Emergency medical services for children.
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Child↗
Biomedical subjects
D Reilly
Publications and source records attributed to D Reilly.
A sequence within the cytoplasmic tail of GpIIb independently activates platelet aggregation and thromboxane synthesis.
All integrin alpha subunits contain a highly conserved KXGFFKR motif in their cytoplasmic domains that plays a crucial role in the regulation of integrin affinity for their ligands. We show that a lipid-modified peptide corresponding to the cytoplasmic region, 989-995, of the platelet integrin subunit glycoprotein GpIIb (alphaIIb), palmitoyl-KVGFFKR (Ppep; 10 microM), but not a similarly modified scrambled peptide (palmitoyl-FKFVRGK), can specifically induce platelet activation and aggregation equivalent to that of strong agonists such as thrombin. Ppep-induced aggregation is also associated with indices of platelet activation including thromboxane A2 (TXA2) synthesis (EC50 = 45 +/- 5 microM), secretion of alpha-granules detected as enhanced surface expression of P-selectin (EC50 = 52 +/- 8 microM), and conformational changes in GpIIb/IIIa measured by the monoclonal antibody, PAC-1 (EC50 = 3.7 +/- 1 microM). The TXA2 receptor antagonist, SQ29548, PGE1, and the ADP scavenger, apyrase, differentially inhibit the aggregation response and TXA2 synthesis in response to Ppep. Similarly, GpIIb/IIIa antagonists (RO-449883 and integrelin), which inhibit aggregation by greater than 90%, have little effect on peptide-induced TXA2 synthesis, suggesting that this event is independent of fibrinogen binding to GpIIb/IIIa. Alanine-stepping of the Ppep sequence identifies GFFK(991-994) as the critical residues in all peptide-mediated events. We conclude that this peptide can imitate the cytoplasmic domain of GpIIb and initiate parallel but independent signaling pathways, one leading to ligand binding and platelet aggregation and the other to intracellular signaling events such as TXA2 synthesis and secretion.
Acetates↗
Update on imaging of orthopedic infections.
Although the diagnosis of infection is only a small part of the orthopedist's job description, it is a important part. This article discusses the fundamentals of orthopedic infections and highlights the refinements on this topic from a radiologic perspective. In addition to reviewing the imaging appearance of musculoskeletal infections in bone and the surrounding soft tissues, we focus on the advantages and disadvantages of five imaging methods: radiography, sonography, CT, scintigraphy, and MR imaging. Finally, we review three specific situations that have garnered substantial attention in recent medical literature: chronic recurrent multifocal osteomyelitis, musculoskeletal infections in AIDS patients, and pedal infections in diabetic patients.
Diabetic Foot↗
A phase I clinical and pharmacokinetic study of the new topoisomerase inhibitor GI147211 given as a 72-h continuous infusion.
GI147211 is a novel, totally synthetic camptothecin with promising preclinical and early clinical activity. This study was designed to determine the maximum tolerated dose of Gl147211 as a 72-h infusion and to describe its pharmacokinetics and pharmacodynamics on this schedule. In a single-arm, rising-dose study in patients with advanced cancer, eight cohorts of three or more patients received 72-h infusions of Gl147211 at doses ranging from 0.25 to 2.5 mg m(-2) day(-1). Forty-four patients received a total of 124 cycles. All patients had refractory tumours and 40 had received prior chemotherapy and/or radiotherapy. Whole-blood Gl147211 lactone, total blood and total concentrations were measured during and over the 12 h following the infusion. Myelosuppression was observed at all dose levels. Neutropenia was dose limiting at 2.0 mg m(-2) day(-1) in minimally pretreated patients, while both neutropenia and thrombocytopenia were limiting at 1.5 mg m(-2) day(-1) in those more heavily pretreated. Phlebitis occurred with infusions through peripheral veins early in this study, necessitating the use of central venous access. Other toxicities included mild nausea and vomiting, fatigue, headache, central venous catheter infections and alopecia. Three partial and two minor responses lasting 8-34+ weeks were noted in patients with ovarian, colon and breast carcinomas and hepatoma. Mean steady-state concentrations of Gl147211 increased with dose over a range of 0.25-1.24 ng ml(-1). The mean terminal elimination half-life was 7.5 h, and the clearance averaged 1074 ml min(-1) m(-2) over the doses studied. The mean fractional excretion of unchanged drug in urine was 0.114. Gl147211 lactone exposure correlated with haematological toxicity. The recommended phase II doses for this regimen are 1.75 mg m(-2) day(-1) and 1.2 mg m(-2) day(-1) for minimally pretreated and heavily pretreated patients respectively. At these doses, steady-state Gl147211 concentrations within the range of those effective in vitro were achieved. Extensive phase II evaluation of this compound and further phase I trials evaluating more prolonged infusions are ongoing.
Adult↗
Solution structure of the fourth metal-binding domain from the Menkes copper-transporting ATPase.
Menkes disease is an X-linked disorder in copper transport that results in death during early childhood. The solution structures of both apo and Ag(I)-bound forms of the fourth metal-binding domain (mbd4) from the Menkes copper-transporting ATPase have been solved. The 72-residue mbd4 has a ferredoxin-like beta alpha beta beta alpha beta fold. Structural differences between the two forms are limited to the metal-binding loop, which is disordered in the apo structure but well ordered in the Ag(I)-bound structure. Ag(I) binds in a linear bicoordinate manner to the two Cys residues of the conserved GMTCxxC motif; Cu(I) likely coordinates in a similar manner. Menkes mbd4 is thus the first bicoordinate copper-binding protein to be characterized structurally. Sequence comparisons with other heavy-metal-binding domains reveal a conserved hydrophobic core and metal-binding motif.
Adenosine Triphosphatases↗
Alcohol education in licensed premises using brief intervention strategies.
AIMS: The aim of this feasibility study is to present a description of a new approach based on alcohol brief intervention strategies to intervene with at-risk drinkers in licensed premises. DESIGN: The data presented are from a convenience sample of tavern patrons. SETTING: The intervention called Operation Drinksafe was conducted on 75 occasions in 67 licensed premises in an Australian rural area. PARTICIPANTS: There were 2761 voluntary participants who were provided with a personal risk assessment of their alcohol use. MEASUREMENTS: The main measurements were the Alcohol Use Disorders Identification Test (AUDIT) combined with a blood alcohol concentration (BAC) reading. FINDINGS: The median AUDIT score was 11.0, with 53% scoring in the hazardous range and 25% in the harmful range. For binge drinking, almost two-thirds (62%) of males drank six or more drinks once a week or more compared to 29% of females. Median BAC was 0.03 mg/100 ml with 11% of males above 0.10. Regression analysis revealed that significant predictors of alcohol-related injury were being aged less than 30 years, forgetting the previous night, having had another person express concern about your drinking, and binge drinking weekly or more. CONCLUSIONS: The program had high acceptance among licensees and their patrons. Patrons who are ready to change their at-risk drinking behaviour may benefit from this type of minimal intervention when presented in an interesting way in their drinking environments.
Adult↗
Long-term cannabis use: characteristics of users in an Australian rural area.
AIM: To investigate the characteristics and patterns of cannabis and other drug use among long-term cannabis users in an Australian rural area. DESIGN: Cross-sectional survey of a "snowball" sample of long-term cannabis users. SETTING: The North Coast of New South Wales is an area with high levels of cannabis cultivation and use, and many long-term users. PARTICIPANTS: The study involved 268 long-term cannabis users who had regularly used cannabis for at least 10 years. MEASUREMENTS: A structured interview schedule obtained information on: demographics, social circumstances, patterns of cannabis and other drug use, contexts of use, perceptions about cannabis and legal involvement. FINDINGS: The mean age of the sample was 36 years and 59% were made. The median length of regular cannabis use was 19 years. Most (94%) used two or more times a week and 60% used daily, with a median of two joints per day. Two-thirds (67%) used cannabis in social settings and two-thirds grew cannabis for their own use. The most common reasons for using cannabis were for relaxation or relief of tension (61%) and enjoyment or to feel good (27%). The most commonly reported negative effects were feelings of anxiety, paranoia, or depression (21%), tiredness, lack of motivation and low energy (21%) and effects of smoke on the respiratory system (18%). The majority drank alcohol (79%) and over one-third were drinking at hazardous levels. Most were current (64%) or ex-tobacco smokers (24%). One-quarter (25%) had been charged with possession of cannabis, 11% for cultivation and 6% for supply, with non-drug offences low (8%or less). Overall, three-quarters (72%) believed that the benefits of cannabis use outweighed the risks, 21% felt there was an even balance, and 7% said cannabis had done them more harm than good. CONCLUSIONS: Among long-term cannabis users in this Australian rural area, cannabis use was an integral part of everyday life and it was primarily used in social situations for the same reasons that alcohol use is used in the wider community.
Adult↗
Patterns and correlates of cannabis dependence among long-term users in an Australian rural area.
AIM: To examine prevalence and correlates of cannabis dependence among long-term cannabis users. DESIGN: A cross-sectional survey of patterns and experiences of cannabis use and dependence. SETTING AND PARTICIPANTS: A snowball sample of 243 long-term cannabis smokers, who were currently smoking 3-4 times a week, were recruited from the New South Wales North Coast, an area long associated with cannabis cultivation and use. MEASUREMENTS: A structured interview was administered, incorporating the following dependence measures: an approximation to a life-time DSM-III-R diagnosis, an approximation to a 12-month ICD-10 diagnosis, and the Severity of Dependence Scale (SDS). FINDINGS: Prevalence of a life-time DSM-III-R diagnosis of cannabis dependence was 57%, while 57% received an ICD-10 dependence diagnosis for the last year. Substantially fewer (15%) of the sample were diagnosed as dependent according to the Severity of Dependence Scale (SDS). Only 26% believed they had a problem with cannabis at least sometimes. There was general concordance between DSM-III-R and ICD-10 measures, but not between these and the SDS. ICD-10 and DSM-III-R dependence diagnoses were modestly correlated with age, life-time illicit drug use and quantity of cannabis use. Principal components analyses of the dependence measures provided little evidence for a unidimensional dependence syndrome for ICD-10 and DSM-III-R criteria. There was strong support for unidimensionality of the SDS. CONCLUSIONS: Symptoms of cannabis dependence were common among these long-term users but only one-quarter perceived that they had a cannabis problem. There was no strong evidence for a unidimensional cannabis dependence syndrome.
Cross-Sectional Studies↗
Lithium vs placebo in cluster headache.
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Bias↗
A phase I and pharmacokinetic study of tallimustine [PNU 152241 (FCE 24517)] in patients with advanced cancer.
Tallimustine [PNU 152241 (FCE 24517)] is a synthetic derivative of the DNA minor groove binder distamycin A, in which the NH2-terminal formyl group is substituted by benzoyl mustard. In this Phase I clinical trial, patients with advanced solid tumors received i.v. bolus injections of tallimustine daily for 3 consecutive days. Patients were treated at six dosage levels of 33.3 micrograms/m2/day to 250 micrograms/m2/day for 3 consecutive days, with courses of therapy repeated every 28 days. Detailed pharmacokinetic blood sampling was performed during the first 3 days of the first course of tallimustine. The plasma samples were analyzed by high-performance liquid chromatography with UV detection. Forty-eight eligible patients were treated at all six dosage levels. The dominant dose-related toxicity of tallimustine was neutropenia, becoming dose limiting at 250 micrograms/m2/day. At this dosage level, one patient experienced febrile neutropenia, and a second patient died on study of indeterminate cause. Thrombocytopenia was not observed, and only 10 patients developed anemia < 8.0 gm/dl. Sporadic elevation of liver enzymes or bilirubin was observed but was not dose related. Pharmacokinetic analysis gave reliable results for 33 patients. For most patients, analysis of the data best fit a three-exponential model. Dose-related increases in areas under the concentration-time curve and end-of-infusion concentrations were observed. There was no significant plasma accumulation of tallimustine over the 3 days of administration. The terminal half-life of tallimustine in individual patients ranged from 6.83 to 39.02 h following the last dose. In summary, the recommended Phase II dosage for tallimustine is 200 micrograms/m2/day for 3 consecutive days every 28 days. Neutropenia is the principal toxicity of this agent at this dosage and schedule.
Adult↗
Role of high-performance liquid chromatographic protein analysis in developing fermentation processes for recombinant human growth hormone, relaxin, antibody fragments and lymphotoxin.
Development of efficient and reliable fermentation processes for protein pharmaceuticals is aided by the availability of accurate quantitative and qualitative product analyses. We have developed a variety of single and dual column chromatographic separations that meet the needs of process development and examples will be provided of how the resulting data has been used to optimize the culture process. For single column methods, reversed-phase chromatography has been the most versatile, permitting the reliable quantitation of many yeast, Chinese hamster ovary (CHO) cell and Escherichia coli-expressed products in the matrix of culture broth or cell extract. Analysis of secreted human growth hormone synthesized in E. coli, along with clipped and unprocessed forms, will be discussed. Another reversed-phase assay for direct analysis of a peptide product (B-chain relaxin) and its degradation products secreted into E. coli fermentation medium has allowed the purification of the responsible protease. Cation-exchange has proven extremely useful for the direct analysis of antibody fragment synthesized in E. coli, allowing the separation and quantitation of the desired Fab' and Fab'2, as well as the unwanted products of glutathione addition and translational read-through. Assay development is often complicated by the presence of host proteins with chromatographic behavior that is similar to that of the product. Commercial instrumentation now permits the facile development of multidimensional chromatographic assays. We show examples of coupled receptor affinity-reversed-phase assays for a mistranslation product and for covalent multimers of E. coli-synthesized lymphotoxin.
Amino Acid Sequence↗
Expression, purification, and characterization of Escherichia coli-derived recombinant human melanoma growth stimulating activity.
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Cell Division↗
Nursing care--who cares?
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Complementary Therapies↗
A novel portable grab sampler for tritiated water vapor.
The purpose of this work was to evaluate the applicability of a recently developed portable high-throughput liquid-absorption air sampler for measuring tritiated water vapor concentrations in nuclear facilities. A procedure for sampling tritiated water with a portable high-throughput liquid-absorption air sampler and measuring its concentration in air was developed with the aid of a theoretical model. Tritium concentrations in the air of a heavy water reactor currently being decommissioned, derived from samples obtained with a portable high-throughput liquid-absorption air sampler, a bubbler, and a cold trap, yielded comparable results, suggesting the applicability of the portable high-throughput liquid-absorption air sampler to tritium grab sampling.
Air Pollutants, Radioactive↗
Solution structure of the E-domain of staphylococcal protein A.
The E-domain of staphylococcal protein A is one of five homologous IgG-binding domains designated E, D, A, B, and C that comprise the extracellular portion of protein A. The E-domain binds tightly to Fc fragments of IgG and binds certain Fv fragments with micromolar affinity. To explore further the structural features of Fc binding by protein A, and as a first step in developing a structural understanding of E-domain/Fv complex formation, we have determined the solution structure of the uncomplexed E-domain using 2D homonuclear and heteronuclear NMR spectroscopy. Complete 1H and 15N resonance assignments were obtained, and the structure was determined from 383 NOE-derived distance restrains, 34 phi and 19 chi 1 dihedral angle restraints, and 54 restraints for 27 H-bonds. 3JH alpha-H beta coupling constants and long-range NOEs involving Phe11 indicate the side chain exists in more than one conformation with differing chi 1 values. NOE restraints that were incompatible with chi 1 = -60 degrees were removed from one set of structure calculations, and those incompatible with chi 1 = 180 degrees were removed from a second set to allow Phe11 to explore both rotamer wells. Thus, two sets of 20 final structures, having no distance or dihedral angle restraint violations greater than 0.12 A or 1.6 degrees, respectively, represent the solution structure of the E-domain. Backbone atomic rms differences with respect to the mean coordinates for each set of 20 structures for residues 8-53 averaged 0.41 +/- 0.06 and 0.35 +/- 0.06 A. No significant differences in the overall structure result from the different orientations of Phe11. The solution structure of the E-domain consists of three alpha-helices that pack together to form a compact helical bundle. A detailed comparison between the E-domain ensembles and the previously determined structure for the B-domain in complex with Fc indicates that only the 180 degrees chi 1 rotamer of Phe11 is competent for binding; the -60 degrees chi 1 rotamer must reorient to 180 degrees to create a cavity that is filled by Ile253 from the CH2 domain of Fc in the Fc-bound complex.
Amino Acid Sequence↗
High-level expression and characterization of a purified 142-residue polypeptide of the prion protein.
The major, and possible only, component of the infectious prion is the scrapie prion protein (PrPSc); the protease resistant core of PrPSc is PrP 27-30, a protein of approximately 142 amino acids. PrPSc is derived from the cellular PrP isoform (PrPC) by a post-transliatonal process in which a profound conformational change occurs. Syrian hamster (SHa) PrP genes of varying length ranging from the N- and C- terminally truncated 90-228 up to the full-length mature protein 23-231 were inserted into various secretion and intracellular expression vectors that were transformed into Escherichia coli deficient for proteases. Maximum expression was obtained for a truncated SHaPrP containing residues 90-231, which correspond to the sequence of PrP 27-30; disruption of the bacteria using a microfluidizer produced the highest yields of this protein designated rPrP. After solubilization of rPrP in 8 M GdnHC1, it was purified by size exclusion chromatography and reversed phase chromatography. During purification the recovery was approximately 50%, and from each liter of E. coli culture, approximately 50 mg of purified rPrP was obtained. Expression of the longer species containing the basic N-terminal region was less successful and was not pursued further. The primary structure of rPrP was verified by Edman sequencing and mass spectrometry, and secondary structure determined by circular dichroism and Fourier transform infrared spectroscopy. When rPrP was purified under reducing conditions, it had a high beta-sheet content and relatively low solubility similar to PrPSc, particularly at pH values > 7. Refolding of rPrP by oxidation to form a disulfide bond between the two Cys residues of this polypeptide produced a soluble protein with a high alpha-helical content similar to PrPC. These multiple conformations of rPrP are reminiscent of the structural plurality that characterizes the naturally occurring PrP isoforms. The high levels of purified rPrP which can now be obtained should facilitate determination of the multiple tertiary structures that Prp can adopt.
Amino Acid Sequence↗
High-resolution solution structure of the EGF-like domain of heregulin-alpha.
The solution structure of the 63-residue heregulin-alpha (HRG-alpha) epidermal growth factor (EGF)-like domain, corresponding to residues 177-239 of HRG-alpha, has been determined to high resolution using data from two-dimensional and three-dimensional homo- and heteronuclear NMR spectroscopy. The structure is based on a total of 887 internuclear distance and dihedral restraints derived from data obtained using unlabeled and uniformly 15N-labeled protein samples, at pH 4.5, 20 degrees C. A total of 20 structures were calculated using a hybrid distance geometry-simulated annealing approach with the program DGII, followed by restrained molecular dynamics using the program DISCOVER. The average maximum violations are 0.12 +/- 0.01 angstroms and 1.4 +/- 0.3 degrees for distance and dihedral restraints, respectively. The backbone (N,C(alpha),C) atomic rms distribution about the mean coordinates for residues 3-23 and 31-49 is 0.29 +/- 0/07 angstroms. The N-and C-terminal residues (1-2 and 50-63) and 24-30 are disordered. Comparison of the HRG-alpha EGF-like domain structure with the previously determined structure of human EGF [Hommel et al. (1992) J. Mol. Biol. 227, 271-282] reveals a high degree of structural similarity; excluding the N-terminal region (residues 1-13), the disordered phi-loop region (residues 24-30) that contains a three-residue insertion in HRG-alpha relative to hEGF, and the disordered C-terminal region (residues 50-63), the C(alpha) alignment between the HRG-alpha and hEGF minimized mean structures has a rms difference of approximately 1 angstrom. In HRG-alpha the N-terminal residues 2-6 form a well-defined beta strand rather than being disordered as found for hEGF. This structural difference correlates with functional data which suggest that the N-terminal region of the HRG-alpha EGF-like domain is responsible for the observed receptor specificity differences between HRG-alpha and EGF.
Amino Acid Sequence↗
The bglX gene located at 47.8 min on the Escherichia coli chromosome encodes a periplasmic beta-glucosidase.
A new Escherichia coli gene, bgIX, encoding a beta-D-glucosidase (EC 3.2.1.21) has been characterized. The bgIX gene is located adjacent to the dld gene at 47.8 min or 2225 kb on the E. coli chromosome. The sequence of a 2.6 kb DNA fragment from this region revealed a large open reading frame encoding a protein of 765 amino acids. The BgIX protein was purified from the periplasm, and amino-terminal sequencing suggests that the mature protein is derived from cleavage of a 20 residue signal peptide. A search of the sequence databases revealed that BgIX is a member of a large family of beta-glucosidases from a variety of bacteria and fungi, and the plant Arabidopsis thaliana. It differs from the other four E. coli phospho-beta-glucosidases in sequence and in its periplasmic rather than cytoplasmic location. The BgIX enzyme has a Km of 18 +/- 1 mM and a Vmax of 3 +/- 0.7 mumol min-1 for the colorimetric substrate o-nitrophenyl beta-D-glucopyranoside.
Amino Acid Sequence↗