Circulating cytokeratin 18 fragment m65-a potential marker of malignancy in colorectal cancer patients.

Soluble cytokeratin 18 fragments (M30, M65) are released from human cancer cells during cell death and hold potential as biomarkers in colorectal cancer characterized by frequent metastatic spread. A total of 62 colorectal cancer and 27 control patients were included in the study. M65 (necrosis and apoptosis) and M30 (apoptosis) were quantified preoperatively (n = 62) and postoperatively (n = 31) using specific enzyme-linked immunosorbent assays. Presence of disseminated tumor cells (DTC) in the bone marrow was assessed by staining of A45-B/B3-positive cells in aspirates. M65 was significantly elevated in patients with International Union against Cancer stage I and IIA tumors compared to controls. A subgroup (19/31) exhibited a significant (p < 0.05) decrease of M65 after tumor surgery (503.9 +/- 230.7 to 342.6 + 94.8 U/l; -32.0 +/- 16.5%), in contrast to 12 patients who revealed higher M65 levels postoperatively (386.5 +/- 128.5 to 519.1 +/- 151 U/l; +37.4 +/- 32.3%). DTC in bone marrow were found in 10% (2/19) of patients with decreasing and 50% (6/12) of the patients with increasing M65 serum concentrations after surgery (p = 0.028). In conclusion, M65 as marker is likely to be valuable to identify patients with a high incidence of systemic disease.

Immunomagnetic CD45 depletion does not improve cytokeratin 20 RT-PCR in colorectal cancer.

BACKGROUND: Cytokeratin 20 reverse transcriptase polymerase chain reaction (CK20 RT-PCR) of blood and bone marrow specimens has been suggested for assessment of hematogenously disseminated tumor cell (DTC) spread in colorectal cancer (CRC) patients. Considerable discrepancies among the studies reported indicate a need for better evaluation procedures. We investigated whether mononucleated cell (MNC) enrichment by Ficoll density gradient centrifugation followed by immunomagnetic depletion of CD45-positive cells (extended enrichment) allows better detection of DTC-associated CK20 mRNA compared to MNC enrichment by Ficoll density gradient centrifugation alone (Ficoll enrichment). METHODS: We analyzed 53 samples [38 peripheral blood (PB), 15 bone marrow (BM)] from 38 CRC patients. Extended enrichment was performed for 30 specimens (PB and BM, n=15 each), and Ficoll enrichment for 23 blood specimens. Total RNA was extracted, reverse-transcribed and analyzed by real-time RT-PCR using a LightCycler instrument. RESULTS: Despite extended enrichment, 10 PB and 8 BM samples could not be analyzed because of low cellular yield. The depletion efficiency of CD45 separation was 2 log. RT-PCR of the housekeeping gene PBGD resulted in high and varied crossing point values (mean 37.1+3.0) for five PB and seven BM specimens. Ficoll enrichment yielded 23 analyzable blood specimens for which the mean crossing point value was 26.7+0.5 in PBGD RT-PCR. CK20 RT-PCR of 23 blood samples (all from Dukes D patients) revealed CK20 transcripts in four cases (17%). CONCLUSIONS: Extended enrichment was not superior to Ficoll enrichment; in fact, the sensitivity was lower. Improvement of the reported CK20 RT-PCR assay of Ficoll-enriched MNC populations is warranted.

A rare case of interstitial pneumonitis after tandem high-dose melphalan conditioning and autologous stem cell transplantation in multiple myeloma.

A 57-yr-old woman with multiple myeloma received an autologous tandem transplant at a 4-month interval. She was conditioned twice with 225 mg/m2 melphalan. After the second transplant, interstitial pneumonitis (IP) ensued. The clinical course was life threatening and mechanical ventilation was required for 32 d. All attempts to identify an infectious agent failed. A presumptive diagnosis of idiopathic IP, possibly related to melphalan toxicity, was made. High-dose methylprednisolone administration led to rapid and durable improvement. Melphalan was employed for conditioning in the tandem setting with an interval of only 3-4 months between two courses or a dose elevation to 225 instead of 200 mg/m2, may have induced IP which responded favorably to methylprednisolone.

Biochemical markers of bone metabolism in bone marrow edema syndrome of the hip.

The aim of this study was to evaluate bone metabolism in patients with bone marrow edema syndrome of the hip. In 37 consecutive patients undergoing core decompression of the femoral head, biochemical markers of bone metabolism were measured in aspirates from cancellous bone and in samples obtained simultaneously from peripheral blood. The diagnosis was made by means of radiographs, magnetic resonance imaging (MRI), and core biopsy specimens. Undecalcified microtome section were available for histopathological evaluation. Bone specific alkaline phosphatase (bone ALP), osteocalcin (OC), procollagen Type I N-terminal propeptide (PINP), and C-terminal cross-linking telopeptide (ICTP) were studied. Mean serum levels of analytes were 13.1 ng/mL (OC), 11.2 ng/mL (bone ALP), 4.7 ng/mL (ICTP), and 38.8 ng/mL (PINP). In samples obtained from cancellous bone, mean concentrations of all markers were elevated significantly. The mean bone to serum ratios for bone ALP and OC were 14.1 (P=0.005) and 4.1 (P=0.002), respectively. For collagen Type I metabolites, bone to serum ratios averaged 16.3 (P=0.001) for ICTP and 9.6 (P=0.001) for PINP. Markers of bone formation correlated with each other in serum as well as in aspirates from cancellous bone. Elevation of all markers in aspirates from cancellous bone pointed at increased bone turnover, which correlated with histopathological findings of irregularly woven bone, osteoid seams, and lining cells. Mean serum concentrations of all markers, however, were not different from healthy individuals and thus did not provide any useful clue in the diagnosis of this disease. The lack of osteonecrotic regions in our specimens, the marked increase of bone turnover in samples obtained from edematous lesions, and the fact that none of the patients developed osteonecrosis of the femoral head so far seem to further support the contention that transient bone marrow edema syndrome of the hip is a distinct clinical entity.

Decreased cutaneous vitamin D-synthesis in heavily melanized individuals: a rare cause for pathologic fractures of the hip.

Painful pathological fractures of the femoral neck and the subtrochanteric region of the femur are reported in two women originating from India. After exclusion of renal or intestinal causes, laboratory data on bone metabolism, scintigraphic and radiographic examinations were characteristic for the presence of secondary hyperparathyroidism. Based on vitamin deficiency and low calcium absorption, disturbed mineralization of bone and increased osteoclastic resorption have apparently led to osteomalacia and subsequent fracturing. Fracture localization necessitated surgical fixation in one patient; conservative treatment including protected weightbearing was effective in the other women. After supplementation of calcium and vitamin D3, levels of parathyroid hormone and scintigraphic alterations returned to normal in both patients. In these two cases, pathological fractures of the hip could be attributed to the presence of secondary hyperparathyroidism based on decreased cutaneous vitamin D synthesis.

Comparison of biochemical markers of bone metabolism in serum and femur aspirates.

In 27 patients undergoing arthroscopy of the knee for treatment of meniscal diseases, biochemical markers of bone metabolism were measured in cancellous bone, and levels were compared with concentrations obtained from peripheral blood. Bone-specific alkaline phosphatase, osteocalcin, and collagen Type I metabolites (procollagen Type I N-terminal peptide and carboxy-terminal cross-linked telopeptide) were studied simultaneously in serum and in the distal femur using a radioimmunoassay. Although levels of bone-specific alkaline phosphatase and osteocalcin did not differ between serum and cancellous bone, concentrations of collagen Type I metabolites were elevated significantly in healthy cancellous bone. The close correlations between bone and serum concentrations confirmed accuracy of results obtained from cancellous bone. The mean bone-to-serum ratio for alkaline phosphatase and osteocalcin was 1.1 and 1.2, respectively. Collagen Type I metabolite ratios of 2.2 (for carboxy-terminal cross-linked telopeptide) and 2.3 (for procollagen Type I N-terminal peptide) indicate that these markers are formed locally and then released into the circulation. Bone seems to be a major contributor of collagen Type I metabolites to the serum pool.