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PubMed1980

Genetic mapping in phage group 2 Staphylococcus aureus.

Two regimens for transformation have been devised for the phage group 2 strains UT0002-19 and UT0017 of Staphylococcus aureus. Strain UT0002-19 produces exfoliative toxin, which is responsible for the clinical manifestations of the staphylococcal scalded skin syndrome, whereas strain UT0017 does not produce exfoliative toxin. A large pool of antibiotic-resistant and auxotrophic mutants from strains UT0002-19 and UT0017 were isolated by using (i) antibiotic gradient plates, (ii) trimethoprim selection, and (iii) nitrosoguanidine mutagenesis, which sometimes was coupled by enrichment with penicillin or methicillin. Transformation frequencies of genetic markers in mutant strains ranged from 10(-6) to 10(-8). Three genetic linkage groups were identified on the strain UT0017 chromosome. The first linkage group was thy-4-lys-5-trp-21-thr-4, the second was pyr-26-nov-9-his-3, and the third consisted of ilv-9 and pen-1, a genetic determinant for beta-lactamase synthesis. Two linkage groups were detected on the strain UT0002-19 CHROMOSOMe. The first was defined as thy-1-lys-5-trp-3-thr-4-ala-8, whereas the second consisted of nov-9 and his-3 markers. A chromosomal locus that controlled synthesis of exfoliative toxin could not be mapped.

Bacterial Toxins↗
PubMed1984

Biochemical and functional alterations in macrophages after thermal injury.

Biochemical and functional measurements of rat pulmonary alveolar macrophages were measured 4 h after 1 10-s, 26 to 28% total body surface area, full-thickness scald burn induced under ether anesthesia. Both phagocytic activity and capacity were significantly decreased to a comparable extent, whereas microbicidal activity was increased almost twofold in macrophages from the burned animals. Concurrent with the decreased phagocytic function was a marked impairment in chemotaxis and random migration of these cells when zymosan-activated serum was used as the chemoattractant. When biochemical parameters were examined, it was demonstrated that, on a per-cell but not total-protein basis, alveolar macrophages from burned animals had elevated levels of RNA, total protein beta-glucuronidase, acid phosphatase, and 5'-nucleotidase. These results raise the possibility that the increased pneumonitis in burned individuals may be due to more complex macrophage dysfunctions than impaired microbicidal activity, as was once thought. Alternatively, the biochemical and functional changes observed may be a reflection of a new population of macrophages appearing in the lungs after thermal injury.

5'-Nucleotidase↗
PubMed2002

Identification of the Staphylococcus aureus etd pathogenicity island which encodes a novel exfoliative toxin, ETD, and EDIN-B.

We identified a novel pathogenicity island in Staphylococcus aureus which contains open reading frames (ORFs) similar to the exfoliative toxin (ET) gene, glutamyl endopeptidase gene, and edin-B gene in tandem and the phage resistance gene, flanked by hsdM, hsdS (restriction and modification system), and IS256. The protein encoded by the ET-like gene showed 40, 59, and 68% amino acid sequence identities with exfoliative toxin A (ETA), exfoliative toxin B (ETB), and Staphylococcus hyicus ETB (ShETB), respectively. When injected into neonatal mice, the recombinant protein derived from the ET-like gene induced exfoliation of the skin with loss of cell-to-cell adhesion in the upper part of the epidermis as observed in histological examinations, just as was found in neonatal mice injected with ETA or ETB. Western blot analysis indicated that the recombinant protein is serologically distinct from ETA and ETB. Therefore, the product encoded by this new ORF is a new ET member produced by S. aureus and is termed ETD. ETD did not induce blisters in 1-day-old chickens. In the skins of mice injected with ETD, cell surface staining of desmoglein 1 (Dsg1), a cadherin type cell-to-cell adhesion molecule in desmosomes, was abolished without affecting that of desmoglein 3 (Dsg3). Furthermore, in vitro incubation of the recombinant extracellular domains of Dsg1 and Dsg3 with the recombinant protein demonstrated that both mouse and human Dsg1, but not Dsg3, were directly cleaved in a dose-dependent manner. These results demonstrate that ETD and ETA induce blister formation by identical pathophysiological mechanisms. Clinical strains positive for edin-B were suggested to be clonally associated, and all edin-B-positive strains tested were positive for etd. Among 18 etd-positive strains, 12 produced ETD extracellularly. Interestingly, these strains are mainly isolated from other sources of infections and not from patients with bullous impetigo or staphylococcal scalded-skin syndrome. This strongly suggests that ETD might play a pathogenic role in a broader spectrum of bacterial infections than previously considered.

Amino Acid Sequence↗
PubMed1986

Cloning and expression of the exfoliative toxin B gene from Staphylococcus aureus.

Exfoliative toxin type B is produced by bacteriophage group II strains of Staphylococcus aureus and is a causative agent of staphylococcal scalded-skin syndrome. In addition to exfoliative toxin B, most isolates also produce a bacteriocin and are immune to the action of the bacteriocin. These phenotypes, as well as resistance to cadmium, were lost after elimination of a 37.5-kilobase plasmid, pRW001, from S. aureus UT0007. Transduction and transformation showed that pRW001 carries the structural genes for four phenotypic characteristics of S. aureus UT0007: (i) exfoliative toxin B production, (ii) bacteriocin production, (iii) bacteriocin immunity, and (iv) resistance to Cd(NO3)2. The exfoliative toxin B structural gene (etb), which is located on a 1.7-kilobase HindIII fragment of pRW001, was cloned in the plasmid pDH5060 and transformed into phage group III S. aureus RN4220. Transformant clones produced extracellular exfoliative toxin B that was biologically active in the neonatal mouse assay. In the Escherichia coli genetic background, the exfoliative toxin B gene was expressed only after being cloned into the positive selection-expression vector pSCC31. The structural gene for cadmium resistance was also isolated on an HindIII fragment of pRW001 cloned in pDH5060. The loci for the exfoliative toxin B gene and the cadmium resistance gene(s) were identified on a restriction map of plasmid pRW001.

Bacterial Toxins↗
PubMed1996

At least two separate gene clusters are involved in albicidin production by Xanthomonas albilineans.

Transposon mutagenesis was used to obtain mutations affecting production of the toxin albicidin in Xanthomonas albilineans, which causes leaf scald disease of sugarcane and is also pathogenic to corn. Transposon Tn5-gusA inserted randomly into genomic DNA of X. albilineans Xa23R1 at a frequency of 10(-4) to 10(-5) per recipient after conjugal transfer from Escherichia coli. Fifty prototrophic mutants defective in albicidin production were isolated from 7,100 Tn5-gusA insertional derivatives tested for toxin production by an antibiosis bioassay. EcoRI fragments containing Tn5 flanking sequences from two mutants (AM15 and AM40) were cloned and used to probe a wild-type Xa23R1 DNA library by colony hybridization. Nine cosmids showed homology to the AM15 probe, and six showed homology to the AM40 probe. Four cosmid clones hybridized to both probes. Forty-five of the 50 defective mutants were restored to albicidin production with two overlapping cosmid clones. Restriction mapping showed that these mutants span a genomic region of about 48 kb. At least one other gene cluster is also involved in albicidin production in Xa23R1. DNA fragments from the 48-kb cluster proved to be very specific to X. albilineans. Some mutants affected in albicidin production retain their ability to colonize sugarcane cultivated in vitro.

Anti-Bacterial Agents↗
PubMed1997

Exfoliative toxin detection using reversed passive latex agglutination: clinical and epidemiologic applications.

A rapid and simple method for detecting exfoliative toxin serotypes A and B from clinical isolates has been developed as a test kit (EXT-RPLA; Denka Seiken Co. Ltd., Niigata, Japan). This method is based on reversed passive latex agglutination. The detection limit of the EXT-RPLA observed for purified exfoliative toxin serotypes A and B was 1 ng/ml. We evaluated the clinical and epidemiologic uses of the EXT-RPLA. A total of 381 isolates of Staphylococcus aureus, 292 from various clinical specimens and 89 from the skin of dermatologic patients, were studied. The EXT-RPLA detected 19 exfoliative toxin producers, including 16 serotype A producers and 3 serotype B producers, but no double producers. The sensitivity and specificity of the EXT-RPLA were confirmed by the newborn mouse bioassay and a PCR assay for the structural genes for exfoliative toxin serotypes A and B (eta and etb, respectively). The overall positivity rate of exfoliative toxin producers was 5.0% (19 of 381), including 16 serotype A isolates and 3 serotype B isolates. Of the 89 isolates from the skin of dermatologic patients, 12 (13.5%) were positive for exfoliative toxin production. Only 2 (1.3%) of the 153 methicillin-resistant S. aureus isolates produced exfoliative toxin, while 17 (7.5%) of the 228 methicillin-sensitive isolates produced exfoliative toxin. The EXT-RPLA assay is a simple and reliable method for detecting exfoliative toxin, and we recommend its use for the rapid diagnosis of staphylococcal scalded skin syndrome. We also recommend its use for detection of this syndrome so that effective control measures can be taken against the spread of this syndrome.

Agglutination Tests↗
PubMed2001

Staphylococcus aureus isolated in cases of impetigo produces both epidermolysin A or B and LukE-LukD in 78% of 131 retrospective and prospective cases.

Clinical symptoms of impetigo and staphylococcal scalded skin syndrome may not only be expressed as the splitting of cell layers within the epidermis but are often accompanied by some localized inflammation. Toxin patterns of Staphylococcus aureus isolates originating from patients with impetigo and also from those with other primary and secondary skin infections in a retrospective isolate collection in France and a prospective isolate collection in French Guiana revealed a significant association (75% of the cases studied) of impetigo with production of at least one of the epidermolysins A and B and the bicomponent leucotoxin LukE-LukD (P < 0.001). However, most of the isolates were able to produce one of the nonubiquitous enterotoxins. Pulsed-field gel electrophoresis (PFGE) of genomic DNA hydrolyzed with SmaI showed a polymorphism of the two groups of isolates despite the fact that endemic clones were suspected in French Guiana and France. The combination of toxin patterns with PFGE fingerprinting may provide further discrimination among isolates defined in a given cluster or a given pulsotype and account for a specific virulence. The new association of toxins with a clinical syndrome may reveal principles of the pathological process.

Adolescent↗
PubMed2005

Sensitive and specific detection of staphylococcal epidermolysins A and B in broth cultures by flow cytometry-assisted multiplex immunoassay.

Two of the most common bacterial skin infections of young infants and children are bullous impetigo due to Staphylococcus aureus and its more acute form, staphylococcal scalded skin syndrome. Epidermolysin A (ETA), ETB and, possibly, ETD are responsible for these diseases, which may appear as epidemics in pediatric patients. We tested the reliability of a flow cytometry-assisted multiplex immunoassay (Bio-Plex system) for the detection of ETA and ETB. The Bio-Plex system was found to be highly specific and highly sensitive for toxin concentrations of between 2 and 80,000 pg/ml. The results of this assay were 100% identical to the results of a PCR-based method. We demonstrated that this test did not generate any cross-reactions with ETD-producing isolates. The level of detection of ETB by this test differed according to culture conditions and from isolate to isolate; these results must be taken into account for diagnostic purposes.

Exfoliatins↗
PubMed2005

Staphylococcus aureus isolates carrying Panton-Valentine leucocidin genes in England and Wales: frequency, characterization, and association with clinical disease.

Staphylococcus aureus isolates carrying the genes that encode for Panton-Valentine leucocidin (PVL), a highly potent toxin, have been responsible for recent outbreaks of severe invasive disease in previously healthy children and adults in the United States of America and Europe. To determine the frequency of PVL-positive isolates sent to the Staphylococcus Reference Unit (United Kingdom) for epidemiological purposes, we tested 515 isolates of S. aureus, and 8 (1.6%) were positive for the PVL locus. A further 470 isolates were selected to explore the association of PVL-positive S. aureus with clinical disease. Of these, 23 (4.9%) were PVL positive and most were associated with skin and soft tissue infections (especially abscesses). The PVL genes were also detected in isolates responsible for community-acquired pneumonia, burn infections, bacteremia, and scalded skin syndrome. Genotyping by pulsed-field gel electrophoresis and multilocus sequence typing revealed that the PVL-positive isolates were from diverse genetic backgrounds, although one prevalent clone of 12 geographically dispersed methicillin-resistant S. aureus (MRSA) isolates was identified (ST80). All 12 isolates were stapylococcal cassette chromosome mec type IVc, had an agr3 allele, and shared a common toxin gene profile (sea-see, seg-sej, eta, etb, and tst negative but etd positive). ST80 strains with similar genetic characteristics have been responsible for community-acquired infections in France and Switzerland. The remaining PVL-positive isolates were mostly methicillin-sensitive S. aureus and belonged to 12 different sequence types, including ST22 and ST30, which are closely related to the most prevalent MRSA clones in United Kingdom hospitals, EMRSA-15 and EMRSA-16, respectively.

Bacterial Toxins↗
PubMed1988

Increasing medical burden of child abuse.

During the decade January 1976 to December 1985, as a result of increased public awareness, the numbers of reported (but not proved) cases of abuse to children living in South Birmingham increased thirty-fold; the incidence of detected and proved cases of abuse increased fourfold. The increase was principally due to bruises, scalds, and neglect. The numbers of serious fractures and brain injuries did not increase. The first case of sexual abuse was reported in 1981 and this was followed by a subsequent rise in the detection rate. The combination of increased reporting, increased incidence of proved abuse, and the necessity for increased supervision of families and intervention has created a serious burden on the child health services that may necessitate a reduction in services to other children. Alternative ways of tackling the problems of child abuse may have to be introduced, but they may have unwelcome consequences.

Child↗
PubMed1988

Measuring severity of injuries to children from home accidents.

The severity of injuries from home accidents was assessed in 402 children under the age of 5 treated in the accident department of an inner London hospital. Our child injury severity scale comprises three grades of severity for six types of injury. There was moderate agreement between the observer and an assessor in categorising the cases. Burns and scalds and poisoning caused more severe injuries than other accidents. A strong correlation was found between the parent's social class and the severity of the accident, but there was no correlation with ethnic group as indicated by the parents' country of birth. The development of a reliable scale of severity is important if programmes of prevention of accidents to children in the home are to be evaluated successfully.

Accidental Falls↗
PubMed1990

The sexually abused battered child.

A total of 130 children were identified in whom both evidence of sexual abuse and non-accidental, non-genital physical injuries (bruises, fractures, scratches, burns and scalds, including failure to thrive) were found. There were 77 girls and 53 boys with mean ages 5.7 and 6.8 years respectively and the peak age between the second and seventh birthdays; this reflects previous reports indicating that physical and sexual abuse predominantly involves young children. Patterns of injury that suggested sexually motivated assault included bruises, scratches, and burns around the lower trunk and genitalia, thighs, buttocks, and upper legs including knees. Pinch and grip marks were found where the child was held. The sexual abuse often involved attempted or achieved penetration of mouth, vagina, or anus, and physical signs were seen relatively more often than in sexually abused children as a whole. Four children died and sexual aggression and child death in the domestic setting may be linked. One in six of 769 physically abused children (16.9%) and one in seven of 949 sexually abused children (13.6%) have suffered both forms of abuse.

Adolescent↗
PubMed2001

Randomised controlled trial assessing the impact of increasing information to health visitors about children's injuries.

AIMS: To assess the effect on health visitor action of providing community health visitors with information on all injury attendances in children under 5 attending an accident and emergency (A&E) department and of providing additional information about each injury. METHODS: Children under 5 years attending the A&E department at Queen's Medical Centre, Nottingham between October 1998 and April 1999 were studied, using a randomised controlled trial with a 2x2 factorial design. All attendances or selected attendances (all children under 1, burns and scalds, poisonings, head injuries, and repeat attendances in preceding six months) were notified. Provision of standard (diagnosis, circumstances surrounding injury, and disposal) or additional information (standard information plus place of injury, number of A&E attendances for injury in previous year, and any information recorded about safety equipment) was noted. RESULTS: Many notifications (56%) do not result in any action. Health visitors were nearly twice as likely to take action of some kind and three times as likely to undertake a home visit on receipt of notifications for selected attendances than on those for all attendances. A greater number of actions per notification were taken on receipt of information about selected attendances. Providing additional information had little effect on the action taken. CONCLUSIONS: A selective policy for notifying community health visitors of child injury attendances at A&E results in a greater proportion of attendances in which the health visitor takes action and in a greater number of actions per attendance being taken. The utility of notifying all injury attendances is questionable, as many do not result in any action.

Accident Prevention↗
PubMed2002

Cross sectional survey of socioeconomic variations in severity and mechanism of childhood injuries in Trent 1992-7.

OBJECTIVE: To determine the relation between morbidity from injury and deprivation for different levels of injury severity and for different injury mechanisms for children aged 0-14 years. DESIGN: Cross sectional survey of routinely collected hospital admission data for injury 1992-7. SETTING: 862 electoral wards in Trent Region. SUBJECTS: 21 587 injury related hospital admissions for children aged 0-4 years and 35 042 admissions for children aged 5-14. MAIN OUTCOME MEASURES: Rate ratios for hospital admission for all injuries, all injuries involving long bone fracture, and all injuries involving long bone fracture requiring an operation; rate ratios for hospital admission for six types of injury mechanism divided by quintiles of the electoral wards' Townsend scores for deprivation. Rate ratios calculated by Poisson regression, with adjustment for distance from nearest hospital admitting patients with injuries, rurality, ethnicity, and percentage of males in each electoral ward. RESULTS: Both total number of admissions for injury and admissions for injuries of higher severity increased with increasing socioeconomic deprivation. These gradients were more marked for 0-4 year old children than 5-14 year olds. In terms of injury mechanisms, the steepest socioeconomic gradients (where the rate for the fifth of electoral wards with the highest deprivation scores was > or =3 times that of the fifth with the lowest scores) were for pedestrian injuries (adjusted rate ratio 3.65 (95% confidence interval 2.94 to 4.54)), burns and scalds (adjusted rate ratio 3.49 (2.81 to 4.34)), and poisoning (adjusted rate ratio 2.98 (2.65 to 3.34)). CONCLUSION: There are steep socioeconomic gradients for injury morbidity including the most common mechanisms of injury. This has implications for targeting injury prevention interventions and resources.

Accidents↗
PubMed1997

Childhood accidents: epidemiology, trends, and prevention.

Accidents are the most common cause of death in children over one year of age. Prevention remains a high priority. We have reviewed the current epidemiology of childhood accidents and their prevention, and made recommendations for the future. In 1992, 559 children died in United Kingdom as a result of an accidents--240 from road traffic accidents and 100 from burns and scalds. Every year 50 children drown. Accidents cause significant disability to children. Many children, up to one in four of the population in urban areas, attend accident and emergency departments, and 5-10% of these are admitted to hospital. Accident risk factors include low social class, psychosocial stress, an unsafe environment, and child developmental disorders. Research has shown that prevention is best achieved by making the child's environment safer, often through legislation. Insufficient resources have been put into both research into childhood injuries and preventive work in communities. Collaboration between health authorities, NHS trusts, local authorities and community networks is vital if success is to be achieved. A national safety agenda for children would focus the attention that this problem deserves.

Accident Prevention↗
PubMed2000

The domestic iron. A danger to young children.

OBJECTIVES: To study the epidemiology of thermal injury caused by the domestic iron in children 5 years old or less. METHODS: Retrospective review of case notes held in the accident and emergency (A&E) department of a large teaching hospital over a 36 month period. Data regarding demographics, site and extent of injury, mechanism of injury and outcome were retrieved. RESULTS: 62 thermal injuries were identified in 59 patients. Of these, 60 were contact burns and two were scalds. The male to female ratio was 2:1. The mean age was 24 months. Fifty five per cent were aged between 1 and 2 years old. The hand was the commonest site of injury (63%) and, of these, two thirds were on the palm. Interestingly 10% occurred on the face. Iron contact burns accounted for 23.5% of all contact burns in this age group over this period. The majority of contact burns were partial thickness and most were less than 1% body surface area. Inadequate supervision is a recurring theme in many of these cases. A suspicion of non-accidental injury was raised in 10 cases and confirmed in nine of these. CONCLUSIONS: Iron burns are common in young children, particularly boys aged between 1 and 2 years old. Most can be treated in the A&E clinic. The potential for serious injury does exist. Non-accidental injury always needs to be considered. Efforts at prevention and increasing public awareness are needed.

Accidents, Home↗
PubMed1995

Determinants of modern health care use by families after a childhood burn in Ghana.

OBJECTIVES: This study examined determinants of modern health care use by families after their child aged 0-5 years sustained a burn injury in the Ashanti Region of Ghana. METHODS: A community based survey of children aged 0-5 years was conducted in 50 enumeration areas in the region. Mothers of all children with scars as evidence of a burn were selected for a follow up interview using a standard questionnaire two to three months later. Determinants of health care use were investigated through a multivariate logistic regression using interview responses from mothers of 617 children for whom report on some treatment was given. RESULTS: Overall, 48% of the burned children were taken to a modern health facility for treatment. Of those taken to a modern health facility, 68% were sent within 24 hours of the burn event. Factors with large adjusted odds ratios for modern health care use included wound infection, burns covering 6% or more of the body surface, and third degree burns. Compared with scalds, children with contact and flame burns were less likely to be taken to a health facility, as were burns to rural children, and those given first aid treatment at home. CONCLUSIONS: It is concluded that families, particularly rural residents, should be educated about appropriate health care seeking practices after a burn.

Adult↗
PubMed2005

Long term effects of a home visit to prevent childhood injury: three year follow up of a randomized trial.

OBJECTIVE: To assess the long term effect of a home safety visit on the rate of home injury. DESIGN: Telephone survey conducted 36 months after participation in a randomized controlled trial of a home safety intervention. A structured interview assessed participant knowledge, beliefs, or practices around injury prevention and the number of injuries requiring medical attention. SETTING: Five pediatric teaching hospitals in four Canadian urban centres. PARTICIPANTS: Children less than 8 years of age presenting to an emergency department with a targeted home injury (fall, scald, burn, poisoning or ingestion, choking, or head injury while riding a bicycle), a non-targeted injury, or a medical illness. RESULTS: We contacted 774 (66%) of the 1172 original participants. A higher proportion of participants in the intervention group (63%) reported that home visits changed their knowledge, beliefs, or practices around the prevention of home injuries compared with those in the non-intervention group (43%; p<0.001). Over the 36 month follow up period the rate of injury visits to the doctor was significantly less for the intervention group (rate ratio = 0.74; 95% CI 0.63 to 0.87), consistent with the original (12 month) study results (rate ratio = 0.69; 95% CI 0.54 to 0.88). However, the effectiveness of the intervention appears to be diminishing with time (rate ratio for the 12-36 month study interval = 0.80; 95% CI 0.64 to 1.00). CONCLUSIONS: A home safety visit was able to demonstrate sustained, but modest, effectiveness of an intervention aimed at improving home safety and reducing injury. This study reinforces the need of home safety programs to focus on passive intervention and a simple well defined message.

Accidents, Home↗