Alpha emitter radium-223 and survival in metastatic prostate cancer.

BACKGROUND: Radium-223 dichloride (radium-223), an alpha emitter, selectively targets bone metastases with alpha particles. We assessed the efficacy and safety of radium-223 as compared with placebo, in addition to the best standard of care, in men with castration-resistant prostate cancer and bone metastases. METHODS: In our phase 3, randomized, double-blind, placebo-controlled study, we randomly assigned 921 patients who had received, were not eligible to receive, or declined docetaxel, in a 2:1 ratio, to receive six injections of radium-223 (at a dose of 50 kBq per kilogram of body weight intravenously) or matching placebo; one injection was administered every 4 weeks. In addition, all patients received the best standard of care. The primary end point was overall survival. The main secondary efficacy end points included time to the first symptomatic skeletal event and various biochemical end points. A prespecified interim analysis, conducted when 314 deaths had occurred, assessed the effect of radium-223 versus placebo on survival. An updated analysis, when 528 deaths had occurred, was performed before crossover from placebo to radium-223. RESULTS: At the interim analysis, which involved 809 patients, radium-223, as compared with placebo, significantly improved overall survival (median, 14.0 months vs. 11.2 months; hazard ratio, 0.70; 95% confidence interval [CI], 0.55 to 0.88; two-sided P=0.002). The updated analysis involving 921 patients confirmed the radium-223 survival benefit (median, 14.9 months vs. 11.3 months; hazard ratio, 0.70; 95% CI, 0.58 to 0.83; P<0.001). Assessments of all main secondary efficacy end points also showed a benefit of radium-233 as compared with placebo. Radium-223 was associated with low myelosuppression rates and fewer adverse events. CONCLUSIONS: In this study, which was terminated for efficacy at the prespecified interim analysis, radium-223 improved overall survival. (Funded by Algeta and Bayer HealthCare Pharmaceuticals; ALSYMPCA ClinicalTrials.gov number, NCT00699751.).

Resistance to the mTOR-inhibitor RAD001 elevates integrin α2- and ß1-triggered motility, migration and invasion of prostate cancer cells.

BACKGROUND: Inhibitors of the mammalian target of rapamycin (mTOR) might become a novel tool to treat advanced prostate cancer. However, chronic drug exposure may trigger resistance, limiting the utility of mTOR inhibitors. METHODS: Metastatic potential of PC3 prostate cancer cells, susceptible (PC3(par)) or resistant (PC3(res)) to the mTOR-inhibitor RAD001 was investigated. Adhesion to vascular endothelium or immobilised collagen, fibronectin and laminin was quantified. Motility, migration and invasion were explored by modified Boyden chamber assay. Integrin α and ß subtypes were analysed by flow cytometry, western blotting and real-time PCR. Integrin-related signalling, EGFr, Akt, p70S6kinase and ERK1/2 activation were determined. RESULTS: Adhesion was reduced, whereas motility, migration and invasion were enhanced in PC3(res). The α2 and ß1 integrin subtypes were dramatically elevated, integrins α1 and α6 were lowered, whereas α5 was nearly lost in PC3(res). Activation of the Akt signalling pathway was strongly upregulated in these cells. Treating PC3(par) cells with RAD001 reduced motility, migration and invasion and deactivated Akt signalling. Blocking studies revealed that α2 and ß1 integrins significantly trigger the motile behaviour of the tumour cells. CONCLUSION: Chronic RAD001 treatment caused resistance development characterised by distinct modification of the integrin-expression profile, driving prostate cancer cells towards high motility.

[Gross haematuria as the primary symptom of extramedullary IgM plasmacytoma of the bladder]. / Makrohämaturie als Primärsymptom eines extramedullären IgM-Plasmozytoms der Harnblase.

Haematuria is the most common clinical symptom of bladder cancer. Besides antibiotic treatment of a probably existing urinary tract infection, ultrasonography of the urinary organs, diagnostic cystoscopy (with biopsy if needed), and radiologic evaluation of the upper urinary tract (intravenous urography, computed tomography or magnetic resonance urography, retrograde pyelography) should be done for further evaluation. Atypical manifestations of systemic diseases with bladder infiltration could feign the clinical appearance of chronic cystitis and hinder determination of the correct diagnosis. The case of a 40-year-old man with recurrent gross haematuria due to extremely rare bladder infiltration through an IgM plasmacytoma is presented.

International foodborne outbreak of Shigella sonnei infection in airline passengers.

During 22-24 August 2004, an outbreak of Shigella sonnei infection affected air travellers who departed from Hawaii. Forty-seven passengers with culture-confirmed shigellosis and 116 probable cases who travelled on 12 flights dispersed to Japan, Australia, 22 US states, and American Samoa. All flights were served by one caterer. Pulsed-field gel electrophoresis of all 29 S. sonnei isolates yielded patterns that matched within one band. Food histories and menu reviews identified raw carrot served onboard as the likely vehicle of infection. Attack rates for diarrhoea on three surveyed flights with confirmed cases were 54% (110/204), 32% (20/63), and 12% (8/67). A total of 2700 meals were served on flights with confirmed cases; using attack rates observed on surveyed flights, we estimated that 300-1500 passengers were infected. This outbreak illustrates the risk of rapid, global spread of illness from a point-source at a major airline hub.

[Preclinical studies on the influence of the tyrosine kinase inhibitor AEE788 on malignant properties of renal cell carcinoma cells]. / Präklinische Studien zum Einfluss des Tyrosinkinaseinhibitors AEE788 auf die Malignität des Nierenzellkarzinoms.

BACKGROUND: Conventional therapeutic approaches to treat advanced renal cell carcinoma (RCC) are of limited benefit. Receptor tyrosine kinase inhibitors (RTKI) may open up novel treatment options. In the present study, the effects of the RTKI AEE788 on the growth and adhesion capacity of RCC cell lines were evaluated in vitro. MATERIALS AND METHODS: RCC cells were treated with AEE788, and alterations of tumor growth and tumor cell interaction with vascular endothelium or extracellular matrix proteins were analyzed. Furthermore, the addition of interferon alpha (IFNalpha) was investigated to see whether it may enhance the anti-tumoral potential of AEE788. RESULTS: AEE788 significantly blocked RCC cell growth and adhesion. Analysis of alpha- and beta-integrins revealed distinct alterations of the receptor expression profile and downregulation of integrin-dependent signaling. Growth-blocking effects were further enhanced when the AEE788-IFNalpha combination protocol was applied. In addition, downregulation of integrin-dependent signaling was more intense in the presence of a combination of AEE788 and IFNalpha than with AEE788 monotherapy. CONCLUSIONS: AEE788 exerts significant anti-tumoral properties, particularly when combined with IFNalpha. AEE788 may therefore be an encouraging compound to treat advanced RCC.

New histone deacetylase inhibitors as potential therapeutic tools for advanced prostate carcinoma.

The anti-epileptic drug valproic acid is also under trial as an anti-cancer agent due to its histone deacetylase (HDAC) inhibitory properties. However, the effects of valproic acid (VPA) are limited and concentrations required for exerting anti-neoplastic effects in vitro may not be reached in tumour patients. In this study, we tested in vitro and in vivo effects of two VPA-derivatives (ACS2, ACS33) on pre-clinical prostate cancer models. PC3 and DU-145 prostate tumour cell lines were treated with various concentrations of ACS2 or ACS33 to perform in vitro cell proliferation 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and to evaluate tumour cell adhesion to endothelial cell monolayers. Analysis of acetylated histones H3 and H4 protein expression was performed by western blotting. In vivo tumour growth was conducted in subcutaneous xenograft mouse models. Tumour sections were assessed by immunohistochemistry for histone H3 acetylation and proliferation. ACS2 and ACS33 significantly up-regulated histone H3 and H4 acetylation in prostate cancer cell lines. In micromolar concentrations both compounds exerted growth arrest in PC3 and DU-145 cells and prevented tumour cell attachment to endothelium. In vivo, ACS33 inhibited the growth of PC3 in subcutaneous xenografts. Immunohistochemistry and western blotting confirmed increased histone H3 acetylation and reduced proliferation. ACS2 and ACS33 represent novel VPA derivatives with superior anti-tumoural activities, compared to the mother compound. This investigation lends support to the clinical testing of ACS2 or ACS33 for the treatment of prostate cancer.

Thalidomide for treatment of severe intestinal bleeding.

Apart from its anti-inflammatory activity, which has been used for the treatment of active Crohn's disease, thalidomide is also a potent inhibitor of angiogenesis. We therefore studied the effect of thalidomide in six patients with severe recurrent intestinal bleeding refractory to standard treatment (three patients with Crohn's disease (CD), three patients with obscure intestinal bleeding; mean of 56 blood transfusions within the last 24 months). Bleeding stopped within two weeks after the start of thalidomide in all patients. Haemoglobin normalised without further transfusions for the whole observation period (mean follow up 33 months) while patients needed a mean of 2.2 (CD) and 3.1 (obscure bleeding) blood units/month in the 12 months before treatment. After three months of thalidomide therapy, serum levels of vascular endothelial growth factor were strongly suppressed compared with pretreatment levels. (CD 818 (82) v 129 (86) pg/ml; obscure bleeding 264 (68) v 50 (25) pg/ml). All six patients reported transient fatigue. Peripheral neuropathy was observed in one patient with CD after nine months and was reversible after lowering the dose to 100 mg daily. These results indicate that thalidomide might be useful for patients with otherwise refractory intestinal bleeding.

Thalidomide reduces tumour necrosis factor alpha and interleukin 12 production in patients with chronic active Crohn's disease.

BACKGROUND: Thalidomide improves clinical symptoms in patients with therapy refractory Crohn's disease, as shown in two recent studies. The mechanism of this effect however is still unknown. Suppression of tumour necrosis factor alpha (TNF-alpha) by thalidomide has been suggested as a possible mechanism. However, effects on other cytokines have not been adequately investigated. AIM: The aim of our study was to investigate the effects of thalidomide on cytokine production in patients with inflammatory bowel disease (IBD). METHODS: Ten patients with therapy refractory IBD (nine Crohn's disease, one ulcerative colitis) received thalidomide 300 mg daily in a 12 week open label study. Production of TNF-alpha, interleukin (IL)-1 beta, IL-6, and IL-12 was investigated in short term cultures of stimulated colonic lamina propria mononuclear cells (LPMC) and peripheral blood monocytes (PBMC) before and after 12 weeks of treatment. LPMC were also cultured with graded doses of thalidomide. RESULTS: Three patients discontinued treatment because of sedative side effects. In the other patients, disease activity decreased significantly, with four patients achieving remission. Production of TNF-alpha and IL-12 decreased during treatment with thalidomide: LPMC (TNF-alpha: 42.3 (8.3) pg/ml v 16.4 (6.3); IL-12: 9.7 (3.3) v 5.0 (2.5); p<0.04) and PBMC (TNF-alpha: 62.8 (14.6) v 22.5 (9.2); p<0.02). Production of IL-1 beta and IL-6 did not change significantly. Culturing of LPMC with thalidomide showed a dose dependent decrease in TNF-alpha and IL-12 production. CONCLUSION: The clinical effects of thalidomide in Crohn's disease may be mediated by reduction of both TNF-alpha and IL-12.

Etomidate versus succinylcholine for intubation in an air medical setting.

The objective was to compare rates of successful endotracheal intubation (ETI) and requirement for multiple ETI attempts in patients receiving etomidate (ETOM) versus succinylcholine (SUX). This retrospective study analyzed adults in whom oral ETI was attempted by a helicopter EMS (HEMS) service between July 1997 to July 1999. Data were from records of the HEMS service, which uses a RN/EMTP crew; analysis was with chi-square and logistic regression (P = .05). ETI was successful in 269 (97.8%) of 275 patients, with multiple attempts occurring in 54 (20.1%) of 269. Success rates for SUX (209 of 213, 98.1%) and ETOM (60 of 62, 96.8%) were similar (P = .62). However, of 60 ETOM patients successfully intubated, 7 (11.7%) required rescue succinylcholine. When these patients are tallied as ETOM failures and SUX successes, resultant success rates for ETOM (86.9%) and SUX (98.2%) are different (P = .001). ETOM patients were more likely (P = .004) than SUX patients to require multiple attempts (33.3% versus 16.3%). ETI success rates were high in patients receiving SUX or ETOM as primary adjuncts for airway control, but initial success was more likely with SUX, and ETOM patients were more likely to require multiple attempts.

Appropriateness of endotracheal tube size and insertion depth in children undergoing air medical transport.

OBJECTIVES: Guidelines for pediatric endotracheal tube (ETT) size and insertion depth are important in the helicopter EMS (HEMS) setting, where intubated patients are frequently transported by a non-physician flight crew providing protocol-based care in an environment noted for limitations in clinical airway assessment. The objectives of this study were to characterize, in a HEMS pediatric population, the frequency of compliance with guideline-recommended ETT size and insertion depth, and to test for association between guideline noncompliance and subsequent receiving hospital adjustment of ETT size or insertion depth. DESIGN: This retrospective review analyzed 216 consecutive pediatric (age <14) scene and interfacility HEMS transports, of patients intubated before or during HEMS transport, by an urban two-helicopter HEMS service providing protocol-based care with a nurse/paramedic crew configuration. Patients were transported to one of three receiving academic pediatric referral centers. Pediatric Advanced Life Support (PALS) criteria for ETT size and insertion depth were used to assess guideline-appropriateness of pediatric ETTs. Receiving hospital records were reviewed to determine if post-transport ETT size or lipline adjustment were associated with guideline-appropriateness of size and lipline during HEMS transport. Univariate (chi-square and Fisher's exact) and multivariate (logistic regression) statistics were used to assess and control for the following covariates: intubator group (physician, flight crew, ground EMS), transport year, sex, age, transport type (scene versus interfacility), and receiving hospital. For all analyses, statistical significance was set at the 0.05 level. RESULTS: The initial ETT size was within 0.5 mm of guideline-recommended sizes in 178 (83.6%) of the 213 patients for whom this data were available. Inappropriate sized ETTs were nearly always (32 of 35, 91.4%) too small. Compared to initial ETTs placed by ground EMS personnel, initial ETTs placed by flight crew or physicians were more likely to be appropriate as defined by guidelines (P = .008 and .032, respectively). Receiving hospitals changed the ETT size in 18 (8.3% of 216) cases. Receiving hospital ETT size change was more likely with later transport year (P = .018) and less likely in patients over 2 years of age (P = .03); there was no significant association between receiving hospital ETT size change and intubator group (P > .22) or guideline-appropriateness of ETT size (P = 0.94). The initial ETT insertion depth was within 1 cm of the guideline-recommended lipline in 86 (43.2%) of the 199 patients for whom this data were available. Inappropriate liplines were almost always (109 of 113, 96.5%) too deep. Compared to initial ETT liplines determined by ground EMS personnel, initial liplines determined by flight crew (P = .007), but not physician (P = .47) were more likely to be appropriate as defined by guidelines. Receiving hospitals changed the ETT insertion depth in 72 (33.3% of 216) cases. Receiving hospital lipline change was more likely (P = .03) in patients older than 2 years of age, but was not associated with intubator group (P = .75) or lipline guideline-appropriateness (P = .35). CONCLUSIONS: As judged by frequently used guidelines, pediatric ETTs are often too small and commonly inserted too deep. However, this retrospective study, limited by lack of clinical correlation for ETT size and insertion depth, failed to find an association between lack of ETT size or lipline guideline compliance and subsequent ETT adjustment at receiving pediatric centers. This study's findings, which should be confirmed with prospective investigation, cast doubt upon the utility of pediatric ETT size/lipline guidelines as strict clinical or quality assurance tools for use in pediatric airway management.

Helicopter emergency medical services roles in disaster operations.

Rotor-wing aircraft have previously proven utility in disaster operations, but recent expert reviewers have identified areas of potential improvement in integration of helicopter emergency medical services (HEMS) resources into disaster planning and management. This paper discusses salient points regarding helicopter operations in disaster management, using prior reports regarding rotor-wing aircraft utilization as a basis upon which to provide a concise review of HEMS operations in disasters.

Higher expression of glucocorticoid receptor in peripheral mononuclear cells in inflammatory bowel disease.

OBJECTIVE: Glucocorticoids are widely used in the treatment of inflammatory bowel disease (IBD). Up- and down-regulated expression of glucocorticoid receptors (GR) has been reported for different chronic inflammatory diseases. The aim of this study was to investigate the expression of GR and their apparent dissociation constant (Kd) in patients with IBD. METHODS: Thirty-nine patients with IBD (22 with ulcerative colitis, 17 with Crohn's disease) and 35 normal controls were studied. Twenty-five patients did not receive steroids, 14 patients were treated with steroids. Peripheral blood mononuclear cells from patients and controls were isolated using the Ficoll-Hypaque gradient and a whole cell [3H]-dexamethasone binding assay and Scatchard plot analysis were performed to assess GR number and the apparent dissociation constant. Results were expressed as mean +/- standard deviation. RESULTS: Normal controls showed an expression of 3,969 +/- 1,555 GR per cell with an apparent dissociation constant of 6.16 +/- 3.8 nmol/L. IBD patients without steroids had a significant increase both in the expression of GR per cell (6,401 +/- 2,344; p < 0.0001; Wilcoxon-Mann-Whitney test) and in the apparent dissociation constant (11.02 +/- 7.57 nmol/L; p = 0.006). Expression of GR in IBD patients was suppressed to normal levels under steroid treatment (4,594 +/- 2,237; p = 0.024), but Kd remained elevated (13.56 +/- 9.05 nmol/L). Plasma cortisol levels were not different between IBD patients and the control group. CONCLUSIONS: Our data show a systemic increase in GR expression and a decrease in the affinity to the GR in IBD, in contrast to other inflammatory diseases such as rheumatoid arthritis and asthma. These changes point towards a systemic character of IBD, which might be considered in a decision between topical and systemic treatment.

A genetic linkage map of the baboon (Papio hamadryas) genome based on human microsatellite polymorphisms.

A first-generation genetic linkage map of the baboon (Papio hamadryas) genome was developed for use in biomedical and evolutionary genetics. Pedigreed baboons (n = 694) were selected from the breeding colony maintained by the Southwest Foundation for Biomedical Research. To facilitate comparison with the human genome, the baboon linkage map consists primarily of human microsatellite loci amplified using published human PCR primers. Genotypes for 325 human microsatellites and 6 novel baboon microsatellites were used in linkage analyses performed with the MultiMap expert system. The resulting sex-averaged meiotic recombination map covers all 20 baboon autosomes, with average spacing among loci of 7.2 cM. Direct comparison among homologous (orthologous) loci reveals that, for 7 human autosomes, locus order is conserved between humans and baboons. For the other 15 autosomes, one or more rearrangements distinguish the two genomes. The total centimorgan distances among homologous markers are 28.0% longer in the human genome than in the baboon, suggesting that rates of recombination may be higher in humans. This baboon linkage map is the first reported for any nonhuman primate species and creates opportunities for mapping quantitative trait loci in baboons, as well as for comparative evolutionary analyses of genome structure.

Frequency and costs of laboratory and radiograph repetition in trauma patients undergoing interfacility transfer.

Receiving trauma centers often duplicate laboratory and radiograph testing performed by referring institutions. Our objective was to quantify frequency and costs of this practice. In this prospective study of 104 consecutive interfacility-transported adult trauma patients flown by an emergency medical service to an urban level I center, flight crew noted which labs and radiographs were done at referring hospitals, which tests were sent with patients, and which were repeated on trauma center arrival. Overall, results from 246 of 283 (86.9%) laboratory tests and 241 of 249 (96.8%) radiographs done at referring hospitals were sent with patients. Repetition of laboratory tests at the receiving hospital was frequent regardless of whether initial results were sent (P = .6 by chi2), and radiograph repetition was unrelated to whether sent films were originals or copies (P = .2 by chi2). For these 104 patients, the receiving hospital charged $66,463 for repetition of work-up done at referring facilities.

Flight crew airway management in four settings: a six-year review.

OBJECTIVE: To analyze flight crew airway management in four different settings (in flight, at trauma scenes, in ambulance, and in referring hospitals) and in two different aircraft used by the same helicopter EMS (HEMS) service. The null hypothesis was that there was no association between practice setting, or aircraft, and airway practices or success rate. METHODS: This retrospective study analyzed all patients in whom advanced airway management was attempted by the HEMS service during the study period October 1991 through October 1997. Data used were from flight records of Boston MedFlight Critical Care Transport Service, which uses a nurse/paramedic crew and had a paralytic-assisted intubation protocol in place. Data were analyzed with chi-square and Fisher's exact testing, risk ratio analysis, and logistic regression. RESULTS: Advanced airway management was attempted in 722 patients, with an airway placed in 705 (97.8%). Intubation success was unrelated to site of airway management (p = 0.14), but patients were more likely to have intubation attempted prior to flight (as opposed to in flight) if the crew were in the AS365N2 Dauphin as compared with the BK-117 (p<0.0001). In addition, patients were 0.77 times as likely (95% confidence interval, 0.68-0.88) to receive paralytic-facilitated intubation if airway management occurred in the hospital setting as compared with other sites. CONCLUSIONS: While HEMS crew airway management success rates are equally high in all practice settings, airway management decision making and practice appear to be significantly influenced by practice setting and aircraft type. These data support contentions that nonphysician HEMS crews can effectively manage airways in a variety of circumstances.

Prehospital and emergency department analgesia for air-transported patients with fractures.

OBJECTIVE: To evaluate prehospital and receiving emergency department (ED) analgesia administration in air-transported patients with isolated fractures. METHODS: The study was a retrospective descriptive analysis of flight and hospital records. Study patients were consecutive adults (not pharmacologically paralyzed) with fractures undergoing scene or interfacility helicopter transport during 1994-1996. The study aeromedical program uses two helicopters staffed by a nurse/paramedic flight crew providing protocol-guided care. The receiving ED was in an urban academic Level I trauma center (annual census 65,000). Primary data collected were timing and amount of prehospital and ED analgesia. Analysis was mainly descriptive, with chi-square and nonparametric methods used to compare patients who did and did not receive intratransport fentanyl. RESULTS: 130 patients with isolated fractures underwent air transport during the study period 1994-1996. Of these, 98 (75.4%) received intratransport fentanyl; 20 of 98 (20.4%) received no analgesia in the receiving ED. Patients who did receive repeat analgesia in the receiving ED (n = 78, 79.6% of those receiving prehospital fentanyl) had a median interval of 42.5 minutes (interquartile range 25-100) between ED arrival and analgesia administration; only 62.8% of these patients received their ED analgesia within 60 minutes of arrival. CONCLUSIONS: Some patients receiving intratransport fentanyl received no ED analgesia, and those who did receive ED analgesia often had administration delays surpassing the clinical half-life of intratransport-administered fentanyl. Further study should investigate whether setting-specific analgesia practice differences reflect true differences in analgesia needs, overmedication by prehospital providers, or undermedication by ED staff.

Prehospital fentanyl analgesia in air-transported pediatric trauma patients.

OBJECTIVE: To review the 5.5-year safety record of a protocol guiding fentanyl administration to pediatric trauma patients undergoing aeromedical transport. METHODS: Retrospective review of an urban aeromedical program's trauma scene responses from October 1991 to March 1997 identified the study population as all pediatric patients (age <15 years) receiving fentanyl for analgesia during air transport. Patients receiving fentanyl concurrently with other agents, eg, paralytics, were not studied. The air transport team consisted of a flight nurse and flight paramedic who provided protocol-driven patient care with off-line medical control. Study patients' flight records were reviewed to determine vital signs (systolic blood pressure [SBP], heart rate [HR], and oxygen saturation [SAT]) before and after fentanyl administration. Postfentanyl vital signs were reviewed for evidence of hemodynamic or ventilatory compromise. Pre- and postfentanyl vital signs were compared with the paired t test (P < 0.05). Flight records were also analyzed for narrative information, eg, naloxone administration and assisted ventilation, indicative of fentanyl side effects. RESULTS: Fentanyl (0.33-5.0 microg/kg) was administered 211 times to 131 patients who had a median age of 6.2 years (0.1-14 years), median Glasgow coma score (GCS) of 9 (3-15), and a mean pediatric trauma score of 8.3+/-2.4. Seventy-nine (60.3%) patients were intubated; these patients received 139 (65.9 %) of the 211 total fentanyl doses. No adverse effects from fentanyl were noted in flight record narratives. The median interval between fentanyl administration and postfentanyl vital sign assessment was 9.5 minutes (1-35 minutes). Median postfentanyl changes in SBP and HR were -4.7 and -2.9%, respectively. No patient became hypotensive after fentanyl administration. In nonintubated patients, mean postfentanyl SAT (99.2+/-1.3%) was not significantly different (P = 0.70) from prefentanyl SAT (99.1+/-1.3%), and no patient was noted to have clinically significant SAT decrement after fentanyl. CONCLUSION: Retrospective review of more than five years of prehospital fentanyl administration revealed no untoward events. Although prospective definitive demonstration of fentanyl's field use is pending, it is reasonable to continue discretionary fentanyl administration to injured pediatric children in pain.

Rapid screening and comparison of human microsatellite markers in baboons: allele size is conserved, but allele number is not.

We have developed a rapid screening method for cross-species amplification of homologous microsatellite loci and studied some of the characteristics of those loci in the species of origin (human) and the test species (baboon). We tested a matrix of 64 combinations of magnesium concentration and annealing temperature, and after screening over 750 markers with that system, we found that 32 of those combinations account for >99% of the loci that could be amplified across species. This number of test conditions can be further reduced to 6 conditions when a touchdown PCR profile is used instead of traditional 3-step PCR. When the test primers are used to amplify batched DNA samples, and fluorescently labeled dNTPs are incorporated into the PCR product, the products from the best set of conditions can be loaded directly onto an automated sequencer, and the number of alleles, allele size range, and approximate allele frequencies can be estimated from one lane per locus. We were able to optimize conditions for automated microsatellite genotyping in baboons for 24.3% of the loci screened, resulting in a panel of over 280 genetic markers suitable for genetic analysis of baboons. Analysis of 139 markers for which we had completed human and baboon genotyping indicated that there was no correlation between numbers of alleles in humans and in baboons, but a high correlation between median allele sizes. There is no indication of directional selection for increased human microsatellite size.