the cheese samples were also observed, with pH ranging from 4.3 to 5.3 and moisture from 49.6 to 58.0 % in the final cheese products. Interestingly, the sensory evaluation also revealed differences among samples, with cheese sample B ranking first based on the total score. Overall, the combination of these analyses highlighted the impact of different starter cultures on the Kopanisti microbiota as well as on the physicochemical and sensory characteristics of the final product.

Keywords: Kopanisti cheese, microbiota, classical microbiological analysis, physicochemical analysis

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Authors: E. Poupelloz, S. Gauffinet

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Ettringite Ca₆Al₂(SO₄)₃(OH)₁₂26H₂O is one of the major hydrates formed during cement hydration. Ettringite forms in Portland cement from the reaction between tricalcium aluminate Ca₃Al₂O₆ and calcium sulfate. Ettringite is also present in calcium sulfoaluminate cement in which it is the major hydrate, formed by the reaction between yeelimite Ca₄(AlO₂)₆SO₄ and calcium sulfate. About the formation of ettringite, numerous results are available in the literature even if some issues are still under discussion. However, almost all published work about ettringite was done on cementitious systems. Yet in cement, hydration reactions are very complex, the result of dissolution-precipitation processes and are submitted to various interactions. Understanding the formation process of a phase alone, here ettringite, is the first step to later understand the much more complex reactions happening in cement. This study is crucial for the comprehension of early cement hydration and physical behavior. Indeed formation of hydrates, in particular, ettringite, will have an influence on the rheological properties of the cement paste and on the need for admixtures. To make progress toward the understanding of existing phenomena, a specific study of nucleation and growth processes of ettringite was conducted. First ettringite nucleation was studied in ionic aqueous solutions, with controlled but different experimental conditions, as different supersaturation degrees (β), different pH or presence of exogenous ions. Through induction time measurements, interfacial ettringite crystals solution energies (γ) were determined. Growth of ettringite in supersaturated solutions was also studied through chain crystallization reactions. Specific BET surface area measurements and Scanning Electron Microscopy observations seemed to prove that growth process is favored over the nucleation process when ettringite crystals are initially present in a solution with a low supersaturation degree. The influence of stirring on ettringite formation was also investigated. Observation was made that intensity and nature of stirring have a high influence on the size of ettringite needles formed. Needle sizes vary from less than 10µm long depending on the stirring to almost 100µm long without any stirring. During all previously mentioned experiments, initially present ions are consumed to form ettringite in such a way that the supersaturation degree with regard to ettringite is decreasing over time. To avoid this phenomenon a device compensating the drop of ion concentrations by adding some more solutions, and therefore always have constant ionic concentrations, was used. This constant β recreates the conditions of the beginning of cement paste hydration, when the dissolution of solid reagents compensates the consumption of ions to form hydrates. This device allowed the determination of the ettringite precipitation rate as a function of the supersaturation degree β. Taking samples at different time during ettringite precipitation and doing BET measurements allowed the determination of the interfacial growth rate of ettringite in m²/s. This work will lead to a better understanding and control of ettringite formation alone and thus during cements hydration. This study will also ultimately define the impact of ettringite formation process on the rheology of cement pastes at early age, which is a crucial parameter from a practical point of view.

Keywords: cement hydration, ettringite, morphology of crystals, nucleation-growth process

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Authors: Damian Graham, Ashleigh H. Hall, Damani R. Sulph, Michael A. James, Shawn B. Vassell

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This paper assesses the design and feasibility of a Materials Recovery Facility (MRF) in Jamaica as a possible green industry approach to the nation’s economic and solid waste management problems. Jamaica is a developing nation that is vulnerable to climate change that can affect its blue economy and tourism on which it is heavily reliant. Jamaica’s National Solid Waste Management Authority (NSWMA) collects only a fraction of all the solid waste produced annually which is then transported to dumpsites. The remainder is either burnt by the population or disposed of illegally. These practices negatively impact the environment, threaten the sustainability of economic growth from blue economy and tourism and its waste management system is predominantly a cost centre. The implementation of an MRF could boost the manufacturing sector, contribute to economic growth, and be a catalyst in creating a green industry with multiple downstream value chains with supply chain linkages. Globally, there is a trend to reuse and recycle that created an international market for recycled solid waste. MRFs enable the efficient sorting of solid waste into desired recoverable materials thus providing a gateway for entrance to the international trading of recycled waste. Research into the current state and effort to improve waste management in Jamaica in contrast with the similar and more advanced territories are outlined. The study explores the concept of green industrialization and its applicability to vulnerable small state economies like Jamaica. The study highlights the possible contributions and benefits derived from MRFs as a seeding factory that can anchor the reverse and forward logistics of other green industries as part of a logistic-cantered economy. Further, the study showcases an engineering economic analysis that assesses the viability of the implementation of an MRF in Jamaica. This research outlines the potential cost of constructing and operating an MRF and provides a realistic cash flow estimate to establish a baseline for profitability. The approach considers quantitative and qualitative data, assumptions, and modelling using industrial engineering tools and techniques that are outlined. Techniques of facility planning, system analysis and operations research with a focus on linear programming techniques are expressed. Approaches to overcome some implementation challenges including policy, technology and public education are detailed. The results of this study present a reasonable judgment of the prospects of incorporating an MRF to improve Jamaica’s solid waste management and contribute to socioeconomic and environmental benefits and an alternate pathway for economic sustainability.

Keywords: engineering-economic analysis, facility design, green industry, MRF, manufacturing, plant layout, solid-waste management, sustainability, waste disposal

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Authors: Saeid Asghari

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The growing interest in sustainability and healthy lifestyles has stimulated the development of solutions that promote mindful consumption and healthier choices. One such solution is the use of transparent graded lanes in product packaging, which enables consumers to visually track their product consumption and encourages portion control. However, the extent to which this packaging affects consumer behavior, trust, and loyalty towards a product or brand, as well as the effectiveness of messaging on the graded lanes, remains unclear. The research aims to examine the impact of transparent graded lanes on consumer behavior, trust, and loyalty towards products or brands in the context of the Janbo chain supermarket in Tehran, Iran, focusing on Ketchup and edible oils containers. A representative sample of 720 respondents is selected using quota sampling based on sex, age, and financial status. The study assesses the effect of messaging on the graded lanes in enhancing consumer recall and recognition of the product at the time of purchase, increasing repeat purchases, and fostering long-term relationships with customers. Furthermore, the potential outcomes of using transparent graded lanes, including the promotion of healthy consumption habits and the reduction of food waste, are also considered. The findings and results can inform the development of effective messaging strategies for graded lanes and suggest ways to enhance consumer engagement with product packaging. Moreover, the study's outcomes can contribute to the broader discourse on sustainable consumption and healthy lifestyles, highlighting the potential role of packaging innovations in promoting these values. We used four theories (social cognitive theory, self-perception theory, nudge theory, and marketing and consumer behavior) to examine the effect of these transparent graded lanes on consumer behavior. The conceptual model integrates the use of transparent graded lanes, consumer behavior, trust and loyalty, messaging, and promotion of healthy consumption habits. The study aims to provide insights into how transparent graded lanes can promote mindful consumption, increase consumer recognition and recall of the product, and foster long-term relationships with customers. Findings suggest that the use of transparent graded lanes on Ketchup and edible oils containers can have a positive impact on consumer behavior, trust, and loyalty towards a product or brand, as well as promote mindful consumption and healthier choices. The messaging on the graded lanes is also found to be effective in promoting recall and recognition of the product at the time of purchase and encouraging repeat purchases. However, the impact of transparent graded lanes may be limited by factors such as cultural norms, personal values, and financial status. Broadly speaking, the investigation provides valuable insights into the potential benefits and challenges of using transparent graded lanes in product packaging, as well as effective strategies for promoting healthy consumption habits and building long-term relationships with customers.

Keywords: packaging customer behavior, purchase, brand loyalty, healthy consumption

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Authors: Hyppolite Aignon, Souleymane Yorou, Martin Ryberg, Anneli Svanholm

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Climate change and human actions cause the extinction of wild mushrooms. In Benin, the diversity of fungi is large and may still contain species new to science but the inventory effort remains low and focuses on particularly edible species (Russula, Lactarius, Lactifluus, and also Amanita). In addition, inventories have started recently and some groups of fungi are not sufficiently sampled, however, the degradation of fungal habitat continues to increase and some species are already disappearing. (Yorou and De Kesel, 2011), however, the degradation of fungi habitat continues to increase and some species may disappear without being known. This genus (Inocybe) overlooked has a worldwide distribution and includes more than 700 species with many undiscovered or poorly known species worldwide and particularly in tropical Africa. It is therefore important to orient the inventory to other genera or important families such as Inocybe (Fungi, Agaricales) in order to highlight their diversity and also to know their phylogenetic positions with a combined approach of gene regions. This study aims to evaluate the species richness and phylogenetic position of Inocybe species and affiliated taxa in West Africa. Thus, in North Benin, we visited the Forest Reserve of Ouémé Supérieur, the Okpara forest and the Alibori Supérieur Forest Reserve. In the center, we targeted the Forest Reserve of Toui-Kilibo. The surveys have been carried during the raining season in the study area meaning from June to October. A total of 24 taxa were collected, photographed and described. The DNA was extracted, the Polymerase Chain Reaction was carried out using primers (ITS1-F, ITS4-B) for Internal transcribed spacer (ITS), (LROR, LWRB, LR7, LR5) for nuclear ribosomal (LSU), (RPB2-f5F, RPB2-b6F, RPB2- b6R2, RPB2-b7R) for RNA polymerase II gene (RPB2) and sequenced. The ITS sequences of the 24 collections of Inocybaceae were edited in Staden and all the sequences were aligned and edited with Aliview v1.17. The sequences were examined by eye for sufficient similarity to be considered the same species. 13 different species were present in the collections. In addition, sequences similar to the ITS sequences of the thirteen final species were searched using BLAST. The nLSU and RPB2 markers for these species have been inserted in a complete alignment, where species from all major Inocybaceae clades as well as from all continents except Antarctica are present. Our new sequences for nLSU and RPB2 have been manually aligned in this dataset. Phylogenetic analysis was performed using the RAxML v7.2.6 maximum likelihood software. Bootstrap replications have been set to 100 and no partitioning of the dataset has been performed. The resulting tree was viewed and edited with FigTree v1.4.3. The preliminary tree resulting from the analysis of maximum likelihood shows us that these species coming from Benin are much diversified and are distributed in four different clades (Inosperma, Inocybe, Mallocybe and Pseudosperma) on the seven clades of Inocybaceae but the phylogeny position of 7 is currently known. This study marks the diversity of Inocybe in Benin and the investigations will continue and a protection plan will be developed in the coming years.

Keywords: Benin, diversity, Inocybe, phylogeny placement

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Authors: Maryam Nazari

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COVID-19 is a respiratory disease triggered by the novel coronavirus, SARS-CoV-2, that has reached pandemic status today. Acute inflammation and immune cells infiltration into lung injuries result in multi-organ failure. The presence of other non-communicable diseases (NCDs) with systemic inflammation derived from COVID-19 may exacerbate the patient's situation and increase the risk for adverse effects and mortality. This pandemic is a novel situation and the scientific community at this time is looking for vaccines or drugs to treat the pathology. One of the biggest challenges is focused on reducing inflammation without compromising the correct immune response of the patient. In this regard, addressing the nutritional factors should not be overlooked not only as a matter of avoiding the presence of NCDs with severe infections but also as an adjunctive way to modulate the inflammatory status of the patients. Despite the pivotal role of nutrition in modifying immune response, due to the novelty of the COVID-19 disease, information about the effects of specific dietary agents is limited in this area. From the macronutrients point of view, protein deficiency (quantity or quality) has negative effects on the number of functional immunoglobulins and gut-associated lymphoid tissue (GALT). High biological value proteins or some amino acids like arginine and glutamine are well known for their ability to augment the immune system. Among lipids, fish oil has the ability to inactivate enveloped viruses, suppress pro-inflammatory prostaglandin production and block platelet-activating factors and their receptors. In addition, protectin D1, which is an Omega-3 PUFAs derivation, is a novel antiviral drug. So it seems that these fatty acids can reduce the severity and/or improve recovery of patients with COVID-19. Carbohydrates with lower glycemic index and fibers are associated with lower levels of inflammatory cytokines (CRP, TNF-α, and IL-6). Short-Chain Fatty acids not only exert a direct anti-inflammatory effect but also provide appropriate gut microbial, which is important in gastrointestinal issues related to COVID-19. From the micronutrients point of view, Vitamins A, C, D, E, iron, magnesium, zinc, selenium and copper play a vital role in the maintenance of immune function. Inadequate status in these nutrients may result in decreased resistance against COVID-19 infection. There are specific bioactive compounds in the diet that interact with the ACE2 receptor, which is the gateway for SARS and SARS-CoV-2, and thus controls the viral infection. Regarding this, the potential benefits of probiotics, resveratrol (a polyphenol found in grape), oleoylethanolamide (derived from oleic acid), and natural peroxisome proliferator-activated receptor γ agonists in foodstuffs (like curcumin, pomegranate, hot pepper) are suggested. Yet, it should be pointed out that most of these results have been reported in animal models and further human studies are needed to be verified.

Keywords: Covid-19, inflammation, nutrition, dietary agents

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Authors: Megan L. Srinivas

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Individuals with underlying rheumatologic diseases such as dermatomyositis may not adequately respond to tuberculin (PPD) skin tests, creating false negative results. These illnesses are frequently treated with immunosuppressive therapy making proper identification of TB infection imperative. A 59-year-old Filipino man was diagnosed with dermatomyositis on the basis of rash, electromyography, and muscle biopsy. He was initially treated with IVIG infusions and transitioned to oral prednisone and mycophenolate. The patient’s symptoms improved on this regimen. Six months after starting mycophenolate, the patient began having fevers, night sweats, and productive cough without hemoptysis. He moved from the Philippines 5 years prior to dermatomyositis diagnosis, denied sick contacts, and was PPD negative both at immigration and immediately prior to starting mycophenolate treatment. A third PPD was negative following the onset of these new symptoms. He was treated for community-acquired pneumonia, but symptoms worsened over 10 days and he developed watery diarrhea and a growing non-tender, non-mobile mass on the left side of his neck. A chest x-ray demonstrated a cavitary lesion in right upper lobe suspicious for TB that had not been present one month earlier. Chest CT corroborated this finding also exhibiting necrotic hilar and paratracheal lymphadenopathy. Neck CT demonstrated the left-sided mass as cervical chain lymphadenopathy. Expectorated sputum and stool samples contained acid-fast bacilli (AFB), cultures showing TB bacteria. Fine-needle biopsy of the neck mass (scrofula) also exhibited AFB. An MRI brain showed nodular enhancement suspected to be a tuberculoma. Mycophenolate was discontinued and dermatomyositis treatment was switched to oral prednisone with a 3-day course of IVIG. The patient’s infection showed sensitivity to standard RIPE (rifampin, isoniazid, pyrazinamide, and ethambutol) treatment. Within a week of starting RIPE, the patient’s diarrhea subsided, scrofula diminished, and symptoms significantly improved. By the end of treatment week 3, the patient’s sputum no longer contained AFB; he was removed from isolation, and was discharged to continue RIPE at home. He was discharged on oral prednisone, which effectively addressed his dermatomyositis. This case illustrates the unreliability of PPD tests in patients with long-term inflammatory diseases such as dermatomyositis. Other immunosuppressive therapies (adalimumab, etanercept, and infliximab) have been affiliated with conversion of latent TB to disseminated TB. Mycophenolate is another immunosuppressive agent with similar mechanistic properties. Thus, it is imperative that patients with long-term inflammatory diseases and high-risk TB factors initiating immunosuppressive therapy receive a TB blood test (such as a quantiferon gold assay) prior to the initiation of therapy to ensure that latent TB is unmasked before it can evolve into a disseminated form of the disease.

Keywords: dermatomyositis, immunosuppressant medications, mycophenolate, disseminated tuberculosis

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Authors: Angelo Corallo, Maria Elena Latino, Marta Menegoli

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The link between man and food plays, in the social and economic system, a central role where cultural and multidisciplinary aspects intertwine: food is not only nutrition, but also communication, culture, politics, environment, science, ethics, fashion. This multi-dimensionality has many implications in the food economy. In recent years, the consumer became more conscious about his food choices, involving a consistent change in consumption models. This change concerns several aspects: awareness of food system issues, employment of socially and environmentally conscious decision-making, food choices based on different characteristics than nutritional ones i.e. origin of food, how it’s produced, and who’s producing it. In this frame the ‘consumption choices’ and the ‘interests of the citizen’ become one part of the others. The figure of the ‘Citizen Consumer’ is born, a responsible and ethically motivated individual to change his lifestyle, achieving the goal of sustainable consumption. Simultaneously the branding, that before was guarantee of the product quality, today is questioned. In order to meet these needs, Agri-Food companies are developing specific product lines that follow two main philosophies: ‘Back to basics’ and ‘Less is more’. However, the issue of ethical behavior does not seem to find an adequate on market offer. Most likely due to a lack of attention on the communication strategy used, very often based on market logic and rarely on ethical one. The label in its classic concept of ‘clean labeling’ can no longer be the only instrument through which to convey product information and its evolution towards a concept of ‘clear label’ is necessary to embrace ethical and transparent concepts in progress the process of democratization of the Food System. The implementation of a voluntary traceability path, relying on the technological models of the Internet of Things or Industry 4.0, would enable the Agri-Food Supply Chain to collect data that, if properly treated, could satisfy the information need of consumers. A change of approach is therefore proposed towards Agri-Food traceability that is no longer intended as a tool to be used to respond to the legislator, but rather as a promotional tool useful to tell the company in a transparent manner and then reach the slice of the market of food citizens. The use of mobile technology can also facilitate this information transfer. However, in order to guarantee maximum efficiency, an appropriate communication model based on the ethical communication principles should be used, which aims to overcome the pipeline communication model, to offer the listener a new way of telling the food product, based on real data collected through processes traceability. The Citizen Consumer is therefore placed at the center of the new model of communication in which he has the opportunity to choose what to know and how. The new label creates a virtual access point capable of telling the product according to different point of views, following the personal interests and offering the possibility to give several content modalities to support different situations and usability.

Keywords: agri food traceability, agri-food transparency, clear label, food system, internet of things

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Authors: Ribeiro A., Vilarinho C., Luisa A., Carvalho J

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The pig meat industry generates large volumes of by- and co-products like blood, bones, skin, trimmings, organs, viscera, and skulls, among others, during slaughtering and meat processing and must be treated and disposed of ecologically. The yield of these by-products has been reported to account for about 10% to 15% of the value of the live animal in developed countries, although animal by-products account for about two-thirds of the animal after slaughter. It was selected for further valorization of the principal wastes produced throughout the value chain of pig meat production: Pig Manure, Pig Bones, Fats, Skins, Pig Hair, Wastewater, Wastewater sludges, and other animal subproducts type III. According to the potential valorization options, these wastes will be converted into Biomethane, Fertilizers (phosphorus and digestate), Hydroxyapatite, and protein hydrolysates (Keratin and Collagen). This work includes comprehensive technical and economic analyses (TEA) for each valorization route or applied technology. Metrics such as Net Present Value (NPV), Internal Rate of Return (IRR), and payback periods were used to evaluate economic feasibility. From this analysis, it can be concluded that, for Biogas Production, the scenarios using pig manure, wastewater sludges and mixed grass and leguminous wastes presented a remarkably high economic feasibility. Scenarios showed high economic feasibility with a positive payback period, NPV, and IRR. The optimal scenario combining pig manure with mixed grass and leguminous wastes had a payback period of 1.2 years and produced 427,6269 m³ of biomethane annually. Regarding the Chemical Extraction of Phosphorous and Nitrogen, results proved that the process is economically unviable due to negative cash flows despite high recovery rates. The TEA of Hydrolysis and Extraction of Keratin Hydrolysates indicate that a unit processing and valorizing 10 tons of pig hair per year for the production of keratin hydrolysate has an NPV of 907,940 €, an IRR of 13.07%, and a Payback period of 5.41 years. All of these indicators suggest a highly potential project to explore in the future. On the opposite, the results of Hydrolysis and Extraction of Collagen Hydrolysates showed a process economically unviable with negative cash flows in all scenarios due to the high-fat content in raw materials. In fact, the results from the valorization of 10 tons of pig skin had a negative cash flow of 453 743,88 €. TEA results of Extraction and purification of Hydroxyapatite from Pig Bones with Pyrolysis indicate that unit processing and valorizing 10 tons of pig bones per year for the production of hydroxyapatite has an NPV of 1 274 819,00 €, an IRR of 65.43%, and a Payback period of 1,5 years over a timeline of 10 years with a discount rate of 10%. These valorization routes, circular economy and bio-refinery approach offer significant contributions to sustainable bio-based operations within the agri-food industry. This approach transforms waste into valuable resources, enhancing both environmental and economic outcomes and contributing to a more sustainable and circular bioeconomy.

Keywords: techno-economic analysis (TEA), pig meat processing wastes, circular economy, bio-refinery

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Authors: Domenico Carmelo Mongelli

Abstract:

The context of this research is that of the Italian reality, which, in order to adapt to the EU Directives that prohibit the production of internal combustion engines in favor of electric mobility from 2035, is extremely concerned about the significant loss of jobs resulting from the difficulty of the automotive industry in converting in such a short time and due to the reticence of potential buyers in the face of such an epochal change. The aim of the research is to evaluate for Italy the potential of the most valid alternative to this transition to electric: leaving the current production of diesel engines unchanged, no longer powered by gasoil, imported and responsible for greenhouse gas emissions, but powered entirely by a nationally produced and eco-sustainable fuel such as biodiesel. Today in Italy, the percentage of biodiesel mixed with gasoil for diesel engines is too low (around 10%); for this reason, this research aims to evaluate the functioning of current diesel engines powered 100% by biodiesel and the ability of the Italian production system to cope to this hypothesis. The research geographically identifies those abandoned lands in Italy, now out of the food market, which is best suited to an energy crop for the final production of biodiesel. The cultivation of oilseeds is identified, which for the Italian agro-industrial reality allows maximizing the agricultural and industrial yields of the transformation of the agricultural product into a final energy product and minimizing the production costs of the entire agro-industrial chain. To achieve this objective, specific databases are used, and energy and economic balances are prepared for the different agricultural product alternatives. Solutions are proposed and tested that allow the optimization of all production phases in both the agronomic and industrial phases. The biodiesel obtained from the most feasible of the alternatives examined is analyzed, and its compatibility with current diesel engines is identified, and from the evaluation of its thermo-fluid-dynamic properties, the engineering measures that allow the perfect functioning of current internal combustion engines are examined. The results deriving from experimental tests on the engine bench are evaluated to evaluate the performance of different engines fueled with biodiesel alone in terms of power, torque, specific consumption and useful thermal efficiency and compared with the performance of engines fueled with the current mixture of fuel on the market. The results deriving from experimental tests on the engine bench are evaluated to evaluate the polluting emissions of engines powered only by biodiesel and compared with current emissions. At this point, we proceed with the simulation of the total replacement of gasoil with biodiesel as a fuel for the current fleet of diesel vehicles in Italy, drawing the necessary conclusions in technological, energy, economic, and environmental terms and in terms of social and employment implications. The results allow us to evaluate the potential advantage of a total replacement of diesel fuel with biodiesel for powering road vehicles with diesel cycle internal combustion engines without significant changes to the current vehicle fleet and without requiring future changes to the automotive industry.

Keywords: biodiesel, economy, engines, environment

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Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

Abstract:

Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

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Authors: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

Abstract:

Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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