Robert L. Karvonen

Synovial thickening detected by MR imaging in osteoarthritis of the knee confirmed by biopsy as synovitis

Previous studies have established the value of magnetic resonance imaging (MRI) in detecting articular changes characteristic of osteoarthritis (OA) of the knee. We have observed some MRI features in OA of the knee presumably indicating synovial thickening. To determine whether these MR features represent chronic synovial inflammation, we studied the knees of nine patients at the mild end of the spectrum of OA of relatively short duration (89%: < or = 4 yr), who were selected because MRI showed anatomical abnormalities compatible with synovial thickening. The painful knee was examined using conventional and weight-bearing radiographs, MRI, and arthroscopy. MR images suggestive of synovial thickening typically appeared in or near the intercondylar region of the knee, in the infrapatellar fat pad, or in the posterior joint margin. The site of an arthroscopic biopsy of the synovial membrane was guided by MRI to the area thought to represent synovial thickening for each patient knee. Pathological examination of these synovial membrane biopsies showed a mild chronic synovitis, and thus a correspondence with the synovial thickening detected by MRI. Our results suggest that MRI can be used to evaluate the extent of synovitis, observed as synovial thickening, in patients with early OA of the knee.

Autoantibodies to Annexin XI-A and Other Autoantigens in the Diagnosis of Breast Cancer

We report on the identification of autoantigens commonly recognized by sera from patients with breast cancer. We selected ten sera from patients with invasive ductal carcinoma (IDC) of the breast with high titer IgG autoantibodies for biopanning of a T7 phage breast cancer cDNA display library. A high throughput method involved the assembly of 938 T7 phages encoding potential breast cancer autoantigens. Microarrays of positive phages were probed with sera from 90 patients with breast cancer [15 patients with ductal carcinoma in situ (DCIS) and 75 patients with IDC of the breast], with 51 non-cancer control sera and with sera from 21 patients with systemic autoimmune diseases. A 12-phage breast cancer predictor group was constructed with phage inserts recognized by sera from patients with breast cancer and not by non-cancer or autoimmune control sera (P < 0.0001). Several autoantigens including annexin XI-A, the p80 subunit of the Ku antigen, ribosomal protein S6, and other unknown autoantigens could significantly discriminate between breast cancer and non-cancer control sera. Biopanning with three different sera led to the cloning of partial cDNA sequences identical to annexin XI-A. IgG autoantibodies reacting with the amino acid 41–74 sequence of annexin XI-A were found in 19% of all women with breast cancer but in 60% of sera from women with DCIS of the breast. In addition, partial sequences identical to annexin XI-A, nucleolar protein interacting with the forkhead-associated (FHA) domain of pKi-67, the KIAA1671 gene product, ribosomal protein S6, cyclin K, elongation factor-2, Grb2-associated protein 2, and other unknown proteins could distinguish DCIS from IDC of the breast and appear to be potential biomarkers for the diagnosis of breast cancer.

MR features of osteoarthritis of the knee

A group of patients with idiopathic osteoarthritis (OA) of the knee was surveyed using weight-bearing radiographs and MR imaging to compare the relative value of these methods in disease evaluation. Fifty-two patients with a clinical and radiological diagnosis of OA of the knee of relatively short duration (87%: < or = 4 yr) were compared to a reference group of 40 age- and sex-comparable subjects with no knee symptoms. All patients had a complete history, physical examination, standard anterior-posterior and lateral weight-bearing radiographs, T1-weighted, and FLASH MR images in both knees. The prevalence of MRI abnormalities was significantly greater in patients with OA of the knee in all radiographic grades (Kellgren and Lawrence) compared to the reference subjects. Significant differences were encountered for synovial thickening (OA, 73%; reference, 0%), synovial fluid (60%; 7%), meniscal degeneration (52%; 7%), osteophytes (67%; 12%), and subchondral bone involvement (65%; 7%), even in the patients at the mild end of the osteoarthritic spectrum, indicating the exquisite sensitivity of MRI compared with weight-bearing radiographs.

Articular cartilage defects of the knee: correlation between magnetic resonance imaging and gross pathology.

Magnetic resonance imaging (MRI) of the knee articular cartilage is possible owing to the contrast provided by different signal intensities of adjacent menisci and subchondral bone. The objective of this study was to determine the accuracy of MRI in quantitatively detecting thinning and focal defects of articular cartilage in vivo. High resolution MRI was performed followed by dissection of the knee within one hour of amputations above the knee of eight patients (62-89 years) with peripheral vascular disease. Articular cartilage was examined for erosions, surface irregularities, and appearance. Mean thicknesses of femoral and tibial articular cartilage sagittal sections from MRI were statistically indistinguishable from matched gross thicknesses. In those joints in which cartilage erosions, thinning, or irregularities were detected by MRI the same defects were apparent by gross examination. Cartilage that appeared normal by MRI had a normal gross appearance by gross examination. Thus high resolution MRI can accurately predict gross articular cartilage appearance and thickness, allowing an objective, quantitative, noninvasive assessment of eroded cartilage.

Antinuclear antibodies as potential markers of lung cancer

There are multiple case reports of antinuclear antibodies (ANAs) in patients with malignancies, yet to date there has not been a systematic survey of ANAs in lung cancer. We have previously reported that autoantibodies to collagen antigens resembling those found in the connective tissue diseases are consistently detected in the sera from lung cancer patients. In this work, we looked for the presence of ANAs in the sera from these same patients. Sera from 64 patients with lung cancer and 64 subjects without a history of cancer were retrospectively tested for reactivity on immunoblots of nuclear extracts of HeLa, small cell carcinoma, squamous cell carcinoma, adenocarcinoma, large cell carcinoma of the lung, and of normal lung cells. Associations were sought between the reactivities on immunoblots and lung cancer cell type, diagnosis, and progression-free survival by the method of classification and regression trees (CARTs). Cross-validated CART analyses indicated that reactivities to certain bands in immunoblots are associated with different types of lung cancer. Some of these autoantibodies were associated with a prolonged survival without disease progression. Our data suggest that autoimmunity is often a prominent feature of lung cancer and that molecular characterization of these antigens may lead to the discovery of proteins with diagnostic and prognostic value.

Proteoglycans from Osteoarthritic Human Articular Cartilage Influence Type II Collagen In Vitro Fibrillogenesis

Collagen fibrils were formed in the presence of dermatan sulfate (DSPG) and high density (HDPG) proteoglycans isolated from human adult knee femoral articular cartilage. Eroded cartilage had a higher percentage of DSPGs in the extracted proteoglycans than normal cartilage (p = .018). The dermatan sulfate proteoglycans (DS-PGI and DS-PGII) were detected in normal and osteoarthritic cartilage. DSPGs compared to HDPG inhibited in vitro collagen fibrillogenesis producing a longer lag phase (p less than .05) and a slower rate of fibril formation (p less than .05). DSPGs from eroded osteoarthritic cartilage alone or in combination with HDPG produced a longer lag phase than DSPGs from normal cartilage alone or in combination with HDPG (p less than .05). The inhibition of fibrillogenesis by DSPGs suggests that collagen fibril formation in vivo may be abnormal due to the influence of molecular changes in proteoglycan as well as an increased proportion of DSPGs occurring in osteoarthritic cartilage. Abnormal fibril formation may produce a weakened cartilage matrix, thus contributing to an accelerated process of cartilage degeneration in osteoarthritis.

Dependence of proteoglycan induced arthritis in BALB/c mice on the development of autoantibodies to high density proteoglycans.

BALB/c mice were immunised with high or low density native human cartilage proteoglycans, or the respective core proteins obtained from chondroitin ABC lyase digestion. Mice injected with high density native proteoglycans developed arthritis whereas mice injected with low density proteoglycans or with core proteins did not. Analysis of the immune response by enzyme linked immunosorbent assay (ELISA) and western blot showed a stronger and more polyspecific response in animals injected with low density proteoglycans compared with mice with arthritis which had been injected with high density proteoglycans. Autoantibodies to mouse high density proteoglycans were only present in mice injected with native human high density proteoglycans, however. The data suggest that an arthritogenic epitope lies within the glycosaminoglycan rich region of the native proteoglycan molecule, which may induce an autoantibody response and subsequently arthritis in BALB/c mice.

A human cartilage metalloproteinase with elastolytic activity.

A metalloproteinase with elastolytic properties found in human fetal and osteoarthritic cartilage could not be detected in normal adult cartilage. During extraction the enzyme appeared to be mostly associated with cartilage proteoglycans, from which it can be separated by ion exchange chromatography. This enzyme migrated during electrophoresis with an apparent molecular weight of 62,000 daltons and was found to be fully activated in the tissue under the study conditions. The enzyme showed a preference for substrates rich in non-polar amino acid residues and was capable of breaking down elastin and casein at neutral pH. The enzyme activity can be inhibited by chelating agents and specific affinity reagents, chloroketones, and is not inhibited by other proteinase inhibitors such as PMSF, aprotinin and alpha-1-antitrypsin. This enzyme may play a significant role in conditions demanding rapid cartilage matrix turnover and/or remodeling, such as normal embryonic development or osteoarthritis.