Aberrant expression of metastasis-inducing proteins in ectopic and matched eutopic endometrium of women with endometriosis: implications for the pathogenesis of endometriosis.

BACKGROUND: Endometriosis is a metastatic disease without obvious tumorigenesis. Expression of S100P, S100A4, osteopontin (OPN) or anterior gradient homologue 2 (AGR2) proteins can induce metastasis but fail to induce tumorigenesis per se. We now explore whether this group of metastasis-inducing proteins (MIPs) are associated with the pathogenesis of endometriosis. METHODS: Eutopic endometrial biopsies were taken from 73 women (35 fertile women without endometriosis and 38 women with surgically diagnosed endometriosis). Ectopic endometriotic lesions were collected from eight of the women with endometriosis. The expression of MIPs at the cellular level was evaluated by immunohistochemistry and the presence of these proteins in the endometrial tissues was verified by western blotting and their gene expression was confirmed by RT-PCR. RESULTS: All four MIPs were immunolocated in the endometrium of control women and S100P, AGR2 and OPN showed a cyclical variation. Proliferative phase eutopic endometrium of both groups showed a similar staining pattern for all MIPs, whereas secretory phase endometrium showed a differential expression between controls and cases. The secretory phase endometrial immunostaining of controls showed weak stromal and perivascular AGR2, and decreased stromal and glandular S100P. In contrast, immunostaining for all MIPs was increased in the late secretory endometrial samples of women with endometriosis and intense immunostaining was seen for S100A4 in the stroma (P< 0.05) and for S100P (P< 0.001) and AGR2 (P< 0.0001) in both glands and stroma (P< 0.001). All active peritoneal endometriotic lesions showed strong immunostaining for each of the MIPs studied. CONCLUSIONS: We propose that these MIPs enhance endometrial cell invasiveness and contribute to the establishment of ectopic endometriotic deposits after retrograde menstruation.

Identification and functional analysis of dual-specific A kinase-anchoring protein-2.

Since the cloning of dual-specificity A kinase-anchoring protein 2 (D-AKAP2), there has been considerable progress in understanding the structural features of this AKAP and its interaction with protein kinase A (PKA). The domain organization of D-AKAP2 is quite unique, containing two tandem, putative RGS domains, a PKA-binding motif, and a PDZ (PSD95/Dlg/ZO1)-binding motif. Although the function of D-AKAP2 has remained elusive, several reports suggest that D-AKAP2 is targeted to cotransporters in the kidney and that it may play a role in regulating transporter activity. In addition, the finding that a single nucleotide polymorphism in the PKA-binding region of D-AKAP2 may contribute to increased morbidity and mortality emphasizes the potential importance of this protein in pathogenesis. The first part of this article focuses on initial efforts to identify and clone D-AKAP2, followed by tissue localization and expression profiles. The latter half of the article focuses on the domain organization of D-AKAP2 and its interaction with PKA. Finally, a comprehensive analysis of the PKA binding motif is described, which has led to the development of novel peptides derived from D-AKAP2 that can be useful tools in probing the function of this AKAP in cellular and animal models.

PKA, PKC, and AKAP localization in and around the neuromuscular junction.

BACKGROUND: One mechanism that directs the action of the second messengers, cAMP and diacylglycerol, is the compartmentalization of protein kinase A (PKA) and protein kinase C (PKC). A-kinase anchoring proteins (AKAPs) can recruit both enzymes to specific subcellular locations via interactions with the various isoforms of each family of kinases. We found previously that a new class of AKAPs, dual-specific AKAPs, denoted D-AKAP1 and D-AKAP2, bind to RIalpha in addition to the RII subunits. RESULTS: Immunohistochemistry and confocal microscopy were used here to determine that D-AKAP1 colocalizes with RIalpha at the postsynaptic membrane of the vertebrate neuromuscular junction (NMJ) and the adjacent muscle, but not in the presynaptic region. The labeling pattern for RIalpha and D-AKAP1 overlapped with mitochondrial staining in the muscle fibers, consistent with our previous work showing D-AKAP1 association with mitochondria in cultured cells. The immunoreactivity of D-AKAP2 was distinct from that of D-AKAP1. We also report here that even though the PKA type II subunits (RIIalpha and RIIbeta) are localized at the NMJ, their patterns are distinctive and differ from the other R and D-AKAP patterns examined. PKCbeta appeared to colocalize with the AKAP, gravin, at the postsynaptic membrane. CONCLUSIONS: The kinases and AKAPs investigated have distinct patterns of colocalization, which suggest a complex arrangement of signaling micro-environments. Because the labeling patterns for RIalpha and D-AKAP 1 are similar in the muscle fibers and at the postsynaptic membrane, it may be that this AKAP anchors RIalpha in these regions. Likewise, gravin may be an anchor of PKCbeta at the NMJ.

Rheumatological symptoms. Will investigation make a difference?

BACKGROUND: Many tests are available for investigation and monitoring of rheumatological symptoms, but their usefulness depends on appropriate choice and correct interpretation of the result. OBJECTIVE: To discuss which investigations are appropriate in assessing a variety of rheumatological symptoms and how to interpret results. DISCUSSION: Few of the tests used in the investigation of musculoskeletal symptoms are 'diagnostic'. False positives and false negatives often occur. It is important to interpret the test in the context of the individual patient. Monitoring of diseases is usually based on clinical findings, with investigations only used if they reliably parallel disease activity, monitor drug toxicity, or where changes in the test are likely to lead to modification of management.

Ninth Asia Pacific League of Associations for Rheumatology (APLAR) Congress, Beijing, China, 21-26 May, 2000.

The Congress covered the broad field of rheumatology, with participants from China, the Asia Pacific League of Associations of Rheumatology (APLAR) region and the rest of the world. The programme consisted of a mix of plenary lectures, concurrent symposia, workshops, free paper sessions and poster presentations. Basic sciences were well represented, with the general theme of inflammatory cytokines being of particular interest. One plenary lecture and a number of other presentations addressed the problem of atherosclerosis and rheumatic diseases. Diseases prominent in the region, such as Behcet's disease and Takayasu's disease, were represented with large series. Other areas of interest were musculoskeletal infections in HIV-positive patients and the management of spondyloarthritis. Although the use of the most recently developed drugs is restricted in the APLAR region because of cost factors, there were symposia on the latest pharmacological advances such as COX-2 technology, leflunomide and anti-tumour necrosis factor (TNF) therapy.

A survey of outcome measurement procedures in routine rheumatology outpatient practice in Australia.

OBJECTIVE: To assess the extent to which quantitative clinical measurement is performed by rheumatologists in the longitudinal followup of patients with rheumatoid arthritis (RA), osteoarthritis (OA), ankylosing spondylitis (AS), and fibromyalgia (FM) in routine outpatient practice in Australia. METHODS: A cross sectional postal survey was conducted using an 18-item self-administered questionnaire sent to Australian Rheumatology Association (ARA) members. RESULTS: Rheumatologists (response rate = 76%, completion rate = 72%) were more likely to longitudinally follow patients with RA and AS than those with OA or FM. There was a high degree of variability in the methods used to monitor patients longitudinally. Many measures used in clinical research were used infrequently in routine clinical practice. In general, the major health status measures surveyed were not used in clinical monitoring. There was a high level of agreement (> 80%) that the characteristics required of an outcome measure for use in clinical practice should include simplicity, brevity, ease of scoring, reliability, validity, and sensitivity to change. CONCLUSION: The majority of Australian rheumatologists perform outcome measurement during the longitudinal followup of their outpatients with RA, AS, OA, and FM. However, the process lacks standardization. High performance health status measures developed for clinical research have not been widely adopted in rheumatology practices. There is agreement on the characteristics required by Australian rheumatologists for measurement procedures used in routine clinical care. Quantitative measurement in clinical practice using standardized procedures is an attainable, but as yet, unrealized opportunity.

Chronic treatment with oestradiol does not alter in vitro LTP in subfield CA1 of the female rat hippocampus.

Population excitatory post-synaptic potentials (pEPSPs) were recorded in vitro from subfield CA1 of the hippocampus of female rats which had been ovariectomized and treated for 14 days with either oil or 17beta-oestradiol (10 microg/day). The currents applied to the Schaffer collateral-commissural input necessary to induce threshold, maximum and 50% maximum pEPSP responses did not differ between groups. Application of trains of pulses (0.1-1 s; 100 Hz) evoked post-tetanic and long-term (> 60 min) potentiation of pEPSP responses, the magnitude of which was related to stimulus duration in both groups. However, the degree of potentiation induced by near-threshold (0.1, 0.15 and 0.2 s) and saturating (1 s) stimuli did not differ between groups. Thus, despite reports that oestradiol can modulate synaptic spine density and glutamatergic and GABAergic components of the inputs to CA1, these data suggest that chronic oestradiol treatment has no effect on either the excitability or induction of LTP in the Schaffer collateral-commissural-CA1 pathway.

Pneumatosis intestinalis and pneumoperitoneum complicating mixed connective tissue disease.

A case of a young woman with a 3-year history of mixed connective tissue disease who developed secondary pneumatosis intestinalis and pneumoperitoneum and died shortly after of rapidly progressive disease is reported. The pathogenesis, treatment and prognosis of this unusual complication in mixed connective tissue disease are discussed.

Leg ulcers in rheumatoid arthritis.

A retrospective study over an eight-year period of 33 episodes of leg ulceration in 26 patients with rheumatoid arthritis requiring inpatient management is reported. the aetiology of the ulcers was found to be multifactorial. The most common factors were venous insufficiency (45.5%), trauma or pressure (45.5%) and arterial insufficiency (36.4%). Vasculitis (18.2%) and Felty's syndrome (12.1%) were less frequent causes, and pyoderma gangrenosum was rare. Most patients had seropositive erosive disease with high rheumatoid factor titres and significant functional impairment; over half were on maintenance corticosteroids. Colonisation of the ulcers by organisms, predominantly Staphylococcus aureus, was common (69.7%). Skin grafting was required in 63.3%, but the rate of complete take was only 42.9% despite multiple attempts. Hospitalisation was prolonged (mean 47.9 days) and the recurrence rate requiring further hospitalisation was 26.9%. The diagnosis of vasculitis and the limited role of biopsy in establishing its presence are discussed.

Pulmonary involvement in Behçet's syndrome.

We present a case of Behcet's syndrome with unusual pulmonary manifestations. A woman presented five years previously with oral, genital and ocular lesions found on biopsy to be consistent with Behcet's syndrome. While on prednisolone, she complained of nonpleuritic chest pain. Clinical features of active Behcet's syndrome were absent. A previously normal chest x-ray film showed multiple intrapulmonary lesions. Laboratory abnormalities were consistent with findings seen in phases of disease activity. Bronchoscopy showed ulceration of the bronchial mucosa, but no evidence of opportunistic infection or neoplasia. Pulmonary angiography excluded vaso-occlusive abnormalities. A provisional diagnosis of pulmonary involvement with Behcet's syndrome was made and the patient responded to further immunosuppressive therapy.

Difficulties in the use of D-penicillamine in the treatment of rheumatoid arthritis.

The difficulties encountered in administering D-penicillamine to 40 patients with rheumatoid arthritis (RA) over a six to 24 month period are recorded. Side-effects were frequent. Proteinuria occurred in 13 patients (33%) mainly in the fourth to the sixth month. Renal biopsies were performed in six patients and all showed light microscopy abnormalities. Electron microscopy performed in five patients revealed subepithelial deposits in all and in addition some had mesangial and subendothelial deposits. Seven patients (17.5%) developed eight episodes of thrombocytopaenia which was quickly reversed on cessation of treatment or reduction in dosage. On the positive side, there was significant improvement in most parameters of disease activity at six, 12 and 18 months compared to the pretreatment levels, but the results at two years were less impressive. Reduction in steroid dosage was considerable and was greater than half the mean pretreatment dose after two years but the absence of a control group makes the full significance of these uncertain. Patients on high and low dosage regimens were compared over a 12 month period of treatment. Although the differences were not statistically significant, withdrawals and side-effects were less frequent in the low dose group.

Diagnosis and follow-up of children referred to a rheumatic disease unit.

Juvenile rheumatoid arthritis (JRA) has become the predominant rheumatic disease of childhood. In a survey of 292 children referred to a rheumatic disease unit over a 5-year period, 94 had JRA and only three had rheumatic fever. At follow-up, of the children with JRA, 70% were found to be in remission with little or no functional disability. Febrile onset of JRA or onset before six years of age were worse prognostic features than pauciarticular or polyarticular onset of occurrence after six years of age. Of 28 children who had either growing pains or psychogenic rheumatism, two were subsequently found to have chondromalacia patellae. None of the other children in this group for whom follow-up information was available had developed organic disease.