Impact of reduced resuscitation fluid on outcomes of children with 10-20% body surface area scalds.

'Permissive hypovolaemia' fluid regimes in adult burn care are suggested to improve outcomes. Effects in paediatric burn care are less well understood. In a retrospective audit, outcomes of children from the South West Children's Burn Centre (SWCBC) less than 16 years of age with scalds of 10-20% burn surface area (BSA) managed with a reduced volume fluid resuscitation regime (post-2007) were compared to (a) an historical local protocol (pre-2007) and (b) current regimes in burn services across England and Wales (E&W). Outcomes included length of stay per percent burn surface area (LOS/%BSA), skin graft requirement and re-admission rates. 92 SWCBC patients and 475 patients treated in 15 other E&W burn services were included. Median LOS/%BSA for patients managed with the reduced fluid regime was 0.27 days: significantly less than pre-2007 and other E&W burn services (0.54 days, 0.50 days, p<0.001). Skin grafting to achieve healing reduced post-2007 compared to pre-2007 and remains comparable with other E&W services. Re-admission rates were comparable between all groups. A reduced fluid regime has significantly shortened LOS/%BSA without compromising burn depth as measured by skin grafting to achieve healing. A prospective trial comparing permissive hypovolaemia to current regimes for moderate paediatric scald injuries would help clarify.

Activation of latent precursors in the hippocampus is dependent on long-term potentiation.

The recent discovery of a large latent population of precursor cells in the dentate gyrus of adult mice led us to investigate whether activation of this population is regulated by synaptic activity, thereby explaining the observation that environmental signals can affect neurogenesis. Using a variety of stimulation protocols, we found that only a long-term potentiation (LTP)-inducing protocol activated the latent precursor pool, leading to increased neurogenesis in the dentate gyrus. LTP induced by high-frequency stimulation (HFS) of the perforant pathway in vivo produced a two-fold increase in the number of neurospheres cultured from the stimulated hippocampus, compared with the unstimulated hippocampus. No increase in neurosphere number or neurogenesis was observed when the HFS failed to induce LTP. These results show that LTP can activate latent neural precursor cells in the adult mouse dentate gyrus, thereby providing a direct mechanism for regulating activity-driven neurogenesis. In the future, it may be possible to utilize such learning- or stimulation-induced neurogenesis to overcome disorders characterized by neuronal loss.

Characterisation of the Dunaliella tertiolecta RbcS genes and their promoter activity in Chlamydomonas reinhardtii.

The availability of highly active homologous promoters and terminators is critical in the development of a transformation system for the unicellular microalga Dunaliella tertiolecta. To facilitate transformation of this species, we isolated and characterised two native ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit genes (RbcS) including flanking sequences. The two non-allelic cDNA sequences share approximately 80% identity and have approximately 60% identity to the RbcS genes of Chlamydomonas reinhardtii. The D. tertiolecta RbcS promoter and 3' untranslated regions were shown to drive expression of the bleomycin resistance gene (ble) in C. reinhardtii. This is the first demonstration of a heterologous algal promoter being used to drive transgene expression in C. reinhardtii. In addition, promoter deletions were shown to further increase transformation efficiency.

Enhanced in vivo tumour response from combination of carboplatin and low-dose c-myc antisense oligonucleotides.

BACKGROUND: Phosphorothioate oligonucleotides ([S]ODNs) contain a modified phosphate backbone. Antisense [S]ODNs targeted to specific oncogenes have been used to varying success in vivo. Carboplatin is a commonly used chemotherapeutic and is associated with chemoresistance in some human tumours. The potential for combined antisense [S]ODNs and carboplatin chemotherapy has only recently been explored in vivo. MATERIALS AND METHODS: This study examines the effect of c-myc antisense oligomers delivered in isolation as naked DNA and in combination with carboplatin upon the growth kinetics of an in vivo transplantable adenocarcinoma using rodents. RESULTS: Tumours treated with a combination of 600 microg of 15-mer c-myc phosphorothioate antisense oligodeoxyribonucleotide and an intravenous administration of carboplatin (3 mg/kg), demonstrated a significant (p<0.05) retardation in tumour growth kinetics relative to a control. Two mismatch antisense controls did not significantly inhibit tumour growth. C-myc protein studies in tumour sections failed to show significant differences in c-myc expression in any of the treated tumours. CONCLUSION: This study demonstrates that carboplatin affects the relative abundance of c-myc and that combination treatment of carboplatin and c-myc phosphorothioate antisense oligonucleotides in vivo results in synergistic tumour retardation.

Varus derotation osteotomy for the treatment of hip subluxation and dislocation in cerebral palsy: statistical analysis in 73 hips.

The purpose of this study is to review the results of varus osteotomy in patients with cerebral palsy and to determine factors that influence the rates of residual hip displacement. A retrospective chart review of 65 patients who underwent 79 varus osteotomies was performed. Preoperative, postoperative and follow-up radiographs were analyzed for routine radiographic measurements, pelvic obliquity, osteonecrosis (avascular necrosis), joint incongruity or degenerative joint disease. The average follow-up was 5.2 years (range, 1.1-18.4 years). At follow-up, 3 hips were dislocated, 19 were subluxated and 57 were stable (72%). Age at surgery and the degree of preoperative hip displacement had significant effects on outcome. The average age at surgery for initially subluxated hips, which were located at follow-up, was 7.2 years. This was significantly younger (P = 0.008) than initially subluxated hips, which were displaced (10 years of age). Subluxated hips at surgery were also more likely to be located at follow-up than dislocated hips.

Effect of surgery on the nontreated hip in severe cerebral palsy.

We studied the fate of the nonoperated hip in 35 patients with cerebral palsy who underwent surgical stabilization for unilateral hip subluxation (24 patients) or dislocation (11 patients). Review of medical records and radiographs was performed and analysis was accomplished on the effect of preoperative and radiographic variables on the radiographic outcome of the nonoperated hip. The average age at surgery was 5.5 years and at follow-up was 9.7 years, with an average follow-up of 4.2 years. Before subsequent surgery (in 15 nonoperated hips) or at follow-up, 10 of the nonoperated hips were dislocated and 16 hips were subluxated. Hips were stable and less likely to have surgery if they had a lower initial migration index and higher center edge angles. We conclude that there are few indications for unilateral hip surgery in patients with diplegia or quadriplegia undergoing initial hip stabilization surgery, especially if any degree of dysplasia is present.

A microsphere-liposome (microplex) vector for targeted gene therapy of cancer. II. In vivo biodistribution study in a solid tumor model.

Cationic liposomes are commonly used for transfection of plasmids into mammalian cells, while microspheres have been traditionally used for selective delivery of anticancer agents into tumor vasculature. We have developed a novel vector, comprised of cationic liposomes electrostatically bound to ion-exchange microspheres (termed 'microplex') for targeted gene therapy of solid tumors. The delivery modes tested in a rat solid tumor model were free plasmids, plasmids bound to microspheres, to liposomes, or to the combination vector. The greatest amount of chloramphenicol acetyltransferase (CAT) reporter gene expression in tumors was achieved using the microplex vector; 3.4-fold compared with free, and 1.8-fold compared with both microspherical and liposomal deliveries (p < 0.01). Tumor-to-normal kidney tissue CAT expression ratios were as follows: free 1.9:1; microspherical 3.7:1; liposomal 1.4:1 and microplexical 2.7:1. Expression between the two types of tissues was significantly different (p < 0.01) for all delivery modes. Microspheres targeted the plasmids to the tumors, while the action of cationic liposomes on cellular membranes allowed more plasmids to breach the cell membrane. This study has proven that the novel microplex vector is capable of selective delivery of genes to tumors and has the potential to target genes in clinical trials.

A Comparison of Three Cytotoxicity Endpoints in the Corneal HCE-T Model.

The prediction of ocular irritation potential from in vitro assays still presents a problem, despite a number of validation trials. A study with coded cosmetic formulations, for which historic in vivo data were available, has been conducted with a human corneal multi-layered model system. This corneal model, the HCE-T model, was developed by using HCE-T cells, a transfected human corneal epithelial cell line. The relative effectiveness of three endpoints that provide a measure of cytotoxicity in the HCE-T model was evaluated. Cell viability immediately after exposure to the test materials was determined by using the MTT and Alamar Blue™ (AB) assays, and, 24 hours later, by using the MTT, AB and lactate assays. Viability measurements with the MTT, AB and lactate assays gave similar dose-response curves at the 24-hour endpoint. One formulation (an anti-dandruff shampoo) caused a less severe drop in viability in assays conducted immediately after the exposure than at the 24-hour time-point. There was little deterioration in viability with the other test materials. The ranking of the test formulations on the basis of relative loss of viability and release of lactate resulted in the same order as for the Modified Maximum Average Draize Test Score. Comparison of the HCE-T model cytotoxicity assay results with historic in vitro data from two different cytotoxicity assays, conducted by using fibroblast monolayer cultures and the same materials, indicated that the multi-layered corneal model had a greater predictive ability. The results of a blind trial with the lactate assay in two laboratories indicated that the techniques required were transferable between laboratories. The lactate results were reproducible between laboratories, even when cultures derived from different passage human corneal cells were tested, provided that the passage number was below 20.

Comparison of nickel-titanium and stainless-steel spreader penetration and accessory cone fit in curved canals.

The purpose of this study was to compare nickel-titanium and stainless-steel spreader penetration in curved canals. Twenty prepared plastic blocks with a 30 degrees curvature were used for each part of the study. In part 1, the force required to insert each spreader to within 1 mm of working length in an empty canal was measured. In part 2, the force required to insert each spreader to within 3 mm of working length was measured in a canal containing a master cone. In part 3, the depth of penetration of each spreader with a master cone in place using a 1.5 kg force was measured. Additionally, in part 3, the depth of penetration of the first accessory cone was measured. The results from part 1 showed that a nickel-titanium spreader required significantly less force than a stainless-steel spreader (0.30 kg vs. 0.59 kg). In part 2, a nickel-titanium spreader required significantly less force than a stainless-steel spreader (1.56 kg vs. 2.42 kg). In part 3, a nickel-titanium spreader penetrated significantly deeper than a stainless-steel spreader (15.0 mm vs. 14.0 mm). There was no significant difference in the depth of penetration of the first accessory cone used after either spreader (0.8 mm vs. 0.7 mm). Therefore, the potential for vertical root fracture in curved canals during lateral condensation may be minimized by using nickel-titanium spreaders.