Significance of fecal deoxycholic acid concentration for colorectal tumor enlargement.

Deoxycholic acid (DCA) has been shown to promote proliferation of colonic carcinoma cells in many fundamental studies. However, no large-scale prospective clinical study providing direct evidence for an association of DCA with progress of colorectal tumor development in humans has been reported to date. To address this question, we conducted a two-step epidemiological study applying enzyme-linked immunosorbent assays to measure fecal cholic acid (CA) and DCA concentrations. Firstly, we compared bile acid concentrations of fecal samples from 366 patients who had multiple colorectal tumors removed endoscopically (tumor group) with those from 24 controls without abnormality in their large intestine (control group). Secondly, the tumor group was followed-up to evaluate the association between fecal bile acid concentrations and recurrence of colorectal tumors four years later. Fecal DCA level in the tumor group were significantly higher than that in the controls, whereas there was no difference in CA levels between the two groups. In the tumor group, a subgroup with high DCA level had higher recurrence risk of large adenomas (> 3 mm) four years later than the low DCA subgroup (odds ratio:1.85, 95% confidence interval: 1.12-3.05). This trend was observed more strongly in the left side colon. In conclusion, a high fecal DCA concentration may be a promoter of colorectal tumor enlargement.

Changes to N-linked oligosaccharide chains of human serum immunoglobulin G and matrix metalloproteinase-2 with cancer progression.

BACKGROUND: Alterations to the sugar chain structure of E-cadherin, a calcium-dependent adhesion molecule, have been shown to influence cancer metastasis. Furthermore, expression of sialyl Le(x) sugar chains on cancer cells has been demonstrated to influence their adhesion to vascular endothelial cells. On the other hand, matrix metalloproteinase-2 (MMP-2) degrades extracellular matrix and is involved in the invasion and metastasis of cancer cells. PATIENTS AND METHODS: N-linked oligosaccharides of human serum immunoglobulin G (IgG) were analyzed in 36 patients with localized or metastatic cancer (12 lung, 12 gastric and 12 prostate cancer) and 10 healthy controls using fluorophore-associated carbohydrate electrophoresis (FACE). MMP-2 levels in the sera were determined by enzyme immunoassay. RESULTS: Fr1 (monogalactosyl IgG oligosaccharide) and Fr2 (digalactosyl IgG oligosaccharides) were significantly decreased (p < 0.001), while Fr4 (agalactosyl IgG oligosaccharides) were significantly increased (p < 0.001) with cancer metastasis. The Fr4/Fr1+Fr2 ratio in localized and metastatic cancer was significantly increased compared to healthy controls (p < 0.001), and was significantly higher in metastatic than localized cancer (p < 0.001). Serum MMP-2 levels in metastatic cancer were significantly higher than in localized cancer (p < 0.001). There was a good correlation between the Fr4/Fr1+Fr2 ratio and serum MMP-2 levels in patients with metastatic cancer (p < 0.0001). CONCLUSION: The analysis of serum IgG N-linked oligosaccharide chain structures by FACE may be an auxiliary indicator of serum tumor markers useful for monitoring cancer progression.

Serum matrix metalloproteinase-2 levels indicate blood-CSF barrier damage in patients with infectious meningitis.

OBJECTIVE: Protein components in cerebrospinal fluid (CSF) are maintained at a specific concentration by a dynamic gradient between the capillary and intrathecal spaces via the blood-cerebrospinal fluid barrier (BCB) in the brain and spinal cord. Permeability to proteins increases when there is structural damage to the BCB. Matrix metalloproteinase-2 (MMP-2; gelatinase A) has been shown to degrade type IV collagen, a major component of the cellular basement membrane. We analyzed alpha2 macroglobulin (alpha2M) indices and evaluated the relationship between alpha2M, as an indicator of BCB permeability, and MMP-2, which degrades the extra-cellular matrix in patients with infectious meningitis. MATERIALS AND METHODS: Albumin levels in CSF or serum were determined by turbidimetric immunoassay, or bromcresol green assay, respectively. alpha2M levels in CSF or serum were measured with enzyme-linked immunosorbent assay, or laser-nephelometry, respectively. Serum MMP-2 levels were determined by enzyme immuno assay. We calculated the alpha2M index, i.e. the ratio of alpha2M (CSF / serum) to albumin (CSF / serum; alpha2M in CSF / alpha2M in serum x albumin in serum / albumin in CSF). RESULTS: alpha2M indices were significantly increased in infectious meningitis compared to healthy controls (p < 0.05). They were highest in bacterial meningitis, and there was a significant difference between viral or mycotic and bacterial meningitis (p < 0.05). Serum MMP-2 levels were increased in infectious meningitis, being highest in bacterial meningitis, where they were significantly different from healthy controls (p < 0.05). There was a significant positive correlation between serum MMP-2 levels and alpha2M indices (r = 0.64, p < 0.0001). CONCLUSION: Markedly increased levels of serum MMP-2 in infectious, especially bacterial, meningitis may reflect the degree of damage to the BCB.

Kinetic analysis of bile acids in the feces of colorectal cancer patients by gas chromatography-mass spectrometry (GC-MS).

PURPOSE: We used gas chromatography mass spectrometry (GC-MS) and reviewed dynamics of bile acids every colorectal cancer occupation part as contrast with a healthy adults after enforcement by all at once analysis of bile acids in feces of a healthy adults and the colorectal cancer patients so that we clarified the details of a possibility of colorectal cancer outbreak participation of allo type bile acids. SUBJECTS: The fecal bile acid measurements were made in a colorectal cancer group consisting of 89 patients and a control group consisting of 103 healthy adults. METHODS: All at once analyzed bile acid in feces of a control group and colorectal cancer group by GC-MS. Student's t-test was used to test for significant differences. RESULTS: As for allo cholic acid (allo CA), allo deoxycholic acid (allo DCA), allo lithocholic acid (allo LCA) and ursodeoxycholic acid (UDCA) which was 4 ingredients of 12 bile acid ingredients out of feces of colorectal cancer group, it showed a significant high level tendency in colorectal cancer group. The following ingredient showed a tendency to significant high level in every colorectal cancer occupation part. Ascending colon: allo DCA and allo LCA. Transverse colon: allo LCA. Descending colon: UDCA. Sigmoid colon: allo DCA and allo LCA. Rectum: allo CA, allo DCA, allo LCA and UDCA. CONCLUSION: The results suggested that the allo type secondary bile acids (allo DCA and allo LCA) are factors that are more strongly involved in colorectal carcinogenesis than DCA or LCA. It was particularly noteworthy that there was a tendency for the allo LCA values to be higher in both sexes (p<0.001-p< 0.005). The results suggested that allo LCA may be a factor involved in colorectal cancer at all sites, except cecum and descending colon. The results suggested that at high values UDCA may be a bodily defense reaction factor that is involved in suppression of carcinogenesis rather than a factor involved in carcinogenesis.

Studies of serum and feces bile acids determination by gas chromatography-mass spectrometry.

Both radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) methods are utilized in the clinical laboratory to measure bile acids in human body fluids. For a more detailed analysis, we attempted simultaneous analysis of serum and feces bile acids using gas chromatography-mass spectrometry (GC-MS) and also investigated the dynamics of bile acids. Serum bile acid composition was examined in 22 healthy adults (79.9 +/- 6.0 years) and 20 colon cancer patients (65.1 +/- 9.5 years). Feces bile acid composition was examined in 20 healthy adults (50.7 +/- 7.6 years) and 20 colon cancer patients (63.6 +/- 8.5 years). The significance of differences was examined by Student's t-test. In both healthy adults and colon cancer patients, the bile acids detected in serum were cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), lithocholic acid (LCA), ursodeoxycholic acid (UDCA), and hyocholic acid (HCA). The following 12 bile acids were detected in feces: CA, allo CA, CDCA, allo CDCA, DCA, allo DCA, LCA, allo LCA, UDCA, HCA, UCA, and CA-6alpha-ol. For allo CA and allo CDCA, no significant differences were observed between the control group and the colon cancer patients. On the other hand, the concentration of allo LCA tended to be higher in the patients (p < 0.05), and the concentration of allo DCA was distinctly higher (p < 0.001) in the colon cancer group, particularly in men. The GC-MS method demonstrated bile acids undetectable by conventional RIA and ELISA. The dynamics suggested association of allo bile acids (DCA and LCA) with colon cancer.

Histopathologic changes in serum bile acid fractions in pressure ulcer patients.

BACKGROUND/AIMS: Bile acids are synthesized in the liver and released into the intestinal tract to aid in digestion and absorption by increasing permeability via alteration of the cell membrane. Bedridden elderly patients typically have pressure ulcers that may be due to both physical local pressure as well as skin cell changes induced by the physiologic effects of bile acids. METHODOLOGY: This study investigated 31 elderly bedridden patients with pressure ulcers (mean age, 81.7 years) and 19 healthy elderly (mean age, 79.7 years). Five serum bile acid fractions were summed to determine total bile acid, and transaminase and cholesterol levels were also measured. RESULTS: Total cholesterol levels were significantly lower (p<0.05) in pressure ulcer patients and transaminase levels were not significantly different between the two groups. The primary bile acids were generally higher and the secondary and tertiary bile acids lower in pressure ulcer patients. In particular, the secondary bile acid deoxycholic acid was significantly higher in all pressure ulcer patients. When analyzed by grade of pressure ulcer, the primary bile acids were significantly lower in pressure ulcer patients. CONCLUSIONS: Secondary bile acid fraction deoxycholic acid measurements may indicate bedridden patients at higher risk for pressure ulcers.

Development of an enzyme-linked immunosorbent assay for fecal bile acid.

It has been suggested that the bile acids in the feces act as a promoter of colon cancer. Among the bile acids, deoxycholic acid (DCA), which is one kind of the secondary bile acid, is said to have strong influence. DCA/cholic acid (CA) ratio in feces is also said to have a diagnostic significance in colon cancer. With this in mind, we created a CA and DCA's monoclonal antibody (MoAb) to measure them through the enzyme linked immunosorbent assay (ELISA) method. Using these MoAb, we were able to measure CA and DCA concentrations with low cross-reaction to other bile acids compared with the method with polyclonal antibody (PoAb). We measured CA and DCA concentrations and calculated the DCA/CA ratios in healthy subjects and patients with colon cancer. All subjects had been screened for colon cancer. We then compared the healthy subjects, the cancer patients before surgery and the same cancer patients after surgery. Cancer patients after surgery had significantly low DCA/CA ratios compared to before surgery, whereas there was no significant difference between healthy subjects and the pre-operative colon cancer patients.