bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis and characteristics of the interpenetrating polymer network hydrogel composed of chitosan and polyallylamine
Authors: Shin Mi‐Seon , Kim Seon Jeong , Park Sang Jun , Lee Yong Hee , Kim Sun I.
Published: October 2002

A temperature‐ and pH‐dependent hydrogel was studied with interpenetrating polymer network (IPN) hydrogels constructed with water‐insoluble chitosan and polyallylamine. Various IPNs were prepared from different weight ratios of chitosan–polyallylamine. Crosslinked‐IPN hydrogels exhibited relatively high equilibrium water content (EWC) in the range 80–83%. The EWC of IPN hydrogels depended on pH and the amount of complex, which is the content of chitosan and polyallylamine. The differential scanning calorimeter (DSC) thermogram of fully swollen IPN hydrogels appeared between 3 to 4 °C. The IPNs exhibited two glass‐transition temperatures (Tgs), indicating the presence of phase separation in the IPNs as exhibited by dielectric analysis (DEA). © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 498–503, 2002
Cationic UV cure kinetics for multifunctional epoxies
Authors: Boey F. , Rath S. K. , Ng A. K. , Abadie M. J. M.
Published: October 2002

Two epoxy monomers with widely different functionalities (f = 2 and f = 8) were selected for the study and comparison of photocuring kinetics, in the presence of two different photoinitiators. It has been observed that for the same photoinitiator, the optimum concentration is lower for the epoxy monomer with a lower functionality. In terms of photoinitiation efficiency, the photoinitiator based on aromatic sulfonium salts has been determined to be a better candidate. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 518–525, 2002
Formation of short‐lived multinucleated giant cells (MGCS) from cultured gilthead seabream macrophages
Authors: Couso Norma , Castro Rosario , Noya Manuel , Obach Alex , Lamas Jesús
Published: October 2002

Monocytes/macrophages obtained from the head kidney and peritoneal cavity of gilthead seabream (Sparus aurata) were cultured using plates from three different manufacturers, and were maintained under different conditions. The effects on the morphology and fusion of monocytes/macrophages of initial cell loading, removal of non‐adherent cells at different times after plating, and addition of serum and antibiotics were evaluated by light microscopy, and transmission (TEM) and scanning (SEM) electron microscopy. Despite variations in adherence, the behaviour and the morphological changes in kidney monocytes/macrophages were similar in all three types of plates. When foetal calf serum (FCS) was added to the incubation medium, most of the cells resembling monocytes/macrophages were connected by cytoplasmic extensions that formed bridges after 24 hr in culture. After 30 hr, the monocytes/macrophages started to fuse, forming multinucleated giant cells (MGCs) which gradually increased in size until the culture was 4–5 days old. After 5 days the MGCs started to die, and after a week most had disappeared from the cultures. Cells incubated with medium without serum showed changes similar to those fed with FCS, but some cells survived for 3 weeks. The addition of fish serum to the medium appeared to accelerate all processes: the monocytes/macrophages and MGCs died after 3 days in culture. Antibiotics had no apparent effect on the cultures. Removal of non‐adherent cells at different times after plating did not appear to affect cell fusion. Coating the wells with extracellular matrix proteins reduced adherence but did not inhibit cell fusion. Curiously, not all macrophages fused with MGCs, and, unlike MGCs, these macrophages phagocytosed sheep red blood cells (SRBCs). Peritoneal macrophages also fused and formed MGCs in culture, similarly to kidney cells. Anat Rec 267:204–212, 2002. © 2002 Wiley‐Liss, Inc.
Sexual dimorphism in digit‐length ratios of laboratory mice
Authors: Brown Windy M. , Finn Christopher J. , Breedlove S. Marc
Published: October 2002

The lengths of the index finger (2D) and ring finger (4D) are sexually dimorphic in humans, and men have a smaller 2D:4D ratio compared to women. Prenatal androgens appear to be important in the development of the 2D:4D sex difference, since it has been reported in children as young as 2 years old, and since humans exposed to supernormal prenatal androgen levels display a smaller 2D:4D ratio. We tested whether another mammalian species in which the process of peripheral sexual differentiation is androgen‐dependent might also show a sex difference in digit ratios. The 2D:4D ratio of adult outbred mice was calculated for both the left and right rear paws. A sex difference was observed in the right rear paw: female mice had a larger 2D:4D ratio than did males. We also found this difference in prepubescent weanling mice. This sex difference is in the same direction as that observed in humans, and suggests that sexual dimorphism in digit length ratios is a feature common to many, if not all, mammals. The mouse may therefore be a useful animal model for studying the factors that influence finger length patterns, which have recently been correlated with several specific behaviors and disease predispositions in humans. Anat Rec 267:231–234, 2002. © 2002 Wiley‐Liss, Inc.
Alterations in carbohydrate and fatty acid levels of Lymantria dispar larvae caused by a microsporidian infection and potential adverse effects on a co‐occurring endoparasitoid, Glyptapanteles liparidis
Authors: Hoch Gernot , Schafellner Christa , Henn Michael W. , Schopf Axel
Published: October 2002

Infection of Lymantria dispar host larvae by the entomopathogenic microsporidium Vairimorpha sp. has a negative impact on the performance of the endoparasitic braconid Glyptapanteles liparidis. To investigate possible causes for this effect, we studied to what extent nutritional host suitability is altered by the microsporidium. Therefore, we analyzed carbohydrates and fatty acids in host larvae after Vairimorpha infection and/or parasitism by G. liparidis. Trehalose levels were significantly reduced in the hemolymph of infected hosts. After day five post infection, it was detected only in traces. Four to six days later, the glycogen resources were depleted in infected larvae. Parasitism by G. liparidis, on the other hand, led to increased hemolymph trehalose levels during the early endoparasitic phase but to a significant decrease at the end of its larval development. No effect of parasitism on the glycogen content was ascertained. Hemolymph levels of the fatty acids analyzed, such as palmitic, stearic, oleic, linoleic, and linolenic acid, were significantly reduced in microsporidia‐infected L. dispar. Vairimorpha sp. develops as an intracellular parasite in the fat body of the host larva and synthesis of trehalose and fatty acids may be disturbed. Moreover, microsporidia may also harness metabolites or energy produced by host cells. We conclude that the microsporidia‐induced decrease in hemolymph carbohydrates and fatty acids adversely affects growth and development of parasitoid larvae. Arch. Insect Biochem. Physiol. 50:109–120, 2002. © 2002 Wiley‐Liss, Inc.
Content and composition of phosphoglycerols and neutral lipids at different developmental stages of the eggs of the codling moth, Cydia pomonella (Lepidoptera: Tortricidae)
Authors: Forte Silvina N. , Ferrero Adriana A. , Alonso Telma S.
Published: October 2002

Phosphoglycerol, triacylglycerol, diacylglycerol, and free fatty acid content was studied in eggs of the codling moth Cydia pomonella at the white, red ring, and black head developmental stages. The composition of total phosphoglycerols and of the three classes of neutral lipids was also analyzed. The highest total lipid content was found in eggs at the white stage, the amount decreasing during development mainly as a result of a diminution in the quantity of phosphoglycerols, which account for approximately 50% of total content at all stages of egg development. The amount of triacylglycerols and free fatty acids changes significantly during development, whereas only minor changes were found in diacyglycerol levels. The total phosphoglycerol acyl composition of eggs at the white and red ring stages is similar, whereas differences are evident at the black head stage of development. Triacylglycerols and free fatty acids are enriched in saturated fatty acids in all analyzed stages. The acyl profile of diacylglycerols is different at each stage. The unsaturation index decreases in diacylglycerols and free fatty acids as a function of egg development. The results of the present paper suggest that triacylglycerols may constitute an important source of energy during the final period of egg development while phosphoglycerols may function as fuel during the beginning. Phosphoglycerols could be precursors for the triacylglycerol biosynthesis that takes place between white and red ring stages. Arch. Insect Biochem. Physiol. 50:121–130, 2002. © 2002 Wiley‐Liss, Inc.
Purification and characterization of a digestive alkaline protease from the larvae of Spilosoma obliqua
Authors: Anwar Adil , Saleemuddin M.
Published: October 2002

A digestive protease from Spilosoma obliqua (Lepidoptera: Arctiidae) fifth instar larval guts was purified and characterized. The protease was purified using ammonium sulfate fractionation, ion‐exchange chromatography, and hemoglobin‐sepharose affinity chromatography. The purification procedure resulted in a 37‐fold increase in the specific activity of the protease. Protease thus obtained was found to be electrophoretically pure under native and denaturing conditions. The purified protease had a molecular mass of 90 kDa as determined by gel filtration, and a pH optimum of 11.0. The purified protease optimally hydrolyzed casein at 50°C. A Km of 2 ×10–6 M was obtained using BApNA as a substrate for the purified alkaline protease. The ability of S. obliqua protease and bovine trypsin to hydrolyze various synthetic substrates (BApNA, BAEE, and BAME), and the inhibition patterns of S. obliqua and bovine trypsin with “classical” trypsin inhibitors are also reported. Arch. Insect Biochem. Physiol. 51:1–12, 2002. © 2002 Wiley‐Liss, Inc.
The beneficial effect of interleukin‐12 on arthritis induced by group B streptococci is mediated by interferon‐γ and interleukin‐10 production
Authors: Puliti Manuela , von Hunolstein Christina , Bistoni Francesco , Mosci Paolo , Orefici Graziella , Tissi Luciana
Published: October 2002

Objective

To assess the effect of interleukin‐12 (IL‐12) administration on the evolution of systemic infection and septic arthritis induced by group B streptococci (GBS) in mice.

Methods

CD1 mice were inoculated intravenously with arthritogenic strain 1/82 of type IV GBS. Exogenous murine IL‐12 was administered intraperitoneally 18 hours or 5 days after infection with 1 × 107 GBS, at doses ranging from 0.5 to 2.5 μg per mouse. Mice were monitored daily for survival and for signs of arthritis. In a subsequent set of experiments, mice were killed at selected times for examination of bacterial clearance, histopathologic changes in the joints, and cytokine production.

Results

IL‐12 administration before the onset of clinical signs had a beneficial effect on GBS‐induced arthritis and was clearly dose‐dependent. The 2.5‐μg dose per mouse totally prevented death from GBS‐induced arthritis. The decrease in pathology was associated with a reduction of the bacterial burden and a change in the cytokine profile. In particular, systemic and joint levels of interferon‐γ (IFNγ) and IL‐10 significantly increased in mice treated with IL‐12, whereas a decrease in IL‐6 and IL‐1β production was observed. The beneficial effects of IL‐12, in terms of the incidence and severity of articular lesions, were reversed by coadministration of anti‐IFNγ or anti–IL‐10–neutralizing antibodies.

Conclusion

The findings of this study demonstrate that IL‐12 is important in controlling the cytokine production that leads to the evolution of GBS‐induced experimental arthritis. The amelioration of articular lesions is mostly attributable to IL‐12–induced IFNγ, but with a relevant participation of IL‐12–induced IL‐10.
Identification of citrullinated rheumatoid arthritis‐specific epitopes in natural filaggrin relevant for antifilaggrin autoantibody detection by line immunoassay
Authors: Union Ann , Meheus Lydie , Humbel René Louis , Conrad Karsten , Steiner Guenter , Moereels Henri , Pottel Hans , Serre Guy , De Keyser Filip
Published: October 2002

Objective

To identify immunodominant epitopes in natural filaggrin that are reactive with antifilaggrin autoantibodies (AFA) in the sera of patients with rheumatoid arthritis (RA) and to explore their use in a diagnostic assay format.

Methods

Based on the results of epitope mapping of human natural filaggrin as well as molecular modeling and computational chemistry, synthetic peptides together with recombinant citrullinated filaggrin were evaluated by a line immunoassay (LIA) for AFA detection. Diagnostic performance was assessed using 336 RA and 253 disease control sera and was compared with that of reference methods.

Results

Several immunoreactive epitopes were identified in natural filaggrin, all of which contained at least 1 citrulline residue. Three antigenic substrates, including 2 synthetic peptides and recombinant citrullinated filaggrin showing maximal reactivity on LIA, were finally selected. Using the 3‐antigen LIA3, overall sensitivity, specificity, and positive predictive value for RA were 65.2%, 98.0%, and 89.1%, respectively, compared with 61.9%, 98.8%, and 92.8% using the 2‐antigen LIA2 (without recombinant protein). Thirty‐seven percent of the rheumatoid factor (RF)‐negative RA samples (30 of 81) were AFA‐positive by LIA2, and 52 of 54 RF‐positive control samples had no AFA detected on LIA2. Higher specificity and sensitivity were obtained by LIA2 versus anti‐RA33 immunoblot, whereas good agreement was observed with antikeratin antibody testing. LIA performed significantly better than AFA immunoblotting using natural filaggrin, at a specificity level of 99% (P = 0.0047).

Conclusion

Citrullinated residues are present in immunoreactive epitopes of natural human filaggrin. AFA can be readily detected by citrullinated peptides in an LIA‐based test, resulting in high specificity and positive predictive value for RA. The LIA could serve as a user‐friendly alternative to existing immunofluorescence tests and AFA immunoblot techniques. Given its complementarity to RF, this test can be a valuable tool in the differential diagnosis of arthritis.
Demethylation of ITGAL (CD11a) regulatory sequences in systemic lupus erythematosus
Authors: Lu Qianjin , Kaplan Mariana , Ray Donna , Ray Doreen , Zacharek Sima , Gutsch David , Richardson Bruce
Published: October 2002

Objective

Inhibition of T cell DNA methylation causes autoreactivity in vitro and a lupus‐like disease in vivo, suggesting that T cell DNA hypomethylation may contribute to autoimmunity. The hypomethylation effects are due, in part, to overexpression of lymphocyte function–associated antigen 1 (LFA‐1) (CD11a/CD18). Importantly, T cells from patients with active lupus have hypomethylated DNA and overexpress LFA‐1 on an autoreactive subset, suggesting that the same mechanism could contribute to human lupus. The present study investigated the nature of the methylation change that affects LFA‐1 expression in vitro and in human lupus.

Methods

Bisulfite sequencing was used to determine the methylation status of the ITGAL promoter and flanking regions in T cells from lupus patients and healthy subjects, and in T cells treated with DNA methylation inhibitors. “Patch” methylation of promoter sequences in reporter constructs was used to determine the functional significance of the methylation changes.

Results

Hypomethylation of specific sequences flanking the ITGAL promoter was seen in T cells from patients with active lupus and in T cells treated with 5‐azacytidine and procainamide. Patch methylation of this region suppressed ITGAL promoter function.

Conclusion

DNA methylation changes occur in specific sequences that regulate LFA‐1 expression in lupus T cells and in the hypomethylation model, indicating that altered methylation of specific genes may play a role in the pathogenesis of lupus.
Quantitative computed tomography of the lumbar spine, not dual x‐ray absorptiometry, is an independent predictor of prevalent vertebral fractures in postmenopausal women with osteopenia receiving long‐term glucocorticoid and hormone‐replacement therapy
Authors: Rehman Q. , Lang T. , Modin G. , Lane N. E.
Published: October 2002

Objective

To determine which measurement of bone mineral density (BMD) predicts vertebral fractures in a cohort of postmenopausal women with glucocorticoid‐induced osteoporosis.

Methods

We recruited 114 subjects into the study. All had osteopenia of the lumbar spine or hip, as demonstrated by dual x‐ray absorptiometry (DXA), and were receiving long‐term glucocorticoids and hormone replacement therapy (HRT). Measurements of BMD by DXA of the lumbar spine, hip (and subregions), and forearm (and subregions), quantitative computed tomography (QCT) of the spine and hip (n = 59), and radiographs of the thoracolumbar spine were performed on all subjects to assess prevalent vertebral fractures. Vertebral fracture prevalence, as determined by morphometry, required a ≥20% (or ≥4‐mm) loss of vertebral body height. Demographic information was obtained by questionnaire. Multiple regression and classification and regression trees (CART) analyses were used to assess predictors of vertebral fracture.

Results

Twenty‐six percent of the study subjects had prevalent fractures. BMD of the lumbar spine, total hip and hip subregions, as measured by QCT, but only the lumbar spine and total hip, as measured by DXA, were significantly associated with prevalent vertebral fractures. However, only lumbar spine BMD as measured by QCT was a significant predictor of vertebral fractures. CART analysis showed that a BMD value <0.065 gm/cm3 was associated with a 7‐fold higher risk of fracture than a BMD value ≥0.065 gm/cm3.

Conclusion

In postmenopausal women with osteoporosis induced by long‐term glucocorticoid treatment who are also receiving HRT, BMD of the lumbar spine as measured by QCT, but not DXA, is an independent predictor of vertebral fractures.
FLICE‐inhibitory protein expression in synovial fibroblasts and at sites of cartilage and bone erosion in rheumatoid arthritis
Authors: Schedel Jörg , Gay Renate E. , Kuenzler Peter , Seemayer Christian , Simmen Beat , Michel Beat A. , Gay Steffen
Published: October 2002

Objective

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by a hyperplastic synovial tissue, inflammatory infiltrates, and a progressive destruction of cartilage and bone. FLICE‐inhibitory protein (FLIP) prevents the association of caspase 8 with FADD and thus exerts an antiapoptotic effect through inhibition of Fas‐mediated apoptosis. We undertook this study to examine the expression of FLIP in RA, osteoarthritic (OA), and normal synovial tissues.

Methods

We investigated the expression of FLIP (long form) in 5 RA, 2 OA, and 2 normal synovial tissue samples. A 393‐bp fragment was amplified from complementary DNA obtained from cultured RA synovial fibroblasts (RASF) by reverse transcription–polymerase chain reaction (RT‐PCR). Using in situ hybridization, the expression of FLIP messenger RNA (mRNA) in paraffin‐embedded synovial tissue sections was investigated semiquantitatively by analyzing the lining layer, the sublining, and sites of invasion. Immunohistochemistry with anti‐CD68 antibodies was performed on serial tissue sections to further characterize the cell types expressing FLIP. In addition, quantitative expression of FLIP was measured by real‐time PCR.

Results

RT‐PCR revealed the expression of FLIP mRNA in all RA and OA samples tested. Using in situ hybridization in synovial tissue, FLIP was detected in all 5 RA samples and in 1 of 2 OA samples, but in neither of the 2 normal control samples. In RA, FLIP expression could be found in both the lining and sublining layers; most importantly, it could also be identified at sites of cartilage invasion and bone destruction. Moreover, quantitative PCR analysis showed 50% higher FLIP expression in RASF than in OASF.

Conclusion

The expression of antiapoptotic FLIP in RA synovial tissue and in synovial fibroblasts suggests the idea of a novel pathway in RA that potentially extends the lifespan of cartilage‐ and bone‐degrading synovial cells, thus contributing to the progression of joint destruction.
One‐year open‐label trial of thalidomide in ankylosing spondylitis
Authors: Huang Feng , Gu Jieruo , Zhao Wei , Zhu Jian , Zhang Jianglin , Yu David Tak Yan
Published: October 2002

Objectives

To test in a large open study whether thalidomide is potentially useful in treating ankylosing spondylitis, and to see if thalidomide induces any change in expression of genes in peripheral blood mononuclear cells (PBMC).

Methods

Thirty male patients with treatment‐refractory ankylosing spondylitis were recruited into a 12‐month open study using thalidomide at a dosage of 200 mg/day. Seven indices were measured as primary outcome measures, and 6 other indices as secondary outcome measures. Transcripts in the PBMC of some of these patients were first screened with microarray, and then measured with reverse transcriptase–polymerase chain reaction.

Results

Twenty‐six patients completed the study. Of these, 80% showed a >20% improvement in 4 of 7 primary indices. Sharp declines in several parameters were noticed at 3–6 months. Nine patients became pain‐free. There was also a statistically significant decrease in tumor necrosis factor α transcripts in the PBMC.

Conclusion

Thalidomide is a reasonably promising drug in treatment‐resistant ankylosing spondylitis.
Mortality in early inflammatory polyarthritis: Cardiovascular mortality is increased in seropositive patients
Authors: Goodson Nicola J. , Wiles Nicola J. , Lunt Mark , Barrett Elizabeth M. , Silman Alan J. , Symmons Deborah P. M.
Published: October 2002

Objective

To determine the degree and causes of any excess mortality observed during the early years of inflammatory polyarthritis (IP).

Methods

Between 1990 and 1994, a total of 1,236 patients were registered with the Norfolk Arthritis Register, a primary care–based inception cohort. All patients were tracked on the National Health Service Central Register for notification of death. The vital status of each patient was determined as of December 31, 1999. Causes of death were coded according to the International Classification of Diseases, Ninth Revision. Expected death rates were calculated using annual death rates for the Norfolk population. Standardized mortality ratios (SMRs) were calculated for all IP patients and for the subgroups of patients who did and did not satisfy the American College of Rheumatology (ACR) 1987 criteria for rheumatoid arthritis (RA) at baseline, as well as for the subgroups who were and were not rheumatoid factor (RF) positive at baseline.

Results

By December 31, 1999, 160 patients (13%; 79 women and 81 men) had died. The median duration of followup in the entire cohort was 6.9 years. Mortality rates were not significantly increased in the entire group of patients with IP or in the subgroup who met the ACR 1987 criteria for RA at baseline. In contrast, RF‐positive patients had an increased rate of death from all causes (SMR in men 1.51, in women 1.41). Cardiovascular disease was the most common cause of death. The majority of the excess mortality in the RF‐positive patients could be attributed to cardiovascular causes (SMR in men 1.34, in women 2.02).

Conclusion

Excess mortality in the early years of IP is confined to patients who are seropositive for RF. While excess cardiovascular mortality has been described in patients with established RA, this is the first report of premature death from heart disease in the early years of IP.
Effect of Wegener's granulomatosis on work disability, need for medical care, and quality of life in patients younger than 40 years at diagnosis
Authors: Reinhold‐Keller Eva , Herlyn Karen , Wagner‐Bastmeyer Rosemarie , Gutfleisch Joachim , Peter Hartmut H. , Raspe Heiner H. , Gross Wolfgang L.
Published: October 2002

Objectives

To evaluate the effects of Wegener's granulomatosis (WG) on employment status, work disability, and need for medical care of 60 consecutive WG patients aged ≤40 years at diagnosis.

Methods

Sixty WG patients (26 male, 34 female) with a median age of 36 years (range 17–48 years) and a median duration of disease of 39 months (range 0–228 months) completed self‐administered questionnaires on hospitalization, medical care, and employment status plus the Medical Outcomes Study‐Short Form‐36 (SF‐36) estimating their health‐related quality of life.

Results

Thirty‐two of the 60 patients reported full‐ or part‐time employment more than 3 years after diagnosis. Only 14 of the 51 patients employed at diagnosis (27%) were currently receiving a permanent work disability pension due to WG. Two additional patients had lost work because of WG. Women who were employed at diagnosis had a nearly 3‐fold higher risk of losing their jobs compared with men (P = 0.0006). There were no differences with regard to age at diagnosis, disease duration, disease severity, or education level between employed and unemployed patients. Employed patients had missed a median of 14 workdays (range 0–18 days) due to WG within the past 12 months. More than half of all patients (33 of 60) had been hospitalized during the previous 12 months because of WG. Ninety‐three percent of all patients had visited their physician once or more per month, more than half of them at least once per week, regardless of employment status, severity of disease, or type of current medication. Unemployed WG patients experienced significant reductions in social and physical function and in their perceived degree of general health as assessed by the SF‐36.

Conclusions

Twenty‐seven percent of WG patients younger than age 40 who were employed at diagnosis received permanent work disability within a disease duration of 39 months. Unemployment is followed by a considerable reduction in disease‐related quality of life compared with employed patients, independent of severity and extent of disease. Furthermore, because patients were followed closely by an interdisciplinary team, a high rate of hospitalization and frequent visits to physicians resulted.
Identification of synovium‐specific homing peptides by in vivo phage display selection
Authors: Lee Lewis , Buckley Christopher , Blades Mark C. , Panayi Gabriel , George Andrew J. T. , Pitzalis Costantino
Published: October 2002

Objective

To identify homing peptides specific for human synovium that could be used as targeting devices for delivering therapeutic/diagnostic agents to human joints.

Methods

Human synovium and skin were transplanted into SCID mice. A disulfide‐constrained 7–amino acid peptide phage display library was injected intravenously into the animals and synovial homing phage recovered from synovial grafts. Following 3–4 cycles of enrichment, DNA sequencing of homing phage clones allowed the identification of specific peptides that were synthesized by a‐fluorenylmethyloxycarbonyl chemistry and used in competitive in vivo assays and immunohistochemistry analyses.

Results

We isolated synovial homing phages displaying specific peptides that distinctively bound to synovial but not skin or mouse microvascular endothelium (MVE). They retained their tissue homing specificity in vivo, independently from the phage component, the original pathology of the transplanted tissue, and the degree of human/murine graft vascularization. One such peptide (CKSTHDRLC) maintained synovial homing specificity both when presented by the phage and as a free synthetic peptide. The synthetic peptide also competed with and inhibited in vivo the binding of the parent phage to the cognate synovial MVE ligand.

Conclusion

This is the first report describing peptides with homing properties specific for human synovial MVE. This was demonstrated using a novel approach targeting human tissues, transplanted into SCID mice, directly by in vivo phage display selection. The identification of such peptides opens the possibility of using these sequences to construct joint‐specific drug delivery systems that may have considerable impact in the treatment of arthritic conditions.
Trends in incidence and mortality in rheumatoid arthritis in Rochester, Minnesota, over a forty‐year period
Authors: Doran Michele F. , Pond Gregory R. , Crowson Cynthia S. , O'Fallon W. Michael , Gabriel Sherine E.
Published: October 2002

Objective

To determine time trends in the epidemiology of rheumatoid arthritis (RA) in a population‐based cohort.

Methods

An inception cohort of residents of Rochester, Minnesota ≥18 years of age who first fulfilled the American College of Rheumatology 1987 criteria between January 1, 1955 and December 31, 1994 (applied retrospectively, as appropriate) was assembled and followed up until January 1, 2000. Incidence rates were estimated and were age‐ and sex‐adjusted to the 1990 white population of the US. A birth cohort analysis was performed, and survival rates over time were examined.

Results

The incidence cohort comprised 609 patients, 445 (73.1%) of whom were female and 164 (26.9%) were male, with a mean age at incidence of 58.0 years. The overall age‐ and sex‐adjusted annual incidence of RA among Rochester, Minnesota, residents ≥18 years of age was 44.6/100,000 population (95% confidence interval 41.0–48.2). While the incidence rate fell progressively over the 4 decades of study, from 61.2/100,000 in 1955–1964, to 32.7/100,000 in 1985–1994, there were indications of cyclical trends over time. Birth cohort analysis showed diminishing incidence rates through successive cohorts following a peak in the 1880–1890 cohorts. Incidence rates increased with age until age 85, but peaked earlier in women than in men. The survival rate in RA patients was significantly lower than the expected rate in the general population (P < 0.001), and no improvement was noted over time.

Conclusion

The secular trends demonstrated in this study population, including the progressive decline in the incidence of RA over the last 40 years, suggest that an environmental factor may play a role in the etiology of RA.
Risk and rationality: the influence of decision domain and perceived outcome control on managers' high‐risk decisions
Authors: Forlani David
Published: October 2002

The issue of when managers will undertake different forms and amounts of risk in their new business creation decisions is still not fully understood. To address one facet of this issue, this paper advances the proposition that decision domain and perceived outcome control interact to impact managers' new‐product introduction decisions. Employing a causal method, a nested experimental design and indicators of risk's probability of loss and magnitude of loss elements, the study's results demonstrate that the manager's level of perceived outcome control combines with decision domain to determine subsequent levels of risk taking that are specific to each element of risk. Implications for theory, including prospect theory, and practice, especially firms wanting to pursue today's high‐risk new‐economy based initiatives, are discussed. Copyright © 2002 John Wiley & Sons, Ltd.
Eliminating recency with self‐review: the case of auditors' ‘going concern’ judgments
Authors: Ashton Robert H. , Kennedy Jane
Published: October 2002

This paper examines the use of self‐review to debias recency. Recency is found in the ‘going concern’ judgments of staff auditors, but is successfully eliminated by the auditor's use of a simple self‐review technique that would be extremely easy to implement in audit practice. Auditors who self‐review are also less inclined to make audit report choices that are inconsistent with their going concern judgments. These results are important because the judgments of staff auditors often determine the type and extent of documentation in audit workpapers and serve as preliminary inputs for senior auditors' judgments and choices. If staff auditors' judgments are affected by recency, the impact of this bias may be impounded in the ultimate judgments and choices of senior auditors. Since biased judgments can expose auditors to significant costs involving extended audit procedures, legal liability and diminished reputation, simple debiasing techniques that reduce this exposure are valuable. The paper also explores some future research needs and other important issues concerning judgment debiasing in applied professional settings. Copyright © 2002 John Wiley & Sons, Ltd.
Gene expression of cytokine receptors in HL60 cells exposed to a 50 Hz magnetic field
Authors: Zhou Jiliang , Li Changlin , Yao Gengdong , Chiang Huai , Chang Zongliang
Published: October 2002

The effects of a 50 Hz extremely low frequency (ELF) sinusoidal magnetic field (MF) on the expression of genes relating to cytokine receptors were studied in HL60 cells. Transcription levels of tumor necrosis factor receptor (TNFR) p55 and p75, interleukin‐6 receptor‐α (IL‐6Rα) and transforming growth factor‐β receptor 1 (TGFβR1) were quantified in cells exposed to an intensity of 0.1 or 0.8 mT for periods ranging from 30 min to 72 h. Cells treated with 10 nM of phorbol 12‐myristate 13‐acetate (PMA) for 8 h served as a positive control. Gene expression values were assessed by the ribonuclease protection assay (RPA) and normalized to those of the noninducible gene GAPDH. The results showed that MF exposure at 0.1 and 0.8 mT for 72 h increased TNFR p75 and IL‐6Rα mRNA expression in HL60 cells. No significant change in gene expression levels of TNFR p55 and TGFβR1 was observed under any of the exposure conditions. In addition, we report here for the first time that IL‐6Rα mRNA expression can be suppressed by PMA in HL60 cells. Bioelectromagnetics 23:339–346, 2002. © 2002 Wiley‐Liss, Inc.
Effects of pulsed electromagnetic field (PEMF) stimulation on bone tissue like formation are dependent on the maturation stages of the osteoblasts
Authors: Diniz Pericles , Shomura Kenji , Soejima Kazuhisa , Ito Gakuji
Published: October 2002

The effects of pulsed electromagnetic field (PEMF, 15 Hz pulse burst, 7 mT peak) stimulation on bone tissue‐like formation on osteoblasts (MC3T3‐E1 cell line) in different stages of maturation were assessed to determine whether the PEMF stimulatory effect on bone tissue‐like formation was associated with the increase in the number of cells and/or with the enhancement of the cellular differentiation. The cellular proliferation (DNA content), differentiation (alkaline phosphatase activity), and bone tissue‐like formation (area of mineralized matrix) were determined at different time points. PEMF treatment of osteoblasts in the active proliferation stage accelerated cellular proliferation, enhanced cellular differentiation, and increased bone tissue‐like formation. PEMF treatment of osteoblasts in the differentiation stage enhanced cellular differentiation and increased bone tissue‐like formation. PEMF treatment of osteoblasts in the mineralization stage decreased bone tissue‐like formation. In conclusion, PEMF had a stimulatory effect on the osteoblasts in the early stages of culture, which increased bone tissue‐like formation. This stimulatory effect was most likely associated with enhancement of the cellular differentiation, but not with the increase in the number of cells. Bioelectromagnetics 23:398–405, 2002. © 2002 Wiley‐Liss, Inc.
Patterning the marginal zone of early ascidian embryos: localized maternal mRNA and inductive interactions
Authors: Nishida Hiroki
Published: October 2002

Early animal embryos are patterned by localized egg cytoplasmic factors and cell interactions. In invertebrate chordate ascidians, larval tail muscle originates from the posterior marginal zone of the early embryo. It has recently been demonstrated that maternal macho‐1 mRNA encoding transcription factor acts as a localized muscle determinant. Other mesodermal tissues such as notochord and mesenchyme are also derived from the vegetal marginal zone. In contrast, formation of these tissues requires induction from endoderm precursors at the 32‐cell stage. FGF–Ras–MAPK signaling is involved in the induction of both tissues. The responsiveness for induction to notochord or mesenchyme depends on the inheritance of localized egg cytoplasmic factors. Previous studies also point to critical roles of directed signaling in polarization of induced cells and in subsequent asymmetric divisions resulting in the formation of two daughter cells with distinct fates. One cell adopts an induced fate, while the other assumes a default fate. A simple model of mesoderm patterning in ascidian embryos is proposed in comparison with that of vertebrates. BioEssays 24:613–624, 2002. © 2002 Wiley Periodicals, Inc.
p53 and ageing: too much of a good thing?
Authors: Kirkwood Thomas B.L.
Published: October 2002

A recent report by Tyner et al.1 suggests that p53 is bad for longevity. Heterozygotic mice carrying a p53 mutation that apparently enhances the stability of the wild‐type protein showed shorter lifespans and faster ageing while also developing fewer tumours. This fits with the idea that cellular ageing is the price paid for better protection against unlimited proliferation of cancer cells. But other work shows that there is a strong positive association between DNA repair‐mediated protection against cancer and ageing. So what are we to make of the new data with regard to overall understanding of the mechanisms of ageing? BioEssays 24:577–579, 2002. © 2002 Wiley Periodicals, Inc.
Radiation‐induced chromosome aberrations: Insights gained from biophysical modeling
Authors: Hlatky Lynn , Sachs Rainer K. , Vazquez Mariel , Cornforth Michael N.
Published: October 2002

Enzymatic misrepair of ionizing‐radiation‐induced DNA damage can produce large‐scale rearrangements of the genome, such as translocations and dicentrics. These and other chromosome exchange aberrations can cause major phenotypic alterations, including cell death, mutation and neoplasia. Exchange formation requires that two (or more) genomic loci come together spatially. Consequently, the surprisingly rich aberration spectra uncovered by recently developed techniques, when combined with biophysically based computer modeling, help characterize large‐scale chromatin architecture in the interphase nucleus. Most results are consistent with a picture whereby chromosomes are mainly confined to territories, chromatin motion is limited, and interchromosomal interactions involve mainly territory surfaces. Aberration spectra and modeling also help characterize DNA repair/misrepair mechanisms. Quantitative results for mammalian cells are best described by a breakage‐and‐reunion model, suggesting that the dominant recombinational mechanism during the G0/G1 phase of the cell cycle is non‐homologous end‐joining of radiogenic DNA double strand breaks. In turn, better mechanistic and quantitative understanding of aberration formation gives new insights into health‐related applications. BioEssays 24:714–723, 2002. © 2002 Wiley Periodicals, Inc.
DNA melting in the presence of fluorescent intercalating oxazole yellow dyes measured with a gel‐based assay
Authors: Bjorndal Michael T. , Fygenson D. Kuchnir
Published: October 2002

We measured the effect of the intercalating oxazole yellow DNA dye quinolinium,4‐[(3‐methyl‐2(3H)‐benzoxazolylidene)methyl]‐1‐[3‐(trimethylammonio)propyl]‐,diiodide (YO‐PRO) and its homodimer (YOYO) on the melting of self‐complementary DNA duplexes using a gel‐based assay. The assay, which requires a self‐complementary DNA sequence, is independent of the optical properties of the molecules in solution. The melting temperature of the DNA is observed to increase in direct proportion to the number of occupied intercalation sites on the DNA, irrespective of whether the dye molecules are in monomer or dimer form. The increase is ∼ 2.5°C for each intercalation site occupied in the presence of 38 mM [Na+], for dye/duplex ratios in which less than 1/5 of the available intercalation sites are occupied. © 2002 Wiley Periodicals, Inc. Biopolymers 65: 40–44, 2002
A two‐step, one‐pot enzymatic synthesis of cephalexin from D‐phenylglycine nitrile
Authors: Wegman Margreth A. , van Langen Luuk M. , van Rantwijk Fred , Sheldon Roger A.
Published: October 2002

A cascade of two enzymatic transformations is employed in a one‐pot synthesis of cephalexin. The nitrile hydratase (from R. rhodochrous MAWE)‐catalyzed hydration of D‐phenylglycine nitrile to the corresponding amide was combined with the penicillin G acylase (penicillin amidohydrolase, E.C. 3.5.1.11)‐catalyzed acylation of 7‐ADCA with the in situ–formed amide to afford a two‐step, one‐pot synthesis of cephalexin. D‐Phenylglycine nitrile appeared to have a remarkable selective inhibitory effect on the penicillin G acylase, resulting in a threefold increase in the synthesis/hydrolysis (S/H) ratio. 1,5‐Dihydroxynaphthalene, when added to the reaction mixture, cocrystallized with cephalexin. The resulting low cephalexin concentration prevented its chemical as well as enzymatic degradation; cephalexin was obtained at 79% yield with an S/H ratio of 7.7. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 356–361, 2002.
Production of lipolyzed butteroil by a calf pregastric esterase immobilized in a hollow fiber reactor: III. Effect of glycerol
Authors: Lessard Louis P. , Hill Charles G.
Published: October 2002

Lipolysis of butter oil in a hollow fiber reactor containing an immobilized calf pregastric esterase was studied at 40°C, a pH of 6.0, and glycerol concentrations of 0, 150, and 500 g/L in the buffer solution. The concentrations of 10 fatty acid species in the lipolyzed product were determined using high‐performance liquid chromatography. The rate of loss of enzyme activity and the relative selectivities of this esterase depended on the glycerol concentration. By contrast, the overall rate of release of fatty acids was not affected by the glycerol concentration. Loss of enzyme activity was modeled using first‐order kinetics. The models for deactivation and reaction kinetics were fit simultaneously to the data. The model was successful in describing the rates of release of all 10 fatty acid species for a range of space times from 0 to 25 h. The parameters of the model were tested for dependence on glycerol concentration. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 323–333, 2002.
Purification of plasmid DNA by an integrated operation comprising tangential flow filtration and nitrocellulose adsorption
Authors: Kendall D. , Lye G. J. , Levy M. S.
Published: October 2002

There is an increasing interest in the development of scaleable and reproducible plasmid DNA purification protocols for vaccine and gene therapy. The use of an integrated unit operation, comprising tangential flow microfiltration coupled with the adsorption of contaminants onto nitrocellulose membranes as a single processing step was examined in this work. Experiments were performed using a custom‐built tangential flow microfiltration rig (membrane area=12.5 cm2). Tangential flow filtration–adsorption of E. coli lysates containing a plasmid product removed most solids (>75%) and decreased chromosomal DNA contamination by 75% w/w. Total plasmid DNA concentration and supercoiled content of the permeate were virtually identical to those of the feed, indicating a recovery yield of 100% (transmission equal to 1). Results were similar for E. coli lysates containing either a 6.9 kb or a 20 kb plasmid. Significant reductions in RNA, endotoxin, and protein levels were also observed. The reproducibility and potential for scale up of this integrated filtration–adsorption operation makes it at attractive option for intermediate‐ to large‐scale pharmaceutical‐grade plasmid processing. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 816–822, 2002.
Computer‐aided real‐time estimation of reaction conversion for lipase‐catalyzed esterification in solvent‐free systems
Authors: Won Keehoon , Jeong Jae Cheol , Lee Sun Bok
Published: October 2002

Real‐time conversion estimation through macroscopic balancing was investigated for enzymatic esterification reactions in a solvent‐free system. In principle, the conversion of ester synthesis can be determined from the amount of water produced by the reaction because water is formed as a by‐product in the same molar ratio as the product. In this study, we show that the water production rate, and thereby the reaction conversion, can be estimated on‐line from measurements of the relative humidity of the inlet and outlet air and the material balances of water in the system. In order to test the performance of the real‐time conversion estimation method, the lipase‐catalyzed esterification reaction of n‐capric acid and n‐decyl alcohol in solvent‐free media was conducted while controlling the water activity at various values. When the reaction conversions estimated on‐line were compared with those analyzed off‐line by gas chromatography, good agreement was obtained: the average mean absolute error was ± 2.4% of the reaction conversion despite the simplicity of the method. The on‐line estimation method presented here requires no expensive or complicated analytical instruments and no sampling of reaction medium. It can be used for monitoring nonaqueous enzymatic reactions where water is produced or consumed during reaction. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 795–803, 2002.
Protein–protein interactions in concentrated electrolyte solutions
Authors: Curtis R. A. , Ulrich J. , Montaser A. , Prausnitz J. M. , Blanch H. W.
Published: October 2002

Protein–protein interactions were measured for ovalbumin and for lysozyme in aqueous salt solutions. Protein–protein interactions are correlated with a proposed potential of mean force equal to the free energy to desolvate the protein surface that is made inaccessible to the solvent due to the protein–protein interaction. This energy is calculated from the surface free energy of the protein that is determined from protein–salt preferential‐interaction parameter measurements. In classical salting‐out behavior, the protein–salt preferential interaction is unfavorable. Because addition of salt raises the surface free energy of the protein according to the surface‐tension increment of the salt, protein–protein attraction increases, leading to a reduction in solubility. When the surface chemistry of proteins is altered by binding of a specific ion, salting‐in is observed when the interactions between (kosmotrope) ion–protein complexes are more repulsive than those between the uncomplexed proteins. However, salting‐out is observed when interactions between (chaotrope) ion–protein complexes are more attractive than those of the uncomplexed proteins. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 367–380, 2002.
Structure‐function relationships in immobilized chymotrypsin catalysis
Authors: Clark Douglas S. , Bailey James E.
Published: October 2002

Specific activities and the amounts of active immobilized enzyme were determined for several different preparations of α‐chymotrypsin immobilized on CNBr‐activated Sepharose 4B. Electron paramagnetic resonance (EPR) spectroscopy of free and immobilized enzyme with a spin label coupled to the active site was used to probe the effects of different immobilization conditions on the immobilized enzyme active site configuration. Specific activity of active enzyme decreased and rotational correlation time of the spin label increased with increasing immobilized enzyme loading. Enzyme immobilized using an intermediate six‐carbon spacer arm exhibited greater specific activity and spin label mobility than directly coupled enzyme. The observed activity changes due to immobilization were completely consistent with corresponding active site structure alterations revealed by EPR spectroscopy. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 539–549, 2002.