TLR-Stimulated Eosinophils Mediate Recruitment and Activation of NK Cells In Vivo.

Eosinophils like many myeloid innate immune cells can provide cytokines and chemokines for the activation of other immune cells upon TLR stimulation. When TLR-stimulated eosinophils were inoculated i.p. into wild-type mice, and NK cells were rapidly recruited and exhibited antitumour cytotoxicity. However, when mice depleted of CD11c+ cells were used, a marked decrease in the number of recruited NK cells was observed. We postulated that CpG or LPS from the injected eosinophils could be transferred to host cells, which in turn could recruit NK cells. However, by inoculating mice deficient in TLR4 or TLR9 with LPS or CpG-stimulated eosinophils respectively, NK cell recruitment was still observed alongside cytotoxicity and IFNγ production. CpG stimulation of eosinophils produced the pro-inflammatory cytokine IL-12 and the chemokine CXCL10, which are important for NK cell activation and recruitment in vivo. To demonstrate the importance of CXCL10 in NK cell recruitment, we found that CpG-stimulated eosinophils pretreated with the gut microbial metabolite butyrate had reduced expression and production of CXCL10 and IL-12 and concomitantly were poor at recruitment of NK cells and inducing IFNγ in NK cells. Therefore, eosinophils like other innate immune cells of myeloid origin can conceivably stimulate NK cell activity. In addition, products of the gut microbiota can be potential inhibitors of NK cell.

Botulinum toxin assessment, intervention and after-care for lower limb spasticity in children with cerebral palsy: international consensus statement.

Botulinum neurotoxin type-A (BoNT-A) has been used in association with other interventions in the management of spasticity in children with cerebral palsy (CP) for almost two decades. This consensus statement is based on an extensive review of the literature by an invited international committee. The use of BoNT-A in the lower limbs of children with spasticity caused by CP is reported using the American Academy of Neurology Classification of Evidence for therapeutic intervention. Randomized clinical trials have been grouped into five areas of management, and the outcomes are presented as treatment recommendations. The assessment of children with CP and evaluation of outcomes following injection of BoNT-A are complex, and therefore, a range of measures and the involvement of a multidisciplinary team is recommended. The committee concludes that injection of BoNT-A in children with CP is generally safe although systemic adverse events may occur, especially in children with more physical limitations (GMFCS V). The recommended dose levels are intermediate between previous consensus statements. The committee further concludes that injection of BoNT-A is effective in the management of lower limb spasticity in children with CP, and when combined with physiotherapy and the use of orthoses, these interventions may improve gait and goal attainment.

Observation of the desorption of mineral nanoparticles from a hybrid Langmuir monolayer: an in situ grazing incidence X-ray (GIXOS) study.

The kinetics and mechanism of the desorption of mineral clay particles originally adsorbed to a phospholipid monolayer has been studied. The desorption was induced by the injection of citric acid solution into the aqueous subphase. To follow the related kinetic processes, grazing incidence diffuse X-ray scattering out of the specular plane (GIXOS) was used with a remarkable time resolution of the order of one minute. From the variations of the electron density profile normal to the surface, the Langmuir desorption isotherm and the related kinetic constants were determined.

Parotid and submandibular botulinum toxin A injections for sialorrhoea in children with cerebral palsy.

The aim of this study was to determine whether botulinum toxin A (BTX-A) injected into the parotid and submandibular salivary glands of children aged 6 to 16 years with cerebral palsy (CP) and relative sialorrhoea significantly decreases their drooling and improves their quality of life. Twenty children (10 males, 10 females; mean age 10y 1mo [SD 3y 8mo]; range 6y 1mo-16y 7mo) with CP, identified as having significant daily drooling (scoring at least 6 on drooling frequency/severity scales) were recruited. BTX-A (2U/kg; maximum 70U) was injected under sedation, using ultrasound guidance (1.4U/kg and 0.6U/kg divided between parotid and submandibular glands respectively). Nineteen children completed the study: 15 with spastic quadriplegia (Gross Motor Function Classification System [GMFCS] Level V); one with dystonia (GMFCS Level V); one with hemiplegia (GMFCS Level II); and two with a mixed pattern of CP (GMFCS Levels III and V). Drooling was assessed by five methods at baseline, and 4 and 12 weeks after injection. Qualitative assessment of drooling frequency and severity scores showed statistically significant reductions at 4 weeks (p<0.001) and 12 weeks (p=0.006). Qualitative assessment of quality of life scores (rated by parents and teachers separately) also significantly improved (p<0.001 and p=0.023 respectively). Quantitative assessments showed that the number of bibs/scarves changed per day was significantly reduced at 4 weeks (p<0.001). There was no side effect from the injections themselves; 89% of parents and children wished for further intrasalivary BTX-A injections in the future. We conclude that percutaneous intrasalivary BTX-A injections into the parotid and submandibular salivary glands can reduce drooling in children with CP and relative sialorrhoea, leading to an improvement in their quality of life.

Inhibition of bacteriophage K proliferation on Staphylococcus aureus in raw bovine milk.

AIMS: To assess the ability of staphylococcal bacteriophage K to inhibit Staphylococcus aureus in raw milk. METHODS AND RESULTS: The ability of bacteriophage (phage) to replicate in milk is important in situations where phage might be used as a therapeutic for bovine mastitis. Phage K was able to replicate normally, leading to elimination of the host culture in milk, which had been previously heat-treated. When raw milk was used under identical conditions, the phages were unable to replicate. Phage adsorption assays were performed and these demonstrated that adsorption of phage was significantly reduced in the raw milk while it was restored in the heat-treated sample (86.50% compared with 99.96% adsorption respectively). When confocal microscopy with a Live/Dead Bac light staining system was employed, it was observed that in raw milk S. aureus formed clusters associated with fat globules, while in heat-treated milk, bacterial agglutination had not occurred. CONCLUSIONS: Raw milk inhibits staphylococcal phage K proliferation. SIGNIFICANCE AND IMPACT OF THE STUDY: This observation has implications for the exploitation of staphylococcal therapeutic phage in milk.

The recombinant phage lysin LysK has a broad spectrum of lytic activity against clinically relevant staphylococci, including methicillin-resistant Staphylococcus aureus.

This study concerns the cloning, characterization, and expression of the lysin (LysK) from staphylococcal phage K in Lactococcus lactis. Lactococcal lysates containing recombinant LysK were found to inhibit a range of different species of staphylococci isolated from bovine and human infection sources, including methicillin-resistant Staphylococcus aureus. LysK thus has potential as an antimicrobial for applications in the prevention and/or treatment of infections caused by staphylococci.

Potential of the polyvalent anti-Staphylococcus bacteriophage K for control of antibiotic-resistant staphylococci from hospitals.

The increasing prevalence of antibiotic-resistant staphylococci has prompted the need for antibacterial controls other than antibiotics. In this study, a lytic bacteriophage (phage K) was assessed in vitro for its ability to inhibit emerging drug-resistant Staphylococcus aureus strains from hospitals and other species of Staphylococcus isolated from bovine infections. In in vitro inhibitory assays, phage K lysed a range of clinically isolated methicillin-resistant S. aureus (MRSA) strains, S. aureus with heterogeneous vancomycin resistance and vancomycin resistance, and teicoplanin-resistant strains. In these assays, 14 of the MRSA strains were initially only weakly sensitive to this phage. However, propagation of phage K on these less-sensitive strains resulted in all 14 being sensitive to the modified phages. The results enforce the principle that, while certain target bacteria may be relatively insensitive to lytic phage, this can be overcome by obtaining modified phage variants from passage of the phage through the insensitive strains. Model in situ hand wash studies using a phage-enriched wash solution resulted in a 100-fold reduction in staphylococcal numbers on human skin by comparison with numbers remaining after washing in phage-free solution. Infusion of the phage into a nonimmunogenic bismuth-based cream resulted in strong anti-Staphylococcus activity from the cream on plates and in broth.

Isolation and characterization of two anti-staphylococcal bacteriophages specific for pathogenic Staphylococcus aureus associated with bovine infections.

AIMS: The aim of this study was to isolate and characterize bacteriophages against bovine Staphylococcus aureus associated with mastitis. METHODS AND RESULTS: We describe the isolation of two anti-staphylococcal phages namely DW2 and CS1 from farmyard slurry. Both phages were characterized by electron microscopy and restriction analysis and shown to belong to the Siphoviridae family. CS1 and DW2 were lytic for representatives of all three clonal groups of Irish mastitis-associated staphylococci. These phages were compared with the previously characterized Myoviridae phage K. Infusion of a cocktail of all three phages at 10(8) PFU ml(-1) into live cow teats resulted in no detectable increase in somatic cell counts in milks indicating that the phages did not irritate the animal. CONCLUSION: Two new anti-staphylococcal phages CS1 and DW2 were isolated and characterized and tested for immunogenicity in animal teats. SIGNIFICANCE AND IMPACT OF THE STUDY: The phages isolated in this study are active against pathogenic S. aureus and may be incorporated into teat-dips or teat-washes as a non-antibiotic prophylaxis against staphylococcal bovine mastitis.

Bromocriptine for the management of autonomic dysfunction after severe traumatic brain injury.

This case report describes a child with severe traumatic brain injury with clinical features of autonomic dysfunction in the immediate post-traumatic period. A history of severe asthma in this child contraindicated the use of beta-blockers, the first line approach, and she was managed with bromocriptine (0.05 mg/kg t.d.s) with good effect.

Personal health records: an evaluation.

A cohort of mothers whose babies were born over one calendar month were followed up eight to 11 months after being given a personal health record for their newborn babies. Eight per cent of mothers lost the records and three more said they had not been given a record while in hospital; a total of 10% of mothers had either lost or misplaced the record. There were no particular demographic characteristics which identified the mothers who were more likely to lose the record. Most parents liked personal health records and used them frequently, as did the community health staff. Most private doctors, however, did not find them useful. Before wider distribution of such records is contemplated health workers should be adequately prepared, especially doctors in the private sector.

Continuous intrathecal baclofen infusion for children with cerebral palsy: a pilot study.

OBJECTIVES: To determine whether continuous intrathecal baclofen infusion (CIBI) would decrease spasticity and improve function in children with spastic cerebral palsy. DESIGN: Prospective study with measurement of changes from baseline assessed at regular intervals for 6 months following the intervention. PATIENTS: Two subjects aged 8 and 9 years with cerebral palsy. Inclusion criteria included severe spasticity and age greater than 4 years. STUDY CENTRE: Department of Rehabilitation, The Children's Hospital at Westmead, Westmead, New South Wales, Australia. RESULTS: Both subjects had clinically significant reductions in lower limb spasticity from a single intrathecal dose of baclofen (screening procedure), and had intrathecal pumps implanted. An intensive physical therapy programme was provided for both subjects in the 1-3-month period after commencing CIBI, in order to maximize functional gains. Modified Ashworth Scale scores remained reduced during the 6-month study period. There was a clinically significant increase in upper limb function for Subject 1. Gross Motor Function Measure scores decreased from 22% to 19% for Subject 1, and increased from 6% to 10% for Subject 2 over the study period. Paediatric Evaluation of Disability Inventory scores for Subject 1 showed a reduction in the level of caregiver assistance required, while Subject 2 showed significant improvement in the functional mobility domain. The major changes noted in the parent questionnaires were reduction in tone, increased range of motion and reduced time taken helping with activities of daily living such as toilet and dressing. No significant side-effect was seen after the screening procedure or after continuous intrathecal infusion. Both subjects' parents felt their child was improved following the intervention. CONCLUSIONS: Children with spastic cerebral palsy can have their spasticity effectively reduced with CIBI. In this study of two children, the clinical improvements were encouraging, and it is proposed that CIBI may be of benefit to those patients whose level of spasticity severely interferes with function. Further studies using multidimensional assessment approaches, with larger numbers of children, are warranted.

Neurological deterioration following head injury: the eyes had it.

A 17-year-old male presented with confusion following a mild head injury. Repeated CT scans of the head were normal. There was a 3 year history of decreased vision, associated with a focal pigmentary retinopathy. On assessment he demonstrated visual agnosia and early dementia. An MRI scan showed symmetrical demyelination of the white matter, particularly of the occipital lobes. The diagnosis of subacute sclerosing panencephalitis (SSPE) was confirmed by the typical EEG findings and the presence of measles antibodies in the CSF. The head injury was the precipitating factor which led to a diagnosis of SSPE. This disease should be considered in young patients who have persisting cognitive dysfunction out of keeping with the severity of the initial trauma. A focal pigmentary retinopathy, especially with macular involvement, should also raise the possibility of SSPE, despite the absence of neurological symptoms initially. We report the longest interval to date between the visual symptoms and onset of neurological signs of SSPE.

Herpes simplex encephalitis in infants.

Two cases are reported of severe encephalitis in infants that were caused by a primary infection with herpes simplex virus type 1. Both patients presented with focal seizures; the clinical diagnosis was confirmed by the rapid appearance of IgM antibody to herpes simplex virus type 1 in cerebrospinal fluid and serum. In spite of the early use of acyclovir, which was administered by the intravenous route, the clinical response was poor and both infants were left with severe neurological impairment.

Genome of staphylococcal phage K: a new lineage of Myoviridae infecting gram-positive bacteria with a low G+C content.

Phage K is a polyvalent phage of the Myoviridae family which is active against a wide range of staphylococci. Phage genome sequencing revealed a linear DNA genome of 127,395 bp, which carries 118 putative open reading frames. The genome is organized in a modular form, encoding modules for lysis, structural proteins, DNA replication, and transcription. Interestingly, the structural module shows high homology to the structural module from Listeria phage A511, suggesting intergenus horizontal transfer. In addition, phage K exhibits the potential to encode proteins necessary for its own replisome, including DNA ligase, primase, helicase, polymerase, RNase H, and DNA binding proteins. Phage K has a complete absence of GATC sites, making it insensitive to restriction enzymes which cleave this sequence. Three introns (lys-I1, pol-I2, and pol-I3) encoding putative endonucleases were located in the genome. Two of these (pol-I2 and pol-I3) were found to interrupt the DNA polymerase gene, while the other (lys-I1) interrupts the lysin gene. Two of the introns encode putative proteins with homology to HNH endonucleases, whereas the other encodes a 270-amino-acid protein which contains two zinc fingers (CX(2)CX(22)CX(2)C and CX(2)CX(23)CX(2)C). The availability of the genome of this highly virulent phage, which is active against infective staphylococci, should provide new insights into the biology and evolution of large broad-spectrum polyvalent phages.

Upper-limb function in Australian children with traumatic brain injury: A controlled, prospective study.

OBJECTIVE: To describe upper-limb function in children with mild and severe traumatic brain injury (TBI), by using both quantitative and qualitative measures. DESIGN: Controlled, prospective cohort study with assessment points initially, at 6 months, and at 2 years after TBI. SETTING: A tertiary pediatric trauma center in Australia. PATIENTS: Fifty-one children, ranging in age up to 14 years, who were consecutive admissions with TBI. On the basis of initial and persisting abnormal coma score and persistence of posttraumatic amnesia, they were assigned to either a mild (n = 26) or a severely injured (n = 25) group. Thirty children admitted with non-TBI trauma were recruited as a control group. MAIN OUTCOME MEASURES: Quantitative measures included Bruininks-Oseretsky Test of Motor Proficiency and Peabody Developmental Motor Scales. Qualitative measures included Brunnstrom Recovery Stages (adapted), categoric scales of muscle tone, grasp used when handwriting, quality of writing product, bilateral activity, and splint use. RESULTS: There was little difference between the groups on the standardized assessments for subjects who could complete the tests. Qualitative measures showed the severe TBI group to have more difficulties with gross arm control, hand control, and hand function. CONCLUSION: Children with severe TBI experience more and persisting difficulties with upper-limb function. It is essential to include both quantitative and qualitative measures in this type of research.

Greenhouse and Field Evaluations of an Autoselective System Based on an Essential Thymidylate Synthase Gene for Improved Maintenance of Plasmid Vectors in Modified Rhizobium meliloti.

The stability of the thy autoselective system, based on an essential thymidylate synthase gene, for enhanced maintenance of plasmid vectors in Rhizobium meliloti was evaluated in the greenhouse and with field-grown alfalfa. The thy autoselective system consists of a free-replicating, broad-host-range plasmid vector containing a copy of the thyA gene from Lactococcus lactis subsp. lactis and a spontaneous mutant of R. meliloti deficient in thymidylate synthase (Thy(sup-)). Under greenhouse conditions, Thy(sup-) rhizobia did not persist in rooting solution alone unless supplemented with thymidine but survived in the presence of the host plant. Nodules formed on alfalfa plants grown in thymidine-free rooting solution and inoculated with Thy(sup-) rhizobia contained only Thy(sup+) revertants. In soil, Thy(sup-) rhizobia were compromised and failed to nodulate alfalfa. Thy(sup-) mutants containing a thy plasmid survived in the rhizosphere and nodulated alfalfa like the wild-type strain. The thy autoselective system was tested in the field with Thy(sup-) strain Rm24T and pPR602, a thy plasmid vector devoid of antibiotic resistance genes and marked with constitutively expressed lacZY. At 80 days after sowing, most rhizobia isolated from the nodules of field-grown alfalfa inoculated with Rm42T(pPR602) contained pPR602. The thy autoselective system proved useful to ensure maintenance of the plasmid vector under greenhouse and field conditions in R. meliloti.

Rabies. A second Australian case, with a long incubation period.

OBJECTIVE: The description of a second case of rabies in Australia, stressing the clinical features and that long incubation periods are possible. CLINICAL FEATURES: A 10-year-old Vietnamese girl presented with fever, shoulder pain, subcutaneous emphysema, swallowing difficulty and agitation. After a period of maniacal behaviour all peripheral and central nervous system function was lost. INTERVENTION AND OUTCOME: Despite maximal intensive care, the patient died. The diagnosis of rabies was made at autopsy. CONCLUSIONS: Rabies occurs in Australia and needs to be considered in the differential diagnosis of acute encephalitis and/or the Guillain-Barré syndrome. Incubation periods of more than six years can occur.

The Westmead Pediatric TBI Multidisciplinary Outcome study: use of functional outcomes data to determine resource prioritization.

OBJECTIVE: To measure functional outcome in the 2 years after traumatic brain injury (TBI) in 2 groups of children and to determine the usefulness of a TBI severity classification system for resource allocation. DESIGN: Prospective inception cohort study with 3 assessment points during the 2 years after trauma. SETTING: Tertiary pediatric trauma center in Sydney, Australia. PARTICIPANTS: Eighty-one consecutive admissions aged 0 to 14 years. Fifty-one were allocated to the Mild (n = 26) or Severe (n = 25) TBI groups, according to preset determinants of severity; 30 admissions with non-TBI trauma constituted the control group. MAIN OUTCOME MEASURES: Standardized psychometric and clinical assessments of cognition, communication and feeding ability, motor performance (ambulation, fine and gross motor), neurologic status, self-care independence, and school/academic performance. RESULTS: Those with Mild TBI severity had no significant deficits at the 2-year data point. In contrast, those in the Severe TBI group demonstrated continued problems with fine motor performance, neurologic status, self care, and school/academic performance. CONCLUSIONS: A classification system has been developed that may be useful in the allocation of children with a TBI, age younger than 15 years, to 1 of 2 severity groups early in their rehabilitation. This classification system may be useful in determining areas of high and low resource prioritization.