Functional Dissection of Streptococcus pyogenes M5 Protein: the Hypervariable Region is Essential for Virulence

The surface-localized M protein of Streptococcus pyogenes is a major virulence factor that inhibits phagocytosis, as determined ex vivo. Because little is known about the role of M protein in vivo we analyzed the contribution of different M protein regions to virulence, using the fibrinogen (Fg)-binding M5 protein and a mouse model of acute invasive infection. This model was suitable, because M5 is required for mouse virulence and binds mouse and human Fg equally well, as shown here. Mixed infection experiments with wild type bacteria demonstrated that mutants lacking the N-terminal hypervariable region (HVR) or the Fg-binding B-repeat region were strongly attenuated, while a mutant lacking the conserved C-repeats was only slightly attenuated. Because the HVR of M5 is not required for phagocytosis resistance, our data imply that this HVR plays a major but unknown role during acute infection. The B-repeat region is required for phagocytosis resistance and specifically binds Fg, suggesting that it promotes virulence by binding Fg. However, B-repeat mutants were attenuated even in Fg-deficient mice, implying that the B-repeats may have a second function, in addition to Fg-binding. These data demonstrate that two distinct M5 regions, including the HVR, are essential to virulence during the early stages of an infection. In particular, our data provide the first in vivo evidence that the HVR of an M protein plays a major role in virulence, focusing interest on the molecular role of this region

Human Complement Regulators C4b-Binding Protein and C1 Esterase Inhibitor Interact with a Novel Outer Surface Protein of Borrelia recurrentis

The spirochete Borrelia recurrentis is the causal agent of louse-borne relapsing fever and is transmitted to humans by the infected body louse Pediculus humanus. We have recently demonstrated that the B. recurrentis surface receptor, HcpA, specifically binds factor H, the regulator of the alternative pathway of complement activation, thereby inhibiting complement mediated bacteriolysis. Here, we show that B. recurrentis spirochetes express another potential outer membrane lipoprotein, termed CihC, and acquire C4b-binding protein (C4bp) and human C1 esterase inhibitor (C1-Inh), the major inhibitors of the classical and lectin pathway of complement activation. A highly homologous receptor for C4bp was also found in the African tick-borne relapsing fever spirochete B. duttonii. Upon its binding to B. recurrentis or recombinant CihC, C4bp retains its functional potential, i.e. facilitating the factor I-mediated degradation of C4b. The additional finding that ectopic expression of CihC in serum sensitive B. burgdorferi significantly increased spirochetal resistance against human complement suggests this receptor to substantially contribute, together with other known strategies, to immune evasion of B. recurrentis

Developing core elements and checklist items for global hospital antimicrobial stewardship programmes:a consensus approach

International audienc

Yersinia enterocolitica Serum Resistance Proteins YadA and Ail Bind the Complement Regulator C4b-Binding Protein

Many pathogens are equipped with factors providing resistance against the bactericidal action of complement. Yersinia enterocolitica, a Gram-negative enteric pathogen with invasive properties, efficiently resists the deleterious action of human complement. The major Y. enterocolitica serum resistance determinants include outer membrane proteins YadA and Ail. Lipopolysaccharide (LPS) O-antigen (O-ag) and outer core (OC) do not contribute directly to complement resistance. The aim of this study was to analyze a possible mechanism whereby Y. enterocolitica could inhibit the antibody-mediated classical pathway of complement activation. We show that Y. enterocolitica serotypes O:3, O:8, and O:9 bind C4b-binding protein (C4bp), an inhibitor of both the classical and lectin pathways of complement. To identify the C4bp receptors on Y. enterocolitica serotype O:3 surface, a set of mutants expressing YadA, Ail, O-ag, and OC in different combinations was tested for the ability to bind C4bp. The studies showed that both YadA and Ail acted as C4bp receptors. Ail-mediated C4bp binding, however, was blocked by the O-ag and OC, and could be observed only with mutants lacking these LPS structures. C4bp bound to Y. enterocolitica was functionally active and participated in the factor I-mediated degradation of C4b. These findings show that Y. enterocolitica uses two proteins, YadA and Ail, to bind C4bp. Binding of C4bp could help Y. enterocolitica to evade complement-mediated clearance in the human host

Virulence of Group A Streptococci Is Enhanced by Human Complement Inhibitors

Streptococcus pyogenes, also known as Group A Streptococcus (GAS), is an important human bacterial pathogen that can cause invasive infections. Once it colonizes its exclusively human host, GAS needs to surmount numerous innate immune defense mechanisms, including opsonization by complement and consequent phagocytosis. Several strains of GAS bind to human-specific complement inhibitors, C4b-binding protein (C4BP) and/or Factor H (FH), to curtail complement C3 (a critical opsonin) deposition. This results in diminished activation of phagocytes and clearance of GAS that may lead to the host being unable to limit the infection. Herein we describe the course of GAS infection in three human complement inhibitor transgenic (tg) mouse models that examined each inhibitor (human C4BP or FH) alone, or the two inhibitors together (C4BPxFH or 'double' tg). GAS infection with strains that bound C4BP and FH resulted in enhanced mortality in each of the three transgenic mouse models compared to infection in wild type mice. In addition, GAS manifested increased virulence in C4BPxFH mice: higher organism burdens and greater elevations of pro-inflammatory cytokines and they died earlier than single transgenic or wt controls. The effects of hu-C4BP and hu-FH were specific for GAS strains that bound these inhibitors because strains that did not bind the inhibitors showed reduced virulence in the 'double' tg mice compared to strains that did bind; mortality was also similar in wild-type and C4BPxFH mice infected by non-binding GAS. Our findings emphasize the importance of binding of complement inhibitors to GAS that results in impaired opsonization and phagocytic killing, which translates to enhanced virulence in a humanized whole animal model. This novel hu-C4BPxFH tg model may prove invaluable in studies of GAS pathogenesis and for developing vaccines and therapeutics that rely on human complement activation for efficacy

Causal mechanisms proposed for the Alcohol Harm Paradox - a systematic review

Background and Aims The Alcohol Harm Paradox (AHP) posits that disadvantaged groups suffer from higher rates of alcohol-related harm compared with advantaged groups, despite reporting similar or lower levels of consumption on average. The causes of this relationship remain unclear. This study aimed to identify explanations proposed for the AHP. Secondary aims were to review the existing evidence for those explanations and investigate whether authors linked explanations to one another. Methods Systematic review. We searched MEDLINE (1946-January 2021), EMBASE (1974 – January 2021) and PsycINFO (1967 – January 2021), supplemented via manual searching of grey literature. Included papers either explored the causes of the AHP or investigated the relationship between alcohol consumption, alcohol-related harm, and socioeconomic position. Papers were set in Organisation for Economic Co-operation and Development high income countries. Explanations extracted for analysis could be evidenced in the empirical results or suggested by researchers in their narrative. Inductive thematic analysis was applied to group explanations. Results Seventy-nine papers met the inclusion criteria and initial coding revealed these papers contained 41 distinct explanations for the AHP. Following inductive thematic analysis, these explanations were grouped into 16 themes within six broad domains: Individual, Lifestyle, Contextual, Disadvantage, Upstream and Artefactual. Explanations related to risk behaviours, which fit within the Lifestyle domain, were the most frequently proposed (n=51) and analysed (n=21). Conclusions While there are many potential explanations for the Alcohol Harm Paradox, most research focuses on risk behaviours while other explanations lack empirical testing

Analysis of magnetic field measurement results for the AGS Booster magnets

Magnetic field measurements have been made on nearly 200 conventional magnets that have been installed in the AGS Booster and its associated transfer lines. The measurements were intended to monitor the quality of the magnets being produced and to check the performance of each magnet before installation. The magnetic measurements effort led to certain improvements in the manufacturing process, which ten subsequently produced very good, very uniform magnets. The integrated dipole fields of the 36 booster dipoles are uniform to 1.5 parts in ten thousand. The magnetic measurements indicate that the quadrupoles were manufactured to an accuracy of 3 ten thousandths of an inch, which is better than we can physically measure. 3 refs., 2 figs., 4 tabs

Power generation from low heat sources

In this chapter a thorough review of the latest research findings on power generation from non-conventional low heat sources is presented. Main discoveries and results of research works ranging from source exploitation technologies to final power production are reported and discussed to offer an overview of their potential. Firstly the concept of low-grade source is presented and the main energy sources in this group (i.e. geothermal energy, solar thermal systems, industrial waste heat and ocean thermal energy) are introduced. Each of them is briefly described and the latest developments and improvements on the technologies for their exploitation are enumerated. Afterwards the state-of-the-art available power cycles for the conversion of low heat into electricity are reported. Only thermal power conversion technologies are presented due to their higher presence in commercial applications and their potential for small scale power generation. Among these technologies, last findings and results on organic Rankine cycles (ORC) and power cycles based on working fluid mixtures (e.g. Kalina cycle) are described. A special emphasis is placed on organic Rankine cycles (ORC) since over the last few years this technology has experienced a significant global growth, boosted by their viable performance and the inherited knowledge from the refrigeration industry. In addition, the suitability of less known technologies such as Stirling cycles and their current development status and perspectives are also commented. After this review it follows an examination of the implementation of these technologies in present power production systems. Discussion will be provided on which are the current barriers that the mentioned technologies are facing for their introduction or during their operation. On the other hand, practical restrictions concerning the availability of suitable technology, environmental requirements or economic viability are stated. Limitations regarding thermodynamic and technological aspects, as well as operational concerns will be considered of special interest. In the final section we deal with the future scenario for the integration of small-scale power generation. Potential solutions for overcoming technology development barriers are presented and directions of current research works on this topic are pointed out

A beam scraper using a linear motor

A beam scraper using a linear motor drive has been developed for use in the AGS at Brookhaven National Laboratory. The device is used to measure beam size by moving a target to a predetermined location and measuring the intercepted beam with nearby loss monitors or by noting the decrease in the circulating beam current. This device has excellent vacuum characteristics, as the motor and sensor coils are outside the vacuum, coupled magnetically to the moving parts which are inside. There are no bellows or dynamic seals required. The position-time profile is controlled by a closed-loop servo system which uses position feedback. 2 refs., 4 figs