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ABSTRACT Group A Streptococcus (GAS) is a human-adapted pathogen responsible for a variety of diseases. The GAS M1UK lineage has contributed significantly to the recently reported increases in scarlet fever and invasive infections. However, the basis for its evolutionary success is not yet fully understood. During the transition to systemic disease, the M1 serotype is known to give rise to spontaneous mutations in the control of virulence two-component regulatory system (CovRS) that confer a fitness advantage during invasive infections. Mutations that inactivate CovS function result in the de-repression of key GAS virulence factors such as streptolysin O (SLO), a pore-forming toxin and major trigger of inflammasome/interleukin-1β-dependent inflammation. Conversely, expression of the streptococcal cysteine protease SpeB, which is required during initial stages of colonization and onset of invasive disease, is typically lost in such mutants. In this study, we identified and characterized a novel covS single nucleotide polymorphism detected in three separate invasive M1UK isolates. The resulting CovSAla318Val mutation caused a significant upregulation of SLO resulting in increased inflammasome activation in human THP-1 macrophages, indicating an enhanced inflammatory potential. Surprisingly, SpeB production was unaffected. Site-directed mutagenesis was performed to assess the impact of this mutation on virulence and global gene expression. We found that the CovSAla318Val mutation led to subtle, virulence-specific changes of the CovRS regulon compared to previously characterized covS mutations, highlighting an unappreciated level of complexity in CovRS-dependent gene regulation. Continued longitudinal surveillance is warranted to determine whether this novel covS mutation will expand in the M1UK lineage.IMPORTANCEThe M1UK lineage of GAS has contributed to a recent global upsurge in scarlet fever and invasive infections. Understanding how GAS can become more virulent is critical for infection control and identifying new treatment approaches. The two-component CovRS system, comprising the sensor kinase CovS and transcription factor CovR, is a central regulator of GAS virulence genes. In the M1 serotype, covRS mutations are associated with an invasive phenotype. Such mutations have not been fully characterized in the M1UK lineage. This study identified a novel covS mutation in invasive Australian M1UK isolates that resulted in a more nuanced virulence gene regulation compared to previously characterized covS mutations. A representative isolate displayed upregulated SLO production and triggered amplified interleukin-1β secretion in infected human macrophages, indicating an enhanced inflammatory potential. These findings underscore the need for comprehensive analyses of covRS mutants to fully elucidate their contribution to M1UK virulence and persistence.

Most existing autoscaling approaches help to avoid violating the performance-related Service Level Objectives (SLO). However, these solutions do not aim to recover the SLO. The proposed novel autoscaling solution covers both SLO violation avoidance and recovery. The SLO failure avoidance part of the solution aims to avoid SLO violations by adjusting autoscaling thresholds based on compliance with SLO. It also dynamically selects the required CPU threshold, cooldown intervals and the number of replicas based on load velocity. The recovery part of the solution aims to recover SLO by additional resource provisioning if SLO is violated due to a delay or resource underestimation. The proposed implementation of the solution targets embarrassingly parallel workloads. It is compared with two autoscaling solutions in five workload scenarios, evaluating the ability of the solution to operate close to the defined SLO values. The results show that the proposed solution provides sufficient resources to support or recover performance-based SLOs. [ABSTRACT FROM AUTHOR]

This article offers a discussion of the intricate relation between the human body and its technological supplementation. It argues that while the physical body may seem to sublimate in the plethora of discourses, it keeps reasserting its materiality via processes of technologization which capture perception and cognition in a constant cycle of disembodiment and re-embodiment. The article offers an analysis of Elizabet Goldring’s work with the Seeing Laser Ophthalmoscope, and her RetinaPrints (a unique form of visualization) to comment on the ways in which lived experiences are inexorably linked to practices of fragmentation, prosthesis and technological mediation.

ABSTRACTExtracellular vesicles (EVs) can be released from gram-positive bacteria and would participate in the delivery of bacterial toxins. Streptococcus pyogenes (group A Streptococcus, GAS) is one of the most common pathogens of monomicrobial necrotizing fasciitis. Spontaneous inactivating mutation in the CovR/CovS two-component regulatory system is related to the increase of EVs production via an unknown mechanism. This study aimed to investigate whether the CovR/CovS-regulated RopB, the transcriptional regulator of GAS exoproteins, would participate in regulating EVs production. Results showed that the size, morphology, and number of EVs released from the wild-type strain and the ropB mutant were similar, suggesting RopB is not involved in controlling EVs production. Nonetheless, RopB-regulated SpeB protease degrades streptolysin O and bacterial proteins in EVs. Although SpeB has crucial roles in modulating protein composition in EVs, the SpeB-positive EVs failed to trigger HaCaT keratinocytes pyroptosis, suggesting that EVs did not deliver SpeB into keratinocytes or the amount of SpeB in EVs was not sufficient to trigger cell pyroptosis. Finally, we identified that EV-associated enolase was resistant to SpeB degradation, and therefore could be utilized as the internal control protein for verifying SLO degradation. This study revealed that RopB would participate in modulating protein composition in EVs via SpeB-dependent protein degradation and suggested that enolase is a potential internal marker for studying GAS EVs.

In dogs, symmetrical lupoid onychodystrophy (SLO) results in nail loss and an abnormal regrowth of the claws. In Bearded Collies, an autoimmune nature has been suggested because certain dog leukocyte antigen (DLA) class II haplotypes are associated with the condition. A genome-wide association study of the Bearded Collie revealed two regions of association that conferred risk for disease: one on canine chromosome (CFA) 12 that encompasses the DLA genes, and one on CFA17. Case-control association was employed on whole genome sequencing data to uncover putative causative variants in SLO within the CFA12 and CFA17 associated regions. Genotype imputation was then employed to refine variants of interest. Although no SLO-associated protein-coding variants were identified on CFA17, multiple variants, many with predicted damaging effects, were identified within potential candidate genes on CFA12. Furthermore, many potentially damaging alleles were fully correlated with the presence of DLA class II risk haplotypes for SLO, suggesting that the variants may reflect DLA class II haplotype association with disease or vice versa. Strong linkage disequilibrium in the region precluded the ability to isolate and assess the individual or combined effect of variants on disease development. Nonetheless, all were predictive of risk for SLO and, with judicious assessment, their application in selective breeding may prove useful to reduce the incidence of SLO in the breed.

The emerging serverless computing has become a captivating paradigm for deploying cloud applications, alleviating developers' concerns about infrastructure resource management by configuring necessary parameters such as latency and memory constraints. Existing resource configuration solutions for cloud-based serverless applications can be broadly classified into modeling based on historical data or a combination of sparse measurements and interpolation/modeling. In pursuit of service response and conserving network bandwidth, platforms have progressively expanded from the traditional cloud to the edge. Compared to cloud platforms, serverless edge platforms often lead to more running overhead due to their limited resources, resulting in undesirable financial costs for developers when using the existing solutions. Meanwhile, it is extremely challenging to handle the heterogeneity of edge platforms, characterized by distinct pricing owing to their varying resource preferences. To tackle these challenges, we propose an adaptive and efficient approach called FireFace, consisting of prediction and decision modules. The prediction module extracts the internal features of all functions within the serverless application and uses this information to predict the execution time of the functions under specific configuration schemes. Based on the prediction module, the decision module analyzes the environment information and uses the Adaptive Particle Swarm Optimization algorithm and Genetic Algorithm Operator (APSO-GA) algorithm to select the most suitable configuration plan for each function, including CPU, memory, and edge platforms. In this way, it is possible to effectively minimize the financial overhead while fulfilling the Service Level Objectives (SLOs). Extensive experimental results show that our prediction model obtains optimal results under all three metrics, and the prediction error rate for real-world serverless applications is in the range of 4.25∼9.51%. Our approach can find the optimal resource configuration scheme for each application, which saves 7.2∼44.8% on average compared to other classic algorithms. Moreover, FireFace exhibits rapid adaptability, efficiently adjusting resource allocation schemes in response to dynamic environments. [ABSTRACT FROM AUTHOR]

BackgroundPrimary hypoadrenocorticism (Addison's disease, AD) and symmetrical lupoid onychodystrophy (SLO) are two clinical conditions with an autoimmune etiology that occur in multiple dog breeds. In man, autoimmunity is associated with polymorphisms in immune-related genes that result in a reduced threshold for, or defective regulation of, T cell activation. The major histocompatibility complex (MHC) class II genes encode molecules that participate in these functions, and polymorphisms within these genes have been associated with autoimmune conditions in dogs and humans. Bearded collies have a relatively high prevalence of autoimmune diseases, particularly AD and SLO. Our study assessed the relationship between particular MHC (dog leukocyte antigen, DLA) class II haplotypes and the two autoimmune diseases most common in this breed. Moreover, five unrelated breeds at increased risk for AD were studied for comparative purposes and analyzed in the context of extant literature.ResultsA single DLA class II three-locus haplotype, determined by sequence-based typing, was associated with increased risk for AD (DLA-DRB1*009:01/DQA1*001:01/DQB1*008:02) in bearded collies. Comparative analysis with the five additional breeds showed limited allele sharing, with DQA1*001:01 and DQB1*002:01 being the only alleles observed in all breeds. A distinct three-locus risk haplotype (DLA-DRB1*001:01/DQA1*001:01/DQB1*002:01) was associated with AD in the West Highland white terrier and Leonberger. Two different risk haplotypes were associated with increased risk for SLO in the bearded collie (DLA-DRB1*018:01/DQA1*001:01/DQB1*002:01 and DLA-DRB1*018:01/DQA1*001:01/ DQB1*008:02).ConclusionTwo-locus DQ haplotypes composed of DLA-DQA1*001:01 in association with DLA-DQB1*002:01 or DLA-DQB1*008:02 make up the four risk haplotypes identified in the present study and are also found in other risk haplotypes previously associated with diabetes mellitus and hypothyroidism across different dog breeds. Our findings build upon previously published data to suggest that this two-locus (DQ) model serves as a good indicator for susceptibility to multiple organ-specific autoimmune diseases in the canine population. However, it is also clear that additional loci are necessary for actual disease expression. Investigation of affected and unaffected dogs carrying these predisposing DQ haplotype signatures may allow for the identification of those additional genetic components that determine autoimmune disease expression and organ specificity.

Symmetrical lupoid onychodystrophy (SLO) is characterized by inflammation of the nail bed and nail sloughing that causes affected dogs considerable pain. Disease etiology remains unclear, although an autoimmune component is suspected. A genome-wide association study on Bearded Collies revealed regions of association on canine chromosomes (CFA) 12 and 17. The large region of association on CFA12 likely consists of two smaller linked regions, both of which are also linked to the dog leukocyte antigen (DLA) class II genes. Dogs homozygous for the alternate allele at the top CFA12 SNP also carried two DLA class II risk haplotypes for SLO, and this locus explained most of the increased risk for disease seen throughout the CFA12 region of association. A stronger peak was seen on CFA17 when analysis was done solely on dogs that carried DLA class II risk haplotypes for SLO. The majority of SLO dogs carried a homozygous alternate genotype on CFA12 and at least one CFA17 risk haplotype. Our findings offer progress toward uncovering the genetic basis of SLO. While the contribution of the CFA17 region remains unclear, both CFA12 and CFA17 regions are significantly associated with SLO disease expression in the Bearded Collie and contain potential candidate genes for this disease.

Studies on the welfare of carriage horses have been addressed during working activity; however, their nocturnal resting behavior remains poorly explored. The aim of our study was to quantify the frequency of negative, positive, and neutral facial expressions (FE) of carriage horses used in tourism displayed under nocturnal resting conditions. Observations were carried out in the tropical city of Mérida, México, from January to March 2021. Nine crossbreed horses, including 3 mares and 6 geldings (400 ± 50 kg, 9-20 years old), were used. Prior to FE observations, an evaluation of resting locations was performed with regard to available surface area, movement restriction, floor and roof conditions, and visual and auditory contact between horses. A 40-point veterinary health examination was also performed on all horses. FE were identified based on the Horse Grimace Scale and EquiFacs coding system. Occurrences of stiffly backwards ears EAD103, orbital tightening AU145, tension above the eye area AU5, prominent strained chewing muscles AU18, strained mouth and pronounced chin AU24+17, and strained nostril and flattening of the profile EAD103+AUH13 were considered negative; ears forward EAD101 and nostril dilation AD38 were considered positive; and right ear forward EAD101R and without movement (WM) were considered neutral. All FE were photographed with a hidden camera (GoProHero3+, 12 megapixels) every minute for at least 8 hours overnight immediately following a randomly selected working day. Good resting location scores were fulfilled by 100% of horses except for good floor and roof conditions (66.6%). Physical health evaluations resulted in 90% of horses with good health scores. The most frequent FE per hour were WM (27.0 ± 5.6), EAD101 (12.8 ± 5.6), and EAD103 (8.9 ± 5.8). Statistical differences (P < 0.0001) were found for mean comparison test between neutral (15.7 ± 3.1 FE/h), positive (6.6 ± 2.9 FE/h), and negative (2.4 ± 1.3 FE/h) FE/h. No significant differences were found for the frequency of neutral, positive, or negative FE/h according to age, body condition score, or health status of the animals, nor according to floor and roof condition of resting locations. The higher frequency of neutral and positive FE compared to negative FE is consistent with the good health scores and generally good conditions of resting locations in the large majority of the horses observed. [ABSTRACT FROM AUTHOR]

This study investigated the current situation of social media adoption in the energy and resources sector, the preferred social media platforms in the energy and resources sector, and the reasons for the differences between these preferred platforms. In this study, quantitative analysis method was adopted, and we used Excel and SPSS for data collection and data analysis. Through the investigation of 262 companies, all companies maintained a strong online presence through corporate websites, email contacts, and various social media platforms. However, the adoption and use of social media platforms differed among countries. Linear regression analysis revealed that companies should constantly update their relevant information on social media platforms to increase their followers, improve their influence and popularity, and achieve better interaction with stakeholders. Through quantitative analysis, it was found that companies in the energy and resources sector should maintain a strong online presence. These companies should choose specific social media platforms according to their target audience. Because some platforms are more suitable for specific purposes, companies should pay attention to the different designs and functions of each social media platform. Future research should focus on the use of social media in B2B marketing by companies operating in the energy and resources sector.

Cloud computing has been consolidated as a support for the vast majority of current and emerging technologies. However, there are some barriers that prevent the exploitation of the full potential of this technology. First, the major cloud providers currently put the onus of implementing the mechanisms that ensure compliance with the desired service levels on cloud consumers. However, consumers do not have the required expertise. Since each cloud provider exports a different set of low‐level metrics, the strategies defined to ensure compliance with the established service‐level agreement (SLA) are bound to a particular cloud provider. This fosters provider lock‐in and prevents consumers from benefiting from the advantages of multi‐cloud environments. This paper presents a solution to the problem of automatically translating SLAs into objectives expressed as metrics that can be measured across multiple cloud providers. First, we propose an intelligent knowledge‐based system capable of automatically translating high‐level SLAs defined by cloud consumers into a set of conditions expressed as vendor‐neutral metrics, providing feedback to cloud consumers (intelligent tutoring system). Secondly, we present the set of vendor‐neutral metrics and explain how they can be measured for the different cloud providers. Finally, we report a validation based on two use cases (IaaS and PaaS) in a multi‐cloud environment formed by leading cloud providers. This evaluation has demonstrated that, thanks to the complementarity of the two solutions, cloud consumers can automatically and transparently exploit the multi‐cloud in many application domains, as endorsed by the cloud experts consulted in the course of this study. [ABSTRACT FROM AUTHOR]

Cloud systems and microservices are becoming powerful tools for businesses. The evidence of the advantages of offering infrastructure, hardware or software as a service (IaaS, PaaS, SaaS) is overwhelming. Microservices and decoupled applications are increasingly popular. These architectures, based on containers, have facilitated the efficient development of complex SaaS applications. A big challenge is to manage and design microservices with a massive range of different facilities, from processing and data storage to computing predictive and prescriptive analytics. Computing providers are mainly based on data centers formed of massive and heterogeneous virtualized systems, which are continuously growing and diversifying over time. Moreover, these systems require integrating into current systems while meeting the Quality of Service (QoS) constraints. The primary purpose of this work is to present an on-premise architecture based on Kubernetes and Docker containers aimed at improving QoS regarding resource usage and service level objectives (SLOs). The main contribution of this proposal is its dynamic autoscaling capabilities to adjust system resources to the current workload while improving QoS.

ABSTRACT Mutation within the Streptococcus pyogenes (Streptococcus group A; Strep A) covR/S regulatory system has been associated with a hypervirulent phenotype resulting from the upregulation of several virulence factors, including the pore-forming toxin, streptolysin O (SLO). In this study, we utilized a range of covR/S mutants, including M1T1 clonal strains (5448 and a covS mutant generated through mouse passage designated 5448AP), to investigate the contribution of SLO to the pathogenesis of covR/S mutant Strep A disease. Up-regulation of slo in 5448AP resulted in increased SLO-mediated hemolysis, decreased dendritic cell (DC) viability post coculture with Strep A, and increased production of tumor necrosis factor (TNF) and monocyte chemoattractant protein 1 (MCP-1) by DCs. Mouse passage of an isogenic 5448 slo-deletion mutant resulted in recovery of several covR/S mutants within the 5448Δslo background. Passage also introduced mutations in non-covR/S genes, but these were considered to have no impact on virulence. Although slo-deficient mutants exhibited the characteristic covR/S-controlled virulence factor upregulation, these mutants caused increased DC viability with reduced inflammatory cytokine production by infected DCs. In vivo, slo expression correlated with decreased DC numbers in infected murine skin and significant bacteremia by 3 days postinfection, with severe pathology at the infection site. Conversely, the absence of slo in the infecting strain (covR/S mutant or wild-type) resulted in detection of DCs in the skin and attenuated virulence in a murine model of pyoderma. slo-sufficient and -deficient covR/S mutants were susceptible to immune clearance mediated by a combination vaccine consisting of a conserved M protein peptide and a peptide from the CXC chemokine protease SpyCEP. IMPORTANCE Streptococcus pyogenes is responsible for significant numbers of invasive and noninvasive infections which cause significant morbidity and mortality globally. Strep A isolates with mutations in the covR/S system display greater propensity to cause severe invasive diseases, which are responsible for more than 163,000 deaths each year. This is due to the upregulation of virulence factors, including the pore-forming toxin streptolysin O. Utilizing covR/S and slo-knockout mutants, we investigated the role of SLO in virulence. We found that SLO alters interactions with host cell populations and increases Strep A viability at sterile sites of the host, such as the blood, and that its absence results in significantly less virulence. This work underscores the importance of SLO in Strep A virulence while highlighting the complex nature of Strep A pathogenesis. This improved insight into host-pathogen interactions will enable a better understanding of host immune evasion mechanisms and inform streptococcal vaccine development programs.

Operation Worthiness Standard (OWS) is one of fisheries surveilance instrument based on Minister Decree of Fisheries and Marine Affairs Number 23 Year 2021. This research aim to identify flow process and procedure of OWS issuance, and its implementation status on fishing vessel at Mayangan fishing port. The method used were direct observation and interview. The data used on this study were primary data and secondary data. Primary data were collected by following the examination of fishing vessel documents and physical inspection. Secondary data were obtained from Surveilance Unit of Marine and Fisheries Resources Surveillance on Probolinggo that was form of record on OWS issuance. Flow process of OWS issuance is starting from departure plan report by fishers on SIPELIKAN which is followed up with document examination and physical inspection. This study found that expired documents and vessel monitoring system (VMS) transmitter were not installed on the large fishing vessel as the cause of rejection of OWS issuance. There were 256 OWS issued during January to June 2022, dominated by OWS for fishing vessels 78.13 %, the rest were collecting vessels. OWS issuance in Mayangan fishing port were dominated by large-scale fishing vessels as much as 62 %, followed by medium-scale vessels 36% and small-scale vessels 2 %. [ABSTRACT FROM AUTHOR]

In lipoxygenases, the topologically conserved C-terminal domain catalyzes the oxidation of polyunsaturated fatty acids, generating an assortment of biologically relevant signaling mediators. Plant and animal lipoxygenases also contain a 100-150-amino acid N-terminal C2-like domain that has been implicated in interactions with isolated fatty acids and at the phospholipid bilayer. These interactions may lead to increased substrate availability and contribute to the regulation of active-site catalysis. Because of a lack of structural information, a molecular understanding of this lipid-protein interaction remains unresolved. Herein, we employed hydrogen-deuterium exchange MS (HDXMS) to spatially resolve changes in protein conformation upon interaction of soybean lipoxygenase with a fatty acid surrogate, oleyl sulfate (OS), previously shown to act at a site separate from the substrate-binding site. Specific, OS-induced conformational changes are detected both at the N-terminal domain and within the substrate portal nearly 30 Å away. Combining previously measured kinetic properties in the presence of OS with its impact on the Kd for linoleic acid substrate binding, we conclude that OS binding brings about an increase in rate constants for both the ingress and egress of substrate. We discuss the role of OS-induced changes in protein flexibility in the context of changes in the mechanism of substrate acquisition.

The measurement of antibodies to vaccine antigens is crucial for research towards a safe and effective vaccine for Streptococcus pyogenes (Strep A). We describe the establishment and detailed characterisation of a four-plex assay to measure IgG to the Strep A vaccine antigens SpyCEP, Slo, SpyAD and GAC using the Luminex multiplex platform. A standard curve was established and characterized to allow the quantification of antigen-specific IgG. Assay specificity, precision, linearity, reproducibility and repeatability were determined via the measurement of antigen-specific IgG from pooled human serum. The assay is highly specific, reproducible and performs well across a large range of antibody concentrations against all four antigens. It is, therefore, suitable for future clinical trials in humans with a four-component vaccine, as well as for seroepidemiological studies to gain insights into naturally occurring immunity. [ABSTRACT FROM AUTHOR]

Non-invasive imaging is an invaluable diagnostic tool in ophthalmology. Two imaging devices, the scanning laser ophthalmoscope (SLO) and spectral domain optical coherence tomography (SDOCT), emerged from the clinical realm to provide research scientists with a real-time view of ocular morphology in living animals. We utilized these two independent imaging modalities in a complementary manner to perform in vivo optical sectioning of the adult zebrafish retina. Due to the very high optical power of the zebrafish lens, the confocal depth of field is narrow, allowing for detailed en face views of specific retinal layers, including the cone mosaic. Moreover, we demonstrate that both native reflectance, as well as fluorescent features observed by SLO, can be combined with axial in-depth information obtained by SDOCT. These imaging approaches can be used to screen for ocular phenotypes and monitor retinal pathology in a non-invasive manner.

The Social License to Operate (SLO), understood as the consent of a wide group of stakeholders to mining activities, has become an important element in the process of obtaining a mining license in recent years. Such a pattern of increasing importance is common, both in Poland and throughout the EU. Therefore, it should be of key importance to prevent society's reluctance to this industry. The article indicates that a very important tool in this matter should be reliable and knowledge-based raw materials education, carried out as early as at the stage of school education. Deficiencies in this respect were revealed through a survey conducted among children and youth up to 15 years of age. The results of the survey were discussed in the context of the development of raw materials education in Poland, which results indicate should focus on emphasizing the benefits of mining, both in terms of the general public and individuals. Shaping and strengthening the mineral raw materials awareness of the inhabitants of Poland is also of key importance for the implementation of standards for the protection of mineral deposits and ensuring the country's resource security. [ABSTRACT FROM AUTHOR]

Formaldehyde reacts with amino acids in living organisms to form toxic intermediates that cause epithelial cell damage. In past epidemiological studies, a statistically significant relationship was found between FA and leukemia risks and occupational inhalation exposure. As a complementary and alternative medicine (CAM) technique or alternative medicine, shark cartilage (SC) and shark liver oil (SLO) are presented as a new and different alternative source in this study. In this study, the toxic effects of formaldehyde (FA) on lung and the protective effects of SC and SLO against these toxins have been investigated. For the experiment, 40 rats were classified as follows: 4, control group (experiment control); 6, the group that received FA but was not treated (treatment control); 15, the group that was given FA and SC for treatment; and the last 15 were the group that was given FA and SLO for treatment. Negative effects of FA on lung were evaluated biochemically, genetically and pathologically. In terms of therapeutic efficacy, SLO appears to be more effective in improving lung injury on the basis of genetic, pathological and biochemical findings, against to FA administration. The toxic effect of FA in lung and the therapeutic effect of SLO and SC were determined and we believe that our experimental model provided the desired goal and success on the basis of our work. [ABSTRACT FROM AUTHOR]

The measurement of antibodies to vaccine antigens is crucial for research towards a safe and effective vaccine for Streptococcus pyogenes (Strep A). We describe the establishment and detailed characterisation of a four-plex assay to measure IgG to the Strep A vaccine antigens SpyCEP, Slo, SpyAD and GAC using the Luminex multiplex platform. A standard curve was established and characterized to allow the quantification of antigen-specific IgG. Assay specificity, precision, linearity, reproducibility and repeatability were determined via the measurement of antigen-specific IgG from pooled human serum. The assay is highly specific, reproducible and performs well across a large range of antibody concentrations against all four antigens. It is, therefore, suitable for future clinical trials in humans with a four-component vaccine, as well as for seroepidemiological studies to gain insights into naturally occurring immunity.

Lifecycle Services Orchestration (LSO) architecture for Metro Ethernet Services was introduced by MEF. In the LSO architecture, the ALLEGRO interface was defined for dynamic service control between customer and Service Provider while the INTERLUDE interface was defined between Orchestrators of two Operators to initiate a technical operation or dynamic service control. In order to address dynamic control of services supported by networks of multiple operators over ALLEGRO and INTERLUDE interfaces, Elastic E-Line Service was introduced by the author in MEF. In addition, Cloud Services Architecture was introduced as an architectural framework for virtual services by the author. In this paper, we describe LSO architecture and Elastic E-Line Service concept, and interactions between various LSO entities for dynamic service provisioning. After that, we describe Cloud Services Architecture and dynamic modifications of Elastic Cloud Services over ALLEGRO and INTERLUDE interfaces, for the first time. [ABSTRACT FROM AUTHOR]

As cloud data centers are dramatically growing, various applications are moved to cloud data centers owing to cost benefits for maintenance and hardware resources. However, latency-critical workloads among them suffer from some problems to fully achieve the cost-effectiveness. The latency-critical workloads should show latencies in a stable manner, to be predicted, for strictly meeting QoSs. However, if they are executed with other workloads to save the cost, they experience QoS violation due to the contention for the hardware resources shared with co-location workloads. In order to guarantee QoSs and to improve the hardware resource utilization, we proposed a memory bandwidth management method with an effective prediction model using machine learning. The prediction model estimates the amount of memory bandwidth that will be allocated to the latency-critical workload based on a REP decision tree. To construct this model, we first collect data and train the model with the data. The generated model can estimate the amount of memory bandwidth for meeting the SLO of the latency-critical workload no matter what batch processing workloads are collocated. The use of our approach achieves up to 99% SLO assurance and improves the server utilization up to 6.8 × on average. [ABSTRACT FROM AUTHOR]

Abstract Background Primary hypoadrenocorticism (Addison’s disease, AD) and symmetrical lupoid onychodystrophy (SLO) are two clinical conditions with an autoimmune etiology that occur in multiple dog breeds. In man, autoimmunity is associated with polymorphisms in immune-related genes that result in a reduced threshold for, or defective regulation of, T cell activation. The major histocompatibility complex (MHC) class II genes encode molecules that participate in these functions, and polymorphisms within these genes have been associated with autoimmune conditions in dogs and humans. Bearded collies have a relatively high prevalence of autoimmune diseases, particularly AD and SLO. Our study assessed the relationship between particular MHC (dog leukocyte antigen, DLA) class II haplotypes and the two autoimmune diseases most common in this breed. Moreover, five unrelated breeds at increased risk for AD were studied for comparative purposes and analyzed in the context of extant literature. Results A single DLA class II three-locus haplotype, determined by sequence-based typing, was associated with increased risk for AD (DLA-DRB1*009:01/DQA1*001:01/DQB1*008:02) in bearded collies. Comparative analysis with the five additional breeds showed limited allele sharing, with DQA1*001:01 and DQB1*002:01 being the only alleles observed in all breeds. A distinct three-locus risk haplotype (DLA-DRB1*001:01/DQA1*001:01/DQB1*002:01) was associated with AD in the West Highland white terrier and Leonberger. Two different risk haplotypes were associated with increased risk for SLO in the bearded collie (DLA-DRB1*018:01/DQA1*001:01/DQB1*002:01 and DLA-DRB1*018:01/DQA1*001:01/ DQB1*008:02). Conclusion Two-locus DQ haplotypes composed of DLA-DQA1*001:01 in association with DLA-DQB1*002:01 or DLA-DQB1*008:02 make up the four risk haplotypes identified in the present study and are also found in other risk haplotypes previously associated with diabetes mellitus and hypothyroidism across different dog breeds. Our findings build upon previously published data to suggest that this two-locus (DQ) model serves as a good indicator for susceptibility to multiple organ-specific autoimmune diseases in the canine population. However, it is also clear that additional loci are necessary for actual disease expression. Investigation of affected and unaffected dogs carrying these predisposing DQ haplotype signatures may allow for the identification of those additional genetic components that determine autoimmune disease expression and organ specificity.

The mouse optic nerve crush (ONC) model has been widely used to study optic neuropathies and central nervous system (CNS) axon injury and repair. Previous histological studies of retinal ganglion cell (RGC) somata in retina and axons in ON demonstrate significant neurodegeneration after ONC, but longitudinal morphological and functional assessment of RGCs in living animals is lacking. It is essential to establish these assays to provide more clinically relevant information for early detection and monitoring the progression of CNS neurodegeneration. Here, we present in vivo data gathered by scanning laser ophthalmoscopy (SLO), optical coherence tomography (OCT), and pattern electroretinogram (PERG) at different time points after ONC in mouse eyes and corresponding histological quantification of the RGC somata and axons. Not surprisingly, direct visualization of RGCs by SLO fundus imaging correlated best with histological quantification of RGC somata and axons. Unexpectedly, OCT did not detect obvious retinal thinning until late time points (14 and 28-days post ONC) and instead detected significant retinal swelling at early time points (1–5 days post-ONC), indicating a characteristic initial retinal response to ON injury. PERG also demonstrated an early RGC functional deficit in response to ONC, before significant RGC death, suggesting that it is highly sensitive to ONC. However, the limited progression of PERG deficits diminished its usefulness as a reliable indicator of RGC degeneration.

The Social License to Operate (SLO), understood as the consent of a wide group of stakeholders to mining activities, has become an important element in the process of obtaining a mining license in recent years. Such a pattern of increasing importance is common, both in Poland and throughout the EU. Therefore, it should be of key importance to prevent society’s reluctance to this industry. The article indicates that a very important tool in this matter should be reliable and knowledge-based raw materials education, carried out as early as at the stage of school education. Deficiencies in this respect were revealed through a survey conducted among children and youth up to 15 years of age. The results of the survey were discussed in the context of the development of raw materials education in Poland, which results indicate should focus on emphasizing the benefits of mining, both in terms of the general public and individuals. Shaping and strengthening the mineral raw materials awareness of the inhabitants of Poland is also of key importance for the implementation of standards for the protection of mineral deposits and ensuring the country’s resource security.

Purpose: To compare the scanning laser ophthalmoscopy (SLO), optical coherence tomography (OCT), and fluorescein angiography (FA) findings in retrievers with a single unilateral circular retinal plaque to those of an English springer spaniel with bilateral retinal dysplasia. Procedures: A retrospective record review identified three dogs with circular retinal plaques that underwent SLO and OCT; in two of the three dogs, FA was also completed. Morphologic changes, lesion measurements, and angiogram characteristics were documented. An English springer spaniel with bilateral retinal dysplasia that had undergone SLO, OCT, and FA was used for comparison. Results: Scanning laser ophthalmoscopy of the retriever dogs revealed circular retinal plaques with a dark periphery located in the tapetal retina. OCT revealed a thickening of the nerve fiber layer corresponding to the circular pattern observed on SLO. Within the circular plaque, the retina was predominantly of normal architecture. FA revealed variable hypofluorescence of both the rim and the center of the circular lesion throughout the early angiogram phases. In the late recirculation phase, small multifocal areas of hyperfluorescence were observed. OCT of geographic retinal dysplasia in the English springer spaniel revealed disorganization of both inner and outer retinal layers, and retinal detachment. Conclusions: Circular plaques observed in the tapetal retina are predominantly formed by a thickening of inner retina, while retinal dysplasia has disorganization of both inner and outer retinal layers. Further etiologic research is needed, including pedigree mapping to determine whether retinal plaques are an acquired or inherited condition. [ABSTRACT FROM AUTHOR]

The mouse optic nerve crush (ONC) model has been widely used to study optic neuropathies and central nervous system (CNS) axon injury and repair. Previous histological studies of retinal ganglion cell (RGC) somata in retina and axons in ON demonstrate significant neurodegeneration after ONC, but longitudinal morphological and functional assessment of RGCs in living animals is lacking. It is essential to establish these assays to provide more clinically relevant information for early detection and monitoring the progression of CNS neurodegeneration. Here, we present in vivo data gathered by scanning laser ophthalmoscopy (SLO), optical coherence tomography (OCT), and pattern electroretinogram (PERG) at different time points after ONC in mouse eyes and corresponding histological quantification of the RGC somata and axons. Not surprisingly, direct visualization of RGCs by SLO fundus imaging correlated best with histological quantification of RGC somata and axons. Unexpectedly, OCT did not detect obvious retinal thinning until late time points (14 and 28-days post ONC) and instead detected significant retinal swelling at early time points (1–5 days post-ONC), indicating a characteristic initial retinal response to ON injury. PERG also demonstrated an early RGC functional deficit in response to ONC, before significant RGC death, suggesting that it is highly sensitive to ONC. However, the limited progression of PERG deficits diminished its usefulness as a reliable indicator of RGC degeneration. [ABSTRACT FROM AUTHOR]

MinK-related peptides (MiRPs) are single-span membrane proteins that assemble with specific voltage-gated K+ (Kv) channel alpha-subunits to establish gating kinetics, unitary conductance, expression level, and pharmacology of the mixed complex. MiRP3 (encoded by the KCNE4 gene) has been shown to alter the behavior of some Kv alpha-subunits in vitro but its natural partners and physiologic functions are unknown. Seeking in vivo partners for MiRP3, immunohistochemistry was used to localize its expression to a unique subcellular site, the apical membrane of renal intercalated cells, where one potassium channel type has been recorded, the calcium- and voltage-gated channel BK. Overlapping staining of these two proteins was found in rabbit intercalated cells, and MiRP3 and BK subunits expressed in tissue culture cells were found to form detergent-stable complexes. Electrophysiologic and biochemical evaluation showed MiRP3 to act on BK to reduce current density in two fashions: shifting the current-voltage relationship to more depolarized voltages in a calcium-dependent fashion ( approximately 10 mV at normal intracellular calcium levels) and accelerating degradation of MiRP3-BK complexes. The findings suggest a role for MiRP3 modulation of BK-dependent urinary potassium excretion.

Geothermal energy is a resource that has the potential for development in many countries around the world. Despite its versatility and economic viability, this resource faces numerous obstacles that hinder its deployment and capacity for achieving a similar market share as other renewable and clean energy sources. Both technical and non-technical barriers can be highly detrimental to the implementation of geothermal projects. A social license to operate (SLO) is a tool that can help the deployment of geothermal energy. As a new concept, SLO is little developed in the business literature but is still being adopted in many industries. Its main challenges reside in its context-dependence and the lack of clear frameworks to utilize it. This paper introduces, in a first known attempt, through a qualitative approach, a conceptual model of the social license to operate in the geothermal energy sector. For its development, three case studies, working group discussion and surveying were conducted in the framework of the H2020 funded CROWDTHERMAL project, which aims to empower EU citizens for direct participation in geothermal projects through crowdfunding. Findings of this paper also drew on existing general conceptual models of the SLO, and experiences from other sectors that have developed their own SLO models. The paper contributes to a more contextualized understanding of the social license within the geothermal sector and sheds the light on practices and challenges that influence the acquisition and maintenance of SLO in geothermal energy projects and initiatives.

Construction programs seeking accreditation from the American Council for Construction Education (ACCE) must now demonstrate student achievement of 20 Student Learning Outcomes (SLO). Part of accomplishing this involves clarifying how the program interprets each SLO. This is problematic because there is no precedent for how this should be done. The overall aim of the research was to establish a model for clarifying the meaning of the 20 SLOs through identifying performance criteria for each SLO. This paper reports on a case study implementing the proposed methodology on a 4-year construction program focused on preparing students for careers in commercial construction management. Details of the multi-phased mixed methods study involving a panel of industry experts to identify the essential performance criteria is presented. The method resulted in identifying performance criteria representing views of industry, alumni, and faculty of the program that aided in preparations for accreditation and provided cause for reflection about strengths, weaknesses, and areas of improvement for the program. [ABSTRACT FROM AUTHOR]

BK Ca2+-activated K+ channels are important regulators of membrane excitability. Multiple regulatory mechanisms tailor BK current properties across tissues, such as alternative splicing, posttranslational modifications, and auxiliary subunits. Another potential mechanism for modulating BK channel activity is genetic variation due to single nucleotide polymorphisms (SNPs). The gene encoding the human BK α subunit, KCNMA1 , contains hundreds of SNPs. However, the variation in BK channel activity due to SNPs is not well studied. Here, we screened the effects of four SNPs (A138V, C495G, N599D, and R800W) on BK currents in HEK293T cells, selected based on predicted protein pathogenicity or disease linkage. We found that the SNPs C495G and R800W had the largest effects on BK currents, affecting the conductance–voltage relationship across multiple Ca2+ conditions in the context of two BK channel splice variants. In symmetrical K+, C495G shifted the V1/2 to more hyperpolarized potentials (by −15 to −20 mV) and accelerated activation, indicating C495G confers some gain-of-function properties. R800W shifted the V1/2 to more depolarized potentials (+15 to +35 mV) and slowed activation, conferring loss-of-function properties. Moreover, the C495G and R800W effects on current properties were found to persist with posttranslational modifications. In contrast, A138V and N599D had smaller and more variable effects on current properties. Neither application of alkaline phosphatase to patches, which results in increased BK channel activity attributed to channel dephosphorylation, nor bidirectional redox modulations completely abrogated SNP effects on BK currents. Lastly, in physiological K+, C495G increased the amplitude of action potential (AP)-evoked BK currents, while R800W had a more limited effect. However, the introduction of R800W in parallel with the epilepsy-linked mutation D434G (D434G/R800W) decreased the amplitude of AP-evoked BK currents compared with D434G alone. These results suggest that in a physiological context, C495G could increase BK activation, while the effects of the loss-of-function SNP R800W could oppose the gain-of-function effects of an epilepsy-linked mutation. Together, these results implicate naturally occurring human genetic variation as a potential modifier of BK channel activity across a variety of conditions. [ABSTRACT FROM AUTHOR]

We present a novel method to image the vasculature of the anterior segment of the eye using a non-invasive optical coherence tomography angiography (OCTA) system with a modified lens system. This system utilises the Heidelberg Spectralis OCTA and a 3D printed lens mount holding a simple biconvex lens in place to allow capture of images in the anterior segment. [ABSTRACT FROM AUTHOR]

Slo3 is a pH-sensitive and weakly voltage-sensitive potassium channel that is essential for male fertility in mouse and whose expression is regarded as sperm-specific. These properties have proposed Slo3 as a candidate target for male contraceptive drugs. Nonetheless, the tissue distribution of Slo3 expression has not been rigorously studied yet. Applying computational and RT-PCR approaches, we identified expression of two short Slo3 isoforms in somatic mouse tissues such as brain, kidney and eye. These isoforms, which seem to result of transcription starting sites between exons 20 and 21, have an identical open reading frame, both encoding the terminal 381 amino acids of the cytosolic Slo3 domain. We corroborated the expression of these isoforms in mouse brain and testis by Western-blot. The complete isoform encoding the Slo3 ion channel was uniquely detected in testis, both at transcript and protein level. Although the functional role of the cytosolic Slo3 isoforms remains to be established, we propose that they may have a functional effect by modulating Slo channels trafficking and/or activity. This study confirms that expression of full-length Slo3 is sperm-specific but warns against developing contraceptive drugs targeting the C-terminal tail of Slo3 channels. [ABSTRACT FROM AUTHOR]

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To provide low-latency and high-throughput guarantees, most large key-value stores keep the data in the memory of many servers. Despite the natural parallelism across lookups, the load imbalance, introduced by heavy skew in the popularity distribution of keys, limits performance. To avoid violating tail latency servicelevel objectives, systems tend to keep server utilization low and organize the data in micro-shards, which provides units of migration and replication for the purpose of load balancing. These techniques reduce the skew but incur additional monitoring, data replication, and consistency maintenance overheads. In this work, we introduce RackOut, a memory pooling technique that leverages the one-sided remote read primitive of emerging rack-scale systems tomitigate load imbalance while respecting service-level objectives. In RackOut, the data are aggregated at rack-scale granularity, with all of the participating servers in the rack jointly servicing all of the rack's micro-shards. We develop a queuing model to evaluate the impact of RackOut at the datacenter scale. In addition, we implement a RackOut proof-of-concept key-value store, evaluate it on two experimental platforms based on RDMA and Scale-Out NUMA, and use these results to validate the model. We devise two distinct approaches to load balancing within a RackOut unit, one based on random selection of nodes--RackOut_static--and another one based on an adaptive load balancing mechanism--RackOut_adaptive. Our results show that RackOut_static increases throughput by up to 6× for RDMA and 8.6× for Scale-Out NUMA compared to a scale-out deployment, while respecting tight tail latency servicelevel objectives. RackOut_adaptive improves the throughput by 30% for workloads with 20% of writes over RackOut_static. [ABSTRACT FROM AUTHOR]

Virtualization technology takes an important place for the cloud service provider (CSP) because of the rapid development of cloud computing. In order to obtain more profits, the CSP need to maximize hardware performance as much as possible while ensuring good user experience. By leveraging information such as the priority and importance of I/O requests, researchers have proposed many methods to improve program performance in Linux kernel. However, the information in virtual machines can get lost during the transfer to the host. We therefore propose an I/O assurance framework based on service-level objective (SLO). We firstly explore the reasons for the loss of information such as I/O request priority, and then present the key issues to be addressed for information delivering. Our framework transmits the SLO information of the threads in the virtual machine to the host machine successfully by expanding Linux kernel, the virtio protocol and KVM I/O virtualization program QEMU. What's more, we implement a scheduler based on SLO. Experimental results verify the feasibility of the proposed framework. The thread with highest priority has a throughput up to 260 KB/s while the throughput of the thread with lowest priority is 10 KB/s. we prove that the SLO information delivered by the proposed framework plays a positive role in the scheduling of the scheduler in the host machine. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Slo1 and Slo3 channels functionally diverged from their ancestral duplication. • Slo1 is strongly conserved, whereas Slo3 has evolved more rapidly. • Slo3 divergence is primarily driven by relaxed selection. • Positive selection favored amino acid changes in Slo3 with functional implications. Abstract Slo1 and Slo3 encode close paralogues of the Slo potassium (K+) channels family. Despite their evolutionary relatedness, Slo1 and Slo3 channels show marked functional differences and evolutionary dynamics. Whereas Slo1 is a highly conserved and widely expressed channel, Slo3 is a rapidly evolving channel restricted to sperm. However, the molecular mechanisms behind the structural-functional differences of Slo1 and Slo3 channels are unknown. In this study, we explored the functional divergence of Slo1 and Slo3 subfamilies in vertebrates and examined the structure-function relationships of our predictions using experimental data. We found that ∼25% of sites between Slo1 and Slo3 underwent altered functional constraints, affecting some residues with important roles in Slo1 channel gating. Because functional divergence was principally generated by accelerated evolution of Slo3 after gene duplication, we explored selective forces behind Slo3 diversification. We observed that Slo3 subjected was principally subjected to relaxation of purifying selection, but we also identified several sites evolving under positive selection in the cytosolic domain of this channel. Concerning Slo1, this channel presented strong purifying selection. Whether residues evolving under different selection in Slo1 and Slo3 are responsible for functional differences observed between these channels, as well as among Slo3 orthologs, remains to be established. [ABSTRACT FROM AUTHOR]

Latency-critical workloads such as web search engines, social networks and finance market applications are sensitive to tail latencies for meeting service level objectives (SLOs). Since unexpected tail latencies are caused by sharing hardware resources with other co-executing workloads, a service provider executes the latency-critical workload alone. Thus, the data center for the latency-critical workloads has exceedingly low hardware resource utilization. For improving hardware resource utilization, the service provider has to co-locate the latency-critical workloads and other batch processing ones. However, because the memory bandwidth cannot be provided in isolation unlike the cores and cache memory, the latency-critical workloads experience poor performance isolation even though the core and cache memory are allocated in isolation to the workloads. To solve this problem, we propose an optimized memory bandwidth management approach for ensuring quality of service (QoS) and high server utilization. By providing isolated shared resources including the memory bandwidth to the latency-critical workload and co-executing batch processing ones, firstly, our proposed approach performs few pre-profilings under the assumption that memory bandwidth contention is the worst with a divide and conquer method. Second, we predict the memory bandwidth to meet the SLO for all queries per seconds (QPSs) based on results of the pre-profilings. Then, our approach allocates the amount of the isolated memory bandwidth that guarantees the SLO to the latency-critical workload and the rest of the memory bandwidth to co-executing batch processing ones. It is experimentally found that our proposed approach can achieve up to 99% SLO assurance and improve the server utilization up to 6.5×. [ABSTRACT FROM AUTHOR]

Retinoblastoma (Rb) is the most frequent primary intraocular tumour in children and, if left untreated, can cause death. Preclinical animal models that mimic molecular, genetic, and cellular features of cancers are essential for studying cancer and searching for promising diagnosis and treatment modalities. There are several models described for Rb, but none of them fully meet our requirements. The aim of this study was to create a novel xenograft-nude mouse-model with broad application possibilities, which closely resembles the clinical observations of Rb patients and which could be used to investigate the development and spread of the tumour by using scanning laser ophthalmoscopy/optical coherence tomography (SLO/OCT) as well as histology methods. We injected human retinoblastoma Y79 cells intravitreally in both eyes of immune-deficient nude mice. The incidences of retinoblastoma as well as growth velocity were analysed 3, 6, 9 and 12 weeks after cell injection in vivo by SLO/OCT as well as ex vivo by electron microscopy (EM) and hematoxylin/eosin (HE) staining. Moreover, internal organs were histologically screened for potentially occurring metastases. Three weeks post-injection, animals developed a retinoblastoma, and after five weeks tumour growth resulted in swelling of the eyes in individual animals, showing a similar phenotype to that of untreated Rb patients at advanced stages of tumour-development. After 12 weeks, 67.5% of all analysed eyes (29 of 42) contained a retinoblastoma. At early stages of Rb development, the SLO/OCT analysis correlated with the histology results. If the tumours were too large, only histological investigations were feasible. The ultrastructural characteristics of the xenograft-tumours were very similar to those described for patient's tumours. In one mouse, brain metastases were observed. Our retinoblastoma mouse model closely resembles the human disease. SLO/OCT can be used for the detection of Rb at early stages of development and could be used for monitoring the success of future therapies.