Pathogenic Streptococcus strains employ novel escape strategy to inhibit bacteriostatic effect mediated by mammalian peptidoglycan recognition protein

Pathogenic streptococcal species are responsible for some of the most lethal and prevalent animal and human infections. Previous reports have identified a candidate pathogenicity island (PAI) in two highly virulent clinical isolates of Streptococcus suis type 2, a causative agent of high‐mortality streptococcal toxic shock syndrome. This PAI contains a type‐IVC secretion system C subgroup (type‐IVC secretion system) that is involved in the secretion of unknown pathogenic effectors that are responsible for streptococcal toxic shock syndrome caused by highly virulent strains of S. suis. Both virulence protein B4 and virulence protein D4 were demonstrated to be key components of this type‐IVC secretion system. In this study, we identify a new PAI family across 3 streptococcal species; Streptococcus genomic island contains type‐IV secretion system, which contains a genomic island type‐IVC secretion system and a novel PPIase molecule, SP1. SP1 is shown to interact with a component of innate immunity, peptidoglycan recognition protein (PGLYRP‐1) and to perturb the PGLYRP‐1‐mediated bacteriostatic effect by interacting with protein PGLYRP‐1. Our study elucidates a novel mechanism by which bacteria escape by components of the innate immune system by secretion of the SP1 protein in pathogenic Streptococci, which then interacts with PGLYRP‐1 from the host. Our results provide potential targets for the development of new antimicrobial drugs against bacteria with resistance to innate host immunity.     

The impact of immunization on competition within Plasmodium infections

Evolutionary theory argues that ecological interactions between pathogens within an infection can be a potent source of selection shaping traits such as virulence, drug resistance, and infectiousness. In humans, malaria infections are frequently genetically diverse, with mixed genotype infections the norm. A wide variety of evidence shows that crowding occurs within infections, with the population densities of individual genotypes suppressed by the presence of others. Public health interventions are expected to impact on levels of immunity experienced by pathogens, indirectly by reducing the rate of acquisition of natural immunity by reducing the force of infection, and directly in the case of vaccination programs. Here we ask how enhanced host immunity affects competitive interactions between malaria parasites within hosts and thus the strength of in-host selection on traits such as virulence. We used a model malaria system, Plasmodium chabaudi in laboratory mice, where it has been previously shown that less virulent parasites are competitively suppressed by more virulent strains, generating within-host selection for increased virulence. We found that immunization with either a recombinant antigen or with live parasites suppressed parasite densities, but that there was no evidence that immunization relieved or exacerbated competitive suppression, or affected the relative frequency of clones within infections. There is thus no reason to think that immunization strengthens or alleviates the potentially very potent selection on parasite traits arising from interactions between pathogen genotypes within infections.     

Use of selective capture of transcribed sequences to identify genes preferentially expressed by Streptococcus suis upon interaction with porcine brain microvascular endothelial cells

By using the selective capture of transcribed sequences (SCOTS) approach, we identified 28 genes preferentially expressed by the major swine pathogen and zoonotic agent Streptococcus suis upon interaction with porcine brain microvascular endothelial cells. Several of these genes may be considered new S. suis candidate virulence factors. Results from this study demonstrate the suitability of SCOTS for the elucidation of gene expression in streptococcal species and may contribute to a better understanding of the pathogenesis of S. suis infections.     

Identification of a lysin associated with a bacteriophage (A25) virulent for group A streptococci.

A phage-associated lysin was found in culture lysates resulting from the propagation of virulent bacteriophage A25 on the group A streptococcal strain designated K56. In contrast to the previously described group C streptococcal phage-associated lysins, A25 phage-associated lysin was more active on chloroform-treated cells, was not phage bound, and was active on some group G and H strains, as well as on group A and C strains. A25 phage-associated lysin had an optimum pH of 6.7 and was inactivated by 10(-3) M p-hydroxymercuribenzoate. Group A cells exposed to penicillin were more susceptible to A25 phage-associated lysin, whereas chloramphenicol-treated cells became resistant to lysis. Release of lipoteichoic acid appeared to precede lysis, and cardiolipin treatment of cells reversed the effects of chloroform and penicillin treatments. These results suggest the possibility that A25 phage-associated lysin may have a mechanism similar to the mechanism of an autolysin or that cell lysis may be due to the activation of an autolysin.     

Use of Selective Capture of Transcribed Sequences To Identify Genes Preferentially Expressed by Streptococcus suis upon Interaction with Porcine Brain Microvascular Endothelial Cells

By using the selective capture of transcribed sequences (SCOTS) approach, we identified 28 genes preferentially expressed by the major swine pathogen and zoonotic agent Streptococcus suis upon interaction with porcine brain microvascular endothelial cells. Several of these genes may be considered new S. suis candidate virulence factors. Results from this study demonstrate the suitability of SCOTS for the elucidation of gene expression in streptococcal species and may contribute to a better understanding of the pathogenesis of S. suis infections.     

Is plasminogen deployed as a Streptococcus pyogenes virulence factor?

Streptococcus pyogenes (group A streptococcus) causes human skin and throat infections as well as highly invasive diseases including necrotizing fasciitis. Group A streptococcal infections and invasive disease have made a resurgence in developed countries during the past two decades. S. pyogenes use multiple pathways for the acquisition and activation of human plasminogen, securing potent proteolytic activity on the bacterial cell surface. Recent experimental evidence using a humanized transgenic mouse model suggests a crucial role for human plasminogen in the dissemination of S. pyogenes in vivo.     

Mixed macromolecular crowding accelerates the refolding of rabbit muscle creatine kinase: implications for protein folding in physiological environments

The effects of four single macromolecular crowding agents, Ficoll 70, dextran 70, polyethylene glycol (PEG) 2000, and calf thymus DNA (CT DNA), and three mixed crowding agents containing both CT DNA and polysaccharide (or PEG 2000) on the refolding of guanidine hydrochloride-denatured rabbit muscle creatine kinase (MM-CK) have been examined by activity assay. When the total concentration of the mixed crowding agent is 100 g/l, in which the weight ratio of CT DNA to Ficoll 70 is 1:9, the refolding yield of MM-CK after refolding for 3 h under these conditions increases 23% compared with that in the presence of 10 g/l CT DNA, 18% compared with 100 g/l Ficoll 70, and 19% compared with that in the absence of crowding agents. A remarkable increase in the refolding yield of MM-CK by a mixed crowding agent containing CT DNA and dextran 70 (or PEG 2000) is also observed. Further folding kinetics analyses show that these three mixed crowding agents remarkably accelerate the refolding of MM-CK, compared with single crowding agents. Aggregation of MM-CK in the presence of any of the three mixed crowding agents is less serious than that in the presence of a single crowding agent at the same concentration but more serious than that in the absence of crowding agents. Both the refolding yield and the refolding rate of MM-CK in mixtures of these agents are increased relative to the individual agents by themselves, indicating that mixed macromolecular crowding agents are more favorable to MM-CK folding and can be used to reflect the physiological environment more accurately than single crowding agents.     

Leucocyte recruitment and molecular fortification of keratinocytes triggered by streptococcal M1 protein

Streptococcus pyogenes of the M1 serotype is commonly associated with invasive streptococcal infections and development of streptococcal toxic shock syndrome. The M1 protein is a powerful inducer of inflammatory responses for several human cell types, but the reason why M1 protein‐related strains is over‐represented in invasive streptococcal diseases is still not understood. This study was undertaken to investigate if soluble M1 protein can aggravate the severity of streptococcal skin infections in respect to inflammation, leucocyte recruitment, and tissue remodelling as seen in patients with cellulitis and necrotizing fasciitis. We found that HaCaT cells are able to recruit activated leucocytes when encountering M1 protein. Neither the bacterial protein nor activated leucocytes caused cell damage on HaCaT cells, instead HaCaT cells responded to the bacterial virulence factor by releasing several proteins protective against bacterial infection and leucocyte responses. However, although not cytotoxic, M1 protein completely abolished wound healing abilities of HaCaT cells. Taken together, our results demonstrate that M1 protein is a critical virulence factor that can augment streptococcal skin infection suggesting that the protein is an interesting target for drug development.     

Drug resistance in Chromobacterium violaceum

Chromobacterium violaceum is a free-living bacterium commonly found in aquatic habitats of tropical and subtropical regions of the world. This bacterium is able to produce a large variety of products of biotechnological and pharmacological use. Although C. violaceum is considered to be non-pathogenic, some cases of severe infections in humans and other animals have been reported. Genomic data on the type strain ATCC 12472(T) has provided a comprehensive basis for detailed studies of pathogenicity, virulence and drug resistance genes. A large number of open reading frames associated with various mechanisms of drug resistance were found, comprising a remarkable feature of this organism. Amongst these, beta-lactam (penicillin and cephalosporin) and multidrug resistance genes (drug efflux pumps) were the most numerous. In addition, genes associated with bacitracin, bicyclomycin, chloramphenicol, kasugamycin, and methylenomycin were also found. It is postulated that these genes contribute to the ability of C. violaceum to compete with other bacteria in the environment, and also may help to explain the common drug resistance phenotypes observed in infections caused by this bacterium.     

Phase changes in the population of group A streptococci and their influence on the course of the epidemic process of tonsillitis

This work presents the data on the complex evaluation of the population of group A streptococci, studied at each of four phases (reservation, epidemic transformation, epidemic spread, reservational transformation) of the course of the epidemic process of streptococcal infection of the respiratory tracts (tonsillitis) in an organized group of adults. The characterization of the phases of the infective agent in accordance with the level of the carrier state, the size of streptococcal foci and the virulence of streptococci is given. Thus, the study shows that the heterogeneity of group A streptococci with respect to their virulence reaches its maximum level at the phases of reservation and epidemic spread and its minimum level at the phases of epidemic and reservational transformation. The size of streptococcal foci in carriers and the virulence of streptococci isolated from them are the inter-related unidirectional signs of the population of the infective agent and, at the same time, the main factors responsible for the phase character of the epidemic process and the morbidity level in tonsillitis.     

Streptococcal L forms and phage. A clinical-epidemiologic study

This study showed that streptococcal L forms could not be isolated from children who were either carriers of group A streptococci or had disease due to this pathogen. It was possible to induce L colony formation in 15 strains of group A. Streptococcal bacteriophages were demonstrated in 20% of group A streptococci isolated from school children who were carriers, but did not have clinical evidence of streptococcal disease, and from 44.9% of children whose physicians considered they had acute streptococcal upper respiratory infections. Lysogeny (bacteriophage) was demonstrated more frequently during 1969-70 when carrier rates were high and from children who had manifest streptococcal disease, suggesting a possible positive relationship between lysogeny, high carrier rates, and infection in the children studied. Lysogeny and erythrogenic toxin production by group A streptococci occurred simultaneously in approximately half of the strains of group A streptococci tested, suggesting that lysogeny is not a sine qua non for erythrogenic toxin production.     

The fundamental contribution of phages to GAS evolution,genome diversification and strain emergence

The human bacterial pathogen group A Streptococcus (GAS) causes many different diseases including pharyngitis, tonsillitis, impetigo, scarlet fever, streptococcal toxic shock syndrome, necrotizing fasciitis and myositis, and the post-infection sequelae glomerulonephritis and rheumatic fever. The frequency and severity of GAS infections increased in the 1980s and 1990s, but the cause of this increase is unknown. Recently, genome sequencing of serotype M1, M3 and M18 strains revealed many new proven or putative virulence factors that are encoded by phages or phage-like elements. Importantly, these genetic elements account for an unexpectedly large proportion of the difference in gene content between the three strains. These new genome-sequencing studies have provided evidence that temporally and geographically distinct epidemics, and the complex array of GAS clinical presentations, might be related in part to the acquisition or evolution of phage-encoded virulence factors. We anticipate that new phage-encoded virulence factors will be identified by sequencing the genomes of additional GAS strains, including organisms non-randomly associated with particular clinical syndromes.     

Listeriolysin S, a novel peptide haemolysin associated with a subset of lineage I Listeria monocytogenes

Streptolysin S (SLS) is a bacteriocin-like haemolytic and cytotoxic virulence factor that plays a key role in the virulence of Group A Streptococcus (GAS), the causative agent of pharyngitis, impetigo, necrotizing fasciitis and streptococcal toxic shock syndrome. Although it has long been thought that SLS and related peptides are produced by GAS and related streptococci only, there is evidence to suggest that a number of the most notorious Gram-positive pathogenic bacteria, including Listeria monocytogenes, Clostridium botulinum and Staphylococcus aureus, produce related peptides. The distribution of the L. monocytogenes cluster is particularly noteworthy in that it is found exclusively among a subset of lineage I strains; i.e., those responsible for the majority of outbreaks of listeriosis. Expression of these genes results in the production of a haemolytic and cytotoxic factor, designated Listeriolysin S, which contributes to virulence of the pathogen as assessed by murine- and human polymorphonuclear neutrophil-based studies. Thus, in the process of establishing the existence of an extended family of SLS-like modified virulence peptides (MVPs), the genetic basis for the enhanced virulence of a proportion of lineage I L. monocytogenes may have been revealed.     

THE EXPERIMENTAL AND CLINICAL USE OF POLYMYXIN,CHLOROMYCETIN, AND AUREOMYCIN

Polymyxin is an effective antibiotic for the treatment of severe infections produced by Ps. aeruginosa, H. pertussis, H. influenzae, E. coli, and A. aerogenes. Its toxicity to date precludes its general use in infections susceptible to its therapeutic effects.Chloromycetin has been demonstrated to be an effective antibiotic agent for the treatment of rickettsial diseases and typhoid fever. It will undoubtedly prove effective in the treatment of other infections produced by certain Gram-negative micro-organisms and viral agents.Aureomycin has been shown to be an active antibiotic agent against rickettsial diseases, primary atypical pneumonia, acute brucellosis, pneumococcal, streptococcal, and staphylococcal infections, urinary tract infections produced by E. coli, A. aerogenes and Strept. fecalis, certain types of infections of the eye, and in subacute bacterial endocarditis when the infecting agent is Strept. fecalis. Its clinical use in forms of extrapulmonary tuberculosis is in a completely experimental stage. It is not recommended in typhoid fever or in infections due to Ps. aeruginosa or P. vulgaris, and it seems to be ineffective in whooping cough.To date, neither chloromycetin nor aureomycin has shown significant signs of systemic toxicity.     

I. Clinical evaluation

A study was undertaken to evaluate the therapy of streptococcal pharyngitis. The compliance of 118 patients with beta-hemolytic streptococcal pharyngitis to follow-up was 72%. Of 74 patients checked by means of urine tests 66 took their oral medication. No differences were detected in the clinical and bacteriological results (>98% streptococcal eradication) after the 7th or 10th day of therapy after taking either cephalexin or penicillin.It was concluded that: (a) for effective surveillance and follow-up special attention should be given to the uncooperative segment of the patient population; (b) a seven-day course of penicillin may be satisfactory in the eradication of BHS from the throat; and (c) cephalexin appears to be an effective alternative to penicillin for the treatment of streptococcal pharyngitis.     

The experimental and clinical use of polymyxin,chloromycetin, and aureomycin

Polymyxin is an effective antibiotic for the treatment of severe infections produced by Ps. aeruginosa, H. pertussis, H. influenzae, E. coli, and A. aerogenes. Its toxicity to date precludes its general use in infections susceptible to its therapeutic effects. Chloromycetin has been demonstrated to be an effective antibiotic agent for the treatment of rickettsial diseases and typhoid fever. It will undoubtedly prove effective in the treatment of other infections produced by certain Gram-negative micro-organisms and viral agents. Aureomycin has been shown to be an active antibiotic agent against rickettsial diseases, primary atypical pneumonia, acute brucellosis, pneumococcal, streptococcal, and staphylococcal infections, urinary tract infections produced by E. coli, A. aerogenes and Strept. fecalis, certain types of infections of the eye, and in subacute bacterial endocarditis when the infecting agent is Strept. fecalis. Its clinical use in forms of extrapulmonary tuberculosis is in a completely experimental stage. It is not recommended in typhoid fever or in infections due to Ps. aeruginosa or P. vulgaris, and it seems to be ineffective in whooping cough. To date, neither chloromycetin nor aureomycin has shown significant signs of systemic toxicity.     

New insight into the diagnosis of fastidious bacterial endocarditis

Sterile blood cultures are noted in one third of patients with infectious endocarditis. Although in half of cases this is due to previous antibiotic therapy, in the other half, the aetiology of culture-negative endocarditis is intracellular bacteria such as Coxiella burnetii or fastidious growing bacteria. Although it was previously considered that the prevalence of such organisms was identical throughout the world, recent investigations on Bartonella endocarditis clearly showed that the aetiology of culture-negative endocarditis is likely to be strongly related to epidemiology of the agent in each country. During the past decade the use of molecular techniques such as PCR with subsequent sequencing to detect or to identify bacteria in valves from patients with infectious endocarditis have considerably improved the aetiological diagnosis. This is especially true in the case of culture-negative endocarditis following earlier antibiotic therapy. However, the fact that DNA remnants of past endocarditis can be detected some time after the acute episode, when the patient has been cured, suggests that the predictive value of these techniques along with the traditional histology and culture need to be evaluated closely.     

CONTROL OF STREPTOCOCCAL THROAT INFECTIONS IN SCHOOLS—A Cooperative Program Followed in Orange County

Attempts to identify streptococcal throat infections on clinical evidence alone do not provide an adequate or reliable index of the prevalence of these infections in the community.Epidemiologic information on streptococcal throat infections based on bacteriological identification permits a more accurate assessment of the situation and more logical and more effective control measures.Recent refinements in laboratory procedures have provided a simple, reliable and relatively inexpensive method for the identification of Group A beta hemolytic streptococci by public health or clinical laboratories.In Orange County a program for the identification of streptococcal throat infections by cooperative action of the medical profession, the health department and the school authorities greatly aided in control of the disease. A voluntary health agency (heart association) made an important contribution toward the success of the control program.     

A study on pathogenicity and mosquito transmission success in the rodent malaria parasite Plasmodium chabaudi adami

We investigated the parasitology, pathogenicity (virulence) and infectivity to mosquitoes of blood infections in mice, of two strains, DS and DK, of the rodent malaria parasite Plasmodium chabaudi adami. Blood infections of DS were found to be highly pathogenic; the asexual parasites in these infections were fast-growing and showed no evidence of selectivity in their infection of host erythrocytes. In contrast to DS, blood infections of DK were much less pathogenic; the asexual parasites were slower-growing and showed a moderate degree of selectivity to a subset of erythrocytes which were not reticulocytes. In both DS and DK infections, infectivity to mosquitoes was highest before the peak of asexual parasitaemia had occurred; usually this did not coincide with the time when gametocyte numbers in the blood were highest. Infections with the pathogenic DS strain in CBA mice produced fewer gametocytes than did the less pathogenic DK strain. The DS strain infections in both CBA and C57 mice were also significantly much less infective to mosquitoes than the DK strain. Investigations by others on the related rodent malaria parasite subspecies, Plasmodium chabaudi chabaudi, have indicated that the mosquito infectivity of blood infections in mice tended to be higher in the more pathogenic (virulent) and lower in the less pathogenic strains of this parasite subspecies. This is the converse of the finding of the present investigation of blood infections of P. c. adami in mice in which a more pathogenic, or virulent, strain (DS) of these parasites was significantly much less infective to mosquitoes than was a less pathogenic strain (DK).     

A review of cardiovascular complications accompanying AIDS

Manifestations of cardiovascular system involvement are not uncommon complications of HIV infection, especially in AIDS patients. However, the frequency of these manifestations is influenced by different variables including: survival prolongation in HIV-infected patients, because of advances in antiretroviral treatment; improvement of immunodepression and reduction in the occurrence of opportunistic infections; adverse effects of some drugs. At present, on the whole cardiovascular complications that are HIV correlated in the western world, including Italy, occur less frequently than in the past. However complications associated with alterations in lipometabolism prevail because they can be promoted by some protease inhibitors in predisposed subjects. The most frequently reported questions and a careful analysis of recent data in the medical literature regarding the most common HIV-correlated cardiovascular complications are discussed in this review.