How to change the oligomeric state of a circular protein assembly: switch from 11-subunit to 12-subunit TRAP suggests a general mechanism

Many critical cellular functions are performed by multisubunit circular protein oligomers whose internal geometry has evolved to meet functional requirements. The subunit number is arguably the most critical parameter of a circular protein assembly, affecting the internal and external diameters of the assembly and often impacting on the protein's function. Although accurate structural information has been obtained for several circular proteins, a lack of accurate information on alternative oligomeric states has prevented engineering such transitions. In this study we used the bacterial transcription regulator TRAP as a model system to investigate the features that define the oligomeric state of a circular protein and to question how the subunit number could be manipulated.We find that while Bacillus subtilis and Bacillus stearothermophilus TRAP form 11-subunit oligomers, the Bacillus halodurans TRAP exclusively forms 12-subunit assemblies. Significantly, the two states of TRAP are related by a simple rigid body rotation of individual subunits around inter-subunit axes. We tested if such a rotation could be induced by insertion or deletion mutations at the subunit interface. Using wild type 11-subunit TRAP, we demonstrate that removal of five C-terminal residues at the outer side of the inter-subunit axis or extension of an amino acid side chain at the opposite, inner side, increased the subunit number from 11 to 12. Our findings are supported by crystal structures of TRAP oligomers and by native mass spectrometry data.The subunit number of the TRAP oligomer can be manipulated by introducing deletion or addition mutations at the subunit interface. An analysis of available and emerging structural data on alternative oligomeric states indicates that the same principles may also apply to the subunit number of other circular assemblies suggesting that the deletion/addition approach could be used generally to engineer transitions between different oligomeric states

Consumer attitudes and preference exploration towards fresh-cut salads using best–worst scaling and latent class analysis

This research explored the preferences and buying habits of a sample of 620 consumers of fresh-cut, ready-to-eat salads. A best–worst scaling approach was used to measure the level of preference stated by individuals regarding 12 attributes for quality (intrinsic, extrinsic and credence) of fresh-cut salads. The experiment was carried out through direct interviews at several large-scale retail outlets in the Turin metropolitan area (north-west of Italy). Out of the total number of questioned consumers, 35% said they did not consume fresh-cut salads. On the contrary, the rest of the involved sample expressed the highest degree of preference towards the freshness/appearance attribute, followed by the expiration date and the brand. On the contrary, attributes such as price, organic certification and food safety did not emerge as discriminating factors in consumer choices. Additionally, five clusters of consumers were identified, whose preferences are related both to purchasing styles and socio-demographic variables. In conclusion, this research has highlighted the positive attitude of consumers towards quality products backed by a brand, providing ideas for companies to improve within this sector and implement strategies to answer the needs of a new segment of consumers, by determining market opportunities that aim to strengthen local brands

Food sustainability perception at universities: Education and demographic features effects

The 2030 Agenda of the United Nations merged in 17 goals the strong need to change the pattern of human life on the planet in a path of strengthening sustainability especially in an era that is widely defined as Anthropocene. The Global Action Program (GAP) on Education and Sustainable Development was adopted based on the power of education and knowledge with the idea of 'green universities' aimed at improving the perception of sustainability for future policy decisions. Based on a Best-Worst (BW) scaling methodological approach, in this study we quantified the preferences of generation Y at University of Turin as they relate to issues explicitly connected to the ordinary consumption of food and the relationship between this and the perception of a sustainable approach. Data show that sustainability definitions belonging to the environmental and policy dimensions were the most closely related to the sustainability concept by the students interviewed while the economic and socio-cultural spheres were the least appreciated. In relation to food issues, students generally don't attribute high value to the assessment of local production. Few but significant differences were found in some specific topics between male and female groups with women perceiving sustainability consistently linked to the concept of local/territory and to the protection of the environment

Diverse mobilized class 1 integrons are common in the chromosomes of pathogenic Pseudomonas aeruginosa clinical isolates

Eleven clinical class 1 integron-containing Pseudomonas aeruginosa isolates from Australia and Uruguay were investigated for the genomic locations of these elements. Several novel class 1 integrons/transposons were found in at least four distinct locations in the chromosome, including genomic islands. These elements seem to be undergoing successful dispersal by lateral gene transfer since integrons were identified across several lineages and more than one clonal line. Copyright © 2012, American Society for Microbiology. All Rights Reserved

Novel anti-inflammatory and chondroprotective effects of the human melanocortin MC1 receptor agonist BMS-470539 dihydrochloride and human melanocortin MC3 receptor agonist PG-990 on lipopolysaccharide activated chondrocytes

Human melanocortin MC1 and MC3 receptors expressed on C-20/A4 chondrocytes exhibit chondroprotective and anti-inflammatory effects when activated by melanocortin peptides. Nearly 9 million people in the UK suffer from osteoarthritis, and bacterial infections play a role in its development. Here, we evaluate the effect of a panel of melanocortin peptides with different selectivity for human melanocortin MC1 (alpha-MSH, BMS-470539 dihydrochloride) and MC3 receptors ([DTrp8]-g-MSH, PG-990) and C-terminal peptide alpha-MSH11-13(KPV), on inhibiting LPS-induced chondrocyte death, pro-inflammatory mediators and induction of anti-inflammatory proteins. C-20/A4 chondrocytes were treated with a panel of melanocortin peptides prophylactically and therapeutically in presence of LPS (0.1 ug/ml). The chondroprotective properties of these peptides determined by cell viability assay, RT-PCR, ELISA for detection of changes in inflammatory markers (IL-6, IL-8 and MMP-1, -3 and -13) and western blotting for expression of the anti-inflammatory protein heme-oxygenase-1. C-20/A4 expressed human melanocortin MC1 and MC3 receptors and melanocortin peptides elevated cAMP. LPS stimulation caused a reduction in C-20/A4 viability, attenuated by the human melanocortin MC1 receptor agonist BMS-470539 dihydrochloride, and MC3 receptor agonists PG-990 and [DTrp8]-g-MSH. Prophylactic and therapeutic regimes of [DTrp8]-g-MSH significantly inhibited LPS-induced modulation of cartilage-damaging IL-6, IL-8, MMPs -1,-3 and -13 mediators both prophylactically and therapeutically, whilst human melanocortin MC1 and MC3 receptor agonists promoted an increase in HO-1 production. In the presence of LPS, activation of human melanocortin MC1 and MC3 receptors provided potent chondroprotection, upregulation of anti-inflammatory proteins and downregulation of inflammatory and proteolytic mediators involved in cartilage degradation, suggesting a new avenue for osteoarthritis treatment

DDoS cyber-incident detection in smart grids

The smart grid (SG) offers potential benefits for utilities, electric generators, and customers alike. However, the prevalence of cyber-attacks targeting the SG emphasizes its dark side. In particular, distributed denial-of-service (DDoS) attacks can affect the communication of different devices, interrupting the SG’s operation. This could have profound implications for the power system, including area blackouts. The problem is that few operational technology tools provide reflective DDoS protection. Furthermore, such tools often fail to classify the types of attacks that have occurred. Defensive capabilities are necessary to identify the footprints of attacks in a timely manner, as they occur, and to make these systems sustainable for delivery of the services as expected. To meet this need for defensive capabilities, we developed a situational awareness tool to detect system compromise by monitoring the indicators of compromise (IOCs) of amplification DDoS attacks. We achieved this aim by finding IOCs and exploring attack footprints to understand the nature of such attacks and their cyber behavior. Finally, an evaluation of our approach against a real dataset of DDoS attack instances indicated that our tool can distinguish and detect different types of amplification DDoS attacks

An FPGA-based flexible demo-board for endoscopic capsule design optimization

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Clinical impact of HSV-1 detection in the lower respiratory tract from hospitalized adult patients

AbstractThe occurrence and clinical impact of herpes simplex virus (HSV) were evaluated in 342 bronchoalveolar lavage specimens from 237 patients. HSV-1 and HSV-2 were detected in 32.1% and <1% of patients, respectively. A significant difference of HSV-1 prevalence and load was found in relation to admission to intensive care unit, mechanical ventilation and mortality within 28 days; in particular, a viral load ≥105 copies/mL bronchoalveolar lavage fluid was significantly associated with critical features. No association was found with immune status or other characteristics. Nine of 21 (42.9%) cases of ventilator-associated pneumonia were positive for HSV-1, with poor outcome in six

Novel anti-inflammatory and chondroprotective effects of the human melanocortin MC1 receptor agonist BMS-470539 dihydrochloride and human melanocortin MC3 receptor agonist PG-990 on lipopolysaccharide activated chondrocytes

Human melanocortin MC1 and MC3 receptors expressed on C-20/A4 chondrocytes exhibit chondroprotective and anti-inflammatory effects when activated by melanocortin peptides. Nearly 9 million people in the UK suffer from osteoarthritis, and bacterial infections play a role in its development. Here, we evaluate the effect of a panel of melanocortin peptides with different selectivity for human melanocortin MC1 (α-MSH, BMS-470539 dihydrochloride) and MC3 ([DTrp8]-γ-MSH, PG-990) receptors and C-terminal peptide α-MSH11-13(KPV), on inhibiting LPS-induced chondrocyte death, pro-inflammatory mediators and induction of anti-inflammatory proteins. C-20/A4 chondrocytes were treated with a panel of melanocortin peptides prophylactically and therapeutically in presence of LPS (0.1 μg/ml). The chondroprotective properties of these peptides determined by cell viability assay, RT-PCR, ELISA for detection of changes in inflammatory markers (IL-6, IL-8 and MMP-1, -3 and -13) and western blotting for expression of the anti-inflammatory protein heme-oxygenase-1. C-20/A4 expressed human melanocortin MC1 and MC3 receptors and melanocortin peptides elevated cAMP. LPS stimulation caused a reduction in C-20/A4 viability, attenuated by the human melanocortin MC1 receptor agonist BMS-470539 dihydrochloride, and MC3 receptor agonists PG-990 and [DTrp8]-γ-MSH. Prophylactic and therapeutic regimes of [DTrp8]-γ-MSH significantly inhibited LPS-induced modulation of cartilage-damaging IL-6, IL-8, MMPs −1,-3 and −13 mediators both prophylactically and therapeutically, whilst human melanocortin MC1 and MC3 receptor agonists promoted an increase in HO-1 production. In the presence of LPS, activation of human melanocortin MC1 and MC3 receptors provided potent chondroprotection, upregulation of anti-inflammatory proteins and downregulation of inflammatory and proteolytic mediators involved in cartilage degradation, suggesting a new avenue for osteoarthritis treatment