bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigation of fire ash residues using mineral magnetism
Authors: Peters C. , Church M. J. , Mitchell C.
Published: August 2002

As part of a wider research programme of experimental archaeology at Calanais Farm, Isle of Lewis, Scotland, a number of experimental hearths were constructed, based on excavated evidence from the Late Iron Age houses at Bostadh, Lewis. Controlled and repeated burning of different fuel sources (well‐humified peat, fibrous upper peat, peat turf and wood) was carried out over a number of burning episodes, each of three days duration. A range of mineral magnetic measurements, including remanences and the variation of susceptibility with high temperature, were taken from the resulting ash samples. The high temperature susceptibility measurements show that the fibrous upper peat and peat turf have a single magnetic component, with a drop in magnetic susceptibility at ca. 600 °C. In comparison the well‐humified peat and wood have one, sometimes two, distinct magnetic components characterized by drops in susceptibility at ca. 330 and ca. 550 °C. Stepwise discriminant analysis was performed on the room temperature magnetic data. A biplot of the resulting two main variables distinguishes the well‐humified peat and wood. Some overlap is observed between the fibrous upper peat and peat turf. Magnetic measurements also were carried out on Iron Age and Medieval hearth, floor and ash spread samples from the multiperiod archaeological site of Guinnerso, on the Isle of Lewis. Comparison was made with the modern ash samples in order to determine if fuel sources could be identified. The high temperature susceptibility curves and the discriminant analysis biplot suggest that for the selected archaeological samples the predominant fuel source was well‐humified peat. Copyright © 2001 John Wiley & Sons, Ltd.
The iatrogenic costs of NSAID therapy: A population study
Authors: Sturkenboom Miriam C. J. M. , Romano Ferdinando , Simon Giorgio , Correa‐Leite Maria L. , Villa Marco , Nicolosi Alfredo , Borgnolo Giulio , Bianchi‐Porro Gabriele , Mannino Salvatore
Published: August 2002

Objective

To estimate the iatrogenic costs of nonsteroidal antiinflammatory drug (NSAID) treatment from the perspective of the Italian National Health Service.

Methods

We conducted a retrospective cohort study using the primary and secondary care claims data registered in the regional health service database in the Friuli‐Venezia Giulia (Italy). The study cohort comprised all persons(265,114) who received at least one prescription for any NSAID between August 1996 and July 1998. The outcomes of interest were the costs of medical interventions for upper gastrointestinal disorders following NSAID treatment (i.e., prescriptions for gastroprotective drugs, hospitalizations, and outpatient diagnostic procedures).

Results

The study population received a total of 660,311 NSAID prescriptions for a cost of 6,587,533 Euros (€) (€0.53 per treatment day). The cost of medical interventions for gastrointestinal events added 58% to the cost of NSAID therapy (€0.31 per NSAID treatment day, up to 64% directly attributable to NSAID use). The iatrogenic costs were generated by 12.4% of the patients, 77% of whom had a positive history of gastrointestinal disorders and 82% of whom were older than 50 years. Co‐prescriptions for gastroprotective drugs accounted for 78.6% of the overall iatrogenic costs. The iatrogenic costs did not differ between cyclooxygenase (COX) nonselective and COX‐2 preferential drugs within strata of age and prior history of gastrointestinal disorders, but were significantly higher for the parenteral NSAIDs than the oral or rectal formulations.

Conclusions

In Italy, the iatrogenic costs of NSAID therapy add 58% to the cost of NSAID treatment; most of the cost is generated by co‐prescriptions of gastroprotective drugs to elderly NSAID users or patients with a history of gastrointestinal disorders.
Comparative bioavailability of two cefadroxil formulations in healthy human volunteers after a single‐dose administration
Authors: Oliveira C.H. , Salmon J. , Sucupira M. , Ilha J. , De Nucci G.
Published: August 2002

Objective. To compare the bioavailability of two cefadroxil capsule (500 mg) formulations (Cefadroxila from Eurofarma Laboratórios Ltd, Brazil, as test formulation and Cefamox® from Bristol–Myers Squibb, Brazil S.A. as reference formulation) in 24 volunteers of both sexes.

Material and methods. The study was conducted open with randomized two‐period crossover design and a 1‐week washout period. Plasma samples were obtained over a 12‐h interval. Cefadroxil concentrations were analysed by combined reversed‐phase liquid chromatography and tandem mass spectrometry (LC‐MS‐MS) with positive ion electrospray ionization using a selected ion monitoring method. From the cefadroxil plasma concentration versus time curves the following pharmacokinetic parameters were obtained: AUClast, AUC0–∞ and Cmax.

Results. Geometric mean of Cefadroxila/Cefamox® 500 mg individual percent ratio was 103.97% for AUClast, 104.08% for AUC0–∞ and 95.23% for Cmax. The 90% confidence intervals (CI) were 98.14–110.16%, 98.37–110.12%, and 85.59–105.96%, respectively.

Conclusion. Since the 90% CI for Cmax, AUClast and AUC0–∞ were within the 80–125% interval proposed by the Food and Drug Administration, it was concluded that the Cefadroxila 500 mg capsule was bioequivalent to the Cefamox® 500 mg capsule, according to both the rate and extent of absorption. Copyright © 2000 John Wiley & Sons, Ltd.
Dose‐dependent pharmacokinetics of a new reversible proton pump inhibitor, DBM‐819, after intravenous and oral administration to rats: hepatic first‐pass effect
Authors: Kim Eun J. , Kim Sun‐O. , Lee Dong H. , Lim Hong , Lee Myung G.
Published: August 2002

The dose‐dependent pharmacokinetic parameters of DBM‐819 were evaluated after intravenous (5, 10 and 20 mg/kg) and oral (10, 20 and 50 mg/kg) administrations of the drug to rats. The hepatic first‐pass effect was also measured after intravenous and intraportal administrations of the drug, 10 mg/kg, to rats. After intravenous administration, the dose‐normalized (based on 5 mg/kg) area under the plasma concentration–time curve from time zero to time infinity, AUC, at 20 mg/kg (27.0 and 45.8 μg min/ml) was significantly greater than that at 5 mg/kg due to saturable metabolism. After oral administration, the dose‐normalized (based on 10 mg/kg) AUC0–12 h at 50 mg/kg (25.1, 18.3 and 49.2 μg min/ml) was significantly greater than those at 10 and 20 mg/kg again due to saturable metabolism. After oral administration of DBM‐819, 10 mg/kg, 2.86% of oral dose was not absorbed and the extent of absolute oral bioavailability (F) was estimated to be 46.7%. After intraportal administration of DBM‐819, 10 mg/kg, the AUC was 51.9% of intravenous administration, suggesting that approximately 48.1% was eliminated by liver (hepatic first‐pass effect). The considerable hepatic first‐pass effect of DBM‐819 was also supported by significantly greater AUC of M3 (3.70 and 6.86 μg min/ml), a metabolite of DBM‐819, after intraportal administration. The AUCs of DBM‐819 were not significantly different (comparable) between intraportal and oral administrations of the drug, 10 mg/kg, suggesting that gastrointestinal first‐pass effect of DBM‐819 was almost negligible in rats. At 10 mg/kg oral dose of DBM‐819, the hepatic first‐pass effect was approximately 48.1%, F was approximately 46.7 and 2.86% was not absorbed from gastrointestinal tract in rats. Copyright © 2001 John Wiley & Sons, Ltd.
Clinical pharmacokinetic studies with INN 00835 (Nemifitide), a novel pentapeptide antidepressant
Authors: Feighner John P. , Nicolau Gabriela , Abajian Henry , Marricco Nadia Cardillo , Morrison John , Sverdlov Lev , Hlavka Joseph , Tonelli George , Spirito Carlo Di , Faria George
Published: August 2002

Nemifitide (4‐fluoro‐L‐phenylalanyl‐trans‐4‐hydroxy‐L‐prolyl‐L‐arginylglycyl‐L‐tryptophanamide ditrifluoroacetate) is a novel antidepressant, currently in phase 2/3 clinical trials. The purpose of our phase 1 clinical trials (conducted over a three year period) was to provide safety and pharmacokinetic data to support its clinical development as an antidepressant drug. Single and multiple doses ranging from 18 to 320 mg were administered subcutaneously to healthy volunteers in five phase 1 studies. Plasma concentrations of unchanged parent drug were determined by a validated LC/MS/MS method in blood samples collected at timepoints between 10 min and 72 h after dosing. Nemifitide was rapidly absorbed (Cmax at 10 min) and eliminated (t1/2 15–30 min) in most subjects. Regression and power model analyses were used to evaluate the data. The results indicate that pharmacokinetic parameters: AUC0−t, AUC 0−∞ and Cmax, were close to dose proportional in the dose range investigated. There was no evidence of systemic accumulation of drug following 5 daily doses. No serious adverse events or clinically significant systemic adverse events occurred at any of the doses investigated in the over 100 subjects dosed in these studies. Drug‐related adverse events were limited to local and transient skin reactions (pain and/or erythema) at the injection site, especially at the high doses administered: 240 and 320 mg. Copyright © 2002 John Wiley & Sons, Ltd.
Chromosomal aberrations in lymphocytes of employees in transformer and generator production exposed to electromagnetic fields and mineral oil
Authors: Skyberg Knut , Hansteen Inger‐Lise , Vistnes Arnt I.
Published: August 2002

The objective was to study the risk of cytogenetic damage among high voltage laboratory workers exposed to electromagnetic fields and mineral oil. This is a cross sectional study of 24 exposed and 24 matched controls in a Norwegian transformer factory. The exposure group included employees in the high voltage laboratory and in the generator soldering department. Electric and magnetic fields and oil mist and vapor were measured. Blood samples were analyzed for chromosomal aberrations in cultured lymphocytes. In addition to conventional cultures, the lymphocytes were also treated with hydroxyurea and caffeine. This procedure inhibits DNA synthesis and repair in vitro, revealing in vivo genotoxic lesions that are repaired during conventional culturing. In conventional cultures, the exposure group and the controls showed similar values for all cytogenetic parameters. In the DNA synthesis‐ and repair‐inhibited cultures, generator welders showed no differences compared to controls. Among high voltage laboratory testers, compared to the controls, the median number of chromatid breaks was doubled (5 vs. 2.5 per 50 cells; P<0.05) the median number of chromosome breaks was 2 vs. 0.5 (P>0.05) and the median number of aberrant cells was 5 vs. 3.5 (P<0.05). Further analysis of the inhibited culture data from this and a previous study indicated that years of exposure and smoking increase the risk of aberrations. We conclude that there was no increase in cytogenetic damage among exposed workers compared to controls in the conventional lymphocyte assay. In inhibited cultures, however, there were indications that electromagnetic fields in combination with mineral oil exposure may produce chromosomal aberrations. Bioelectromagnetics 22:150–160, 2001. © 2001 Wiley‐Liss, Inc.
Spontaneous conformational change within the prion protein—implications for disease pathogenesis?
Authors: Jackson Graham S.
Published: August 2002

A recent paper by Leclerc et al(1) describes how recombinant hamster prion protein can undergo a spontaneous change in conformation to a structure that has features in common with PrPSc. Structural change in the host prion protein, PrPC to an insoluble and aggregated form with increased β‐sheet content (PrPSc) is central to the pathology of prion diseases.(2) A detailed understanding of the nature of these conformational changes will increase our knowledge of the molecular basis of prion pathology. These findings may have implications for how the disease is initiated and provide a format for further investigation. BioEssays 23:772–774, 2001. © 2001 John Wiley & Sons, Inc.
A post hoc analysis of multiple‐stimulus preference assessment results
Authors: Graff Richard B. , Ciccone Francis J.
Published: August 2002

Fifteen individuals with developmental disabilities participated in multiple‐stimulus without replacement preference assessments (seven assessment sessions, seven trials each). Twelve of 15 individuals completed a second set of preference assessments 6 months later. Post hoc analyses indicated that in 25 of 27 cases, the same most preferred stimulus was identified in the first three trials as the full seven trials. Additionally, five sessions of three trials each were sufficient to identify the most highly preferred stimulus in 22 of 27 instances. Copyright © 2002 John Wiley & Sons, Ltd.
Ordering in aqueous polysaccharide solutions. I. Dielectric relaxation in aqueous solutions of a triple‐helical polysaccharide schizophyllan
Authors: Hayashi Yoshihito , Shinyashiki Naoki , Yagihara Shin , Yoshiba Kazuto , Teramoto Akio , Nakamura Naotake , Miyazaki Yuji , Sorai Michio , Wang Qi
Published: August 2002

Deuterium oxide solutions of a triple‐helical polysaccharide schizophyllan, undergoing an order–disorder transition centered around 17°C, were studied by the time‐domain reflectometry (TDR) to obtain dielectric dispersions in the solution and frozen states. In the solution state, the dispersion below the transition temperature is resolved in three dispersions (relaxation times at 0°C) ascribed to side chain glucose residue (1; 102 ns), structured water (s; 2.0 ns) and bulk water (h), respectively, from low to high frequencies. Bulk water is divided into slow water (h2; 0.04 ns) and free or pure water (h1; 0.02 ns). Above the transition temperature structured water almost disappears and is compensated by slow water. Structured water is similar to bound water for proteins but different from it because of this transition behavior. Another dispersion (l) seen at the lowest frequency is assigned to the rotation of side‐chain glucose residue coupled with hydrated water. Parts of this dispersion and structured water are suggested to constitute bound water. In the frozen state were observed a major dispersion (h; 0.14 ns) and a minor one (m; 28 ns), which were ascribed to considerably mobile and less mobile waters. They are similar to but not exactly the same as that for unfreezable water in bovine serum albumin solutions argued by Miura et al. (Biopolymers, 1995, Vol. 36, p. 9). Water is molded into different structures by the triple helix. © 2002 John Wiley & Sons, Inc. Biopolymers 63: 21–31, 2002
Glucose‐based optimization of CHO‐cell perfusion cultures
Authors: Dowd Jason E. , Kwok K. Ezra , Piret James M.
Published: August 2002

Perfusion cultures of CHO cells producing t‐PA were performed using acoustic filter cell retention. A robust off‐line glucose analysis and predictive control protocol was developed to maintain the process within approximately 0.5 mM of the glucose set point, without the need for a more fallible on‐line sensor. Glucose usage (the difference between the inlet and reactor glucose concentrations) provided an easily measured indicator of overall medium utilization for mapping acceptable ranges of operation, including the edge of failure. Earlier onset of perfusion with a ramping glucose set point (1.5 mM/d) resulted in improved growth and consistency during the perfusion culture start‐up. At steady state, the t‐PA concentration variability increased gradually with increasing glucose usage up to approximately 22 mM, then up to 24 mM the variability increased threefold. Peak t‐PA concentrations of over 90 mg/L were obtained by controlling at a glucose usage of approximately 24 mM, but these t‐PA levels were not sustainable for more than 3 days. A consistent t‐PA concentration of 40 mg/L was obtained at a glucose usage of 21.5 mM. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 252–256, 2001.
Water‐based nanoparticulate polymeric system for protein delivery
Authors: Prokop Ales , Holland Celia A. , Kozlov Evgenii , Moore Billy , Tanner Robert D.
Published: August 2002

This article features a new production technology for nanoparticles comprised of multicomponent polymeric complexes that are candidates for delivery vehicles of biological molecules such as proteins and drugs. Biocompatible and mostly natural polymers are fabricated into thermodynamically stable nanoparticles insoluble in water and buffered media, in the absence of organic solvents, using two types of processing: batch and continuous. Careful choice of construction materials and the superposition of several interacting principles during their production allow for the customization of the physicochemical properties of the structures. Detailed experiments in batch and continuous systems allowed time‐dependent stoichiometric characterization of the production process and an understanding of fundamental assembly principles of such supramolecular structures. Continuous‐flow production is shown to provide more consistent data in terms of product quality and consistency, with further possibility of process development and commercialization. The development of nanoparticles using the described methodology is expected to lead to a flexible nanoparticle drug delivery system for medical applications, which has particular bearing to the slow release of drugs, antigens (for vaccine design), and genes (for gene therapy). Several chemistries of particles are presented. © John Wiley & Sons, Inc. Biotechnol Bioeng 75: 228–232, 2001.
A kinetic model describing cell growth and production of highly active, recombinant ice nucleation protein in Escherichia coli
Authors: Palaiomylitou M. A. , Matis K. A. , Zouboulis A. I. , Kyriakidis D. A.
Published: August 2002

A structured kinetic model, which describes the production of the recombinant ice nucleation protein in different conditions, was applied. The model parameters were estimated based on the variation of the specific growth rate and the intracellular product concentration during cultivation. The equations employed relate the cellular plasmid content or plasmid copy number with the cloned‐gene expression; these correlations were successfully tested on the experimental data. The optimal nutrient conditions for the growth of Escherichia coli expressing the inaZ gene of Pseudomonas syringae were determined for the production of active ice nucleation protein. The kinetics of the cultures expressing the inaZ gene were studied in a bioreactor at different growth temperatures and nutrient conditions. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 321–332, 2002.
Monitoring of chemical and enzymatic hydrolysis of water‐soluble proteins using flow‐injection analysis with fluorescence detection and an aqueous eluant containing 2‐p‐toluidinylnaphthalene‐6‐sulfonate as the fluorescent probe
Authors: Sentandreu E. , Carbonell J. V. , Sendra J. M.
Published: August 2002

The exposed hydrophobicity of proteins, which is due to the hydrophobic regions located on their surfaces, enhances the fluorescence intensity of the probe 2‐p‐toluidinylnaphthalene‐6‐sulfonate (2,6‐TNS) by the formation of a complex. During the hydrolysis of a protein, the average exposed hydrophobicity of the substrate continuously changes with incubation time, and these changes are immediately reflected by a corresponding change in the fluorescence intensity of the 2,6‐TNS/substrate complex. Therefore, 2,6‐TNS seems to be a good probe to monitor the course of the depolymerization processes of proteins. In this work, bovine serum albumin and α‐casein have been hydrolyzed both chemically and enzymatically, and the course of the reactions is monitored by using flow‐injection analysis (FIA) with fluorescence detection and a buffered aqueous eluant containing 2,6‐TNS as the fluorescent probe. Results indicate that the time evolution of the fluorescence intensity of the 2,6‐TNS/substrate complex can be correlated with the initial concentration of the parent protein, in mass per unit volume, the hydrolytic activity added, and the time evolution of the mean chain length of the substrate. In addition, because the time elapsed between injection of the sample into the FIA system and measurement of the corresponding fluorescence intensity is only a few seconds, this methodology could be a useful tool for on‐line monitoring of processes for the production of protein hydrolysates. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 829–833, 2002.
Glutamate synthesis via photoreduction of NADP+ by photostable chlorophyllide coupled with polyethylene‐glycol
Authors: Asada Hideharu , Itoh Tetsuji , Kodera Yoh , Matsushima Ayako , Hiroto Misao , Nishimura Hiroyuki , Inada Yuji
Published: August 2002

Chlorophyllide a was coupled with α‐(3‐aminopropyl)‐ω‐methoxypoly(oxyethylene) (PEG–NH2) to form a PEG–chlorophyllide conjugate through an acid–amide bond. The conjugate catalyzed the reduction of methylviologen in the presence of 2‐mercaptoethanol. It also catalyzed the photoreduction of NADP+ or NAD+ in the presence of ascorbate as an electron donor and ferredoxin‐NADP+ reductase as the coupling enzyme. Utilizing the reducing power of NADPH generated by PEG‐chlorophyllide conjugate under illumination, glutamate was synthesized from 2‐oxoglutarate and NH4+ in the presence of glutamate dehydrogenase. PEG‐chlorophyllide conjugate was quite stable toward light illumination compared with chlorophyll a. The increase in the molecular weight of PEG in the PEG‐chlorophyllide conjugates was accompanied by the enhancement of photostability of the conjugate and also by the increased solubility in the aqueous solution. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 76: 86–90, 2001.
Role of organic solvents on Pa‐hydroxynitrile lyase interfacial activity and stability
Authors: Hickel Andrea , Radke Clayton J. , Blanch Harvey W.
Published: August 2002

Catalytic activity and adsorption of Pa‐hydroxynitrile lyase (Pa‐Hnl) was investigated at various organic solvent/water interfaces. We focused on the role of solvent polarity in promoting activity and stability in two‐phase systems, specifically for the solvents heptane, dibutyl ether (DBE), diisopropyl ether (DIPE), butylmethyl ether (BME), and methyl tert‐butyl ether (MTBE). Enzyme activity towards mandelonitrile cleavage was determined in a recycle reactor with a well‐defined interfacial area as described by Hickel, et al. 1999. Here the recycle reactor was modified to permit exchange of the aqueous phase. With this modification, irreversibility of enzyme adsorption was determined as a function of the adsorption time at the interface. Irreversibility of enzyme adsorption was also investigated by measuring the surface pressure of a sessile‐drop upon washout. We find that Pa‐Hnl exhibits the highest stability but the lowest initial catalytic activity at the aqueous/organic solvent interface with the most polar organic solvents. Thus, DIPE and MTBE display no loss in enzyme activity over a period of several hours. However, the more apolar the solvent is the higher the initial Pa‐Hnl activity, but the faster the loss of activity. Dynamic tensiometry reveals that Pa‐Hnl adsorbs more strongly at the interface of the more apolar solvents. Surprisingly, Pa‐Hnl develops some irreversible adsorption after 30 min at the DIPE/water interface, but does not lose catalytic activity. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 18–28, 2001.
Production of a recombinant hybrid molecule of cholera toxin‐B‐subunit and proteolipid‐protein‐peptide for the treatment of experimental encephalomyelitis
Authors: Yuki Yoshikazu , Byun Youngjin , Fujita Mitsugu , Izutani Wakako , Suzuki Toru , Udaka Shigezo , Fujihashi Kohtaro , McGhee Jerry R. , Kiyono Hiroshi
Published: August 2002

Mucosal administration of experimental autoimmune encephalomyelitis (EAE)‐specific autoantigens can reduce the onset of disease. To examine whether cholera toxin‐B‐subunit (CTB)‐conjugated EAE‐specific T‐cell epitope can reduce development of the autoimmune disease in mice, we produced a recombinant hybrid molecule of CTB fusion protein linked with proteolipid‐protein (PLP)‐peptide139‐151(C140S) at levels up to 0.1 gram per liter culture media in Bacillus brevis as a secretion‐expression system. Amino acid sequencing and GM1‐receptor binding assay showed that this expression system produced a uniformed recombinant hybrid protein. EAE was induced in SJL/J mice by systemic administration with the PLP‐peptide. When nasally immunized 5 times with 70 μg rCTB PLP‐peptide hybrid protein, mice showed a significantly suppressed development of ongoing EAE and an inhibition of both the PLP‐peptide‐specific delayed‐type hypersensitivity (DTH) responses and leukocyte infiltration into the spinal cord. In contrast, all mice given the PLP‐peptide alone or the PLP‐peptide with the free form of CTB did not suppress the development of EAE and DTH responses. These results suggest that nasal treatment with the recombinant B. brevis‐derived hybrid protein of CTB and autoantigen peptide could prove useful in the control of multiple sclerosis. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 62–69, 2001.
A new method for the generation of patterned protein films by encapsulation in arrays of thermally evaporated lipids
Authors: Gole Anand , Sastry Murali
Published: August 2002

In this article we demonstrate a versatile method for the generation of patterned protein films by encapsulation in arrays of the lipids, octadecylamine (ODA, cationic), and arachidic acid (AA, anionic). A simple 2 × 2 array of ODA and AA was vacuum deposited on different substrates using appropriate masks. Thereafter, the enzymes pepsin and fungal protease as well as the heme‐proteins cytochrome c and hemoglobin were encapsulated in the different elements of the array by sequential immersion (combined with judicious masking) of the array elements in the different protein solutions. The proteins are incorporated into the lipid elements by electrostatic interaction between charged amino acid residues on the protein surface and charged functional groups in the lipid matrix. This procedure leads to spatially distinct regions of the different proteins on one substrate and shows promise for single‐chip multianalyte immunoassay/multiplex, high‐throughput biosensor and catalysis applications. Fourier transform infrared spectroscopy (FTIR) was used to monitor the incorporation of the proteins in the different elements of the array as well as to ascertain whether intermixing of the proteins in a particular array element had occurred. The heme‐protein composite regions were further characterized using UV‐VIS spectroscopy. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 172–178, 2001.
Improved glycosylation of a foreign protein by Tn‐5B1‐4 cells engineered to express mammalian glycosyltransferases
Authors: Breitbach Kevin , Jarvis Donald L.
Published: August 2002

The major advantages of using the baculovirus–insect cell system for recombinant protein production are its ability to produce large amounts of recombinant proteins and its ability to provide eucaryotic modifications, such as glycosylation. However, the glycans linked to recombinant glycoproteins produced by this system typically differ from those found on native mammalian products. This is an important problem because glycans on mammalian glycoproteins can influence their functions in many different ways. The inability of baculovirus‐infected insect cells to produce glycans identical to those found on native mammalian glycoproteins is due, in part, to the absence of functional levels of certain glycosyltransferases in insect cells. Thus, the purpose of this study was to engineer these activities into Tn‐5B1‐4, an established insect cell line that is widely used as a host for baculovirus‐mediated protein production. Expression plasmids were constructed in which cDNAs encoding mammalian β1,4‐galactosyltransferase and α2,6‐sialyltransferase were placed under the transcriptional control of a baculovirus immediate early promoter. These plasmids were then used to isolate two different transgenic Tn‐5B1‐4 derivatives and the biological and biochemical properties of these cell lines were examined. The results show that both of the engineered insect cell lines have improved glycoprotein‐processing capabilities, relative to the parental cell line. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 230–239, 2001.
Novel kinetic analysis of enzymatic dipeptide synthesis: Effect of pH and substrates on thermolysin catalysis
Authors: Murakami Yoshihiko , Chiba Kazuyuki , Oda Takaya , Hirata Akira
Published: August 2002

The point of maximum activity is specific to a particular substrate‐enzyme system but may vary with different substrates and the same enzyme. The specificity of enzymes has, however, been generally reported only at their “optimal” pH. In this article, we introduce the Michaelis‐Menten equation taking pH into account, and apply it to the pH‐activity profile of the thermolysin‐catalyzed dipeptide synthesis. It has been reported to date that the pH‐activity profile of thermolysin follows a bell‐shaped curve with a maximal activity at or near pH 7.0. The profiles obtained in this study, however, indicated that the optimal pH varied from 5.8 (for F‐AspPheOMe) to 7.3 (for Z‐ArgPheOMe), and the order of thermolysin activity was greatly dependent on the pH of reaction media. We have succeeded in evaluating the substrates‐induced change of the dissociation states of the active site of thermolysin using the hydrophobicity of substrates. We have obtained apparent kinetic parameters which are independent of the pH of reaction media. The apparent specificity of thermolysin which were independent of pH of the reaction media was in order L‐Leu > L‐Asp > L‐Arg > L‐Ala > L‐Gly > L‐Val and Z > Boc = F at P1 and P2 positions, respectively. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 74: 406–415, 2001.
Novel surface tagging technology for selection of complex proliferation‐controlled mammalian cell phenotypes
Authors: Schlatter Stefan , Bailey James E. , Fussenegger Martin
Published: August 2002

Regulated overexpression of the cyclin dependent kinase inhibitor p27 enables biphasic production processes which consist of a nonproducing expansion phase followed by an extended proliferation‐arrested production phase. During the growth‐arrested production phase proliferation‐competent mutants emerge as a consequence of genetic drift and strong counterselection. Here, we evaluate the use of cell surface markers for ex vivo selection of growth‐arrested phenotypes by magnetic or FACS‐mediated cell sorting. Multigene metabolic engineering resulted in a Chinese hamster ovary‐ (CHO) derived cell line CHO‐SS1015, which expresses the model product protein SEAP (secreted alkaline phosphatase), the human cyclindependent kinase inhibitor p27, and a membrane‐anchored multidomain surface marker Hook in a tricistronic tetracycline‐repressible manner. In the absence of tetracycline in the cell culture medium, p27 mediated a G1‐phase‐specific cell‐cycle arrest of CHO‐SS1015 and resulted in a fivefold increase in SEAP production compared to proliferation‐competent control cells. Concomitant expression of Hook enabled FACS‐ or magnetic‐based selection of CHO‐SS1015 cells from various mixed populations. Surface selection of engineered cells will likely become important for biopharmaceutical manufacturing and for in vivo maintenance of treated cells in gene therapy and tissue engineering. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 597–606; 2001.
Determination and pharmacokinetic study of 1‐p‐(3.3‐dimethyl‐1‐triazeno) benzoic acid in cancer patients by capillary gas chromatography
Authors: Li Ai‐Qun , Zhao Xiang‐Lan
Published: August 2002

A capillary gas chromatographic method was developed for determining 1‐p‐(3.3‐dimethyl‐1‐triazeno) benzoic acid in the plasma and urine of cancer patients under pharmacokinetic study. The drug was extracted with ethyl acetate and methylated with diazomethane. Octadelane (10 µg/ml) was added as internal standard. The separation was carried out on an OV‐1 quartz capillary column, 15 m × 0.32 mm (0.52 µm), with high‐purity nitrogen as carrier gas and flame ionization detector (FID) as detector. The column temperature was held at 130°C for 9 min and then programed to 240°C, at a rate of 35°C/min. The temperature of both injector and detector was 260°C. The standard curve was linear from 0.4 to 40 µg/mL in plasma, and from 0.8 to 20 µg/mL in urine, with correlation coefficients of 0.9979 and 0.9932. The relative standard deviations (RSD) were less than 9.7%. The minimum recovery of this method was 81.8%. This method was applied to the pharmacokinetic studies of 1‐p‐(3.3‐dimethyl‐1‐triazeno) benzoic acid in cancer patients after a single dose (i.v.) of 160, 420 or 760 mg/m2 was administered. They all conformed to the two‐compartment open model and showed linear pharmacokinetics. The excretion of this drug in the urine was minimal. Copyright © 2001 John Wiley & Sons, Ltd.

Abbreviations used:

AUC

area under the curve

DTIC

dacarbazine

MTD

maximum tolerance dosage

P‐COOH‐DMT

1‐p‐(3.3‐dimethyl‐1‐triazeno) benzoic acid

RSD

relative standard deviation.
Validation study of assay method for DX‐8951 and its metabolite in human plasma and urine by high‐performance liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry
Authors: Oguma Toshihiro , Konno Tomomi , Inaba Atsuhiro , Nakaoka Minoru
Published: August 2002

A new liquid chromatographic/mass spectrometric assay has been developed for the determination of DX‐8951, a new anti‐tumor drug, and its 4‐hydroxymethyl metabolite (UM‐1) in human plasma and urine. Solid‐phase extractions were used for sample preparation. A gradient reverse‐phase HPLC separation was developed with mobile phases consisting of trifluoroacetic acid and methanol. The detection was conducted using atmospheric pressure chemical ionization tandem mass spectrometry in the selected reaction monitoring mode. A structural analog, camptothecin (CPT), was used as the internal standard. The assay was validated for the determination of DX‐8951 and UM‐1 in human plasma and urine. The lower limits of quantitation of DX‐8951 and UM‐1 were 0.1 ng/mL in plasma and 1 ng/mL in urine. The method showed a satisfactory sensitivity, precision, accuracy, recovery and selectivity. Copyright © 2001 John Wiley & Sons, Ltd.

Abbreviations used:

CID

Collision‐induced dissociation

CPT

camptothecin

SRM

selective reaction monitoring

TFA

trifluoro‐acetic acid

UM‐1

4‐hydroxy methyl metabolite
William Hunter's casts of the gravid uterus at the University of Glasgow
Authors: McCulloch N.A. , Russell D. , McDonald S.W.
Published: August 2002

The Hunterian Collection at the University of Glasgow possesses 11 plaster casts showing the pregnant uterus. Three correspond to Plates I, IV, and VI of Hunter's The Anatomy of the Gravid Uterus Exhibited in Figures (1774), progressive stages of dissection of the same specimen. A further three casts show consecutive stages of dissection of a uterus containing a fetus presenting by the breech. The other specimens show a normal pregnancy at about 6 months, a normal full‐term uterus with the fetal head becoming engaged, a breech presentation with placenta previa and the umbilical cord around the fetal neck, and an obstructed labor with distended bladder and colon. The 10 on display show realistic coloring and are mounted on black wooden stands. An 11th specimen, amateurishly painted, is not on display. The casts differ in their style; some show only the abdomen, pelvis, perineum, and thighs, others show the full torso. They also differ in the amount of detail shown. The first three casts show the cut femur and muscles at the transected ends of the thighs, also shown in some of the plates in the Gravid Uterus. Although these features enhance the artistic impact of both the engravings and the casts, the authors are unconvinced that Hunter deliberately used them to achieve this, as has been claimed. Clin. Anat. 14:210–217, 2001. © 2001 Wiley‐Liss, Inc.
Variations of the prominences of the bony palate and their relationship to complete dentures in Korean skulls
Authors: Lee S.P. , Paik K.S. , Kim M.K.
Published: August 2002

The osteological and morphological variations of the prominences in the bony palate of 160 Korean skulls were studied. The frequency of the occurrence of the posterior palatine crest, located on the posterior border of the greater palatine foramen, was 13.8%. Palatal ridges were observed commonly in the skulls; however, the smooth type, which has no palatal ridges in the palate, was shown in 14.7% of cases, and palatal spines were observed in 33.8%. The prevalence of palatal tubercles was 11.6%, and all were found in the molar region. The palatine torus was found in 18.8% of cases and the most common type was along the median palatine suture from the incisive foramen to the posterior border of the palatine bone (63.3%). No significant differences between sexes or sides were found in the posterior palatine crest, palatal ridges, and palatal tubercle. However, the sex distribution of the palatine torus was significantly different (P < 0.05). These results would be helpful clinically in fabricating maxillary complete dentures for edentulous patients. Clin. Anat. 14:324–329, 2001. © 2001 Wiley‐Liss, Inc.
Developing an early intervention programme to prevent child maltreatment
Authors: Naughton Aideen , Heath Alan
Published: August 2002

This paper describes the development and implementation of a programme to prevent child maltreatment in a multicultural, inner city community setting with very high rates of physical abuse and neglect. The rationale and philosophy underpinning the prevention programme is outlined. It has a unique approach, combining several features present to varying degrees in other home visiting programmes within a well‐structured framework. This includes the identification of vulnerable parents during the antenatal period, the use of generic health professionals (health visitors) in identification and intervention with families, an ‘active engagement’ strategy, the development of specialist non‐stigmatizing clinics for parents and the creation of a consultation service for generic workers. A variety of interventions are briefly described which are used to support vulnerable families. These include antenatal parenting workshops; baby massage; dedicated crying, sleeping and feeding clinics; and clinics to help parents manage toddler behaviour.Evaluation over the first 5 years of these clinics shows almost 75% of cases have successful outcomes. Copyright © 2001 John Wiley & Sons, Ltd.
Use of small stents for rehabilitation of hypoplastic pulmonary arteries in pulmonary atresia with ventricular septal defect
Authors: Vranicar Mark , Teitel David F. , Moore Phillip
Published: August 2002

Branch pulmonary artery stenosis frequently occurs in pulmonary atresia with ventricular septal defect (PA/VSD). Balloon dilation alone is often unsuccessful in patients with severely hypoplastic pulmonary arteries with residual stenoses after surgical repair. In an attempt to promote distal pulmonary artery growth, 17 stents were placed in 12 severely stenotic pulmonary artery lesions in 10 patients with PA/VSD. All had prior surgery, including pulmonary artery repair, right ventricle to pulmonary artery homograft, and, in 6 of 10, closure of VSD. Median age at stent placement was 16.8 months (range, 13.2–56). Stents were placed using 3.0, 3.5, or 4.0 mm balloons in all but one lesion, in which a 7 mm balloon was used. Following stent placement, there was an increase in the lesion diameter from 1.5 to 3.4 mm (P < 0.05) and an increase in flow to the affected lung from 27% to 34% (P < 0.05). Repeat catheterization 2 to 6 months after stenting in six patients revealed complete occlusion in two of eight lesions. In the other six vessels, there was an increase in distal vessel diameter from 2.96 to 3.94 mm (P < 0.05) even though four had severe restenosis requiring restenting. Two patients underwent surgical pulmonary artery reconstruction and stent removal because of adequate distal vessel growth. Stenting of hypoplastic pulmonary arteries in PA/VSD results in immediate improvement in vessel size and blood flow. Stent restenosis is common although distal vessel growth can be achieved. Stenting of these lesions should be reserved only for those patients unresponsive to other interventions. Cathet Cardiovasc Intervent 2002;55:78–82. © 2002 Wiley‐Liss, Inc.
Intracoronary beta‐radiation of de novo coronary lesions using a 186Re liquid‐filled balloon system: Six‐month results from a clinical feasibility study
Authors: Coussement Patrick K. , Stella Pieter , Vanbilloen Hubert , Verbruggen Alfons , van Rijk Peter , Hoekstra A. , Van Limbergen Erik , de Jaegere Peter , De Scheerder Ivan
Published: August 2002

Vascular brachytherapy has shown to be effective for in‐stent restenosis, but efficacy in de novo lesions remains uncertain. We evaluated feasibility and outcome of intracoronary beta‐radiation therapy in de novo coronary lesions using a 186Re liquid‐filled balloon system. Thirty‐three patients received 20 Gy 186Re beta‐radiation immediately after balloon angioplasty. The 6‐month restenosis rate was 41% (12/29) and restenosis was located within the target lesion in eight patients and at the edges of the injured and irradiated segment, outside the target lesion, in four patients. At 6 months, four patients (12%), all stented during the initial procedure, had experienced a late (> 30 days) total occlusion. Intracoronary beta‐radiation therapy of de novo coronary lesions using 186Re is technically feasible. No reduction in restenosis was observed. The high incidence of late total occlusions may have been prevented by avoiding new stent implantation and prolonging double antiplatelet therapy. Cathet Cardiovasc Intervent 2002;55:28–36. © 2002 Wiley‐Liss, Inc.
Closure of an iatrogenic aortocoronary arteriovenous fistula: Transcatheter balloon embolization following failed coil embolization and salvage of coils that migrated into the coronary venous system
Authors: Maier Lars S. , Buchwald Arnd B. , Ehlers Björn , Rühmkorf Klaus , Scholz Karl H.
Published: August 2002

We report a 50‐year‐old patient with successful percutaneous closure of a large inadvertent surgical aortocoronary arteriovenous fistula (shunt flow: 1.8 L/min). With initial embolization of multiple coils, no lasting occlusion of the large fistula could be achieved. Above that, two coils migrated into the coronary venous system. Following rescue of the migrated coils through a retrograde coronary sinus approach, the fistula was occluded using a detachable balloon. Follow‐up angiograms confirmed successful closure of the fistula. In contrast to coil embolization, use of a detachable balloon seems to be the appropriate technique for percutaneous closure of such fistulas. Cathet Cardiovasc Intervent 2002;55:109–112. © 2002 Wiley‐Liss, Inc.
Use of a 0.052″ Gianturco coil to embolize a persistent right superior vena cava following extracardiac total cavopulmonary connection
Authors: Tomita Hideshi , Ishikawa Yuichi , Hasegawa Satoshi , Ono Yasuo , Yamada Osamu , Kimura Kohji , Uemura Hideki , Yagihara Toshikatsu , Echigo Shigeyuki
Published: August 2002

A persistent right superior vena following extracardiac total cavopulmonary connection was occluded using a 0.052″ Gianturco coil combined with a 3 Fr biopsy forceps. Controlled delivery of a 0.052″ Gianturco coil is a safe and effective procedure to occlude a large anomalous vessel other than a large persistent arterial duct. Cathet Cardiovasc Intervent 2001;52:481–483. © 2001 Wiley‐Liss, Inc.
An unusual case of groin discomfort
Authors: Tuli Ajay , Lim Sung S. , Zafari A. Maziar
Published: August 2002

Groin complications after cardiac catheterizations are common. With the increasing use of mechanical hemostatic devices, cardiologists must be alert to a wide array of potential problems. We report an unusual complication after the use of a closure device. Cathet Cardiovasc Intervent 2001;52:484–485. © 2001 Wiley‐Liss, Inc.
Successful stenting of a complex inferior vena cava stenosis using a modified sharp recanulization technique
Authors: Patel Tejas , Shah Sanjay , Sanghvi Kintur , Fonseca Keith
Published: August 2002

We describe a case of a 30‐year‐old male who presented with features of noncirrhotic portal hypertension, who was diagnosed to have inferior vena cava (IVC) obstruction. IVC angiogram and ultrasound study revealed a long‐segment (36 mm long), chronic total thrombotic occlusion that was dilated and stented with a satisfactory end result. The unique feature of this case is a modified sharp recanulization technique involving the use of Brokenborough (septal puncture) needle and Mullin dilator to create a track in such a long, chronic total occlusion under simultaneous ultrasound and fluoroscopic guidance. Cathet Cardiovasc Intervent 2001;52:492–495. © 2001 Wiley‐Liss, Inc.
Laser angioplasty in a right coronary artery stent restenosis with the guidance of left internal mammary artery contrast injection: Case report
Authors: Okmen Ertan , Cakmak Mahmut , Celik Seden , Yapici Fikri , Erdinler Izzet , Ozturk Recep , Cam Nese
Published: August 2002

A 43‐years‐old male who underwent right coronary artery stent implantation 6 months ago presented with total occlusion of right coronary artery at proximal part of the stent without antegrade flow. We successfully used the left internal mammary artery contrast injections for guidance in all stages of the right coronary laser angioplasty intervention. Cathet Cardiovasc Intervent 2001;53:71–74. © 2001 Wiley‐Liss, Inc.
Noninvasive detection of coronary lesions by multislice computed tomography: Results of the new age pilot trial
Authors: Schroeder Stephen , Kopp Andreas F. , Baumbach Andreas , Kuettner Axel , Herdeg Christian , Rosenberger Albert , Selbmann Hans‐K. , Claussen Claus D. , Oberhoff Martin , Karsch Karl R.
Published: August 2002

The reliable noninvasive assessment of coronary artery disease would constitute an important step forward in clinical cardiology. The aim of the New Age pilot trial was to evaluate the diagnostic accuracy of multislice computed tomography (MSCT) in determining coronary lesions. As a gold standard for in vivo plaque detection, intracoronary ultrasound (ICUS) was used. Forty plaques were detected by ICUS in 15 target vessels (LAD, n = 8; RCA, n = 7) in patients assigned for ICUS‐guided PTCA. Preinterventional MSCT was performed in all patients and the results were compared to ICUS with regard to lesion detection and quantification. According to ICUS results, the 40 plaques were divided into three groups: group I, mild lesions < 50% (n = 14; 44.36% ± 5.77%); group II, intermediate lesions 50%–75% (n = 12; 59.18% ± 9.39%); and group III, severe lesions > 75% (n = 14; 91.47% ± 3.68%). All MSCT scans showed sufficient image quality for analysis. Thirty of 40 (75%) plaques were detected by MSCT in a first blinded session. After unblinding the ICUS results, the remaining 10 (25%) plaques could be identified. Lesion severity was classified correctly in 34 of 40 (85%) plaques. Plaque calcifications were diagnosed correctly in 16 of 19 (84.2%) plaques. Quantification of vessel size revealed a good correlation to the ICUS results (r2 0.68; P = 0.004). Noninvasive MSCT angiography showed good diagnostic accuracy with regard to lesion detection and quantification of vessel size. The overall good image quality, makes this new technology a promising modality, which might become an alternative diagnostic approach in patients with known or suspected coronary artery disease. Cathet Cardiovasc Intervent 2001;53:352–358. © 2001 Wiley‐Liss, Inc.