bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tyrosine‐based “Activatable Pro‐Tag”: Enzyme‐catalyzed protein capture and release
Authors: Lewandowski Angela T. , Small David A. , Chen Tianhong , Payne Gregory F. , Bentley William E. 
Published: February 2006

AbstractProtein recovery is often achieved by a series of capture and release steps that often involve chromatographic binding and elution. We report an alternative, non‐chromatographic, capture and release approach that employs enzymes and the stimuli‐responsive polysaccharide chitosan. We capture our protein using the enzyme tyrosinase that oxidizes accessible tyrosine residues of the protein and “activates” these residues for covalent capture (i.e., conjugation) onto chitosan. Using fusions of green fluorescent protein (GFP) we observed that: (i) enzymatic activation is required for protein capture to chitosan; and (ii) capture is enhanced (approximately five‐fold) by engineering the protein to have a penta‐tyrosine fusion tag that provides additional accessible tyrosine residues for enzymatic activation. Because the fusion tag appears to be the primary site for capture, and capture requires activation, we designate penta‐tyrosine as a “pro‐tag.” The captured GFP‐chitosan conjugate possesses the pH‐responsive solubility that is characteristic of chitosan. We exploit this pH‐responsive solubility to facilitate purification of the captured protein. Two enzymatic methods were explored to release the captured GFP from the chitosan conjugate. The first method employs enterokinase (EK) to cleave the protein at an engineered EK‐cleavage site. The second method employs chitosanase to hydrolyze the chitosan backbone. Using GFP as a model protein, we demonstrated that enzymatic capture and release provides a simple, non‐chromatographic means to recover proteins directly from cell lysates. © 2006 Wiley Periodicals, Inc.
A population-based study of the incidence, management and prognosis of hepatic metastases from colorectal cancer
Authors: Leporrier J , Maurel J , Chiche L , Bara S , Segol P , Launoy G 
Published: March 2006

AbstractBackgroundThe aim of this population-based study was to evaluate the incidence, management and prognosis of patients with hepatic metastases related to colorectal cancer using data from the Digestive Cancer Registry of Calvados, France.MethodsOf 1325 patients with colorectal cancer registered between January 1994 and December 1999, 358 developed hepatic metastases. Logistic regression was used to analyse prognostic factors. Survival analysis was carried out with Cox's proportional hazards model.ResultsSome 18·8 per cent of patients had synchronous metastases, while 29·3 per cent developed metastases at 3 years. Of patients with hepatic metastases, 17·3 per cent had a surgical resection, 40·2 per cent were treated with palliative chemotherapy and 42·5 per cent had symptomatic treatment. Factors associated with receiving symptomatic treatment only were age over 75 years and more than one metastasis, but not place of treatment. Median survival after a diagnosis of hepatic metastases was 10·7 (range 4·6–23·1) months. Significant adverse prognostic factors were: age over 75 years (P = 0·001), lymph node invasion of primary tumour (P = 0·024), bilateral distribution of metastases (P = 0·001), other metastases (P = 0·004) and symptomatic treatment only (P = 0·041).ConclusionDespite improvement in treatment for hepatic metastases, age and extent of disease remain limiting factors for surgical resection and palliative chemotherapy.
Prognosis of small thyroid cancer in patients with Graves' disease
Authors: Kikuchi S , Noguchi S , Yamashita H , Uchino S , Kawamoto H 
Published: March 2006

Abstract

Background
To find the best ways to follow up patients with small thyroid cancer (STC; 1 cm or less in diameter) and concomitant Graves' disease, this study examined whether such patients had the same excellent prognosis as those with STC without Graves' disease.

Methods
Between 1970 and 1996, 2199 patients were diagnosed as having STC by pathology after thyroidectomy. Of those, 509 patients (33 males and 476 females, mean age 43·5 years) underwent thyroidectomy for Graves' disease. Control patients with STC without Graves' disease were matched for age, sex, treatment year and tumour size (33 males and 476 females, mean age 44·0 years).

Results
Patients with STC and Graves' disease had a longer disease-free survival than those with STC alone (99 and 93 per cent at 20 years' follow-up, respectively; P &lt; 0·001). The Cox's proportional hazard analysis showed that concomitant Graves' disease and age at surgery are more significant factors for predicting disease-free survival than surgical procedures.

Conclusion
Patients who undergo thyroidectomy for Graves' disease and are found to have STC have an excellent prognosis and longer disease-free survival than patients with STC alone.
Randomized clinical trial to determine the effect of nasogastric drainage on tracheal acid aspiration following oesophagectomy
Authors: Shackcloth M J , McCarron E , Kendall J , Russell G N , Pennefather S H , Tran J , Page R D 
Published: March 2006

Abstract

Background
The aim of this study was to investigate tracheal acid aspiration after oesophagectomy and to determine whether it is influenced by nasogastric (NG) drainage.

Methods
Thirty-four patients undergoing oesophagectomy were randomized to one of three methods of NG drainage: a single-lumen tube with free drainage and 4-hourly aspiration, a sump-type tube on continuous suction drainage, or no NG tube. A tracheal pH probe was used to collect information on acid aspiration for 48 h after surgery. A pH &lt; 5·5 was considered abnormal (normal pH 6·8–7·2). Total time with tracheal pH &lt; 5·5, number of reflux episodes and longest reflux time were compared between groups.

Results
There was significant and persistent tracheal acid aspiration in all patients. Patients with a sump-type tube had a significantly shorter total time with tracheal pH &lt; 5·5 than those in the other groups (sump-type tube versus single-lumen tube, P = 0·0069; sump-type tube versus no tube, P = 0·0071). Patients randomized to no NG tube experienced more respiratory complications after surgery than those who had either single-lumen or sump-type tubes (seven of 12 versus four of 22 patients; P = 0·023). Insertion of a NG tube was necessary in the first week after surgery in seven of 12 patients in this group.

Conclusion
Routine NG drainage after oesophagectomy is necessary. A sump-type NG tube is better at preventing tracheal acid aspiration and may reduce the incidence of respiratory complications.
Selective cyclo-oxygenase 2 inhibition affects ileal but not colonic anastomotic healing in the early postoperative period
Authors: de Hingh I H J T , van Goor H , de Man B M , Lomme R M L M , Bleichrodt R P , Hendriks T 
Published: March 2006

AbstractBackgroundSelective cyclo-oxygenase 2 (COX-2) inhibitors are increasingly prescribed in the perioperative period. Recent recognition of a possible role for COX-2 in wound healing has raised concerns about the safety of their use in surgical practice. Therefore, the influence of celecoxib, a selective COX-2 inhibitor, on early anastomotic healing was investigated.MethodsCelecoxib, in doses of 15, 50 or 200 mg per kg per day, was given daily from the day before operation onwards to male Wistar rats that received both ileal and colonic anastomoses. Anastomotic strength was assessed by measuring the breaking strength and bursting pressure on the third day after operation. A second group received a dose of 50 mg per kg per day and a colonic anastomosis only, and healing was assessed on the third and fifth day after surgery.ResultsExpression of COX-2 protein was upregulated in the anastomotic area. Administration of celecoxib, at all doses tested, resulted in a significantly higher ileal dehiscence rate than in control rats (P = 0·002). In contrast, colonic anastomoses healed normally within the same animals. The latter was confirmed in rats with colonic anastomoses only.ConclusionIn this model, administration of the COX-2 inhibitor celecoxib affected ileal but not colonic anastomotic healing in the early postoperative period.
Decontamination of organochlorine pesticides in Radix Codonopsis by supercritical fluid extractions and determination by gas chromatography
Authors: Zhao Chunjie , Hao Guiming , Li Huanxin , Luo Xu , Chen Yingjie 
Published: February 2006

AbstractA method involving depuration of 12 organochlorine pesticides (OCPs) from Radix codonopsis was developed using supercritical fluid extraction (SFE). The pesticides investigated in the study included α‐, β‐, γ‐ and δ‐benzene hexachloride, PCNB (pentachloro‐nitrobenzene), PCA (pentachloroaniline), HEPT (heptachlor), MPCPS (methyl‐pentachlorophenyl sulfide), pp′‐DDE [1,1‐dichloro‐2, 2‐bis(p‐chlorophenyl) ethylene], op′‐DDT [1,1,1‐trichloro‐2‐(o‐chlorophenyl)‐2‐(p‐chlorophenyl)ethane], pp′‐DDD [1,1‐dichloro‐2‐2‐bis(p‐chlorophenyl)ethane] and pp′‐DDT [1,1,1‐trichloro‐2,2‐bis(p‐chlorophenyl)ethane]. A series of experiments was conducted to optimize the final extraction conditions as follows: pure CO2, extraction pressure 15 MPa, extraction temperature 60°C, extraction time 20 min and flow rate 55 mL/h. A GC method with electron capture detection was employed for the determination of the OCPs in Radix codonopsis. An HPLC method was developed for the quantitative determination of active constituents. SFE was used to remove the organochlorine pesticide from Radix codonopsis. The results showed that at least 93.5% of the organochlorine pesticide residues in the herb sample were removed by SPE, while 95.0% of the active constituent marker (atractylenoide III) remained. Copyright © 2006 John Wiley & Sons, Ltd.
Determination of erythromycin A in rat plasma and urine by high‐performance liquid chromatography with chemiluminescence detection using Tris(2,2′‐bipyridine)ruthenium(II)
Authors: Hori Toshihiko , Hashimoto Hiroshi , Konishi Masaharu 
Published: February 2006

AbstractA simple and sensitive method was developed for the determination of erythromycin A (EA), decladinosyl erythromycin A (dClEA) and erythromycin B (EB) in rat plasma and urine by high‐performance liquid chromatography with electrogenerated chemiluminescence detection using Tris(2,2′‐bipyridine)ruthenium(II). The recovery rates of EA, dClEA and EB were 97, 94 and 85% from rat plasma and 89, 83 and 93% from rat urine, respectively. The calibration curves were linear over the concentration ranges 0.05–5 µg/mL for plasma and 0.5–50 µg/mL for urine. The precision and accuracy for all analytes in rat plasma were ≤9.0 and −6.3–7.2%, and those in urine were ≤9.4% and −6.1–7.6%, respectively. This method proved to be a powerful tool for determination of EA, dClEA and EB concentrations in samples from rats. Copyright © 2006 John Wiley & Sons, Ltd.
A structure–activity relationship study of compounds with antihistamine activity
Authors: Brzezińska Elżbieta , Kośka Grażyna 
Published: February 2006

AbstractA structure–activity relationship (SAR) analysis of H1‐, H2‐ and H3‐antihistamine activity was carried out and chromatographic data of 2‐[2‐(phenylamino)thiazol‐4‐yl]ethanamine, 2‐(2‐benzyl‐4‐thiazolyl)ethanamine, 2‐(2‐benzhydrylthiazol4‐yl)ethanamine, 2‐(1‐piperazinyl‐ and 2‐(hexahydro‐1H‐1,4‐diazepin‐1‐yl)benzothiazole, 2‐(1‐piperazinyl)benzothiazole, 2‐[4‐(1‐alkyl)piperidinyl]benzothiazole, 2‐(N,N′,N′‐dimethylalkyl‐1,2‐ethanediamino)benzothiazole, 2[1‐(4‐aminopiperidinyl)]benzothiazole, 2‐[2‐phenyl‐4‐thiazolyl]ethanamine derivatives and selected H1‐ and H2‐antihistamine drugs were obtained. NP TLC and RP2 TLC plates (silica gel NP 60F254 and silica gel RP2 60F254 silanized precoated), impregnated with a solution of aspartic acid (l‐Asp) and a solution of an analogue of aspartic acid (propionic acid), were used in two developing solvents as H1‐, H2‐ and H3‐antihistaminic interaction models. The lipophilicity data of the examined compounds were obtained and used in the SAR assay. Biochromatographic tests using TLC plates impregnated with solutions of asparic acid or propionic acid were found to be a source of useful data for the qualitative analysis of compounds with different structures, demonstrating activity to histamine H1‐, H2‐ and H3‐receptors. The four presented discriminant models based on biochromatographic studies are an efficient tool in the SAR analysis for initial prediction of compound activity direction within histamine receptors. Copyright © 2006 John Wiley & Sons, Ltd.
Pharmacokinetic study of p‐coumaric acid in mouse after oral administration of extract of Ananas comosus L. leaves
Authors: Meng Zhen , Wang Wei , Xing Dong‐Ming , Lei Fan , Lan Jia‐Qi , Du Li‐Jun 
Published: February 2006

AbstractQuantification of p‐coumaric acid in mouse plasma following oral administration of Ananas comosus L. leaves was achieved by reversed‐phase high‐performance liquid chromatography using a mobile phase of water–acetonitrile (82:18, v/v) and UV detection at 310 nm. The method was linear (determination coefficient, r2 = 0.9997) within the tested range (0.04–1.28 µg/mL). Intra‐ and inter‐day precision coefficients of variation and accuracy bias were acceptable (maximal CV value was 4.06% for intra‐day and 4.19% for inter‐day) over the entire range. The recoveries were 90.63, 97.98 and 100.01% for concentrations of 0.04, 0.32 and 1.28 µg/mL, respectively. This is a very rapid, sensitive and economical way to determine p‐coumaric acid concentration in mouse plasma after oral administration of A. comosus leaves. The concentration–time curve was fitted to the one‐compartment model. This is the first time that p‐coumaric acid extracted from A. comosus leaves was detected by HPLC‐UV method and its pharmacokinetic characteristic was comprehensively studied. Copyright © 2006 John Wiley & Sons, Ltd.
Sensitive determination of 4‐(4‐chlorophenyl)‐4‐hydroxypiperidine, a metabolite of haloperidol, in a rat biological sample by HPLC with fluorescence detection after pre‐column derivatization using 4‐fluoro‐7‐nitro‐2,1,3‐benzoxadiazole
Authors: Higashi Yasuhiko , Nakamura Shota , Fujii Youichi 
Published: February 2006

Abstract4‐(4‐Chlorophenyl)‐4‐hydroxypiperidine (CPHP), one of the metabolites of haloperidol, is considered to exhibit brain toxicity. CPHP concentrations in plasma and tissue homogenates (each 200 µL) from rats were analyzed by HPLC fluorescence detection after pre‐column derivatization with 4‐fluoro‐7‐nitro‐2,1,3‐benzoxadiazole (NBD‐F). After basic extraction of the samples with benzene, the derivatization with NBD‐F was conducted in borate buffer (pH 8.0) at 60°C for 3 min. Mexiletine was carried through the procedure as an internal standard. The regression equation for CPHP showed a good linearity in the range of 0.03–1 µg/mL with a detection limit of 0.008 µg/mL. The coefficient of variation was less than 11.6%. Plasma concentration–time courses of CPHP after intraperitoneal or per oral administration of CPHP, haloperidol or reduced haloperidol were examined, and the pharmacokinetic parameters were estimated. Additionally, CPHP levels in various tissues at 8 h after intraperitoneal administration of these compounds were compared. The method was simple and sensitive, useful for determination of CPHP in rat biological samples using as little as 200 µL of sample volume and could be applied for pharmacokinetic study. Copyright © 2006 John Wiley & Sons, Ltd.
Polyurethane foam chips combined with liquid chromatography in the determination of unmetabolized polycyclic aromatic hydrocarbons excreted in human urine
Authors: Buratti Marina , Pellegrino Oronzo , Valla Carla , Rubino Federico Maria , Verduci Cinzia , Colombi Antonio 
Published: February 2006

AbstractA method suitable for the determination of unmetabolized polycyclic aromatic hydrocarbons (PAHs) excreted at trace levels (ng/L) in human urine for the monitoring of exposure of the general population to PAH contamination was developed. PAHs were determined, after enrichment by solid‐phase extraction on polyurethane foam (PUF) chips, by HPLC with fluorescence detection. Different parameters affecting analyte extraction to the PUF, including urine salting‐out and organic additives, and optimization of conditions for clean‐up and desorption have been investigated. Optimized conditions were 40 mL acidified urine sample, added with magnesium sulfate, tetrahydrofuran and a 2 cm3 PUF chip, and extracted by shaking at 30 rpm for 1 h at ambient temperature. Desorption was performed, after a clean‐up step with diluted sodium hydroxide, using a small amount of diethyl ether. The recovery of PAH congeners from spiked urines was >90% in the 2–100 ng/L range; the detection limit was 0.1–0.5 ng/L, depending on the considered PAH congener; day‐to‐day precision, at 50 ng/L native PAH content, was CV = 10–20%. The proposed technique provides a simple, economical and effective procedure for the determination of trace amounts of unmetabolized PAHs excreted in human urine spot samples. Copyright © 2006 John Wiley & Sons, Ltd.
A further study on the combined use of internal standard and isotope‐labeled derivatization reagent for expansion of linear dynamic ranges in liquid chromatography–electrospray mass spectrometry
Authors: Tsukamoto Yuhki , Santa Tomofumi , Yoshida Hiroo , Miyano Hiroshi , Fukushima Takeshi , Hirayama Kazuo , Imai Kazuhiro , Funatsu Takashi 
Published: February 2006

AbstractThe combined use of a so‐called internal standard and the isotope‐labeled derivatization reagent for the quantification of analytes for liquid chromatography–mass spectrometry (LC/MS) was further studied. The sample solution (containing the analytes and an internal standard) was derivatized with the light form of the derivatization reagent, 7‐(N,N‐dimethylaminosulfonyl)‐4‐(aminoethyl)piperazino‐2,1,3‐benzoxadiazole (DBD‐PZ‐NH2) or 7‐(N,N‐dimethylaminosulfonyl)‐4‐piperazino‐2,1,3‐benzoxadiazole (DBD‐PZ). A standard solution of the analytes (containing an internal standard) was derivatized with the isotope (d6)‐labeled derivatization reagent, DBD‐PZ‐NH2 (D) or DBD‐PZ (D), and served as the isotope‐labeled internal standards. The peak heights of the targeted analytes derivatives in the sample solution were corrected using those of the internal standard and the heavy form derivatives of the standards, and the calibration curves were constructed. The curve bending of the calibration curves caused by the ion suppression at the ion source was suppressed and the linear dynamic ranges of the calibration curves were expanded. The derivatives of DBD‐PZ‐NH2 were about 10 times more sensitively detected than those of DBD‐PZ derivatives and, therefore, DBD‐PZ‐NH2 might be suitable for sensitive detection. Copyright © 2006 John Wiley & Sons, Ltd.
Laboratory Evolution of P450 BM‐3 for Mediated Electron Transfer
Authors: Nazor Jovana , Schwaneberg Ulrich 
Published: March 2006

AbstractPreparative synthesis with P450 monooxygenases is hampered in cell‐free systems by the requirement for cofactors such as NAD(P)H as reduction equivalents. A validated medium‐throughput screening system was designed for improving P450 monooxygenases by mediated electron transfer with zinc/cobalt(III)sepulchrate (Zn/CoIIIsep) as an alternative and cost‐effective cofactor system. The monooxygenase P450 BM‐3 F87A was used as a model system for developing the screening system in a 96‐well format. A coefficient of variation of less than 10 % was achieved under optimized screening conditions. The mediator evolution screen was validated by comparing the activity of P450 BM‐3 to P450 BM‐3 F87A and by screening a saturation mutagenesis library at amino acid position R47. For mediated electron transfer, two double mutants P450 BM‐3(F87A R47F) and P450 BM‐3 (F87A R47Y) were identified with a two–threefold increased catalytic efficiency (up to 32 μM−1 min−1 for P450 BM‐3(F87A R47F) and 34 μM−1 min−1 for P450 BM‐3 (F87A R47Y)) compared to P450 BM‐3 F87A. The kinetic constants of the double mutants are, in contrast to those of P450 BM‐3 F87A, dependent on CoIIIsep concentration in the presence of NADPH. kcat increases from 145 min−1 (0.25 mM CoIIIsep) to 197 min−1 (0.5 mM CoIIIsep), and Km decreases simultaneously from 7.0 μM to 3.7 μM, for P450 BM‐3 (F87A R47F). For P450 BM‐3 (F87A R47Y), kcat increases from 138 min−1 (0.25 mM CoIIIsep) up to 187 min−1 (0.5 mM CoIIIsep), and Km decreases from 8.2 μM to 4.2 μM. Due to lower Km values, the catalytic efficiencies were improved six times for P450 BM‐3 (F87A R47F) and three times for P450 BM‐3 (F87A R47Y), when comparing catalytic efficiencies of the mediated electron‐transfer system to the natural reduction equivalent NADPH.
Covert sensitization revisited: six case studies
Authors: Kraft Tom , Kraft David 
Published: February 2006

AbstractThis is an up to date study in which covert sensitization – a technique that seems to have gone into disrepute since the 1970s – is employed to treat a variety of maladaptive behaviours. The following six case studies illustrate the value of covert sensitization for the treatment of alcoholism, nail tearing, cigarette smoking, cannabis smoking, overeating and chocolate addiction. The treatment focuses on the craving rather than the actual carrying out of the unwanted behaviour. This study shows that covert sensitization is a rapid and cost effective form of treatment: many patients are able to eliminate the unwanted behaviour in a small number of sessions. Copyright © 2005 British Society of Experimental & Clinical Hypnosis. Published by John Wiley & Sons, Ltd.
Differing roles of imagination and hypnosis in self‐regulation of eating behaviour
Authors: Hutchinson‐Phillips Susan , Jamieson Graham A , Gow Kathryn 
Published: February 2006

AbstractThis study investigates the role of hypnosis‐like and imagery processes in the self‐regulation of eating behavior. Eighty participants were assessed on measures of eating behaviour (EAT), body image (CDRS), trait measures of hypnotic ability (HGSHS:A) and imaginative ability (CIS) as well as process measures (ICMI, SIPI and PCI) spanning both domains. Imagination and hypnosis measures show distinct patterns of relationships with eating behaviour and body image measures. Results emphasize the importance of assessing process as well as trait or ability measures and the utility of the Phenomenology of Consciousness Inventory (PCI) to this approach. Therapeutic implications are drawn for integrating hypnosis and imagination procedures in cognitive behaviour therapy (CBT) of eating disorders. Present findings should be confirmed in a larger sample and the emerging model tested utilizing Structural Equation Modelling. Copyright © 2005 British Society of Experimental & Clinical Hypnosis. Published by John Wiley & Sons, Ltd.
Psychogenic pain: a study using multidimensional scaling
Authors: Whalley Matthew G. , Oakley David A. 
Published: February 2006

AbstractHypnotic suggestions designed to induce a sensation of pain were given to 11 highly hypnotizable participants and produced the intended effect in six of them. Subjective aspects of the pain experiences were investigated using conventional pain questionnaires and a multidimensional scaling technique. The implications are discussed in terms of a model of consciousness and the etiology of chronic pain. Copyright © 2003 British Society of Experimental and Clinical Hypnosis