bluuebunny/crossref_metadata_2025_split huggingface dataset:
Water‐based nanoparticulate polymeric system for protein delivery
Authors: Prokop Ales , Holland Celia A. , Kozlov Evgenii , Moore Billy , Tanner Robert D. 
Published: August 2002

AbstractThis 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

AbstractA 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

AbstractThe 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

AbstractChlorophyllide 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

AbstractCatalytic 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

AbstractMucosal 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

AbstractIn 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

AbstractThe 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

AbstractThe 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

AbstractRegulated 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

AbstractA 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

AbstractA 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

AbstractThe 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

AbstractThe 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

AbstractThis 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.