Laparoscopic restorative proctocolectomy ileal pouch anal anastomosis: How I do it?

Surgery for ulcerative colitis is a major and complex colorectal surgery. Laparoscopy benefits these patients with better outcomes in context of cosmesis, pain and early recovery, especially in young patients. For surgeons, it is a better tool for improving vision and magnification in deep cavities. This is not the simple extension of the laparoscopy training. Starting from preoperative preparation to post operative care there are wide variations as compared to open surgery. There are also many variations in steps of laparoscopic surgery. It involves left colon, right colon and rectal mobilisation, low division of rectum, pouch creation and anastomosis of pouch to rectum. Over many years after standardisation of this technique, it takes same operative time as open surgery at our centre. So we present our standardized technique of laparoscopic assisted restorative proctocolectomy and ileal pouch anal anastomosis (IPAA).

Kinetics of reaction of nitrogen dioxide with dihydrorhodamine and the reaction of the dihydrorhodamine radical with oxygen: implications for quantifying peroxynitrite formation in cells.

Dihydrorhodamine 123 (RhH2) has been used to detect 'reactive nitrogen species', including peroxynitrite and its radical decomposition products, peroxynitrite probably oxidizing RhH2 to rhodamine (Rh) via radical products rather than directly. In this study, the radical intermediate (RhH(.)) was generated by pulse radiolysis, and shown to react with oxygen with a rate constant k approximately 7 x 10(8) M(-1) s(-1). This fast reaction was exploited in experiments observing Rh being formed slowly (k approximately 4-7 x 10(5) M(-1) s(-1)) from oxidation of RhH2 by nitrogen dioxide in a rate-limiting step, >1000-fold slower than the corresponding oxidation by carbonate radicals. The time-dependent uptake of RhH2 into mammalian cells was measured, with average intracellular levels reaching only approximately 10 microM with the protocol used. The combination of low loading and relatively low reactivity of oxidants towards RhH2 compared to competing cellular nucleophiles suggests rather a small fraction of peroxynitrite-derived radicals (mainly CO3(.-)) may be scavenged intracellularly by RhH2.

The roles of thiol-derived radicals in the use of 2',7'-dichlorodihydrofluorescein as a probe for oxidative stress.

2',7'-Dichlorodihydrofluorescein (DCFH2) is one of the most widely used probes for detecting intracellular oxidative stress, but requires a catalyst to be oxidized by hydrogen peroxide or superoxide and reacts nonspecifically with oxidizing radicals. Thiyl radicals are produced when many radicals are "repaired" by thiols, but are oxidizing agents and thus potentially capable of oxidizing DCFH2. The aim of this study was to investigate the reactivity of thiol-derived radicals toward DCFH2 and its oxidized, fluorescent form 2',7'-dichlorofluorescein (DCF). Thiyl radicals derived from oxidation of glutathione (GSH) or cysteine (CysSH) oxidized DCFH2 with rate constants at pH 7.4 of approximately 4 or approximately 2x10(7) M(-1) s(-1), respectively. Both the rates of oxidation and the yields of DCF were pH-dependent. Glutathione-derived radicals interacted with DCF, resulting in the formation of DCFH* absorbing at 390 nm and loss of fluorescence; in contrast, cysteine-derived radicals did not cause any depletion of DCF fluorescence. We postulate that the observed apparent difference in reactivity between GS* and CysS* toward DCF is related to the formation of carbon-centered, reducing radicals from base-catalyzed isomerization of GS*. DCF formation from interaction of DCFH2 with GS* was inhibited by oxygen in a concentration-dependent manner over the physiological range. These data indicate that in applying DCFH2 to measure oxidizing radicals in biological systems, we have to consider not only the initial competition between thiols and DCFH2 for the oxidizing radicals, but also subsequent reactions of thiol-derived radicals, together with variables--including pH and oxygen concentration--which control thiyl radical chemistry.

The effect of intranasal administration of remifentanil on intubating conditions and airway response after sevoflurane induction of anesthesia in children.

BACKGROUND: Intubation without the use of muscle relaxants in children is frequently done before IV access is secured. In this randomized controlled trial, we compared intubating conditions and airway response to intubation (coughing and/or movement) after sevoflurane induction in children at 2 and 3 min after the administration of intranasal remifentanil (4 mcg/kg) or saline. METHODS: One hundred eighty-eight children, 1-7-yr old, were studied. Nasal remifentanil (4 mcg/kg) or saline was administered 1 min after an 8% sevoflurane N2O induction. The sevoflurane concentration was then reduced to 5% in oxygen, and ventilation assisted/controlled. An anesthesiologist blinded to treatment assignment used a validated score to evaluate the conditions for laryngoscopy and response to intubation. Blood samples for determination of remifentanil blood concentrations were collected from 17 children at baseline, 2, 3, 4, and 10 min after nasal administration of remifentanil. RESULTS: Good or excellent intubating conditions were achieved at 2 min (after the remifentanil bolus) in 68.2% and at 3 min in 91.7% of the children who received intranasal remifentanil versus 37% and 23% in children who received placebo (P<0.01). The mean remifentanil plasma concentrations (+/-sd) at 2, 3, 4, and 10 min were 1.0 (0.60), 1.47 (0.52), 1.70 (0.46), and 1.16 (0.36) ng/mL, respectively. Peak plasma concentration was observed at 3.47 min. There were no complications associated with the use of nasal remifentanil. CONCLUSIONS: Nasal administration of remifentanil produces good-to-excellent intubating conditions in 2-3 min after sevoflurane induction of anesthesia.

Clinical spectrum and histopathologic features of chronic hepatitis C infection in children.

OBJECTIVE: To define the natural history and outcomes of children infected with hepatitis C virus (HCV) at birth or in early childhood. STUDY DESIGN: This retrospective, prospective study identified 60 HCV-infected children through a transfusion look-back program (group 1) and by referrals (group 2). Perinatal/transfusion history, clinical course, and laboratory studies were correlated with findings from 42 liver biopsy specimens. RESULTS: Mean age at infection was 7.1 months, and duration of infection 13.4 years. The sources of infection were blood transfusion (68%), perinatal transmission (13%), and both (7%). Most patients were asymptomatic; three referral patients had advanced liver disease at presentation. Mean alanine aminotransferase level was normal in 25%, 1 to 3 times normal in 62%, and greater than 3 times normal in 13%. Liver biopsy specimens showed minimal to mild inflammation in 71%, absent or minimal fibrosis in 88%, and bridging fibrosis in 12%. Age at infection and serum gamma-glutamyltranspeptidase correlated with fibrosis; serum alanine aminotransferase correlated with inflammation unless complicated by comorbidity. Repeat biopsies within 1 to 4 years in four patients showed no significant progression in three and cirrhosis in one. Two patients died after liver transplantation. CONCLUSIONS: Children with chronic HCV infection are generally asymptomatic. By 13 years after infection, 12% of patients had significant fibrosis. Patients enrolled by referral had more severe liver disease than those identified through the look-back program, demonstrating the importance of selection bias in assessing the long-term outcome of HCV infection.

Synthesis and biological properties of bioreductively targeted nitrothienyl prodrugs of combretastatin A-4.

Nitrothienylprop-2-yl ether formation on the 3'-phenolic position of combretastatin A-4 (1) abolishes the cytotoxicity and tubulin polymerization-inhibitory effects of the drug. 5-Nitrothiophene derivatives of 1 were synthesized following model kinetic studies with analogous coumarin derivatives, and of these, compound 13 represents a promising new lead in bioreductively targeted cytotoxic anticancer therapies. In this compound, optimized gem-dimethyl alpha-carbon substitution enhances both the aerobic metabolic stability and the efficiency of hypoxia-mediated drug release. Only the gem-substituted derivative 13 released 1 under anoxia in either in vitro whole-cell experiments or supersomal suspensions. The rate of release of 1 from the radical anions of these prodrugs is enhanced by greater methyl substitution on the alpha-carbon. Cellular and supersomal studies showed that this alpha-substitution pattern controls the useful range of oxygen concentrations over which 1 can be effectively released by the prodrug.

Classical positioning decreases the cross-sectional area of the subclavian vein.

BACKGROUND: The classic position for subclavian venipuncture is the Trendelenberg position, with the head turned away and placement of a shoulder roll (SR). The purpose of this study was to determine whether this position results in the largest cross-sectional area (CSA) of the subclavian vein (SV). METHODS: Adult volunteers underwent ultrasound imaging of the right SV in supine and Trendelenberg positioning in the following four head and shoulder positions: head neutral with the chin midline (NL), head turned away (TA), head neutral with an SR, and head TA with an SR (TA/SR). The mean CSA of the SV in each position was calculated. Statistical significance was determined using Student's t, Wilcoxon signed rank, and Bonferroni test. RESULTS: Eighteen adults participated in the study. Trendelenberg positioning significantly increased the CSA of the SV in all positions except NL compared to supine positioning (P < .03). An SR significantly decreased SV CSA in all positions. The largest SV CSA was obtained in the TA/Trendelenberg position (1.41 +/- .06 cm(2)). The classic positioning for subclavian cannulation, TA/SR/Trendelenberg, resulted in a significantly smaller CSA than TA/Trendelenberg position (1.27 +/- .06 cm(2), 15% reduction, P < .01). CONCLUSIONS: The classic recommended maneuvers of turning the head and placing an SR significantly reduce the CSA of the SV. Positioning patients in Trendelenberg with the head turned away without an SR optimizes SV size. Positioning patients in this manner may serve to reduce the morbidity associated with percutaneous access of the SV.

Peer review interrater concordance of scientific abstracts: a study of anesthesiology subspecialty and component societies.

Abstracts presented at anesthesiology subspeciality and component society meetings are chosen by peer review. We assessed this process by examining selection criteria and determining interrater concordance. For the societies studied, the level of reviewer agreement ranged from poor to moderate, i.e., slightly better than by chance alone. We hypothesize that having clearer evaluation criteria, scoring systems with interval scales, and assessment based on quality can strengthen the peer review process.

Excitatory amino acids and magnesium sulfate in neonatal asphyxia.

OBJECTIVE: The excitatory amino acids (EAA); glutamate and aspartate are released into the cerebrospinal fluids (CSF) of asphyxiated newborns. The objectives of this study were: (a) to examine the relation of the concentration of EAA in the CSF with the degree of brain injury, (b) To determine the time of the release of these EAA into the CSF, and (c) to detect the effect of magnesium sulfate (MgSO(4)) on their levels. DESIGNS AND METHODS. A randomized controlled trial was conducted on 47 full term asphyxiated newborns. Twenty three infants received an intravenous 10% solution of MgSO(4) at a dose of 250 mg/kg within the first 24h of life while the other 24 newborns received isotonic saline (0.9%) of an equal volume. Levels of glutamate and aspartate were measured before and 72 h after giving the trial solution. Results. In the study population (n=47) both glutamate and aspartate were significantly elevated in infants with higher grades of HIE compared to those with lower grades (P=0.013 and 0.031, respectively). Compared to baseline level, glutamate decreased significantly over time in placebo group (-8.28+/-14.26, P=0.025) and in MgSO(4) group (-14.39+/-18.72, P=0.005). Glutamate concentration did not differ between groups when measured at baseline (29.26+/-16.31 vs. 31.27+/-22.62, P=0.82) and at 72 h (19.28+/-15.63 vs. 19.6+/-16.54, P=0.87). The change in aspartate concentration over time was not significant in placebo group (-0.45+/-1.96, P=0.34) or in MgSO(4) group (-0.7+/-3.19, P=0.37). Aspartate did not differ between groups when measured at baseline (3.52+/-2.4 vs. 3.92+/-2.59, P=0.49) or at 72 h (2.79+/-1.24 vs. 3.05+/-2.48, P=0.92). Conclusions. The EAA; glutamate and aspartate are released in the CSF of asphyxiated newborns immediately after birth and declined by 72 h. Their initial concentrations correlated with the severity of HIE. Postnatal administration of MgSO(4) did not alter the levels of these 2 EAA.

Male sex and intraventricular hemorrhage.

BACKGROUND: Neonatal mortality and morbidity are sex biased in low birth weight infants. The "Y chromosome effect" has been suggested to be responsible for these maturational differences. OBJECTIVE: To examine the association of sex and neonatal outcomes. DESIGN AND METHODS: A retrospective observational study. Data on all low birth weight infants who survived for >48 hrs were analyzed. Neonatal outcomes were compared between male and female infants. A regression model was used to detect the influence of sex on outcomes after controlling for confounders. Analysis was repeated after stratification of infants into three groups: group A (<1000 g), group B (1000-1499 g), and group C (1500-2499 g). RESULTS: A total of 833 infants were included in this study; 419 female infants and 414 male infants. Male infants had an increased rate of overall intraventricular hemorrhage (IVH) (12.2% vs. 7.2%, p = .02) and IVH grades 3-4 (4.8% vs. 2.3%, p = .04). In addition, male infants had higher bilirubin levels (10.19 +/- 3.1 mg/dL vs. 9.32 +/- 2.94 mg/dL, p = .001). In a regression model, male sex continued to have significant influence on IVH, IVH grades 3-4, death, and bilirubin. In group A, male infants had a significantly increased prevalence of death (regression coefficient, 1.82 +/- 0.65; p = .005) that could not be explained by the increased prevalence of IVH (p = .18) in regression analysis. In group B, male sex was significantly associated with a higher bilirubin level (regression coefficient, 0.94 + 0.3; p = .002). In bivariate analyses, IVH and IVH grades 3-4 were significantly higher in male compared with female infants (19.8% vs. 3.9%, p < .0001) and (8.5% vs. 0.97%, p = .02), respectively, but these differences lost significance in multiple-regression analysis. In group C, male sex positively influenced the prevalence of IVH (regression coefficient, 1.7 +/- 0.57; p = .003). Bilirubin measured higher in male infants (11.38 +/- 2.87 mg/dL vs. 10.19 +/- 3.22 mg/dL, p = .0004), but the difference lost significance in regression analysis (regression coefficient, 0.21 +/- 0.31; p = .5). CONCLUSIONS: Bilirubin, IVH, and death were significantly higher in male infants. In subgroup analysis, significance was retained in group A (<1000 g). Whether a single biological factor is responsible for these differences or perhaps a multi-causal process involving a complex interaction of physiologic, environmental, and pathologic responses needs to be further addressed in future research.

Reactivity of 2',7'-dichlorodihydrofluorescein and dihydrorhodamine 123 and their oxidized forms toward carbonate, nitrogen dioxide, and hydroxyl radicals.

The aim of this study was to investigate the oxidation of two common fluorescent probes, dichlorodihydrofluorescein (DCFH2) and dihydrorhodamine (DHR), and their oxidized forms, dichlorofluorescein and rhodamine, by the radical products of peroxynitrite chemistry, *OH, NO2*, and CO3*-. At pH 8.0-8.2, rate constants for the interaction of carbonate radical with probes were estimated to be 2.6 x 10(8) x M(-1) s(-1) for DCFH2 and 6.7 x 10(8) M(-1) s(-1) for DHR. Nitrogen dioxide interacted more slowly than carbonate radical with these probes: the rate constant for the interaction between NO2* and DCFH2 was estimated as 1.3 x 10(7) M(-1) s(-1). Oxidation of DHR by nitrogen dioxide led to the production of rhodamine, but the kinetics of these reactions were complex. Hydroxyl radical interacted with both probes with rate constants close to the diffusion-controlled limit. We also found that oxidized forms of these fluorescent probes reacted rapidly with carbonate, nitrogen dioxide, and hydroxyl radicals. These data suggest that probe oxidation may often be in competition with reaction of the radicals with cellular antioxidants.

Does gender affect neonatal hyperbilirubinemia in low-birth-weight infants?

BACKGROUND: Neonatal mortality and morbidity are gender-biased in low-birth-weight (LBW) infants. The male disadvantage theory has been suggested to be responsible for these maturational differences. OBJECTIVE: To examine the impact of gender on neonatal hyperbilirubinemia. DESIGN/METHODS: A retrospective observational study. Data on all LBW infants admitted to George Washington University neonatal intensive care unit and surviving for >48 hrs from January 1992 to March 2003 were analyzed. Males and females were compared for gestational age, birth weight, race, Apgar scores at 1 and 5 mins, peak bilirubin levels, sepsis, and intraventricular hemorrhage (IVH). Significant differences were entered in a regression model to detect the influence of gender on bilirubin (Bili). Analysis was repeated after stratification of infants into: group A, <1000 g; group B, 1000-1499 g; and group C, 1500-2499 g. RESULTS: A total of 840 infants were included in this study. When comparing males (n = 407) with females (n = 433), significant differences were detected in birth weight (1,539 +/- 541 vs. 1,428 +/- 549 g; p = .003), IVH (14.2% vs. 9%; p = .025), and Bili (10.1 +/- 3.0 vs. 9.2 +/- 2.8 mg%; p < .001). No differences were detected in gestational age, sepsis, or Apgar 1 and 5. Difference in Bili for the entire group remained significant in the regression model (regression coefficient [RC] = 0.79 +/- 0.22; p < .001). In subgroup analyses: group A Bili (8.4 +/- 2.3 vs. 8.0 +/- 2.0; p = .14) and group B Bili (9.0 +/- 2.1 vs. 9.2 +/- 2.2; p = .51) did not differ in bivariate or multivariate analyses. In group C, Bili was (11.3 +/- 3.1 vs. 10.1 +/- 3.3; p < .001) and remained the only significant difference in the regression model (RC = 1.19 +/- 0.37; p = .001). CONCLUSIONS: Bili in LBW infants is significantly higher in males when compared with females. After stratification to birth weight subgroups, significance is retained in the 1500- to 2499-g group after logistic regression analysis. Bili levels in infants <1500 g are influenced more significantly by factors other than gender, such as sepsis and IVH.

Thrombopoietin level is increased in the serum of asphyxiated neonates: a prospective controlled study.

BACKGROUND: Thrombopoietin (TPO) is a growth factor that controls platelet production. Despite the known association of chronic hypoxia and acute asphyxia with hematologic changes, TPO had not been studied in neonatal asphyxia. OBJECTIVE: To assess TPO concentrations in the serum of asphyxiated and nonasphyxiated neonates, and examine any correlation with the severity of asphyxia. DESIGN/METHODS: This prospective study was carried out on 32 asphyxiated neonates and 30 control subjects admitted at Cairo University Medical Center. Asphyxia was defined if two of the following were found: (1) Apgar score /=-10 and (3) clinical evidence of perinatal asphyxia. Encephalopathy was classified clinically according to Sarnat's stages during the first day of life. Platelet count and TPO level (pg/ml) were measured at 1st, 3rd and 7th day of life. RESULTS: : TPO measured on the first day of life did not differ between cases and controls (900.2+/-526.4 vs 726.6+/-441.9 pg/ml, p=0.2). It increased on the 3rd day of life and was significantly higher in asphyxiated infants compared to controls (1291.4+/-627.9 vs 885.5+/-400.3 pg/ml, respectively; p=0.004). This difference remained significant in a logistic regression model controlling for birth weight, sex and mode of delivery (regression coefficient=476.9+/-146.8; p=0.002). In asphyxiated infants (n=32), encephalopathy was classified as mild (n=17), moderate (n=10) and severe (n=5). TPO correlated with the degree of clinical severity on the 7th day of life (r=0.59, p=0.003). TPO did not differ between survivors (n=24) and nonsurvivors (n=8) within the asphyxia group (1197.1+/-596.8 vs 1613.1+/-605.9 pg/ml; p=0.09). Platelet counts correlated negatively with TPO measured on day 1 (r=-0.415; p=0.02), day 3 (r=-0.64; p=0.001) and day 7 (r=-0.562; p=0.007). CONCLUSIONS: TPO increased and correlated with severity of asphyxia at 3 and 7 days of life. It correlated negatively with the platelet count at all times.

Immunohistochemical analyses of MUC5AC mucin expression in sinus mucosa of children with sinusitis and controls.

OBJECTIVES: The purpose of this study was to analyze MUC5AC protein expression in sinus mucosal specimens of children with and without chronic sinusitis. METHODS: Morphometric, histologic, and immunohistochemical analyses were carried out on sinus mucosa of 7 children with chronic sinusitis and 6 children without sinusitis. RESULTS: MUC5AC protein was expressed in a subset of goblet cells in the surface epithelium, but not in the submucosal glands in either pediatric population. The number of goblet cells that expressed MUC5AC mucin was not significantly different in patients with and without chronic sinusitis. All specimens had similar numbers of goblet cells in the surface epithelium. CONCLUSIONS: The data demonstrate that neither goblet cell hyperplasia nor increased MUC5AC expression occurs in the sinus mucosa of children with chronic sinusitis. This suggests that in contrast to asthma, in which goblet cell hyperplasia is present in the lower respiratory tract, mucus hypersecretion in pediatric chronic sinusitis may involve other secretory cells, eg, submucosal glandular cells, and mucins secreted by these glandular cells.

Novel Trocarless, Scarless Technique for Left Lobe Liver Retraction in Laparoscopic Upper Gastrointestinal Surgeries: Simple, Cost-effective and with Better Cosmesis.

Surgeons always look for ways to reduce the size and number of ports in laparoscopy, where in today's era, we have single-incision laparoscopic surgery (SILS). While doing so, principal 'adequate exposure' should not be compromised. For upper gastrointestinal laparoscopic surgeries, we have adopted a novel technique for retraction of the left lobe of liver, which is described here. Device can be made both single sling and double sling, with help of an infant feeding tube and any routinely used suture material. Placement of device does not require any incision, special energy source, or instrument. It can help in SILS. Detailed technique is described in the text. Operative times did not change significantly. Exposure was excellent. No special instruments or energy devices are required; thus, it is cost-effective. Reducing one port for liver retraction gives better cosmetic results. No liver injury due to the device was noticed in any of the cases. This technique is simpler and cheaper and also gives reasonable cosmetic results compared to other techniques described in the literature.

Oxidation of tetrahydrobiopterin by biological radicals and scavenging of the trihydrobiopterin radical by ascorbate.

One-electron oxidation of (6R)-5,6,7,8-tetrahydrobiopterin (H(4)B) by the azide radical generates the radical cation (H(4)B(*)(+)) which rapidly deprotonates at physiological pH to give the neutral trihydrobiopterin radical (H(3)B(*)); pK(a) (H(4)B(*)(+) <==> H(3)B(*) + H(+)) = (5.2 +/- 0.1). In the absence of ascorbate both the H(4)B(*)(+) and H(3)B(*) radicals undergo disproportionation to form quinonoid dihydrobiopterin (qH(2)B) and the parent H(4)B with rate constants k(H(4)B(*)(+) + H(4)B(*)(+)) = 6.5 x 10(3) M(-1) s(-1) and k(H(3)B(*) + H(3)B(*)) = 9.3 x 10(4) M(-1) s(-1), respectively. The H(3)B(*) radical is scavenged by ascorbate (AscH(-)) with an estimated rate constant of k(H(3)B(*) + AscH(-)) similar 1.7 x 10(5) M(-1) s(-1). At physiological pH the pterin rapidly scavenges a range of biological oxidants often associated with cellular oxidative stress and nitric oxide synthase (NOS) dysfunction including hydroxyl ((*)OH), nitrogen dioxide (NO(2)(*)), glutathione thiyl (GS(*)), and carbonate (CO(3)(*-)) radicals. Without exception these radicals react appreciably faster with H(4)B than with AscH(-) with k(*OH + H(4)B) = 8.8 x 10(9) M(-1) s(-1), k(NO(2)(*) + H(4)B) = 9.4 x 10(8) M(-1) s(-1), k(CO(3)(*-) + H(4)B) = 4.6 x 10(9) M(-1) s(-1), and k(GS(*) + H(4)B) = 1.1 x 10(9) M(-1) s(-1), respectively. The glutathione disulfide radical anion (GSSG(*-)) rapidly reduces the pterin to the tetrahydrobiopterin radical anion (H(4)B(*-)) with a rate constant of k(GSSG(*-) + H(4)B) similar 4.5 x 10(8) M(-1) s(-1). The results are discussed in the context of the general antioxidant properties of the pterin and the redox role played by H(4)B in NOS catalysis.

Modifying rates of reductive elimination of leaving groups from indolequinone prodrugs: a key factor in controlling hypoxia-selective drug release.

3-(4-Methylcoumarin-7-yloxy)methylindole-4,7-diones were synthesised as model prodrugs in order to investigate the correlation between rates of reductive elimination from the (indolyl-3-yl)methyl position with reductive metabolism by hypoxic tumor cells and NADPH: cytochrome P450. Rates of elimination of the chromophore/fluorophore (7-hydroxy-4-methylcoumarin) following one-electron reduction of indolequinones to their semiquinone radicals (Q*-) was measured by pulse radiolysis utilising spectrophotometric and fluorometric detection. Incorporation of a thienyl or methyl substituent at the (indol-3-yl)CHR-position (where R=thienyl or methyl adjacent to the phenolic ether linking bond) significantly shortened the half-life of reductive elimination from 87 to 6 and 2 ms, respectively. Elimination from the methyl substituted analogue can thus compete effectively with the reaction of the semiquinone radical with oxygen at levels typically present in tumours (half-life approximately 1.8 ms at 0.5% O2). Chemical kinetic predictions were confirmed by metabolism in breast tumour MCF-7 cells between 0-2.1% O2. Rates of reductive release of the fluorophore from the non-fluorescent parent indolequinones (R=H, Me, thienyl) were similar under anoxia ( approximately 1.7 nmol coumarinmin(-1)mg protein(-1)) reflecting the similarity in one-electron reduction potential. Whereas coumarin release from the indolequinone (R=H) was completely inhibited above 0.5% O2, the enhanced rate of reductive elimination when R=thienyl or Me increased the metabolic rate of release to approximately 0.35 and 0.7 nmol coumarinmin(-1)mg protein(-1), respectively at 0.5% O2; complete inhibition occurring by 2.1% O2. Similar 'oxygen profiles' of release were observed with NADPH: cytochrome P450 reductase. In conclusion, it is possible to modify rates of reductive elimination from indolequinones to control the release of drugs over a range of tumour hypoxia.

Risk of resistant infections with Enterobacteriaceae in hospitalized neonates.

OBJECTIVE: To determine the risk factors associated with progression from colonization to infection with health care-associated antimicrobial-nonsusceptible Enterobacteriaceae (ANE) in critically ill neonates. STUDY DESIGN: During a 3-year period (1998 to 2000), surveillance rectal cultures were performed on neonates admitted to our Level III neonatal intensive care unit after a cluster of four cases of ANE infection were identified in 1998. ANE were defined as members of the Enterobacteriaceae family that exhibited nonsusceptibility to ceftazidime or laboratory evidence of extended spectrum beta-lactamase (ESBL) production. RESULTS: A total of 1,710 patients were admitted to the neonatal intensive care unit during the study period. Of the 1,710 patients 300 (18%) were excluded from the risk factor analysis. Of the 1,410 remaining neonates the incidence of health care-associated ANE colonization was 17% (240 of 1,410 patients), and 14% of the colonized patients (34 of 240 patients) developed ANE infections. Of the 206 ANE-colonized patients who did not develop disease, 60 (29%) harbored ESBL-producing isolates. Of the 34 ANE-infected patients, 14 (41%) yielded growth of ESBL-producing isolates. Multiple logistic regression analysis revealed that colonized neonates with very low birth weights (<1,000 g) and those who had received prolonged exposures to antimicrobial agents were at increased risk of ANE infections. CONCLUSIONS: Colonization with ANE places hospitalized neonates at risk for development of systemic infections. Very low birth weight (<1,000 g) and prolonged exposure to antimicrobial agents were the only two independent risk factors associated with ANE infection.

Recalibration of the pediatric risk of admission score using a multi-institutional sample.

STUDY OBJECTIVE: Case-mix adjustment is a critical component of quality assessment and benchmarking. The Pediatric Risk of Admission (PRISA) score is composed of descriptive, physiologic, and diagnostic variables that provide a probability of hospital admission as an index of severity. The score was developed and validated in a single tertiary pediatric hospital emergency department (ED) after exclusion of children with minor injuries and illnesses. We provide a multi-institutional recalibration and validation of the PRISA score and test its performance in 4 additional EDs, including patients with minor injuries and illnesses. METHODS: Masked, photocopied, randomly selected medical records of ED patients from 2000 were abstracted and were used to test the performance (discrimination and calibration) of the original PRISA score. This sample differed from the original PRISA sample by including 5 hospitals and including patients with minor injuries and minor illnesses. Independent variables included components of acute and chronic history, physiologic variables, and 3 ED therapies. The dependent variable was hospital admission. PRISA was then recalibrated as needed by using an 80% development sample and a 20% validation sample. Area under the curve and the Hosmer-Lemeshow goodness-of-fit test were used to measure, respectively, discrimination and calibration of the PRISA score after recalibration. We then applied the recalibrated PRISA score to secondary outcomes to test construct validity. We reasoned that a valid measure of ED severity should also be associated with the secondary outcomes of mandatory admissions (admissions using > or =1 inpatient resources) and ICU admissions. RESULTS: The recalibrated PRISA score performed well in all deciles of predicted probability of admission. The area under the curve was 0.81 and the calibration was good (Hosmer-Lemeshow 10.658; df=8; P=.222) for the development sample, and the area under the curve was 0.785 with excellent calibration (Hosmer-Lemeshow 8.341; df=9; P=.500) for the validation sample. The overall development sample had 423.9 admissions predicted and 423 observed; the validation sample had 112.1 predicted and 110 observed. CONCLUSION: The PRISA score has been recalibrated and performs well in EDs of tertiary pediatric hospitals. Comparison with this benchmark may allow individual EDs to improve their performance and may provide insight into best practices.

Increases in serum levels of troponin I are associated with cardiac dysfunction and disease severity in pediatric patients with septic shock.

OBJECTIVES: Myocardial cell injury may contribute to cardiac dysfunction in septic shock. Troponin I is a biochemical marker of myocardial cell injury and death. We hypothesized that troponin I is increased in pediatric patients with septic shock and correlates with cardiac dysfunction and disease severity. DESIGN: Prospective, observational study. SETTING: Children's medical center. PATIENTS: Twenty-three patients with septic shock and cardiovascular failure were enrolled. MEASUREMENTS AND MAIN RESULTS: Serum troponin I was measured at admission and serially over 72 hrs. Within 24 hrs of study enrollment, echocardiograms were performed to determine left ventricular ejection fraction, systolic fractional shortening, heart rate corrected mean velocity of circumferential fiber shortening, and end-systolic wall stress. Requirement for inotropic support (stratified as low, moderate, or high), number of organ system failures, and other demographic data (including Pediatric Risk of Mortality III) were collected. Troponin I was increased on admission in 13 of 23 patients (57%) and at 12 hrs in ten of 22 patients (46%). In all cases, troponin I was maximal within 12 hrs of admission. Admission troponin I was inversely correlated to ejection fraction and fractional shortening and directly correlated to wall stress. Patients who had increased admission troponin I had lower heart rate corrected mean velocity of circumferential fiber shortening (preload and heart rate independent measure of left ventricular systolic function) and higher wall stress (measure of afterload) compared with patients with normal troponin I. Admission troponin I correlated with Pediatric Risk of Mortality III and organ system failure but did not correlate with requirement for inotropic support. CONCLUSIONS: Troponin I was increased in >50% of septic children early in their illness. Increased admission troponin I was associated with decreased measures of systolic cardiac function, as measured by echocardiography, and correlated with severity of illness. Early myocardial cell injury may contribute to the development of subsequent organ failure in septic shock, and measuring troponin I on admission may be helpful in assessing severity of sepsis.