José L. Granda

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.

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.

Does Emotional Disclosure About Stress Improve Health in Rheumatoid Arthritis? Randomized, Controlled Trials of Written and Spoken Disclosure

&NA; Studies of the effects of disclosing stressful experiences among patients with rheumatoid arthritis (RA) have yielded inconsistent findings, perhaps due to different disclosure methods – writing or speaking – and various methodological limitations. We randomized adults with RA to a writing (n = 88) or speaking (to a recorder) sample (n = 93), and within each sample, to either disclosure or 1 of 2 control groups (positive or neutral events), which conducted four 20‐minute, at‐home sessions. Follow‐up evaluations at 1, 3, and 6 months included self‐reported, behavioral, physiological, and blinded physician‐assessed outcomes. In both writing and speaking samples, the disclosure and control groups were comparably credible, and the linguistic content differed as expected. Covariance analyses at each follow‐up point indicated that written disclosure had minimal effects compared with combined controls – only pain was reduced at 1 and 6 months, but no other outcomes improved. Spoken disclosure led to faster walking speed at 3 months, and reduced pain, swollen joints, and physician‐rated disease activity at 6 months, but there were no effects on other outcomes. Latent growth curve modeling examined differences in the trajectory of change over follow‐up. Written disclosure improved affective pain and walking speed; spoken disclosure showed only a marginal benefit on sensory pain. In both analyses, the few benefits of disclosure occurred relative to both positive and neutral control groups. We conclude that both written and spoken disclosure have modest benefits for patients with RA, particularly at 6 months, but these effects are limited in scope and consistency. Both written and spoken methods of emotional disclosure have some health benefits for patients with rheumatoid arthritis, but the effects are limited.

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.

Use of a medication control officer to reduce bias in a clinical trial: lessons learned from the scleroderma lung study:

Background Scleroderma Lung Study (SLS) was designed to evaluate the efficacy and safety of oral cyclophosphamide (CYC) versus placebo taken for 1 year for scleroderma-associated interstitial lung disease. An independent medication control officer (MCO), usually a physician, at each center was assigned to monitor laboratory and clinical toxicity of study medication and regulate its dosing based on these results. By having an MCO who watched and managed toxicity, the study investigators were free to care for study patients and to assess study outcomes without the potential bias of knowing toxicity data (toxicity from cyclophosphamide is distinctive — cytopenias and hematuria in particular). Purpose To assess the usefulness of an MCO, whose chief role was to maintain safety while retaining the blinding in the clinical trial. Methods Patients had safety laboratory testing every 2—4 weeks and results were sent directly to the MCO within 2 days of the test. Other clinical adverse events (AEs) were reported by the patient to a nurse coordinator who reported them to the MCO who then managed the AEs to preserve the blinding of investigators caring for the patients. The MCO was provided pre-determined algorithms for dose adjustments of test medication based on the presence and severity of laboratory abnormalities. Results Safety monitoring by the MCO was effective in the early detection of drug toxicity with provision of appropriate medical intervention on a timely basis. At the same time, investigator blinding appeared to be maintained. Limitations The testing of MCO effectiveness in maintaining blinding and consistency was not defined as an a priori hypothesis and thus complete data relating to the efficacy of the MCO were not collected in a prospective fashion. Conclusion An MCO and pre-specified monitoring and dosing guidelines, coupled with uniform pre-specified responses to AEs, may be used effectively to preserve investigator blinding and provide consistency in response to AEs in a clinical trial setting, even when AEs of the test medication are distinctive. Clinical Trials 2010; 7: 85—89. http://ctj.sagepub.com