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Vagotomy modifies but does not eliminate the increase in body temperature following burn injury in rats.

INTRODUCTION: Rats with burn injury to > or = 50% of the BSA demonstrate a significant increase in body temperature (T(B)) during light hours, and decrease of circadian variation in T(B). This study investigated the hypothesis that part or all of the signal for increased T(B) in rats with burn injury is transmitted to the central nervous system by way of afferent vagal fibers. METHODS: Four groups of animals were studied: Burn-Sham Vagotomy; Control-Sham Vagotomy; Burn-Vagotomy and Control-Vagotomy. Anesthetized animals had bilateral subdiaphragmatic vagotomy (VagX) or sham VagX performed, and AM radio transmitters were implanted in the peritoneal cavity for the purpose of recording T(B) and activity index (AI). Following a one week recovery period, burn animals received scald burns equal to 50.3 +/- 2.5% BSA. RESULTS: The main effect of VagX upon T(B) was a small but significant reduction in T(B) during dark hours. At least 87% of this decrease in post-burn T(B) was also present for the control vagotomy group. Vagotomy had no effect on the increase in T(B) for the burn groups during light hours. During dark hours, VagX significantly reduced T(B) in burns and controls, and burn injury significantly reduced activity. CONCLUSIONS: One may speculate that afferent vagal signals are partially responsible for maintenance of T(B), with or without burn injury, and that the major signal for increase in T(B) for animals with burn injury is not a neural one via afferent vagal pathways.

Afferent Pathways↗

Role of mast cells in the pathogenesis of postburn inflammatory response: reactive oxygen species as mast cell stimulators.

Thermal trauma has a direct effect on mast cells, triggering the secretion of histamine. This secretion leads to an enhanced xanthine oxidase activity and an increased production of reactive oxygen species (ROS), the latter being produced after burns through differing mechanisms. As ROS have been shown to have deleterious effects on cellular membranes, a lesion of the mast cell membrane could close the circle of autoinjury due to the vasoactive actions of mast cell mediators. Our studies were designed to assess the potentiality of ROS as stimulators of mast cell degranulation after burns by comparing two groups of rats treated, respectively, with SOD and saline solution after a scald injury. Plasma levels of tryptase and histamine were analyzed as markers of mast cell activity. A comparison of the mean increases of tryptase between baseline and 3-h postburn levels in the two groups shows significant differences (p < 0.001) (control: 0.13+/-0.04, SOD: 0.03+/-0.01). When comparing the mean increases between the baseline and 3 h postburn levels of histamine in the two groups, significant differences were also found (p < 0.001) (control group: 2.70+/-0.57. SOD group: 1.22+/-0.32). The lower levels of histamine and tryptase induced by SOD provides indirect evidence that ROS are involved in the process, causing the release of such mediators by mast cells, which may in turn suggest that ROS can act as stimulators of mast cell degranulation in burns.

Animals↗

Effectiveness of human amnion preserved long-term in glycerol as a temporary biological dressing.

Human amnion as a temporary biological wound dressing has remained a beneficial and cost-effective means of treating burns in developing countries. The aim of this study was to determine whether human amnion that has undergone long-term preservation in glycerol is an effective biological dressing compared to fresh amnion and glycerol-preserved human skin. Samples of human amnion and skin were preserved in sterile containers of 85% glycerol at 4 degrees C for over a year. Dorsal full-thickness or split-thickness skin wounds were produced in rats. The defects were divided into four areas, each of which was covered with preserved amnion, fresh amnion, preserved skin, or left uncovered as a control. The materials on the wounds were evaluated macroscopically and microscopically after 2, 4, 7, 10 and 14 days. The primary take or adherence of the grafts on full-thickness wounds was evaluated at 4 and 7 days, and material performance was scored based on several macroscopic and microscopic criteria. The bacteria levels reducing effect of the materials were examined by quantitative bacteriology in heavily infected full-thickness scald burn wounds of rats. Qualitative cultures confirmed that the storage conditions the materials were subjected to for over a year were aseptic and that the amnion and skin had maintained their characteristic properties. All materials were found effective on partial-thickness rat wounds as a cover under which re-epithelialization was completed by 7 days. The preserved skin performed better than either preserved or fresh amnion on full-thickness wounds but the performance of preserved amnion was comparable to that of fresh amnion. Glycerol-preserved amnion was found to be as effective as fresh amnion or skin in terms of decreasing bacterial levels in infected rat burn wounds. Amnion stored in glycerol is reliable and effective for a long period of time. Amnion banking could provide an unlimited quantity of biologic dressing for burn treatment at low cost, a factor that is particularly important in developing countries.

Amnion↗

Ten years epidemiological study of paediatric burns in Manipal, India.

Three hundred and nine children of burns injuries treated over last 10 years (1989-1998) in Kasturba Hospital, Manipal (India) were studied retrospectively and were analysed for incidence, severity, extent, causes, risk factors and overall mortality. Children of age < 5 years were affected more than children of age > 5 years (76.1 vs. 23.9%). Females were affected more than males (74.1 vs. 25.9%). Most of the children received burn injuries in the range of 0 to 20% BSA (63.1%). Scald (72.5%) followed by flame (22.7%) and electrical burn (3.2%) were most common cause of burn injuries. Overall paediatric burn mortality was 7.4%.

Age Distribution↗

Cooling the burn wound: evaluation of different modalites.

A study was undertaken to investigate the cooling and healing effect of different modalities: Melaleuca Alternifolia Hydrogel (Levtrade International (Pty) Ltd.) was compared with tap water as a coolant following application onto a fresh deep partial thickness hot water burn in a porcine model. Four identical circular scalds were created on the backs of 10 pigs. One wound was not treated and served as a control. The other 3 wounds were either cooled with tap water (15 degrees C) or had Melaleuca Hydrogel dressing applied immediately, or after a 30 min delay. Intradermal temperatures were monitored in all wounds: preburn, during the burn and at regular intervals for 1 h. The wounds were biopsied for histological assessment. These samples were repeated at 24 h and 3 weeks. The mean decrease in final temperature at 1 h was in comparison to the preburn temperature; control +0.44 degrees C (i.e. a temperature increase); water -7.82 degrees C; Melaleuca Hydrogel -3.87 degrees C; Melaleuca Hydrogel after 30 min delay -2.67 degrees C. Clinical and histological assessment at 21 days indicated more rapid healing in both the Melaleuca Hydrogel and water-cooled burns compared with the untreated controls. Effective cooling of the burn wound and an increased rate of wound healing was achieved by both repeated tap water compresses and by immediate or delayed application of Melaleuca Hydrogel. Cooling is an effective means to reduce tissue damage and increase wound healing.

Administration, Topical↗

Analysis of the genes flanking xabB: a methyltransferase gene is involved in albicidin biosynthesis in Xanthomonas albilineans.

Transposon mutagenesis and complementation studies previously identified a gene (xabB) for a large (526kDa) polyketide-peptide synthase required for biosynthesis of albicidin antibiotics and phytotoxins in the sugarcane leaf scald pathogen Xanthomonas albilineans. A cistron immediately downstream from xabB encodes a polypeptide of 343aa containing three conserved motifs characteristic of a family of S-adenosyl-L-methionine (SAM)-dependent O-methyltransferases. Insertional mutagenesis and complementation indicate that the product of this cistron (designated xabC) is essential for albicidin production, and that there is no other required downstream cistron. The xabB promoter region is bidirectional, and insertional mutagenesis of the first open reading frame (ORF) in the divergent gene also blocks albicidin biosynthesis. This divergent ORF (designated thp) encodes a protein of 239aa displaying high similarity to several IS21-like transposition helper proteins. The thp cistron is not located in a recognizable transposon, and is probably a remnant from a past transposition event that may have contributed to the development of the albicidin biosynthetic gene cluster. Failure of 'in trans' complementation of thp indicates that a downstream cistron transcribed with thp is required for albicidin biosynthesis.

Amino Acid Sequence↗

Albicidin antibiotic and phytotoxin biosynthesis in Xanthomonas albilineans requires a phosphopantetheinyl transferase gene.

Xanthomonas albilineans produces a family of highly potent antibiotics and phytotoxins called albicidins, which are key pathogenesis factors in the systemic development of leaf scald disease of sugarcane. A gene (xabA) required for albicidin biosynthesis encodes a peptide of 278 amino acids, including the signature sequence motifs for phosphopantetheinyl transferases (PPTases) that activate polyketide and non-ribosomal peptide synthetases. The Escherichia coli entD gene, which encodes a PPTase involved in biosynthesis of enterobactin (a siderophore), restored biosynthesis of albicidin (a DNA replication inhibitor) in X. albilineans Tox- LS156 (xabA::Tn5). We conclude that XabA is a PPTase required for post-translational activation of synthetases in the albicidin biosynthetic pathway. This is the first antibiotic or toxin biosynthesis gene characterized from the genus Xanthomonas, and the first demonstration that antibiotic synthetases in the Pseudomonadaceae, like those in the Enterobacteriaceae and in Gram-positive bacteria, can require activation by a PPTase. Coupled with the recent demonstration of a separate albicidin biosynthetic gene cluster, the results indicate the possibility for overproduction of albicidins,which allows better understanding and application of these potent inhibitors of prokaryote DNA replication.

Amino Acid Sequence↗

Genetic analysis of exfoliative toxin A-producing Staphylococcus aureus isolated from mastitic cow's milk.

Exfoliative toxin A (ETA), produced by Staphylococcus aureus, is the causative agent of staphylococcal scalded-skin syndrome (SSSS) in children. Recently, we reported that ETA was detected by reverse passive latex agglutination in three isolates of S. aureus from cow's milk, but that these ETA-positive isolates did not cause the so-called Nikolsky sign in neonatal mice. In this study, therefore, the eta gene encoding ETA and regulatory genes of these bovine isolates were analyzed by the polymerase chain reaction (PCR) and sequencing. The eta gene was amplified from three bovine isolates by PCR and their resulting nucleotide sequences found to correspond to the eta gene from the human isolate, except for three nucleotides in the upstream region of the eta open reading frame (ORF). An accessory gene regulator (agr), which is a global regulatory locus, was detected in these bovine isolates by PCR amplification. In addition, the ORF (J-4), located 120 bp upstream from the eta ORF of the human isolate, was also amplified from these bovine isolates, with their nucleotide sequences differing at 32 positions from the human isolate. Bovine and human ORF J-4 equally enhanced production of ETA in the recombinants of the eta gene, suggesting that the variation in bovine ORF J-4 may be not be the cause of the difference in amount of ETA produced by bovine and human isolates.

Animals↗

Phage conversion of exfoliative toxin A in Staphylococcus aureus isolated from cows with mastitis.

An exfoliative toxin produced by Staphylococcus aureus is the causative agent of staphylococcal scalded-skin syndrome (SSSS) in young children. Recently, we reported that only few isolates of S. aureus from bovine mastitis contained the eta gene encoding exfoliative toxin A (ETA) and produced ETA in vitro. In this study, we isolated temperate phages from two ETA-positive bovine isolates of S. aureus by treatment with mitomycin C. Polymerase chain reaction (PCR) assay of the phage genomes suggested that the temperate phages carried the structural gene for ETA. Moreover, the nucleotide sequence analysis of the PCR products revealed that the eta gene was located very close to an amidase gene on the phage genomes. The nucleotide sequence for the amidase gene of the bovine phage (bovine phi ETA) differed at nine positions from that of the amidase gene of phi ETA from a human isolate reported by Yamaguchi et al. [Mol. Microbiol. 38 (2000) 694], suggesting that eta-converting phages are heterogeneous. Bovine phi ETA had a head with a hexagonal outline and a non-contractile and flexible tail. Bovine phi ETA was able to lysogenize ETA-negative bovine isolates of S. aureus, and the lysogenized S. aureus isolates had the ability to produce ETA. These results suggest the possibility of horizontal transmission of the eta gene by temperate bacteriophages among bovine isolates of S. aureus.

Amidohydrolases↗

16S rDNA sequence analysis of Xylella fastidiosa strains.

The 16S rDNA encoding the small subunit ribosomal RNA were amplified by PCR, cloned, and sequenced from 16 strains of Xylella fastidiosa originating from nine different hosts. In pair-wise comparisons, X. fastidiosa strains showed a maximum variation of 1.0% or 14 nucleotide positions. When all 16 sequences were considered as a set, 54 variable positions were found. Analysis of the sequence data indicated that the X. fastdiosa strains formed three rDNA groups. Group one includes Pierce's disease and mulberry leaf scorch strains; Group two, periwinkle wilt, plum leaf scald, phony peach, oak leaf scorch, and elm leaf scorch strains; and Group three, citrus variegated chlorosis and coffee leaf scorch strains. All X. fastidiosa strains exhibited significantly higher levels of sequence heterogeneity (63 to 83 nucleotide positions) when compared to species from Xanthomonas and Stenotrophomonas. Our data demonstrate that 16S rDNA sequence data could provide valuable information for future classification of X. fastidiosa at the sub-species level.

DNA, Ribosomal↗

Ethanol exacerbates T cell dysfunction after thermal injury.

To understand the mechanism of suppressed immunity following alcohol consumption and thermal injury, we analyzed T cell functions in a mouse model of acute alcohol exposure and burn injury. Mice with blood alcohol levels at approximately 100 mg/dl were given a 15% scald or sham injury. Mice were sacrificed 48 h after injury. Our data demonstrated a 20-25% decrease in Con A-mediated splenic T cell proliferation (p<0.01) and 45-50% decrease in interleukin-2 (IL-2) production (p<0.01) following burn injury compared to the T cells from sham animals. A further decrease in the proliferation (25-30%) and IL-2 production (40-45%) was detected in T cells derived from burned animals receiving alcohol as compared to burn alone. No significant change in the proliferation and IL-2 production was observed in splenic T cells derived from sham-injured mice regardless of alcohol exposure. Additionally, there was no demonstrable difference in splenocyte apoptosis in any treatment group. These results suggest that alcohol consumption prior to burn injury causes a greater decrease in T cell proliferation and IL-2 production compared to either burn or alcohol injury alone that may further attenuate the cell-mediated immunity and thus enhance susceptibility to infection.

Alcohol Drinking↗

Effects of growth hormone and insulin-like growth factor-1 on protein metabolism, gut morphology, and cell-mediated immunity in burned rats.

The effects of recombinant human growth hormone (GH) and insulin-like growth factor-1 (IGF-1) were investigated in burned rats. Sprague-Dawley rats were fed exclusively by total parenteral nutrition and were subjected to 20% third-degree scald burns. The rats were then divided into the following three groups: (1) the GH group received GH at a dose of 1 IU.kg-1.d-1 for 2d (n = 10); (2) the IGF group received IGF-1 at a dose of 4 mg.kg-1.d-1 for 2d (n = 19); and (3) the control group received saline (n = 17). Cumulative nitrogen balance increased significantly in the GH (P < 0.01) and IGF (P < 0.01) groups as compared with the control group. There were no differences in nitrogen balance between the GH and IGF groups. Blood glucose was decreased in the IGF group (P < 0.01) and increased in the GH group (P < 0.05) as compared with the control group. The intestinal villus height and wall thickness of the GH and IGF groups were significantly greater than those of the control group. Delayed-type hypersensitivity was enhanced in both the GH and the IGF groups as compared with the control group (both P < 0.01). Furthermore, the increase in the IGF group was significantly greater than that in the GH group (P < 0.05). It was concluded that both GH and IGF-1 improve protein metabolism and immune responsiveness, as well as promote proliferation of the intestinal mucosa.

Animals↗

Effects of insulin-like growth factor-1 on endotoxin translocation in burned rats receiving total parenteral nutrition.

The purpose of this study was to investigate the effect of insulin-like growth factor-1 (IGF-1) on the translocation of endotoxin from the gut of burned rats. Twenty-eight male Sprague-Dawley rats (7-wk-old) were subjected to 20% full-thickness scald burns and were divided into two groups. One group received 4 mg.kg-1.d-1 of IGF-1 (IGF-1 group, n = 14), and the other received saline (control group, n = 13). All rats were fed exclusively by total parenteral nutrition (TPN). On the second postburn day, rats were killed. The amount of endotoxin in the liver and spleen were measured. RNA from the terminal ileum was extracted, and Northern blot analyses of alpha-tubulin, beta-actin, cell division cycle-2 (cdc2), and immunoglobulin-A (IgA) were performed. Nitrogen balance was improved (p < 0.001), and the wet weight of intestine and its mucosa were increased significantly in the burned rats that received IGF-1. Gene expression of alpha-tubulin and beta-actin were not changed. Cdc2 was elevated (P < 0.05), but IgA was decreased (P < 0.05) in the IGF-1 group. Levels of endotoxin in the liver and spleen were significantly reduced (P<0.05) by the administration of IGF-1. A negative correlation between the levels of endotoxin in the liver and the weight of the intestinal mucosa was observed. In conclusion, IGF-1 improved nitrogen balance, promoted the proliferation of intestinal mucosa and reduced the translocation of endotoxin.

Actins↗

Effect of intravenous omega-6 and omega-3 fat emulsions on nitrogen retention and protein kinetics in burned rats.

The effect of omega-3 fat emulsion on nitrogen retention and kinetics in relation to fatty acid profile were investigated in burned rats receiving total parenteral nutrition (TPN). A fat emulsion of a structured symmetrical triacylglycerol containing only eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (2:1) was prepared. Sprague-Dawley rats were fed by fat-free chow for 2 wk. Then rats were fed exclusively with one of three types of TPN for 7 d. Animals in group C received fat-free TPN (n = 11). Group omega 6 received safflower oil fat emulsion, which accounted for 20% of total caloric intake (n = 11). Group omega 3 received fat emulsion containing only EPA and DHA (1% of total calories, n = 11), in addition to safflower oil emulsion (19% of total calories). On day 5, each rat was subjected to 20% full-thickness scald burns. Rats were sacrificed under ether anesthesia 48 h after burning. The rats in group C became deficient in omega-6 essential fatty acids. Cumulative nitrogen balance was decreased significantly in group omega 6. The rates of whole-body protein synthesis were increased significantly in both groups omega 6 and omega 3. In omega 6, however, the rates of whole-body protein breakdown were increased significantly. In conclusion, the rates of whole-body protein breakdown increased and nitrogen retention was aggravated significantly in animals administered the safflower oil emulsion. Significant increases of urinary excretion of total catecholamine were also observed. Prostaglandin E2 and thromboxane B2 concentrations were not significantly different among three groups. Supplementation with the new omega-3 fat emulsion, however, improved protein metabolism in burned rats receiving TPN.

Animals↗

Role of protein kinase C in cyclic AMP-mediated suppression of T-lymphocyte activation following burn injury.

Major burn injury induces T-lymphocyte dysfunction. Previous studies suggest that prostaglandin (PG) E2, which is elevated post-burn, is the causative factor via a cyclic AMP-dependent process. The present study was conducted to elucidate the mechanism by which cAMP induces T-lymphocyte dysfunction following burn injury. Splenocytes were isolated from mice 7 days after receiving a scald burn covering 25% of their total body surface or sham procedure. ConA-induced proliferation by splenocytes from burned mice was significantly suppressed. Macrophage depletion of the splenocyte cultures abrogated the suppression. Concanavalin A-stimulated proliferation by macrophage-depleted splenocytes was suppressed by PGE2 and dibutyryl cAMP in both groups. The IC50 of these cAMP-elevating agents, however, was approximately 100-fold lower for cells from burned mice, indicating an increased sensitivity to cAMP. PGE2 did not suppress PMA/Ca2+ ionophore-induced T-lymphocyte activation. Addition of PMA to ConA-stimulated cultures prevented the suppression of proliferative responses by PGE2, whereas Ca2+ ionophore had no effect. Thus, the suppression of T-lymphocyte activation following burn injury is macrophage-dependent, related to an increased sensitivity to cAMP and due to an uncoupling of cell surface receptors from protein kinase C activation.

Animals↗

[Staphylococcal and streptococcal pediatric toxic syndrome from 1998 to 2000. Data from the National Center for Staphylococcal Toxemia].

The clinical and microbial settings of staphylococcal and streptococcal toxemia in pediatric patients were investigated by the French National Reference Center for Staphylococcal Toxemia. From 1998 to 2000, the number of cases was low in regard to the usual putative incidence of these toxemia; this low incidence was probably linked to the passive collection of cases. The most significant finding was the evidence of skin infections as the source of the majorities of staphylococcal toxic shock syndrome and staphylococcal scarlet fever as described for streptococcal toxic shock syndrome or nosocomial suppurative infections. Moreover, most of scalded skin syndrome were from pediatric patients and were exceptional in adults. For other syndromes, no significant original findings were observed.

Adolescent↗