Short- and long-term outcome of linear morphoea in children.

BACKGROUND: Linear morphoea (LM) is a rare fibrosing disorder of the limbs or the face that may cause functional disability and severe aesthetic sequelae. Despite a wide range of therapeutics reported for LM, there is currently a lack of consensus on the optimal therapy. Little is known about the long-term outcome of this disease. OBJECTIVES: To describe the short- and long-term outcome of a large series of patients with LM acquired in childhood. METHODS: A retrospective chart review of 52 paediatric patients with LM seen in our centre during a 20-year span (1990-2010) and a telephone survey in 2011 to assess the long-term outcome of these patients. RESULTS: Limbs were affected twice as often as the face, with a higher proportion of female patients. Stabilization was obtained after a mean disease duration of 5·4 years. Patients sometimes experienced long stretches of disease quiescence followed by reactivation; 31% of patients reported active disease after 10 years. All but one patient had aesthetic sequelae, and 38% had functional limitations. The effectiveness of methotrexate and systemic corticosteroids was apparent in the short term. CONCLUSIONS: LM needs prolonged monitoring as the disease can have very long periods of quiescence followed by reactivation. The combination of methotrexate and systemic corticosteroids was effective in the early stages of the disease but did not seem to prevent long-standing active disease or relapse in the long term.

The development of bone mineral lateralization in the arms.

UNLABELLED: Bone mineral content (BMC) is known to be greater in the dominant arm after the age of 8 years. We studied a group of children and found that BMC sidedness gradually increased up to the age of 6 years and then remained stable into late adolescence. INTRODUCTION: Bone mineral content (BMC) exhibits sidedness in the arms after the age of 8 years, but it is not known whether BMC is greater in the dominant arm from birth or whether lateralization develops in early childhood. To address this, we examined bone mineral status in relation to handedness and age. METHODS: Subjects (N = 158) were children recently initiating glucocorticoids for underlying disease (leukemia 43 %, rheumatic conditions 39 %, nephrotic syndrome 18 %). Handedness was determined by questionnaire and BMC by dual-energy X-ray absorptiometry. RESULTS: Median age was 7.2 years (range, 1.5 to 17.0 years), 49 % was male, and the spine BMD Z-score was -0.9 (SD, 1.3). By linear regression, BMC sidedness in the arms was significantly related to age (r = 0.294, p = 0.0005). Breakpoint analysis revealed two lines with a knot at 6.0 years (95 % CI, 4.5-7.5 years). The formula for the first line was: dominant:nondominant arm BMC ratio = 0.029 × age [in years] + 0.850 (r = 0.323, p = 0.017). The slope of the second line was not different from 0 (p = 0.332), while the slopes for the two lines were significantly different (p = 0.027). CONCLUSIONS: These results show that arm BMC sidedness in this patient group develops up to age 6 years and then remains stable into late adolescence. This temporal profile is consistent with mechanical stimulation of the skeleton in response to asymmetrical muscle use as handedness becomes manifest.

Prevalent vertebral fractures among children initiating glucocorticoid therapy for the treatment of rheumatic disorders.

OBJECTIVE: Vertebral fractures are an under-recognized problem in children with inflammatory disorders. We studied spine health among 134 children (87 girls) with rheumatic conditions (median age 10 years) within 30 days of initiating glucocorticoid therapy. METHODS: Children were categorized as follows: juvenile dermatomyositis (n = 30), juvenile idiopathic arthritis (n = 28), systemic lupus erythematosus and related conditions (n = 26), systemic arthritis (n = 22), systemic vasculitis (n = 16), and other conditions (n = 12). Thoracolumbar spine radiograph and dual x-ray absorptiometry for lumbar spine (L-spine) areal bone mineral density (BMD) were performed within 30 days of glucocorticoid initiation. Genant semiquantitative grading was used for vertebral morphometry. Second metacarpal morphometry was carried out on a hand radiograph. Clinical factors including disease and physical activity, calcium and vitamin D intake, cumulative glucocorticoid dose, underlying diagnosis, L-spine BMD Z score, and back pain were analyzed for association with vertebral fracture. RESULTS: Thirteen vertebral fractures were noted in 9 children (7%). Of these, 6 patients had a single vertebral fracture and 3 had 2-3 fractures. Fractures were clustered in the mid-thoracic region (69%). Three vertebral fractures (23%) were moderate (grade 2); the others were mild (grade 1). For the entire cohort, mean +/- SD L-spine BMD Z score was significantly different from zero (-0.55 +/- 1.2, P < 0.001) despite a mean height Z score that was similar to the healthy average (0.02 +/- 1.0, P = 0.825). Back pain was highly associated with increased odds for fracture (odds ratio 10.6 [95% confidence interval 2.1-53.8], P = 0.004). CONCLUSION: In pediatric rheumatic conditions, vertebral fractures can be present prior to prolonged glucocorticoid exposure.

Immunological and clinical differences between juvenile and adult onset of systemic lupus erythematosus.

INTRODUCTION: Systemic lupus erythematosus (SLE) in children usually follows a more severe course than in adults, but sometimes in the previous studies reported there are many confounding factors. OBJECTIVE: To analyse the immunological and clinical characteristics of SLE juvenile onset and SLE adult onset. METHODS: We studied 179 patients with SLE, 49 patients were aged 6-18 yrs at onset of disease. Anti-dsDNA antibodies were detected by radioimmunoassay and antibodies to extractable nuclear antigens (ENA): anti-nRNP, anti-Sm, anti-Ro/SS-A and anti-La/SS-B antibodies by ELISA, counterimmuno-electrophoresis and immunoblotting. RESULTS: Juvenile-onset SLE shows a higher frequency of cutaneous vasculitis (44.8% vs 27.6%; P < 0.05), seizures (18.3% vs 7.6%; P < 0.05) nephropathy (67.3% vs 48.4%; P < 0.025), and discoid lupus erythematosus (26.5% vs 13.8%; P < 0.05). The incidence of articular manifestations is lower than in adults (85.7% vs 96.1%; P < 0.025). No significant differences were found between the two groups in relation with the prevalence of antinuclear antibodies. CONCLUSIONS: Juvenile-onset SLE has more frequent neurological and renal manifestations than adult-onset SLE, but immunological markers are similar in both groups. These features suggest the most severe clinical manifestations in the juvenile-onset SLE group are not related with the presence of studied antibodies by different methods.

The relationship of antiphospholipid antibodies to thromboembolic events in pediatric patients with systemic lupus erythematosus: a cross-sectional study.

The purpose of this study was to evaluate pediatric patients with systemic lupus erythematosus (SLE) to determine 1) the incidence of thrombosis, 2) the incidence of antiphospholipid antibodies, and 3) whether there is an association between the presence of antiphospholipid antibodies and thrombosis. We performed a cross-sectional cohort study in 59 consecutive SLE patients who had been managed at rheumatology clinics in two pediatric hospitals. A history, questionnaire, and chart review were completed by the study nurse blinded to laboratory results. Only the thrombotic events that could be substantiated by review of radiographic tests were accepted. The presence of antiphospholipid antibodies was determined by prospective analysis for a lupus anticoagulant and anticardiolipin antibodies on two separate occasions at least 3 mo apart. Patients were considered to be positive if one or more tests were positive on both occasions. Thirteen thrombotic events occurred in 10 of the 59 patients (17%). Fourteen patients (24%) were classified as positive for lupus anticoagulant, and 19 patients (27%) were classified as positive for anticardiolipin antibodies. A significant relationship between the presence of a lupus anticoagulant and a thrombotic event was shown: odds ratio 28.7 (95% confidence interval 4.03-138.2, p < 0.001). A nonsignificant trend was seen for the presence of an anticardiolipin antibody and a thrombotic event: odds ratio 2.12 (95% confidence interval 0.71-22.8, p=0.08). We conclude that in pediatric patients with SLE: 1) a significant proportion of patients have thrombotic events, 2) a significant proportion of patients have antiphospholipid antibodies, and 3) there is a significant relationship between the presence of a lupus anticoagulant and thrombotic events.

Induction of simian virus 40 transplantation immunity in hamsters by gel-purified SV40 large T antigen.

Simian virus 40 (SV40) large T antigen and p53 cellular protein were isolated from an SV40-transformed hamster cell line by immunoprecipitation with anti-T sera and purified by sodium dodecyl sulfate-gel electrophoresis. These two protein were tested in hamsters for the presence of SV40 transplantation rejection antigenic sites by in vivo transplantation rejection assay. The large T antigen immunized the hamsters against a challenge of SV40 tumor cells and the protected animals generated cytotoxic spleen cells. Hamsters immunized with the p53 cellular protein were not protected against SV40-induced tumor but there was some delay in the appearance of tumor.

Spleen cell populations in normal and tumor-bearing hamsters.

Spleen cell subpopulations from normal and tumor-bearing hamsters (TBH) were quantified using Petri dishes coated with specific antibodies, and by flow cytometric immunofluorescence analysis. The relative numbers of T cells (Thy + cells) decreased, by both methods, as a function of tumor growth, while the number of B cells (bearing surface Ig) increased. Cells without T or B markers (null cells) were more numerous in the spleen of TBH and a large number of them expressed the receptor for the Fc fragment of IgG. Splenic cells were also sorted according to their light-scattering properties, and electron microscopic analysis was performed on the sorted fractions. It showed the presence of secreting plasmocytes and activated macrophages in the spleen of TBH.

Variations in the levels of IgG1 and IgG2 subclasses in the sera of normal, immunized and tumor-bearing hamsters.

The concentration of IgG1 and IgG2 subclasses in the sera of hamsters bearing tumors of different origins were compared to that of normal serum and to that of sera of animals rendered resistant to tumor take by immunization with viable SV40-transformed cells. In the sera of hamsters bearing tumors induced by virus-transformed cells an augmentation of IgG2 and a diminution of IgG1 was observed during the development of the tumor compared to sera of normal hamsters. In the sera of animals bearing tumors induced by methylcholanthrene ( MCH2 ) or by spontaneously transformed cells ( EHB ) the level of IgG2 was almost normal but IgG1 was barely detectable, especially in MCH2 tumors. On the other hand, the sera of animals immunized with virus-transformed cells showed a slight increase in both IgGs, but only that of IgG2 was significant. Antibody activity was tested in the sera as well as in the IgG1 and IgG2 fractions of the sera of hamsters immunized or bearing tumors induced by SV40 transformed cells. Both sera and the subclasses showed antibody activity, the activity being more pronounced in the IgG2 fraction than in the IgG1 fraction.

Immunologic and enzymatic studies of two breeding lines of NZB mice that differ in chromosome breakage.

Two sublines of NZB/BI mice were developed by selective matings according to chromosome breakage frequencies. These sublines--HB, a line with high chromosome breakages, and LB, a line with low or normal breakage rates--were studied in regard to the age of the animals as it related to two different aspects. The first aspect was immunologic: A decreased response to T-cell mitogens was found in old NZB mice, but this response was more pronounced in HB mice. The response to the B-cell mitogen (lipopolysaccharide) was increased in both sublines as compared to that in BALB/c mice. The percentages of IgG-positive and theta-positive spleen cells were evaluated in both sublines: Some increase in IgG-positive cells was observed in the spleens of 2- to 8-month-old NZB mice and a slight decrease was seen after age 8 months. The percentage of theta-positive cells diminished according to the age of the mice, and the decrease occurred earlier in HB than in LB mice. The second aspect studied was enzymatic and concerned the levels of DNA alpha- and beta-polymerases and terminal DNA nucleotidyltransferase in the thymuses and spleens of these animals. The major finding noted was an augmentation of 100-200% in the terminal DNA nucleotidyltransferase levels in HB thymuses by comparison with LB thymuses. The levels of both polymerases were increased in spleen cells of HB mice as compared to those of LB mice.

In vivo studies of spleen lymphoid cells implicated in antitumor immunity in hamsters.

Hamsters were immunized with repeated and progressive doses of strongly antigenic SV40-transformed cells (ZDCl25) and these animals rejected a tumor graft. According to the in vivo Winn test, spleen cells from such immunized animals could be used to transfer the antitumor immunity and protect naive recipients. To characterize the immune effector cells, different techniques of fractionation were used. In all cases the best protection was obtained by whole cell populations, even after reconstitution. The "T-depleted' fraction was more active than the T cell fraction. As shown by the treatment with specific antisera, part of the active population bore surface IgG. The protective activity could be the result of cooperation between different cell populations, one of which, unknown at present, could be the promotor of the transferred immunity and of the tumor neutralization effect.

Nude Syrian hamsters: some immunological and histological characteristics.

Nude mutants appeared in our colony of Syrian hamsters. They were hairless and just had rudiments of thymus. Only 3.5% of splenic cells were killed by rabbit anti-hamster thymocytes serum + C' whereas 55.5% of these cells were lyzed by an anti-hamster IgG + C' and 60-70% fixed the fluorescent protein A, but could not respond either to B-cell mitogens or to rat cell mitogens. The natural cytotoxic activity of the spleen cells from nude hamsters was evaluated in comparison with the same activity expressed by spleen cells of golden Syrian hamsters.

Isolation of two immunoglobulin G subclasses, IgG2 and IgG1, from hamster serum using protein A-sepharose.

The isolation of hamster immunoglobulin classes and subclasses by affinity chromatography on protein A and selective elution was studied using 0.1 M phosphate buffer, pH 8. The IgG fraction was completely absorbed, while IgM did not bind. Sequential application of buffers of decreasing pH allowed the elution of pure IgG2 (eluted at pH 6) and IgG1 (eluted at pH 5). Both subclasses were fully recovered. IgG2 could be subfractionated into 2 peaks eluted respectively at pH 6.5 and 6. Immunodiffusion of the whole IgG2 fraction against anti-hamster immunoglobulin serum gave 2 precipitation lines. One of these lines was missing in the pH 6.5 fraction. Until now only 2 IgG subclasses have been described and these results suggest heterogeneity of hamster IgG2.

Evolution of DNA polymerase alpha, beta and terminal deoxynucleotidyl transferase in hamster lymphoid populations during the development of different types of tumors.

We followed the evolution of DNA polymerase alpha, beta and terminal deoxynucleotidyl transferase (TdT) in the thymus, spleen and bone marrow of tumor bearing hamsters. Tumors were induced by spontaneously transformed fibroblasts (EHB), by SV40 (ZD) or by 20-methylcholanthrene (MCH2) transformed fibroblasts. We have shown that, in the thymus, the TdT activity of the various tumor bearing hamsters is generally inferior to the TdT activity of the control. In the spleen of animals bearing viral or spontaneously induced (ZD or EHB) tumors, the TdT activities were higher than in the controls. Moreover, DNA polymerase alpha and DNA polymerase beta activities in the spleen of the EHB tumor bearing hamsters were higher than in the controls. This fact, however, was not observed in the two other kinds of tumor, possibly because EHB tumors were growing much faster and led to earlier changes in DNA polymerases activities, as well as in spleen size and cellular populations. Finally, in the bone marrow, TdT, polymerase alpha and beta of ZD and EHB tumor bearers reached much higher activities than in the controls; for the MCH2 tumor bearing hamsters, no difference with the controls could be observed.

[Natural cytotoxic activity in different experimental conditions in hamsters (author's transl)]. / Activité cytotoxique naturelle dans différentes conditions expérimentales chez le hamster.

Lymphoid cells from normal hamsters from our inbred Z strain were evaluated for their natural cytotoxic activity against YAC cells (mouse lymphoma) and ZDc125 and EHB cells (cultured syngeneic hamster fibroblasts) employing the 51Cr-release cytotoxic essay. Natural cytotoxic reactivity was high in spleen, intermediate in peritoneal cells and very low in the thymus. Natural cytotoxic response was present in newborn animals, reaches a maximal level within 4-8 months and declines very slowly thereafter. Treatment of hamsters with a single dose of cyclophosphamide (Cy) augmented the natural cytotoxic reactivity 2 to 3 days after injection; this activity was suppressed thereafter and reappeared 20-25 days after the Cy injection. Growth of transplantable ZDc125 or EHB tumour in syngeneic hamsters inhibited the natural cytotoxic activity as soon as the tumour was palpable. Preincubation of target cells in sera of tumour-bearing animals diminished the level of natural cytotoxic activity of spleen lymphoid cells. One can postulate that this could be due to the presence of antibodies or immune complexes in those sera.

Cyclophosphamide induction of splenic suppressor cells in hamsters.

Injection of hamsters with a single sublethal dose of cyclophosphamide induced splenic atrophy followed by considerable hypertrophy. During the splenomegaly phase, the in vitro reactivity of spleen cells to concanavalin A (ConA) or protein A (ProtA) was decreased. The spleens of 6-8-month old animals contained cells able to suppress the in vitro reactivity of normal lymphocytes to ConA or ProtA. These cells were absent in 2-month old animals, and some suppressor activity could be shown in 3-month old hamsters. Suppressor activity was abolished by removal of adherent cells and by silica treatment, but antisera against thymocytes or IgG were ineffective.

Chromosome damaging agent of low molecular weight in the serum of New Zealand black mice.

Two substrains of NZB mice were developed by selective matings according to chromosome breakage frequencies in direct bone marrow preparations: HB, a line with increased chromosome breakage, and LB, a line with low or normal breakage rates. The serum of NZB mice from the HB strain contains a chromosome breaking factor that produces chromosome abnormalities in human lymphocytes. This clastogenic substance could not be detected in serum of NZB line. Nor does it exist in serum of Balb, C3H, C57 B1, Swiss, or AKR mice, in which no chromosomal breakage was observed in bone marrow. The breakage factor has a molecular weight between 1000 and 10,000 daltons.

Circulating immune complexes in sera of hamsters bearing simian virus 40-induced tumours.

Reported previously in certain human carcinomas and rat and mouse experimental tumor systems, circulating immune complexes (IC) have now been detected in Syrian hamsters bearing tumors produced by injection with syngeneic TSV5Cl2 cells. IC were detected by the Raji cell radioimmune assay, adapted for use in hamster sera. A novel feature of this test is the use of a stable covalently linked hamster immunoglobulin G aggregate as the reaction standard. Stable over six months on storage at -70 degrees and showing no tendency to form precipitates on thawing or during test procedures, this preparation greatly facilitated quantitation of hamster IC by Raji cells. Seven of 19 sera of hamsters bearing SV40-induced tumors from 60 to 128 days had IC concentrations exceeding 40 microgram aggregated hamster immunoglobulin G equivalents per ml, as contrasted to IC levels of less than 25 microgram for 19 age-matched normal hamsters. There appeared to be no significant correlation between IC levels and tumor weight or duration of tumor within the hamster host. The results suggest a complex relationship between IC and a number of factors connected with tumor growth.