Serum and Cerebrospinal Fluid Testing in Optic Neuropathy Patients with Malignant Tumors.

PURPOSE: To evaluate the value of serum and cerebrospinal fluid (CSF) testing in optic neuropathy (ON) patients with malignant tumors. METHODS: Fourteen patients clinically diagnosed as ON with malignant tumors but without intracranial or orbital mass in MRI were included in this study. Detailed medical records including medical history, complete ophthalmic examination, colour fundus photography, visual field test, orbital MRI examination, serum and CSF testing data were collected and analyzed. The diagnosis of paraneoplastic optic neuropathy (PON) based on the 2004 recommended criteria of the paraneoplastic syndrome- Euronetwork consortium for paraneoplastic neurological disorders, and current adaption for neuropathies. All patients underwent serum tests for pathogens and autoantibodies including antinuclear antibodies, anticardiolipin antibodies, antineutrophil cytoplasmic antibodies, AQP4-Ab and MOG-Ab, as well as CSF tests for malignant cells under microscope. Serum paraneoplastic antibodies were detected in PON patients. Monkey cerebellar tissue-based assay was used to detect unknown serum anti-neuron antibodies in PON patients with negative paraneoplastic antibody testing results. RESULTS: Fourteen ON patients were classified as four groups based on their clinical and MRI characteristics, as well as serum and CSF testing results: [1] definite PON, 6 cases (11 eyes); [2] possible PON, 3 case (5 eyes); [3] meningeal carcinomatosis-associated optic neuropathy (MCON), 4 cases (6 eyes); [4] infiltrative optic neuropathy (ION), 2 cases (2 eyes). Malignant cells were found under microscope in CSF samples from MCON and ION patients, contrast to no malignant cells in CSF samples from PON cases. All 14 ON patients with malignant tumors showed negative results in serum tests for pathogens and autoantibodies. Serum paraneoplastic antibodies were tested in PON patients, anti- CV2, anti-Yo, and anti- amphiphysin were detected positive in 2, 1, and 1 case, respectively, in definite PON group, whereas no serum paraneoplastic antibody detected in possible PON group. Two unknown serum antineuronal antibodies (an anti- Purkinje cell antibody and an anti-granular cell antibody) were detected using monkey cerebellar tissue-based assay in 2 of 5 PON patients with negative paraneoplastic antibody test results. CONCLUSIONS: Serum and CSF tests are of great importance in differentiating different subtypes of ON with malignant tumors. Current diagnosis of PON still depends on combination of clinical and MRI manifestations, as well as serum and CSF tests. Tissue-based assay may help to detect new biomarkers for ON etiology and diagnosis.

Dual-color proteomic profiling of complex samples with a microarray of 810 cancer-related antibodies.

Antibody microarrays have the potential to enable comprehensive proteomic analysis of small amounts of sample material. Here, protocols are presented for the production, quality assessment, and reproducible application of antibody microarrays in a two-color mode with an array of 1,800 features, representing 810 antibodies that were directed at 741 cancer-related proteins. In addition to measures of array quality, we implemented indicators for the accuracy and significance of dual-color detection. Dual-color measurements outperform a single-color approach concerning assay reproducibility and discriminative power. In the analysis of serum samples, depletion of high-abundance proteins did not improve technical assay quality. On the contrary, depletion introduced a strong bias in protein representation. In an initial study, we demonstrated the applicability of the protocols to proteins derived from urine samples. We identified differences between urine samples from pancreatic cancer patients and healthy subjects and between sexes. This study demonstrates that biomedically relevant data can be produced. As demonstrated by the thorough quality analysis, the dual-color antibody array approach proved to be competitive with other proteomic techniques and comparable in performance to transcriptional microarray analyses.

Role of uCyt+ in the detection and surveillance of urothelial carcinoma.

OBJECTIVES: To test the clinical value and role of uCyt+ as a noninvasive tool for the detection and surveillance of urothelial carcinoma. METHODS: Included in this prospective study were 235 patients (mean age 71.5 years, range 32 to 86). Of these, 98 patients had signs and symptoms suggestive of bladder cancer and 137 patients were being followed up after complete transurethral resection of superficial urothelial cancer (UC). All patients underwent urinary cytology and the uCyt+ test performed on ThinPrep (thin layer). All underwent subsequent cystoscopy and evaluation of any suspicious lesion by biopsy. RESULTS: A total of 102 patients had histologically proven UC. In the group of patients with signs and symptoms suspicious of UC, the sensitivity of cytology increased from 5% for G1 to 84.6% for G3 tumors; for uCyt+, it was 85% for G1, 100% for G2, and 92.3% for G3 tumors. Combining cytology and uCyt+, the sensitivity was 85% for G1 and 100% for G2 and G3. In the group of follow-up patients, the sensitivity of cytology increased from 4.3% for G1 to 94.4% for G3 tumors; for uCyt+, it was 78.2% for G1, 70% for G2, and 94.4% for G3 tumors. Combining both tests, the sensitivity was 78.2% for G1, 90% for G2, and 100% for G3. CONCLUSIONS: The uCyt+ is a valid test in the detection of UC of all grades and stages. It improves the sensitivity of cytology in low-grade tumors. The two tests combined may be a highly sensitive method to detect UC early in detection and surveillance.

Urine antibody against human cancer antigen NY-ESO-1.

NY-ESO-1 is one of the most immunogenic tumor antigens known to date. Spontaneous humoral and cellular immune responses against NY-ESO-1 are detected in a substantial proportion of patients with NY-ESO-1 positive cancers. NY-ESO-1 serum antibody is dependent on the presence of NY-ESO-1+ cancer cells, and antibody titers correlate with the clinical development of the disease. NY-ESO-1 serum antibody is associated with detectable NY-ESO-1-specific CD8+ T cell reactivity. High titers of NY-ESO-1 serum antibodies are found in patients with advanced NY-ESO-1+ malignancies. Urine samples of seropositive patients with normal kidney function were tested for NY-ESO-1 antibody by Western blotting and enzyme-linked immunosorbent assay (ELISA). Antibodies to NY-ESO-1 were found in the urine of patients whose NY-ESO-1 serum antibody titers were 1:10,000 or higher by Western blotting. In patients with weak (positive at 1:250, negative at 1:1,000) or no reactivity, urine antibody was not detectable. No urine NY-ESO-1 antibody was found in patients without detectable NY-ESO-1 serum antibody. Our results show that urine analysis for NY-ESO-1 antibody identifies patients with strong NY-ESO-1 immunity. Urine antibody detection may also be of value in the monitoring of spontaneous and vaccine-induced immunity against other defined tumor antigens.

Urinary monoclonal free light chains in primary Sjögren's syndrome: an aid to the diagnosis of malignant lymphoma.

Three patients, two with typical primary Sjögren's syndrome (SS) and the third with several features of SS, including abnormal sialography and reduced tear secretion, developed B cell non-Hodgkin's lymphoma (NHL) of parotid or lung, or both. Isoelectric focusing of concentrated urine specimens in agarose, followed by immunofixation, demonstrated the presence in each patient's urine of monoclonal free light chains of the same class as that shown on the tumour cells. In one patient the level of urinary free light chains was monitored and found to correlate with disease activity. Similar techniques showed no monoclonal light chains in the urine from a further 26 cases of SS with no clinical evidence of lymphoma. The detection of monoclonal urinary free light chains may provide an early diagnostic clue to the development of lymphoma in patients with SS and be a means of tumour monitoring.

Lymphoma in Sjögren's syndrome: urinary monoclonal free light chains as a diagnostic aid and a means of tumour monitoring.

Non Hodgkins lymphoma (NHL) is reported to be at least 40 times more common in Sjögren's syndrome (SS). Diagnosis may be difficult as blood and bone marrow haematology can remain normal, with no evidence of a serum paraprotein band or Bence-Jones proteinuria by routine electrophoresis. Using the technique of isoelectric focusing in agarose, followed by immunofixation, monoclonal free light chains can be found in the urine of 44% and 74% respectively of patients with NHL and B cell chronic lymphocytic leukaemia, but not in normal individuals. Three patients, two with typical severe primary SS and the third with several features of SS including abnormal sialography and reduced tear secretion, developed B cell NHL of parotid and/or lung. Using the above method on concentrated urine specimens, monoclonal free light chains of the same class as that demonstrated on the tumour cells were found to be present in each patient's urine. In one patient the level of urinary free light chains was monitored and found to correlate with disease activity. Using similar techniques no monoclonal light chains could be found in the urine from a further 10 cases of primary SS and 18 cases of SS secondary to rheumatoid arthritis, all of whom had no clinical evidence of lymphoma. Screening of SS patients' urine by the method described for monoclonal urinary free light chains may provide an early diagnostic clue to the development of lymphoma and be a means of tumour monitoring.

Monoclonal immunoglobulin light chain in urine of patients with B lymphocytic disease: its source and use as a diagnostic aid.

The presence of tumour-related monoclonal light chain has been sought in urine as an immunochemical aid in the diagnosis of B lymphocytic neoplasms. The technique of isoelectric focusing in agarose followed by immunofixation has been applied to concentrated urines from 41 patients. In chronic lymphocytic leukaemia involving neoplastic B lymphocytes, monoclonal light chain was detected in 14 out of 19 patients investigated. For 2 of the positive cases (one kappa light chain type and one lambda light chain type) the urinary light chains were compared directly with culture fluids obtained after incubation of the corresponding neoplastic cells obtained from the patient's peripheral blood: identity of the light chains from urine and cells was established by isoelectric focusing demonstrating for both patients that the tumour cells were the source of the urinary light chain. In patients with non-Hodgkin's lymphoma involving neoplastic B lymphocytes, urinary monoclonal light chains were found in 7/16 of those studied. Such light chains were not detected in 11 control subjects, in 3 patients with true histiocytic tumours or in 2 patients with enlarged reactive lymph nodes. The technique is simple to perform and provides information for diagnosis and possibly monitoring of B cell neoplasms.

[Content of 3-hydroxyanthranilic acid antigens and antibodies in mice in early stages of hepatocarcinogenesis]. / Soderzhanie 3-OAK-antigenov i antitel u myshei na rannykh stadiiakh gepatokatserogeneza.

By means of a qualitative and quantitative method of precipitation it was found possible to record 3-OAA-antigens in mice receiving ortho-aminoazotoluene. the authors obtained some additional data indicating a similar kinetics of binding for exogenous and endogenous carcinogen. The elaborated method of quantitation of 3-OAA-antigens in the urine may be used for studying the kinetics of this index in cancer patients.