Antiproliferative Effects of Short-chain Fatty Acids on Human Colorectal Cancer Cells via Gene Expression Inhibition.

BACKGROUND/AIM: Short-chain fatty acids (SCFAs) inhibit human colorectal cancer cell growth and tumorigenicity. We investigated the mechanism of the anti-proliferative effects of SCFAs on human colorectal cancer cells by examining their effects on gene expression. MATERIALS AND METHODS: The DLD-1 cell line was cultured with different SCFAs. Gene groups whose expression levels decreased to <50% or increased >50% compared to untreated cells and the signalling pathways responsible for DLD-1 cell growth inhibition were identified and analyzed. RESULTS: Genes whose expression levels decreased to ≤50% (791 genes) showed remarkable changes in gene function compared to genes whose expression levels increased ≥50%. These genes encode proteins involved in DNA replication and cell cycle/proliferation that contribute to major pathways responsible for suppression of colorectal carcinogenesis pathways. CONCLUSION: SCFAs inhibited the expression of genes encoding proteins involved in DNA replication and cell cycle/proliferation of human colorectal cancer cells and exerted antiproliferative activity via different pathways.

Intake of Bifidobacterium longum and Fructo-oligosaccharides prevents Colorectal Carcinogenesis.

INTRODUCTION: We aimed to investigate the effects of intake of yogurt containing Bifidobacterium longum (BB536-y) and fructo-oligosaccharides (FOS) in preventing colorectal carcinogenesis in healthy subjects, and the preventive effects of short-chain fatty acids (SCFA), whose production was enhanced by the intake of BB536-y and FOS, in human colon cancer cell lines. MATERIALS AND METHODS: The subjects were 27 healthy persons who were divided into a group taking yogurt containing BB536 (BB536-y group; n = 14) and a group taking yogurt containing BB536 and FOS (BB536-y with FOS group; n = 13) once a day for 5 weeks. The feces were sampled before and after the intake to analyze the amount of SCFA in the feces and the profile of intestinal flora, such as putrefactive bacteria and Bacteroides fragilis enterotoxin (ETBF). Subsequently, human colon cancer cell lines (DLD-1 cells, WirDr cells) were cultured in the presence of SCFA (butyric acid, isobutyric acid, acetic acid) in order to evaluate the cell growth-inhibitory activity of SCFA (WST-8 assay) by calculating the IC50 value from the dose-response curve. RESULTS: Intake of BB536-y increased the total amount of SCFA in the feces and significantly suppressed the detection rate of ETBF and growth of putrefactive bacteria. Intake of BB536-y with FOS was associated with a higher Bifidobacterium detection rate than that of BB536-y alone. The contents of butyric acid, isobutyric acid, and acetic acid, namely, of SCFA, were also decreased. Analysis of the results of culture of DLD-1 cells and WirDr cells in the presence of butyric acid, isobutyric acid, and acetic acid revealed that each of the substances showed significant cell growth-inhibitory activity, with the activity being the highest for butyric acid, followed by that for isobutyric acid and acetic acid. CONCLUSION: These findings suggest that intake of both BB536-y and BB536-y with FOS prevents colorectal carcinogenesis.How to cite this article: Ohara T, Suzutani T. Intake of Bifidobacterium longum and Fructo-oligosaccharides prevents Colorectal Carcinogenesis. Euroasian J Hepato-Gastroenterol 2018;8(1):11-17.

Acute inflammatory biomarkers in cerebrospinal fluid as indicators of blood cerebrospinal fluid barrier damage in Japanese subjects with infectious meningitis.

BACKGROUND: The blood-cerebrospinal fluid barrier (BCB) has selectivity for protein components with different molecular weights. Protein components in the cerebrospinal fluid (CSF) change when the BCB is damaged. We calculated the alpha2 macroglobulin (alpha2M) index as an indicator of BCB permeability from the point of view of molecular weight and evaluated the relationship between the alpha2M index and CSF concentrations of the inflammatory biomarkers interleukin-6 (IL-6), C-reactive protein (CRP), and serum amyloid A (SAA) in Japanese subjects with infectious meningitis, in order to determine the clinical significance of those inflammatory biomarkers in CSF. METHODS: IL-6, CRP, and SAA levels in CSF and serum were measured using various methods. The alpha2M index was calculated as the ratio of alpha2M (CSF/serum) to albumin (CSF/serum). RESULTS: CSF IL-6 levels were higher than serum IL-6 levels in 16 patients with infectious meningitis. The difference in CSF IL-6 and CRP levels between mycotic or bacterial meningitis cases and healthy controls and in CSF SAA levels between all infectious meningitis cases and healthy controls were significant. There was a significant positive correlation between CSF levels of CRP or SAA and alpha2M indices. CONCLUSIONS: Markedly increased levels of IL-6 in the CSF of patients with infectious meningitis may reflect the degree of intrathecal inflammation. On the other hand, increased CSF levels of CRP in patients with infectious meningitis, particularly mycotic or bacterial meningitis, and SAA in patients with all infectious meningitis may reflect the degree of damage to the BCB.

Laparoscopy-assisted total gastrectomy for advanced gastric cancer with carcinomatous ascites after S1 plus cisplatin chemotherapy: a case report.

A 29-year-old man with a type 4 tumor, in the lower third of the stomach, and carcinomatous ascites was diagnosed by aspiration cytology of the ascitic fluid. Curative resection was considered impossible, and S1 (120 mg/d) and cisplatin (90 mg/d) were given for 21 days in 1 course. The cancer lesion showed marked remission (partial response), and the ascites completely disappeared after the fourth course. Twenty-five days after completion of the S1 treatment, laparoscopy-assisted total gastrectomy was performed. Histopathological examination showed no remnant cancer cells in the resected specimen and no lymph node metastases. The tumor was replaced with fibrosis having a granulomatous change. The patient's postoperative course was uneventful. The patient was continued with S1 monotherapy after surgery, and no signs of recurrence or metastases have been seen on any examination 12 months after the surgery.

Lipopolysaccharide induced connective tissue growth factor gene expression in human bronchial epithelial cells.

BACKGROUND AND OBJECTIVE: Connective tissue growth factor (CTGF) is up-regulated in the lungs of patients with chronic obstructive pulmonary disease (COPD). Cigarette smoke and repeated bacterial infections, both of which are rich sources of LPS, are major causes of COPD. The high levels of LPS in lung epithelial lining fluid also suggest that it may have a considerable impact on the airway epithelium, in terms of cytokine and growth factor production. The aim of this study was to clarify the mechanism of LPS-induced CTGF expression in bronchial epithelial cells. METHODS: The expression and transcriptional regulation of the CTGF gene were assessed using the cultured human bronchial epithelial cell line, BEAS-2B. RESULTS: LPS significantly up-regulated CTGF mRNA expression in a dose-dependent fashion, with 100 microg/mL LPS causing a twofold increase after 2 h. CTGF protein expression was also up-regulated by LPS after 8 h. Transforming growth factor-beta1 mRNA expression was not changed by LPS treatment. A pharmacological inhibitor of nuclear factor (NF)-kappaB, MG132, inhibited LPS-induced CTGF mRNA expression. Furthermore, luciferase assays demonstrated that deletion of base pairs -253 to -53 from the CTGF promoter, where the Smad and proximal NF-kappaB binding sites are located, decreased the induction of CTGF by LPS. After stimulation with LPS, the p65 subunit of NF-kappaB was shown to be bound to the CTGF promoter in vitro and in situ. CONCLUSIONS: LPS directly induced CTGF expression in bronchial epithelial cells, independently of transforming growth factor-beta1, suggesting a possible mechanism for airway remodelling in COPD that is induced by smoking and repeated bacterial infections.

CT scan findings of emphysema predict mortality in COPD.

BACKGROUND: Emphysematous change as assessed by CT imaging has been reported to correlate with COPD prognostic factors such as FEV(1) and diffusing capacity of the lung for carbon monoxide (Dlco). However, few studies have assessed the relationship between CT scan assessment and COPD mortality from mild to severe stages of the disease. In this study, we analyzed this relationship in patients with various stages of COPD. METHODS: Two hundred and fifty-one outpatients with stable COPD were included in the study. CT scan and pulmonary function tests were performed at study entry in a single institution. The percentage of low attenuation area was measured to quantitatively evaluate emphysematous change with a custom-made software. Prognostic data also were collected, and the median follow-up time was 8 years. RESULTS: Of the 251 patients, 79 died, with 40 classified as respiratory deaths not involving lung cancer. Univariate Cox analysis revealed that emphysematous change as assessed by CT scan, lung function, age, or BMI were significantly correlated with mortality. Multivariate analysis revealed that emphysematous change as assessed by CT scan had the best association with mortality. CONCLUSIONS: Emphysematous change as assessed by CT scan predicts respiratory mortality in outpatients with various stages of COPD.

Possibility of preventing colorectal carcinogenesis with probiotics.

BACKGROUND/AIMS: There have been no reports on the relationship between the analyses of the intestinal flora of colorectal cancer patients and colorectal carcinogenesis. In this study we investigated the differences between the intestinal flora of colorectal cancer patients and healthy subjects and assessed the possibility of using probiotics to prevent colorectal carcinogenesis. METHODOLOGY: The subjects were 10 colorectal cancer patients and 20 healthy persons. A stool specimen and peripheral blood specimen were collected from the patients and 10 of the healthy subjects to analyze their intestinal flora and measure natural killer (NK) cell activity and IL-1 beta in their blood. Probiotics (Lactobacillus gasseri OLL2716: LG21) was then administered once daily to 10 of the healthy subjects for 12 weeks. Samples were collected after 4 weeks, 8 weeks, and 12 weeks of administration, and the same examinations were performed. RESULTS: The Lactobacillus detection rate was significantly higher in the healthy group than in the colorectal cancer group, and the total Clostridium perfringens was higher in the colorectal cancer group. The stool pH of the colorectal cancer group indicated alkalosis, and the total amount of short-chain fatty acids in the stools tended to be lower than in the healthy group. After ingestion of the probiotic, the Lactobacillus detection rate increased, a decrease in the total amount of Clostridium perfringens was seen, fecal pH indicated acidosis, synthesis of fecal putrefaction products was inhibited, and an increase in the short-chain fatty acid isobutyric acid was observed. The blood IL-1 beta and NK cell activity values were significantly higher from the 4th week onward than the values before ingestion of probiotics. CONCLUSIONS: A deterioration of the intestinal environment was observed in the colorectal cancer patients in comparison to the healthy controls, and the intestinal environment improved when probiotics was taken. These findings suggest the possibility of preventing colorectal carcinoma with probiotics.

[Pre- and probiotics increase host-cell immunological competence, improve bowel movement, and prevent the onset of colon cancer--an analysis based on movements of intestinal microbiota].

The activation of host immunological competence through improvement of the intestinal environment by pre and probiotics has been reported. NK cell activity, the bactericidal phagocytic activities of neutrophils in peripheral blood, and bowel movements and short chain fatty acids (SCFAs) in intestinal microbiota increase after the administration of pre- and probiotics. SCFAs shift to acidosis of the intestinal environment and advance apoptosis. Furthermore, SCFAs promote intestinal peristaltic movements through SCFA receptors such as GPT 41 and GPR43, located in the intestinal epithelium. It is known that the acceleration of intestinal apoptosis prevents the onset of colon cancer. Improvement of the intestinal environment leads to an increase in host-cell immunological competence, bowel movements, and the prevention of colon cancer.

New double-stapling technique for esophagojejunostomy and esophagogastrostomy in gastric cancer surgery, using a peroral intraluminal approach with a digital stapling system.

In the abdominal-transhiatal approach for resection of adenocarcinoma of the cardia or subcardia, and in laparoscopy-assisted total gastrectomy (LATG), the use of a circular stapling device has potential problems with the placement of the purse-string suture and insertion of the anvil of the instrument. We describe a new double-stapling technique for esophagojejunostomy and esophagogastrostomy, using a peroral intraluminal approach with a digital stapling system, a flexible shaft remote-control stapler - the Surg-ASSIST and Power Circular Stapler 21 mm (PCS). The overtube of the flexible shaft of the PCS is prepared with a nylon tie and secured to a nasogastric (NG) tube. The flexible shaft is manually advanced down the esophagus with guidance by pulling the NG tube from the abdominal cavity side. The trocar of the flexible shaft is removed from the stump of the abdominal esophagus and connected to the anvil and they are approximated; the stapler device is then fired to form a double-stapled esophagojejunostomy and esophagogastrostomy. Our peroral intraluminal approach does not require a suturing technique, and it can make anastomosis after resection for carcinoma of the esophagogastric junction and after LATG safe and simple.

Single nucleotide polymorphism typing of the human toll-like receptor 4 gene at the 2-kb upstream region of the 5' untranslated region: New enclosure strategy for the risk grouping of poorly-differentiated gastric adenocarcinoma patients.

To date, no enclosure method for risk grouping patients with poorly-differentiated gastric adenocarcinoma has been identified. We examined the relationship between mutations in toll-like receptor 4 (TLR4) and patients with poorly-differentiated gastric adenocarcinoma. Genomic DNA was extracted from the peripheral blood of 38 patients, 20 with well-differentiated and 18 with poorly-differentiated gastric cancer, from 25 patients with colorectal cancer and from 10 healthy volunteers. The polymorphism of TLR4 up to the 2-kb upstream region of the 5' untranslated region (UTR) was analyzed. The results revealed the presence of single nucleotide polymorphisms (SNPs) only among patients with poorly-differentiated gastric adenocarcinoma. SNPs were found at 3 sites: -2081, -2026 and -1601 in 12, 15 and 15 of the 18 cases of poorly-differentiated gastric adenocarcinoma, respectively. The results of the determination of a consensus among the base sequences of the core promoter, basal promoter and upstream promoter elements reveal that the variant sites were present in the TLR4 mRNA promoter region, suggesting that they were biologically significant variations. Polymorphism analysis of the upstream region of the 5' UTR of TLR4 may be a useful new enclosure strategy for the risk grouping of poorly-differentiated gastric adenocarcinoma patients.

A patient with adenocarcinoma of the jejunum successfully diagnosed by preoperative endoscopy for the small intestine.

In this study, the authors report the case of a 35-year-old man diagnosed preoperatively as having adenocarcinoma of the jejunum using a conventional endoscopy, usually used for the examination of the large intestine.

High incidence of Dieulafoy's lesions in upper gastrointestinal bleeding associated with polyarteritis--clinical examination of patients of polyarteritis nodosa with rapidly progressive glomerulonephritis.

BACKGROUND/AIMS: Polyarteritis nodosa (PN) has been classified into polyarteritis (PA) and microscopic polyarteritis (MA) histologically. To clarify of the characteristics of upper gastrointestinal bleeding lesions in PN, we investigated the patients of PN with rapidly progressive glomerulonephritis (RPGN) presenting with upper gastrointestinal bleeding. METHODOLOGY: The subjects of this study were 21 patients of PN with RPGN (PA: 11, MA: 10) who presented with upper gastrointestinal bleeding. The bleeding lesions and their locations were examined endoscopically in the study subjects, and the relationship of the bleeding to the severity of renal failure, the necessity of hemodialysis (HD), presence/ absence of H. pylori infection and the gender of the patients were analyzed. RESULTS: The bleeding lesions were endoscopically identified as esophageal ulcers in 2 cases, gastric ulcers in 15 cases and duodenal ulcers in 4 cases, respectively. In 10 of the 15 cases with gastric ulcers, the ulcer assumed the form of Dieulafoy's lesions affecting the gastric body, and the underlying disease was PA in all the 10 cases. In the remaining 5 cases of gastric ulcers and 2 cases of esophageal ulcer with underlying MA or 4 cases of duodenal ulcers, in whom assumed the bleeding form of oozing from the marginal zone of ulcers. In all of the 4 cases of duodenal ulcers, and the 1 case with underlying PA and the other cases with MA, no correlation was found between the onset of the upper gastrointestinal bleeding and the severity of renal failure or the necessity for HD, presence of H. pylori infection, or the gender of the patients. CONCLUSIONS: Dieulafoy's lesions are the most frequent sources of upper gastrointestinal bleeding in cases of PA.

Prognostic potential of a PSA complex in sera of prostate cancer patients with alpha2-macroglobulin deficiency.

We previously reported on a number of cases of metastatic prostate cancer (PCa) in which serum alpha2-macroglobulin (alpha2M) levels were markedly decreased to less than 20 mg/dl (alpha2M deficiency). In order to elucidate the relative proportions of free and a prostate-specific antigen (PSA) complex in PCa patients with alpha2M deficiency, we have assessed serum alpha2M and total PSA levels, and ratios of free PSA to total PSA (F/T ratios) at each stage of PCa. Moreover, the PSA reactivity profile was determined on fractionated serum specimens of PCa patients using high-performance liquid chromatography (HPLC) using a TSKG-3000 SWXL column. Measurement of alpha2M concentration was performed by laser-nephelometry. PSA levels were determined by enzyme immunoassay, free PSA by radioimmunoassay. In those PCa patients with alpha2M deficiency, serum alpha2M and F/T ratios were lower, whereas PSA levels were higher when compared with those PCa patients without alpha2M deficiency (P<0.05). PSA elution profiles on HPLC columns revealed two major peaks. The proportion of PSA-antichymotrypsin (PSA-ACT) increased, whereas the proportion of free PSA decreased in PCa patients with alpha2M deficiency as compared with those PCa patients without alpha2M deficiency. F/T ratios were significantly lower in PCa patients with alpha2M deficiency than in those PCa patients without alpha2M deficiency. PSA-ACT and F/T ratio may be useful for monitoring bone metastasis in PCa.

Relationship between pulmonary emphysema and osteoporosis assessed by CT in patients with COPD.

BACKGROUND: Osteoporosis is one of the important systemic features of COPD. Although COPD itself is regarded as one risk factor for osteoporosis, the relationship between the extent of emphysema and reduced bone density is still unclear. Our first aim was therefore to measure vertebral bone density and the percentage of low-attenuation area (LAA%) in the lungs using chest CT scans in COPD patients. Our second aim was to investigate the relationships among CT scan measurements, anthropometric parameters, and pulmonary function. METHODS: Chest CT scans and pulmonary function tests were performed in 65 male patients with COPD. Using CT images, the CT scan density of the thoracic and lumbar vertebrae (T4, T7, T10, and L1) and the LAA% were measured quantitatively, and their correlations were analyzed. RESULTS: Linear regression analyses revealed that LAA% had a significant negative correlation with bone mineral density (BMD) [r = -0.522]. In addition, multiple regression analysis showed that only LAA% and body mass index (BMI) were predictive of BMD among age, BMI, smoking index, FEV(1), arterial blood gas, and LAA%. CONCLUSIONS: The extent of pulmonary emphysema significantly correlated with reduced bone density. Our study suggested that COPD itself could be a risk factor for osteoporosis and that chest CT scanning is useful for the management of COPD as a systemic disease.

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.

Analysis of the differences in structural chromosomal aberrations of the gastric mucosa between H. pylori positive and negative gastric cancer patients: involvement of H. pylori in the onset of gastric cancer and examination of the mechanism in gastric carcinogenesis following H. pylori eradication.

Gene mutations are essential to carcinogenesis. If an evident difference is observed in gastric mucosal chromosomal structure aberrations between H. pylori (Hp)-negative and Hp-positive gastric cancer patients, it may be interpreted as suggesting the involvement of Hp in gene mutations. This study was undertaken to compare chromosomal structural aberrations between Hp-negative and Hp-positive gastric cancer patients and to evaluate the effects of Hp eradication on chromosomal structures in clinical cases. The subjects of this study were 40 patients with gastric cancer divided into four groups: Group A was composed of 12 patients with Hp-negative gastric cancer (well-differentiated gastric cancer in 5 cases and poorly-differentiated in 7 cases), Group B of 8 patients with Hp-negative gastric cancer following Hp eradication (well-differentiated in 4 case and poorly-differentiated in 4 cases), Group C of 13 patients with Hp-positive gastric cancer (well-differentiated in 7 cases and poorly-differentiated in 6 cases) and Group D of 7 patients with gastric cancer (well-differentiated in 5 cases and poorly-differentiated in 2 cases) undergoing Hp eradication at subtotal gastrectomy. In each of the groups A, B and C, the structural chromosomal aberration such as loss of heterozygosity (LOH) and microsatellite instability (MSI) was analyzed. In Group D, changes in structural chromosomal aberrations after Hp eradication as compared to the pre-eradication structures were also analyzed. LOH and MSI were examined by PCR, using DNA extracted from the cancer-affected and intact gastric mucosal tissue specimens from each patient. In A, B and C groups, structural chromosomal aberrations were noted, and these aberrations tended to be more marked in cases of poorly-differentiated gastric cancer in each group. In terms of structural chromosomal aberrations, there was no marked difference between Group A and either Group B or C. Hp eradication resulted in no change in chromosomal structure as compared to the pre-eradication structure in Group D. These results suggest the possibility that Hp eradication does not affect chromosomal structures and Hp is involved in gastric carcinogenesis as an additive environmental factor rather than as a factor acting at the gene level.

Comparison of airway dimensions in different anatomic locations on chest CT in patients with COPD.

OBJECTIVE: It is not well known whether there is heterogeneity in the airway dimensions at different anatomic locations in individual patients with COPD. The first objective was to compare airway dimensions of the basal segment bronchus between COPD patients and healthy controls. The second and third objectives were to compare the airway dimensions in two anatomic locations, and to investigate the relationship between CT measurements and pulmonary function among COPD patients. METHODS: Thirty males with COPD (aged 68.7 +/- 8.1 years) and 18 healthy males (aged 64.9 +/- 14.0 years) were enrolled in the study. COPD was diagnosed according to the criteria of the Global Initiative for Obstructive Lung Disease Workshop Report. Pulmonary function tests and CT scans were performed on all subjects. Airway dimensions and lung attenuation were automatically determined using methods that were validated with a phantom. RESULTS: Age, smoking index and height did not significantly differ between the COPD patients and healthy controls. The COPD patients had a significantly thicker airway wall than healthy controls. Among the COPD patients, there were no significant differences in the airway dimensions of bronchi in different segments; however, the airway and lung attenuation measurements of the lower lung field were more strongly correlated with FEV(1) than those of the upper lung field. CONCLUSION: Patients with COPD had no significant heterogeneity in airway dimensions at different anatomic locations. The airway and lung attenuation measurements of the lower lung field were more strongly correlated with airflow limitation than those of the upper lung field.

Heterozygous Thr 135 Ala polymorphism at leucine-rich repeat (LRR) in genomic DNA of toll-like receptor 4 in patients with poorly-differentiated gastric adenocarcinomas.

The genomic DNA of toll-like receptor (TLR) 2, TLR4, radioprotective 105, TLR6, and TLR9 were examined for mutations in 48 patients with gastric cancer. Of these, 22 had well-differentiated and 20 had poorly-differentiated adenocarcinomas, the latter group including 10 with signet ring cell carcinomas. The remaining 6 had gastric adenomas. Ten healthy volunteers with no family history of malignant diseases served as controls. DNA was extracted from peripheral blood and subjected to electrophoresis using PCR oligonucleotide primers. The resultant gel was analyzed with a DNA sequencer. None of the healthy volunteers, patients with gastric adenomas or those with well-differentiated gastric adenocarcinomas showed mutations. However, 8 of the 20 with poorly-differentiated gastric adenocarcinoma showed heterozygosity at the 135th position of the amino acid sequence of TLR4, and a mutation from threonine to alanine was found at this site. Analysis of the entire available amino acid sequence of TLR4 revealed that this mutation occurred at a leucine-rich repeat corresponding to one of its extracellular components. This suggests a disturbance in the protein phosphorylation reaction of TLR4, and that this disturbance is related to the development of poorly-differentiated gastric adenocarcinomas.

Relationship between N-linked oligosaccharide chains of human serum immunoglobulin G and serum tumor markers with non-small cell lung cancer progression.

BACKGROUND: It has been demonstrated that increased expression of the sialyl Le(x) sugar chains on cancer cells influences cellular adhesion to vascular-endothelial cells. Therefore, it was thought that alterations in the sugar chain structure of E-cadherin, a calcium dependent adhesion molecule, influence the metastasis of cancer cells. MATERIALS AND METHODS: In this study, N-linked oligosaccharides of human serum immunoglobulin G (IgG) were analyzed in 12 patients with non-small cell lung cancer (NSCLC) (6 localized cancer: 3 adenocarcinomas and 3 squamous cell carcinomas; 6 metastatic cancer: 3 adenocarcinomas and 3 squamous cell carcinomas) and 10 healthy controls using fluorophore-associated carbohydrate electrophoresis (FACE). The relationship between changes in sugar chain structure and serum concentrations of carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA21-1) were evaluated. CEA levels in the sera were determined using an enzyme immunoassay, and CYFRA21-1 levels were determined using an enzyme chemiluminescent immunoassay. RESULTS: Fri (monogalactosyl IgG oligosaccharides) and Fr2 (digalactosyl IgG oligosaccharides) decreased, while Fr4 (agalactosyl IgG oligosaccharides) significantly increased (p < 0.01-0.05) with NSCLC progression. The Fr4/Fr1+Fr2 ratio increased with NSCLC progression, and the ratios in localized and metastatic NSCLC were significantly higher than in healthy controls (p < 0.01 and p < 0.01, respectively). There was a strong correlation between serum CEA levels and Fr4 (r = 0.91) and a significant correlation between serum CEA levels and the Fr4/Fr1+Fr2 ratio (r = 0.83, p < 0.05) in patients with lung adenocarcinoma. There was a significant correlation between serum CYFRA21-1 levels and Fr4 (r = 0.88, p < 0.001) and a positive correlation between serum CYFRA21-1 levels and the Fr4/Fr1+Fr2 ratio (r = 0.38) in patients with lung squamous cell carcinoma. CONCLUSION: The analysis of serum IgG N-linked oligosaccharide chain structures by FACE may be an auxiliary indicator of serum tumor markers for monitoring NSCLC progression.

Investigation of the possibility of human-beta defensin 2 (hBD2) as a molecular marker of gastric mucosal inflammation.

BACKGROUND/AIMS: It has previously been reported that human-beta defensin 2 (hBD2) has a physiological role as a proinflammatory mediator in gastric mucosal inflammation as well as an antimicrobial peptide for Helicobacter pylori (Hp). The present study was conducted to evaluate the possibility of hBD2 as a molecular marker of gastric mucosal inflammation. METHODOLOGY: The subjects were 40 A1 to S2 gastric ulcer patients, with or without Hp infection. Biopsy specimens of the mucosa were obtained endoscopically before and after the administration of lansoprazole (LPZ) (15mg/day) or famotidine (FAM) (40 mg/day or 20 mg/day), consecutively, and each set of samples was divided into two groups; one group was subjected to RT-PCR to assess the expression of hBD2, and the other was subjected to immunohistochemical analysis for evaluating the expression of CD68. RESULTS: The expression of hBD2 was observed through the stage of gastric ulcer, from A1 to S1, regardless of the presence or absence of Hp infection, both before and after LPZ or FAM administration, and its intensity of expression decreasing as the number of CD68-positive cells decreased. The number of CD68-positive cells deceased as the severity of the ulcer increased from stage A1 to S2, regardless of the presence/absence of Hp infection. CD68-positive cells could hardly be observed in stage S2 gastric ulcers, in which hBD2 expression was also only scarcely noted. CONCLUSIONS: These results suggested the possibility that hBD2 may be a molecular marker of gastric mucosal inflammation, irrespective of the presence/absence of Hp infection.