bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic insights into familial tumors of the nervous system
Authors: Melean German , Sestini Roberta , Ammannati Franco , Papi Laura
Published: July 2004

Nervous system tumors represent unique neoplasms that arise within the central and peripheral nervous system. While the vast majority of nervous system neoplasm occur sporadically, most of the adult and pediatric forms have a hereditary equivalent. In a little over a decade, we have seen a tremendous increase in knowledge of the primary genetic basis of many of the familial cancer syndromes that involve the nervous system, syndromes that are mostly inherited as autosomal dominant traits. In this review, we discuss the most recent findings on the genetic basis of hereditary nervous system tumors. The identification of genes associated with familial cancer syndromes has in some families enabled a “molecular diagnosis” that complements clinical assessment and allows directed cancer surveillance for those individuals determined to be at‐risk for disease. © 2004 Wiley‐Liss, Inc.
Longitudinal analysis of deciduous tooth emergence: IV. Covariate effects in Japanese children
Authors: Holman Darryl J. , Yamaguchi Kyoko
Published: July 2004

Emergence of the deciduous teeth is generally considered to be robust to moderate environmental insults, malnutrition, and disease. Consequently, deciduous tooth emergence has been used to assess growth and development and for age estimation in children. In this paper, we examine the way in which nutritional status and other covariates affect deciduous tooth emergence in a sample of 114 Japanese children born in Tokyo in 1914 and 1924. Parametric survival analysis was used to quantify the effects of nutritional status, breastfeeding behavior, and sex on the hazard of deciduous tooth emergence. Children of poor nutritional status exhibited significantly delayed emergence of all deciduous teeth, with effects that ranged from 14–29% increases in mean emergence times. Children of medium nutritional status exhibited increases in mean emergence times of 5–9% for the canines and lower molars, and 13–17% for the incisors. Partial breastfeeding had no effect on tooth emergence, but children who were not breastfed at all showed delayed emergence of the upper incisors. No significant sex differences in emergence were found. The findings contradict the idea that moderate malnutrition has little effect on deciduous tooth emergence. Furthermore, nutritional differences may account for some of the observed differences among populations in the timing of tooth emergence. Am J Phys Anthropol, 2004. © 2004 Wiley‐Liss, Inc.
Continuity or colonization in Anglo‐Saxon England? Isotope evidence for mobility, subsistence practice, and status at West Heslerton
Authors: Montgomery Janet , Evans Jane A. , Powlesland Dominic , Roberts Charlotte A.
Published: July 2004

The adventus Saxonum is a crucial event in English protohistory. Scholars from a range of disciplines dispute the scale and demographic profile of the purported colonizing population. The 5th–7th century burial ground at West Heslerton, North Yorkshire, is one of the few Anglian cemeteries where an associated settlement site has been identified and subjected to extensive multidisciplinary postexcavation study. Skeletal and grave good evidence has been used to indicate the presence of Scandinavian settlers. A small, preliminary study using lead and strontium isotope analysis of tooth enamel, mineralized in early childhood, from Neolithic/Early Bronze Age (n = 8), Iron Age (n = 2), and Early Anglo‐Saxon (n = 32) skeletons, was carried out to directly investigate this hypothesis. Results suggest that lead provides dissimilar types of information in different time periods. In post‐Roman England, it appears to reflect the level of exposure to circulated anthropogenic rather than natural geological lead, thus being a cultural rather than geographical marker. Consequently, only strontium provides mobility evidence among the Anglian population, whereas both isotope systems do so in pre‐Roman periods. Strontium data imply the presence of two groups: one of “local” and one of “nonlocal” origin, but more work is required to define the limits of local variation and identify immigrants with confidence. Correlations with traditional archaeological evidence are inconclusive. While the majority of juveniles and prehistoric individuals fall within the “local” group, both groups contain juveniles, and adults of both sexes. There is thus no clear support for the exclusively male, military‐elite invasion model at this site. Am J Phys Anthropol 126:123–138, 2005. © 2004 Wiley‐Liss, Inc.
Paleopathological study of hallux valgus
Authors: Mays S.A.
Published: July 2004

Hallux valgus is the abnormal lateral deviation of the great toe. The principal cause is biomechanical, specifically the habitual use of footwear which constricts the toes. In this study, descriptions of the anatomical changes of hallux valgus from published cadaveric and clinical studies were used to generate criteria for identifying the condition in ancient skeletal remains. The value of systematic scoring of hallux valgus in paleopathology is illustrated using two British skeletal series, one dating from the earlier and one from the later Medieval period. It was found that hallux valgus was restricted to later Medieval burials. This appears consistent with archaeological and historical evidence for a rise in popularity, during the late Medieval period (at least among the richer social classes), of narrow, pointed shoes which would have constricted the toes. Am J Phys Anthropol 126:139–149, 2005. © 2004 Wiley‐Liss, Inc.
Changing intracellular compartmentalization of β‐galactosidase in the ROSA26 reporter mouse during embryonic development: A light‐ and electron‐microscopic study
Authors: Aoyama Naoyoshi , Molin Daniel G.M. , Mentink Monica M.T. , Koerten Henk K. , de Ruiter Marco C. , Gittenberger‐de Groot Adriana C. , Poelmann Robert E.
Published: July 2004

The beta‐geo (LacZ) reporter gene encodes for β‐galactosidase (β‐gal) in all cells of the ROSA26 mouse during embryonic development. As such, β‐gal activity constitutes an excellent marker for in situ labeling of expressing cells. However, the intracellular distribution of β‐gal differs between cells, and changes during embryonic development. Therefore, we studied LacZ‐encoded β‐gal using light and electron microscopy in the heart, lung, liver, and small intestine on days 13 and 16 of gestation, and the kidney on day 16 of gestation in ROSA26 mice. The Bluo‐gal method was carried out under standardized conditions, including fixation, washing, and incubation procedures. Intracellular β‐gal staining is encountered in a combination of membranous compartments, including the nuclear envelope, the endoplasmic reticulum, and the plasma membrane. Its exact localization depends on the cell type and is regulated during development. Therefore, one must take the compartmental transition of intracellular β‐gal staining into consideration when interpreting results obtained from experiments using ROSA26 mice. © 2004 Wiley‐Liss, Inc.
Localization of estrogen and androgen receptors in male reproductive tissues of mice and rats
Authors: Yamashita Shuji
Published: July 2004

Using immunohistochemical methods, we studied the cell‐type‐ and species‐specific expressions of estrogen receptor (ER) isoforms (ERα and ERβ) and androgen receptors (ARs) in the male reproductive tract and accessory sex glands of mature mice and rats. ERα and ERβ showed cell‐type‐ and species‐specific distributions, respectively. In contrast, AR was localized in the epithelial and stroma cells of all tissues examined in this study, in both species. In mice, the epithelial cells of the ductuli efferentes showed a strong ERα‐immunoreaction, and those of the caput epididymis, coagulating glands, and prostate also exhibited a positive reaction. Stroma cells, except in the ductuli efferentes, showed a positive ERα‐immunostaining. In rats, ERα was detected in very few cell types: the epithelial cells of the ductuli efferentes showed a strong reaction, and the stroma cells of the ampullary and urethral glands exhibited a weak reaction. ERβ was localized in the epithelial cells of the prostate in mice, while the reaction was faint or negative in both the epithelial and stroma cells of other tissues. In rats, the ERβ‐immunoreaction was strongest in the epithelial cells of the ventral prostate. The epithelial cells of the corpus and cauda epididymis, ductus deferens, and urethral glands, and the stroma cells of the urethral glands were also positively ERβ‐immunostained. Almost the same AR distribution pattern was observed in both species. In particular, strong AR‐immunostaining was present in the epithelial cells of the caput and corpus epididymis, seminal vesicle, and ventral prostate. These results indicate that species and tissues differences should be taken into careful consideration in assessing the physiological and pharmacological effects of sex steroids (particularly estrogens) on the reproductive tissues of male rodents. © 2004 Wiley‐Liss, Inc.
Adult human mylohyoid muscle fibers express slow‐tonic, α‐cardiac, and developmental myosin heavy‐chain isoforms
Authors: Mu Liancai , Su Hungxi , Wang Jun , Han Yingshi , Sanders Ira
Published: July 2004

Some adult cranial muscles have been reported to contain unusual myosin heavy‐chain (MHC) isoforms (i.e., slow‐tonic, α‐cardiac, embryonic, and neonatal), which exhibit distinct contractile properties. In this study, adult human mylohyoid (MH) muscles obtained from autopsies were investigated to detect the unusual MHC isoforms. For comparison, the biceps brachii and masseter muscles of the same subjects were also examined. Serial cross‐sections from the muscles studied were incubated with a panel of isoform‐specific anti‐MHC monoclonal antibodies that distinguish major and unusual MHC isoforms. On average, the slow type I and fast type II MHC‐containing fibers in the MH muscle accounted for 54% and 46% of the fibers, respectively. In contrast to limb and trunk muscles, the adult human MH muscle was characterized by a large proportion of hybrid fibers (85%) and a small percentage of pure fibers (15%; P < 0.01). Of the fast fiber types, the proportion of the type IIa MHC‐containing fibers (92%) was much greater than that of the type IIx MHC‐containing fibers (8%; P < 0.01). Our data demonstrated that the adult human MH fibers expressed the unusual MHC isoforms that were also identified in the masseter, but not in the biceps brachii. These isoforms were demonstrated by immunocytochemistry and confirmed by electrophoretic immunoblotting. Fiber‐to‐fiber comparisons showed that the unusual MHC isoforms were coexpressed with the major MHC isoforms (i.e., MHCI, IIa, and IIx), thus forming various major/unusual (or m/u) MHC hybrid fiber types. Interestingly, the unusual MHC isoforms were expressed in a fiber type‐specific manner. The slow‐tonic and α‐cardiac MHC isoforms were coexpressed predominantly with slow type I MHC isoform, whereas the developmental MHC isoforms (i.e., embryonic and neonatal) coexisted primarily with fast type IIa MHC isoform. There were no MH fibers that expressed exclusively unusual MHC isoforms. Approximately 81% of the slow type I MHC‐containing fibers expressed slow‐tonic and α‐cardiac MHC isoforms, whereas 80% of the fast type IIa MHC‐containing fibers expressed neonatal MHC isoform. The m/u hybrid fibers (82% of the total fiber population) were found to constitute the predominant fiber types in the adult human MH muscle. At least seven m/u MHC hybrid fiber types were identified in the adult human MH muscle. The most common m/u hybrid fiber types were found to be the MHCI/slow‐tonic/α‐cardiac and MHCIIa/neonatal, which accounted for 39% and 33% of the total fiber population, respectively. The multiplicity of MHC isoforms in the adult MH fibers is believed to be related to embryonic origin, innervation pattern, and unique functional requirements. © 2004 Wiley‐Liss, Inc.
Remodeling capacity of calcified fibrocartilage of the hip
Authors: Bloebaum Roy D. , Kopp Derinna V.
Published: July 2004

It is well known that the incidence of hip fractures increases exponentially with age and that hip fractures can be a major cause of morbidity and morality among elderly humans; this has prompted substantial research on hip fractures. The majority of research on hip fractures has focused on morphological changes of the proximal femur with age. Recently calcified fibrocartilage in the proximal femur has been shown to increase in fractional area with age and can ultimately make up to 60% of the fractional area of the cortex. However, the capacity of the tissue to remodel and heal is currently unknown. The purpose of the present study was to determine the remodeling capacity of calcified fibrocartilage on the proximal femur compared to the underlying cortical bone. The remodeling capacity of calcified fibrocartilage and cortical bone was assessed in adult female sheep by means of tetracycline labeling. The number of double and single labels within each tissue type was quantified and analyzed with a paired t‐test. The data showed very few labels in the calcified fibrocartilage compared to the cortical bone. This indicated that calcified fibrocartilage lacked a capacity to remodel in a manner similar to bone. The results of this investigation demonstrate that while the underlying cortical bone was actively remodeling, the calcified fibrocartilage appeared to have no remodeling capacity similar to that of cortical bone. © 2004 Wiley‐Liss, Inc.
Alveolar bone Sharpey fibers of the rat incisor in normal and altered functional conditions examined by scanning electron microscopy
Authors: Silva Miralva A.J. , Merzel José
Published: July 2004

The morphology and the area density of Sharpey fibers in the socket of the rat incisor under normo‐, hyper‐, and hypofunctional conditions were evaluated using scanning electron microscopy. Sharpey fibers appeared either as dome‐shaped projections, when highly mineralized, or as depressions when less mineralized. Near the alveolar crest, most of the fibers were fully mineralized and arranged in compact longitudinal rows. Toward the basal end of the socket, the rows became interrupted, forming islets of gradually smaller size and number. The density of the Sharpey fibers was higher (P < 0.01) in the mesial and distal faces than in the lingual face in most of the socket length. In normofunctional conditions, in all faces the density decreased 70 to 90 times from the crestal toward the basal region of the socket (P < 0.01). The degree of mineralization of the Sharpey fibers also decreased steadily in the same direction, indicating that, for support, the periodontal ligament matures from basal to incisal and is fully developed only in the crestal region. In hyper‐ and hypofunctional conditions, the same distribution was observed. The area density of the Sharpey fibers in the hyperfunctional condition showed a slight increase at the basal levels of the socket mesial and distal faces (P < 0.01 or P < 0.05). In hypofunctional incisors, the density decreased significantly (P < 0.01) at the mesial and distal faces in all regions of the socket, and at the lingual face, the decrease (P < 0.05) was restricted to the incisal regions. This may be one of the factors for the weakening of the periodontal ligament in hypofunctional incisors. © 2004 Wiley‐Liss, Inc.
Human T cell clones specific for heterogeneous nuclear ribonucleoprotein A2 autoantigen from connective tissue disease patients assist in autoantibody production
Authors: Greidinger Eric L. , Gazitt Tal , Jaimes Kimberly F. , Hoffman Robert W.
Published: July 2004

Objective

To identify and characterize human T cells reactive with heterogeneous nuclear RNP A2 (hnRNP A2) antigen, and to determine the ability of hnRNP‐reactive T cells to assist in the production of human autoantibodies.

Methods

T cells from patients with high serum levels of anti‐hnRNP IgG autoantibody were stimulated with an hnRNP recombinant fusion protein, and the cells were cloned by limiting dilution. The surface phenotype and cytokine profiles of the T cells were examined by flow cytometry and enzyme‐linked immunosorbent assay (ELISA), respectively. T cell clones were cultured with highly purified autologous B cells, and the ability of T cells to enhance autoantibody production under a variety of conditions was measured by ELISA.

Results

Human T cells reactive with hnRNP antigen were cloned from 2 patients with systemic lupus erythematosus (SLE) and 1 patient with mixed connective tissue disease (MCTD). The T cells were CD4+ and had a Th1‐like functional phenotype. In coculture in vitro with autologous B cells, T cell clones augmented anti‐hnRNP autoantibody production and did so without the need for direct T cell–B cell contact.

Conclusion

This study provides direct evidence for a role of anti‐hnRNP–reactive T cells in autoantibody production in SLE and MCTD. These findings support the notion that hnRNP‐reactive T cells play a role in the pathogenesis of these diseases.
A randomized trial of parenteral methotrexate comparing an intermediate dose with a higher dose in children with juvenile idiopathic arthritis who failed to respond to standard doses of methotrexate
Authors: Ruperto Nicolino , Murray Kevin J. , Gerloni Valeria , Wulffraat Nico , Feitosa De Oliveira Sheila Knupp , Falcini Fernanda , Dolezalova Pavla , Alessio Maria , Burgos‐Vargas Ruben , Corona Fabrizia , Vesely Richard , Foster Helen , Davidson Joyce , Zulian Francesco , Asplin Line , Baildam Eileen , Consuegra Julia Garcia , Ozdogan Huri , Saurenmann Rotraud , Joos Rik , Pistorio Angela , Woo Pat , Martini Alberto
Published: July 2004

Objective

To compare the safety and efficacy of parenteral methotrexate (MTX) at an intermediate dosage (15 mg/m2/week) versus a higher dosage (30 mg/m2/week) in patients with polyarticular‐course juvenile idiopathic arthritis (JIA) who failed to improve while receiving standard dosages of MTX (8–12.5 mg/m2/week).

Methods

In the screening phase, 595 patients who were newly started on a standard dose of MTX were followed up for 6 months. Subsequently, the nonresponders, defined according to the American College of Rheumatology (ACR) pediatric 30% improvement criteria (pediatric 30), were randomized to receive an intermediate dose or higher dose of parenteral MTX for an additional 6 months. Improvement in the screening and randomization phase was defined by the ACR pediatric 30 response, as well as by the 50% and 70% response levels (ACR pediatric 50 and ACR pediatric 70, respectively).

Results

In the screening phase, after receiving standard doses of MTX, 430 patients (72%) improved according to the ACR pediatric 30, while 360 (61%) met the ACR pediatric 50 and 225 (38%) met the ACR pediatric 70; among these patients, 69 (12%) also met the definition of complete disease control. Of the 133 nonresponders, 80 were randomized to receive an intermediate dose or higher dose of MTX. In the randomization phase, the ACR pediatric 30 response rate was 25 of 40 children (62.5%) in the intermediate‐dose group versus 23 of 40 children (57.5%) in the higher‐dose group. An ACR pediatric 50 response rate was attained by 23 patients (57.5%) receiving an intermediate dose versus 22 (55%) in the higher‐dose group. An ACR pediatric 70 response rate was seen in 18 children (45%) receiving an intermediate dose versus 19 (47.5%) receiving a higher dose. Five children (12.5%) in the intermediate‐dose group versus 4 (10%) receiving the higher dose of MTX also met the definition of complete disease control. None of the intergroup differences in response rate were significant. There were no significant differences in the frequency of adverse events or laboratory abnormalities between the 2 randomized groups.

Conclusion

This study shows that the plateau of efficacy of MTX in JIA is reached with parenteral administration of 15 mg/m2/week and that a further increase in dosage is not associated with any additional therapeutic benefit. MTX should be administered for up to 9–12 months to appreciate its full therapeutic effect.
Medical care expenditures and earnings losses of persons with arthritis and other rheumatic conditions in the United States in 1997: Total and incremental estimates
Authors: Yelin Edward , Cisternas Miriam G. , Pasta David J. , Trupin Laura , Murphy Louise , Helmick Charles G.
Published: July 2004

Objective

To provide estimates of the total medical care expenditures and earnings losses associated with arthritis and other rheumatic conditions (AORC), as well as the increment in such costs specifically attributable to these conditions, in the US in 1997.

Methods

The estimates were derived from the 1997 Medical Expenditures Panel Survey (MEPS), a national probability sample of 14,147 households including 34,551 persons, of whom 4,776 self‐reported arthritis. After weighting, those who self‐reported AORC represent 38.4 million persons. We tabulated all medical care expenditures of the adult MEPS respondents, stratified by arthritis and comorbidity status, and then used regression techniques to estimate the increment in health care expenditures attributable to AORC, after taking comorbidity, demographic characteristics, and insurance status into account. Using the same methods, we also estimated the magnitude of the earnings losses sustained by persons of working ages (18–64 years) who had AORC.

Results

Persons with AORC incurred mean total medical care expenditures of $4,865 (total $186.9 billion). The largest components of these expenditures were inpatient care (39%), ambulatory care (29%), and prescriptions (14%). The mean increment in medical care expenditures specifically attributable to AORC among those ages 18 years and older was $1,391 (total ∼$51.1 billion). Persons with AORC ages 18–64 years earned $3,812 less on average than did other persons of these ages (total $82.4 billion). Of this average, $1,579 was attributable to the AORC (total $35.1 billion).

Conclusion

In 1997, persons with AORC incurred direct and indirect costs of $269.3 billion, of which $86.2 billion was attributable to these conditions.
Anti–tumor necrosis factor therapy in patients with difficult to treat Takayasu arteritis
Authors: Hoffman Gary S. , Merkel Peter A. , Brasington Richard D. , Lenschow Deborah J. , Liang Patrick
Published: July 2004

Objective

Granulomatous inflammation is a typical feature of Takayasu arteritis (TA), and tumor necrosis factor (TNF) is important in the formation of granulomas. In this study, we assessed therapy with anti‐TNF agents in patients with TA that was not controlled by glucocorticoid therapy or other immunosuppressants.

Methods

We conducted an open‐label trial of anti‐TNF therapy at 3 academic medical centers over a period of 4.25 years. Fifteen patients with active, relapsing TA (median 6 years) were selected. Seven received etanercept (later changed to infliximab in 3 patients), and 8 received infliximab. Relapses had occurred in all patients while they were receiving glucocorticoids and, in 13 patients, additional immunosuppressive drugs. No other agents were added to the treatment regimen concurrently with anti‐TNF. If patients were receiving cytotoxic agents, the dosage was not increased. Clinical symptoms were recorded, and physical examinations, laboratory studies, and serial magnetic resonance imaging were performed.

Results

The median daily dose of prednisone required to maintain remission prior to anti‐TNF therapy was 20 mg. Ten of the 15 patients achieved complete remission that was sustained for 1–3.3 years without glucocorticoid therapy. Four patients achieved partial remission, with a >50% reduction in the glucocorticoid requirement. At a median of 12 months of followup, the median dose of prednisone was 0. Therapy failed in 1 patient. In 9 of the 14 responders, an increase in the anti‐TNF dosage was required to sustain remission. Two relapses occurred during periods when anti‐TNF therapy (etanercept) was interrupted, but remission was reestablished upon reinstitution of therapy.

Conclusion

In this pilot study of relapsing TA, addition of anti‐TNF therapy resulted in improvement in 14 of 15 patients and sustained remission in 10 of 15 patients, who were able to discontinue glucocorticoid therapy. Anti‐TNF may be a useful adjunct to glucocorticoids in the treatment of TA. Our results justify a randomized, controlled clinical trial of anti‐TNF therapy for TA.
Elevation of activated protein C in synovial joints in rheumatoid arthritis and its correlation with matrix metalloproteinase 2
Authors: Buisson‐Legendre Nathalie , Smith Sue , March Lyn , Jackson Christopher
Published: July 2004

Objective

To investigate the involvement of the anticoagulant serine protease activated protein C (APC) in tissue remodeling in rheumatoid arthritis (RA).

Methods

PC/APC, matrix metalloproteinase 2 (MMP‐2), and MMP‐9 were detected in synovial fluid by Western blotting, and their antigen levels were quantified by enzyme‐linked immunosorbent assay in patients with osteoarthritis (OA) or RA. Enzymatic activity of MMP‐2 was assayed using a specific fluorogenic substrate. We developed an improved assay to measure APC activity in synovial fluid utilizing a chromogenic substrate following immunoprecipitation with a specific PC/APC antibody. PC/APC and MMP‐2 were localized by immunohistochemistry in RA, OA, and normal synovial tissues.

Results

Synovial fluid analysis demonstrated that APC is present in both RA and OA synovial fluid, with APC activity being markedly higher in RA (mean ± SEM 462 ± 112 ng/ml versus 136 ± 42 ng/ml; P < 0.02). A correlation (r2 = 0.61) was found between APC and MMP‐2 activity levels in RA patients, but not in OA patients. Immunohistochemical studies of synovial sections showed colocalization of APC and MMP‐2 in endothelial and synovial lining cells. Additionally, APC and MMP‐2 coimmunoprecipitated with an anti‐PC/APC antibody.

Conclusion

Our results show, for the first time, that APC and MMP‐2 are coordinately up‐regulated and tightly bound in RA synovial fluid and colocalized in synovia. Their association suggests that APC may modulate MMP‐2 activity in RA.
Macrophage release of transforming growth factor β1 during resolution of monosodium urate monohydrate crystal–induced inflammation
Authors: Yagnik Darshna R. , Evans Betsy J. , Florey Oliver , Mason Justin C. , Landis R. Clive , Haskard Dorian O.
Published: July 2004

Objective

It has previously been shown that as monocytes differentiate into macrophages, they lose the ability to secrete proinflammatory cytokines in response to monosodium urate monohydrate (MSU) crystals. The purpose of this study was to investigate whether MSU crystals induce macrophages to secrete antiinflammatory factor instead.

Methods

Human monocyte or macrophage isolates were prepared from samples obtained from healthy volunteer donors either by differentiation of blood monocytes in vitro or by collecting cells from skin blisters during the early or late phase of the dermal inflammatory response to cantharidin. Monocyte or macrophage isolates were then incubated with MSU crystals for 24 hours, and culture supernatants were assayed for candidate antiinflammatory mediators (by enzyme‐linked immunosorbent assay) and for the capacity to activate or suppress endothelial cell E‐selectin expression and secondary neutrophil recruitment under shear flow.

Results

Analysis of supernatants from in vitro–differentiated macrophages revealed that transforming growth factor β1 (TGFβ1) was induced following MSU crystal stimulation (mean ± SEM 1.50 ± 0.24 ng/ml/106 cells), but there was no evidence of interleukin‐10 (IL‐10), IL‐1 receptor antagonist, or tumor necrosis factor (TNF) receptor p55 release. Macrophage TGFβ1 significantly suppressed endothelial cell E‐selectin expression and secondary neutrophil capture on endothelial monolayers stimulated with supernatants from MSU‐treated monocytes. Leukocytes isolated from resolving (40‐hour) skin blisters similarly elaborated TGFβ1 when challenged with MSU crystals (0.66 ± 1.3 ng/ml/105 CD14+ cells). In contrast, cells isolated from acute (16‐hour) skin blisters secreted TNFα (0.49 ± 0.08 ng/ml/105 CD14+ cells) but no detectable TGFβ1.

Conclusion

These data provide further support for the concept that differentiated macrophages play a protective role in the pathophysiology of gout, and they identify macrophage TGFβ1 as a mediator of paracrine suppression during the resolution phase of inflammation.
Enhanced intracellular replication of Salmonella enteritidis in HLA–B27–expressing human monocytic cells: Dependency on glutamic acid at position 45 in the B pocket of HLA–B27
Authors: Penttinen Markus A. , Heiskanen Kaisa M. , Mohapatra Rajashree , DeLay Monica L. , Colbert Robert A. , Sistonen Lea , Granfors Kaisa
Published: July 2004

Objective

To reveal the cause of the impaired elimination of Salmonella enteritidis in HLA–B27–transfected human monocytic cells and to study whether the B pocket of HLA–B27 contributes to these modulatory effects.

Methods

Stable U937 cell transfectants expressing HLA–A2, B27, or different forms of B27 with amino acid substitutions in the B pocket were prepared. Mock‐transfected cells were prepared using the antibiotic resistance vector (pSV2neo) alone. Cells were differentiated, infected with S enteritidis, and the number of live intracellular S enteritidis organisms was determined using the colony‐forming unit method. To visualize intracellular S enteritidis, the bacteria were transformed with green fluorescent protein (GFP), and studied by confocal microscopy.

Results

Cells expressing wild‐type HLA–B27 were more permissive of intracellular replication of S enteritidis compared with mock‐transfected or A2‐transfected controls. Cells expressing B27 with an altered B pocket composition having either 6 amino acid substitutions (B27.A2B; substitutions H9F, T24A, E45M, I66K, C67V, and K70H) or a single substitution (B27.E45M) were no longer permissive of S enteritidis replication. In contrast, cells expressing B27 with the single substitution of F for H at position 9 (B27.H9F) retained their permissiveness. Studies using GFP‐transformed S enteritidis confirmed that the increase in the amount of intracellular bacteria in B27‐expressing cells was due to replication of the bacteria.

Conclusion

Our data indicate that HLA–B27 expression modulates the host–microbe interaction that results in an impaired capacity of monocytes to resist intracellular replication of S enteritidis. The phenotype is dependent on glutamic acid at position 45 in the B pocket and, thus, may be due to properties of the B27 heavy chain that are related to this residue. The ability of HLA–B27 to confer susceptibility to Salmonella‐triggered reactive arthritis may occur, at least in part, through these modulatory effects.
Cytochrome P450 pharmacogenetics as a predictor of toxicity and clinical response to pulse cyclophosphamide in lupus nephritis
Authors: Takada Kazuki , Arefayene Million , Desta Zeruesenay , Yarboro Cheryl H. , Boumpas Dimitrios T. , Balow James E. , Flockhart David A. , Illei Gabor G.
Published: July 2004

Objective

Pulse cyclophosphamide is the treatment of choice for severe lupus nephritis. However, not all patients respond to this therapy, and gonadal toxicity is of particular concern. Cyclophosphamide is a prodrug that requires activation by cytochrome P450 (CYP) enzymes. We conducted a retrospective cohort study to test whether genetic polymorphisms of these enzymes are associated with the toxicity of, and clinical response to, cyclophosphamide in patients with lupus nephritis.

Methods

Sixty‐two patients with proliferative lupus nephritis treated with cyclophosphamide were genotyped for common variant alleles of CYP2B6, 2C19, 2C9, and 3A5. We examined the association between these genotypes and the following clinical end points: development of premature ovarian failure, end‐stage renal disease (ESRD), doubling of serum creatinine level, and achievement of complete renal response.

Results

The observed frequencies of the variant alleles CYP2B6*5, CYP2C19*2, CYP2C9*2, and CYP3A5*3 were 12.1%, 25.0%, 4.0%, and 75.8%, respectively. Patients who were either heterozygous or homozygous for CYP2C19*2 had a significantly lower risk of developing premature ovarian failure (relative risk 0.10; 95% confidence interval 0.02–0.52), after adjustment for age and total number of cyclophosphamide pulses received. In a survival analysis, patients homozygous for CYP2B6*5 (n = 3) or CYP2C19*2 (n = 4) had a higher probability of reaching ESRD (P = 0.0005) and of doubling the creatinine level (P = 0.0005) as well as a trend toward a lower probability of achieving a complete renal response (P = 0.051).

Conclusion

Determination of selected cytochrome P450 enzyme genotypes may be valuable for predicting the risk of premature ovarian failure in lupus nephritis patients treated with cyclophosphamide. The association of these genotypes with renal response needs further validation.
Antileukoproteinase: Modulation of neutrophil function and therapeutic effects on anti–type II collagen antibody–induced arthritis
Authors: Sehnert Bettina , Cavcic Andrea , Böhm Beate , Kalden Joachim R. , Nandakumar Kutty Selva , Holmdahl Rikard , Burkhardt Harald
Published: July 2004

Objective

Antileukoproteinase (ALP) is a physiologic inhibitor of granulocytic serine proteases. The present study was undertaken to investigate its therapeutic benefit in an antibody‐transfer model of erosive polyarthritis and to elucidate its potential to interfere with immune complex–dependent inflammatory pathways.

Methods

Arthritis development was induced in male (BALB/c × B10.Q)F1 mice by intravenous injection of two monoclonal antibodies specific for type II collagen and was quantified by clinical scoring and histopathology. Arthritis severity was assessed in a cohort of mice under systemic treatment with recombinant human ALP (daily doses of 0.1 mg for 5 days starting immediately after disease induction) in comparison with untreated controls. Concomitantly, functional assays (phagocytosis, oxidative burst, fluorescence‐activated cell sorting analysis of integrin expression) were performed on neutrophils upon in vitro stimulation by IgG‐coated latex beads.

Results

ALP treatment reduced arthritis incidence and severity and had a protective effect against cartilage and bone erosion. ALP inhibited the conversion of the leukocyte β2 integrins into an active conformation upon Fc receptor stimulation of granulocytes. ALP bound to the actin‐bundling protein L‐plastin and down‐modulated filamentous actin assembly in response to stimulation with IgG‐coated latex beads in granulocytes. ALP exerted additional inhibitory effects on neutrophil functions associated with cytoskeletal reorganization, such as phagocytosis and oxidative burst.

Conclusion

In addition to its antiprotease activity, ALP exerts a variety of blocking effects on neutrophil functions, probably due to modulation of cytoskeletal changes, that may contribute to this inhibitor's antiarthritis potential and qualify it as a multifunctional regulator of inflammatory responses.
First‐degree heart block in the fetus of an anti‐SSA/Ro–positive mother: Reversal after a short course of dexamethasone treatment
Authors: Vesel Samo , Mazić Uroš , Blejec Tanja , Podnar Tomaž
Published: July 2004

Isolated congenital heart block is almost invariably associated with the presence of antibodies to SSA/Ro and SSB/La antigens in the maternal circulation. Once established, third‐degree congenital heart block is permanent. However, a lesser degree of autoantibody‐associated heart block in a fetus can be reversed if it is recognized and treated early enough with fluorinated glucocorticosteroids. The only method available clinically for the recognition of first‐degree heart block in a fetus is measurement of the mechanical PR interval by pulsed Doppler echocardiography. This is the first report of a fetus in whom a diagnosis of first‐degree heart block and the consequent decision to intervene were based solely on this technique. In addition, the first‐degree heart block resolved completely after only 2 weeks of dexamethasone treatment, and the heart rhythm remained stable throughout the remainder of the pregnancy despite early discontinuation of therapy due to oligohydramnios.
Articular hypermobility is a protective factor for hand osteoarthritis
Authors: Kraus V. B. , Li Y.‐J. , Martin E. R. , Jordan J. M. , Renner J. B. , Doherty M. , Wilson A. G. , Moskowitz R. , Hochberg M. , Loeser R. , Hooper M. , Sundseth Scott
Published: July 2004

Objective

Very few studies have evaluated the association of articular hypermobility and radiographic osteoarthritis (OA) in humans. We assessed hypermobility and its relationship to radiographic hand OA in a family‐based study.

Methods

A total of 1,043 individuals were enrolled in the multicenter Genetics of Generalized Osteoarthritis study, in which families were required to have 2 siblings with radiographic OA involving ≥3 joints (distributed bilaterally) of the distal interphalangeal (DIP), proximal interphalangeal (PIP), or carpometacarpal (CMC) joint groups, and OA in at least one DIP joint. Radiographic OA was defined as a score of ≥2 on the Kellgren/Lawrence scale in one or more joints within the group. The Beighton criteria for assessment of hypermobility were recorded on a 0–9‐point scale. Hypermobility was defined as a Beighton score of ≥4, a threshold generally used to establish a clinical diagnosis of joint laxity. A threshold of ≥2 was also evaluated to assess lesser degrees of hypermobility. The Beighton score for the present was calculated based on clinical examination, and that for the past was based on recall of childhood hypermobility in the first 2 decades of life. The association of hypermobility and radiographic OA of the PIP, CMC, and metacarpophalangeal joints was evaluated in all participants and in men and women separately. Multiple logistic regression was used to examine the relationship of hypermobility with radiographic OA in each joint group, after adjusting for age and sex. The association of hypermobility and DIP OA was not evaluated, because evidence of DIP OA was required for study inclusion.

Results

Using a threshold Beighton score of 4, 3.7% of individuals were classified as hypermobile based on the present examination, and 7.4% were classified as hypermobile based on the past assessment. A significant negative association between present hypermobility and age was observed. In persons with hypermobility, the odds of OA in PIP joints was lower (for present, odds ratio [OR] 0.34, 95% confidence interval [95% CI] 0.16–0.71; for past, OR 0.43, 95% CI 0.24–0.78). Similar results were obtained using a threshold Beighton score of 2. The lower odds of PIP OA with hypermobility were significant after adjusting for sex and age (for present, OR 0.44, 95% CI 0.20–0.94; for past, OR 0.48, 95% CI 0.26–0.87).

Conclusion

This study demonstrated a joint‐protective effect of hypermobility for radiographic OA of PIP joints. In contrast to previous studies showing an association of hypermobility and CMC OA, in this cohort there was no evidence for increased odds of OA in any joint group of the hand in association with articular hypermobility.
Absence of antibodies to cyclic citrullinated peptide in sera of patients with hepatitis C virus infection and cryoglobulinemia
Authors: Wener Mark H. , Hutchinson Kathleen , Morishima Chihiro , Gretch David R.
Published: July 2004

Objective

To determine if antibodies to cyclic citrullinated peptide (anti‐CCP) are found in chronic hepatitis C virus (HCV) infection.

Methods

Rheumatoid factor (RF) and anti‐CCP were measured in sera from 50 patients with HCV infection but without cryoglobulinemia, sera from 29 patients with mixed cryoglobulinemia (including 13 with rheumatic symptoms and 5 with arthritis), and sera from 20 normal blood donors. Anti‐CCP was measured by second‐generation enzyme‐linked immunosorbent assay (ELISA).

Results

No sera with elevated anti‐CCP were found in patients with HCV infection without cryoglobulinemia, and in that population, the maximum anti‐CCP was 10 units, well below the positive cutoff of 20 units. Positive findings on RF testing >13 IU/ml were present in 22 (44%) of the HCV patients, with RF >50 IU/ml in 8 (16%) and a maximum RF of 526 IU/ml. Of the cryoglobulinemia patients, 22 (76%) had positive results on tests for RF, including 18 (62%) with RF >50 IU/ml and a maximum RF of 5,540 IU/ml. Two (6.9%) of the cryoglobulinemia patients had borderline‐positive findings on tests for anti‐CCP (25 units and 37 units), which were false‐positive results caused by nonspecific binding in the ELISA. No association between the RF and the anti‐CCP concentrations was found.

Conclusion

Whereas RF was frequent in patients with HCV infection with and without cryoglobulinemia, anti‐CCP was not observed in patients with uncomplicated HCV infection. Borderline‐positive anti‐CCP results were observed infrequently in patients with mixed cryoglobulinemia and were caused by nonspecific binding to plastic. Measurement of anti‐CCP may help in diagnosing RA in patients with chronic HCV infection.
Targeting osteoclasts with zoledronic acid prevents bone destruction in collagen‐induced arthritis
Authors: Sims Natalie A. , Green Jonathan R. , Glatt Markus , Schlict Stephen , Martin T. John , Gillespie Matthew T. , Romas Evan
Published: July 2004

Objective

To study the effect of zoledronic acid (ZA) on synovial inflammation, structural joint damage, and bone metabolism in rats during the effector phase of collagen‐induced arthritis (CIA).

Methods

CIA was induced in female dark agouti rats. At the clinical onset of CIA, rats were assigned to treatment with vehicle or single subcutaneous doses of ZA (1.0, 10, 50, or 100 μg/kg). Clinical signs in all 4 paws were scored on a daily basis. After 2 weeks, the joints in the hind paws were assessed using plain radiographs, microfocal computed tomography (micro‐CT), histologic scoring, and histomorphometry, and the serum levels of type I collagen crosslinks were measured by enzyme‐linked immunosorbent assay.

Results

Although ZA mildly exacerbated synovitis, it effectively suppressed structural joint damage. At doses of ≥10 μg/kg, ZA significantly reduced radiographic bone erosions, Larsen scores, and juxtaarticular trabecular bone loss as quantified by micro‐CT. ZA prevented increased type I collagen (bone) breakdown in CIA and diminished histologic scores of focal bone erosion by up to 80%. Increases in the percentage of eroded surface, osteoclast surface, and osteoclast numbers associated with CIA were prevented by ZA, even though synovitis scores were unchanged.

Conclusion

Single doses (≥10 μg/kg) of ZA strikingly reduced focal bone erosions and juxtaarticular trabecular bone loss, although synovitis was mildly exacerbated. Targeting osteoclasts with ZA may therefore be an effective strategy for preventing structural joint damage in rheumatoid arthritis.
Zoledronic acid protects against local and systemic bone loss in tumor necrosis factor–mediated arthritis
Authors: Herrak Petra , Görtz Birgit , Hayer Silvia , Redlich Kurt , Reiter Erika , Gasser Jürg , Bergmeister Helga , Kollias Giorgos , Smolen Josef S. , Schett Georg
Published: July 2004

Objective

Increased osteoclast activity is a key factor in bone loss in rheumatoid arthritis (RA). This suggests that osteoclast‐targeted therapies could effectively prevent skeletal damage in patients with RA. Zoledronic acid (ZA) is one of the most potent agents for blocking osteoclast function. We therefore investigated whether ZA can inhibit the bone loss associated with chronic inflammatory conditions.

Methods

Human tumor necrosis factor (TNF)–transgenic (hTNFtg) mice, which develop severe destructive arthritis as well as osteoporosis, were treated with phosphate buffered saline, single or repeated doses of ZA, calcitonin, or anti‐TNF, at the onset of arthritis.

Results

Synovial inflammation was not affected by ZA. In contrast, bone erosion was retarded by a single dose of ZA (−60%) and was almost completely blocked by repeated administration of ZA (−95%). Cartilage damage was partly inhibited, and synovial osteoclast counts were significantly reduced with ZA treatment. Systemic bone mass dramatically increased in hTNFtg mice after administration of ZA, which was attributable to an increase in trabecular number and connectivity. In addition, bone resorption parameters were significantly lowered after administration of ZA. Calcitonin had no effect on synovial inflammation, bone erosion, cartilage damage, or systemic bone mass. Anti‐TNF entirely blocked synovial inflammation, bone erosion, synovial osteoclast formation, and cartilage damage but had only minor effects on systemic bone mass.

Conclusion

ZA appears to be an effective tool for protecting bone from arthritic damage. In addition to their role in antiinflammatory drug therapy, modern bisphosphonates are promising candidates for maintaining joint integrity and reversing systemic bone loss in patients with arthritis.
Endothelial precursor cells in the synovial tissue of patients with rheumatoid arthritis and osteoarthritis
Authors: Rüger Beate , Giurea Alexander , Wanivenhaus Axel H. , Zehetgruber Harald , Hollemann David , Yanagida Genya , Groger Marion , Petzelbauer Peter , Smolen Josef S. , Hoecker Paul , Fischer Michael B.
Published: July 2004

Objective

To find evidence for the presence of endothelial precursor cells, which can induce new vessel formation, in the synovial tissue of patients with rheumatoid arthritis (RA) and osteoarthritis (OA).

Methods

Precursor cells in the synovial tissue of 18 RA patients and 15 OA patients were identified by immunohistochemistry, morphometric analysis, and confocal laser scanning microscopy using the following phenotype markers: CD31, CD34, STRO‐1, CD133, vascular endothelial growth factor receptor 2 (VEGFR‐2), and CXCR4. The presence of CD31, CD34, CD133, VEGFR‐2, and CXCR4 messenger RNA in the synovial tissue was determined by reverse transcriptase–polymerase chain reaction, and the message for CXCR4 was quantified by an RNase protection assay.

Results

A population of cells that expressed CD34 on their surface but lacked the endothelial cell marker CD31 was found in the synovial tissue of RA and OA patients. CD34+,CD31− cells were detected in close proximity to STRO‐1+ and CD133+ cells, forming cell clusters in the sublining area of the synovial membrane. Within these cell clusters, CD34+,CD31− precursor cells were located on the inside surrounded by STRO‐1+ cells and with CD133+ cells on the outside. CD34+ precursor cells in the cell layer expressed high levels of the chemokine receptor CXCR4, while VEGFR‐2 was expressed on CD34+ and CD133+ cells, and α‐smooth muscle actin was expressed on STRO‐1+ cells.

Conclusion

The presence of endothelial precursor cells in the synovial tissue of RA and OA patients provides evidence for vasculogenesis induced by precursor cells that arise in situ or from circulating progenitors.
Unknown Repeated Trials in the Unrelated Question Randomized Response Model
Authors: Singh Housila P. , Mathur N.
Published: July 2004

This paper proposes a modified randomization device for collecting information on sensitive issues. The estimator based on the suggested strategy is found to be unbiased for population proportion and is better than the Greenberg et. al.'s (1969) estimator. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)