Nuclear expression of mu-opioid receptors in a human mesothelial cell line.

1 Possibly acting via mu-opioid receptors (MORs), morphine inhibits the formation of experimentally induced postoperative abdominal adhesions in rats. Mesothelial cells may participate in adhesion formation by secreting mediators that interfere negatively with fibrinolysis. Morphine may prevent adhesions by inhibiting the release of pro-adhesion mediators from mesothelial cells. This study aimed to investigate whether human mesothelial cells express MOR-1; if so, such could constitute a site of action for morphine in adhesion prevention. 2 Cells from Met-5A, a human mesothelial cell line were seeded and prepared for immunocytochemistry and Western blotting. 3 Immunocytochemistry showed MOR-1 expression in mesothelial cells, predominantly in the nuclei. Western blotting showed two bands (c. 35 and 50 kDa) which correspond to those obtained with a control lysate from cells known to express MORs. In addition, we found MOR-1 expression with nuclear and cytoplasmatic localization in biopsies from human abdominal adhesions. 4 The current findings may suggest that morphine could interact directly with mesothelial cells via MOR-1 receptors, and thereby modulate adhesion formation, possibly by interfering with the release of pro-adhesion factors from these cells.

Expression of the endogenous, nicotinic acetylcholine receptor ligand, SLURP-1, in human colon cancer.

1. Secreted mammalian Ly-6/urokinase plasminogen activator receptor-related protein-1 (SLURP-1) is a recently discovered endogenous ligand at the alpha7 subunit of the nicotinic acetylcholine receptors. Previous reports have shown that SLURP-1 is expressed in normal human keratinocytes seemingly with a pro-apoptotic function. Conversely, such expression was markedly attenuated in transformed cells and it was suggested that the molecule could convey protection against malignant transformation. 2. In this study, we demonstrated the mRNA expression (by RT-PCR) and protein expression (by Western blotting and immunocytochemistry) of SLURP-1 in the human colon cancer cell line, HT-29. 3. Furthermore, we demonstrated the expression of SLURP-1 (by immunohistochemistry) in tumour cells of human colon cancer tissue, and, to a greater extent, in immune and smooth muscle cells of adjacent, macroscopically tumour-free colon tissue. 4. The current findings suggest that SLURP-1 participates in the regulation of gut immune functions and motility, as well as possibly playing a role in colon carcinogenesis/cancer progression.

P2Y2- and P2Y4 purinergic receptors are over-expressed in human colon cancer.

1. A characteristic of cancer is altered signal transduction leading to uninhibited growth. Adenosine-5'-triphosphate (ATP), a natural ligand at P2X- and P2Y purinergic receptors may regulate cell growth in non-neoplastic, as well as neoplastic tissues. In the human colon cancer cell line, HT-29, we previously demonstrated the expression of purinergic receptors of the P2Y(2)- and P2Y(4) subclasses. 2. The aim of the current study was to investigate whether these two purinergic receptors are expressed also in human colon cancer, and, if so, how such expression is related to that in tumour-free colonic tissue. 3. The immunohistochemical findings of both P2Y(2)- and P2Y(4) receptors in the tumours from three patients, prompted us to conduct an investigation of a consecutive series of patients utilizing Western blotting for protein detection and densitometry for quantitation. 4. Both P2Y(2)- and P2Y(4) purinergic receptors could be identified in tumour-free tissue, and both were significantly over-expressed in each of the 10 colon cancers.

Demonstration of P2Y4 purinergic receptors in the HT-29 human colon cancer cell line.

1 The aim of the current study was to investigate the existence of P 2 Y(4) purinergic receptors in the HT-29 human colon cancer cell line. 2 We utilized Western blots and immunocytochemistry for the analysis. 3 Western blotting demonstrated two bands that could not be found after the antibody had been preabsorbed with the control peptide, suggesting that both bands are related to the P 2 Y(4) purinergic receptor. 4 Immunocytochemistry showed immunoreactivity for the P 2 Y(4) purinergic receptor localized in the cytoplasm of the HT-29 cells. 5 This is the first demonstration of the protein expression of P 2 Y(4) purinergic receptors in a human colon cancer cell line.

Demonstration of functional receptors for noradrenaline and adenosine-5'-triphosphate, but not for prostaglandin E2, in HT-29 human colon cancer cell line.

1. The aim of the current study was to investigate in HT-29 human colon cancer cell line, the existence of functional receptors for the signalling molecules, noradrenaline (NA), prostaglandin E2 (PGE2), and adenosine-5'-triphosphate (ATP). 2. We utilized microphysiometry, which monitors on-line extracellular acidification rate (ECAR) as a measure of cellular metabolic activity, and how this variable is altered by signalling molecules. 3. Challenge with NA (5.9 microM) resulted in an increase in ECAR by approximately 24% of basal. 4. PGE2 (0.0284, 0.284 and 2.84 microM) hardly affected ECAR. 5. ATP (100 microM) elicited a biphasic effect on ECAR (increase and decrease in ECAR by about 58 and 10% of basal, respectively). 6. HT-29 cells were shown to express COX-2 by immunocytochemistry. 7. These data suggest the presence of functional receptors for NA and ATP, but not for PGE2 in HT-29 human colon cancer cell line.

Long-term outcome after colectomy in severe idiopathic constipation.

OBJECTIVE: The specific aim of the investigation was to assess the long-term results of subtotal colectomy with ileorectal anastomosis in patients with severe idiopathic constipation. PATIENTS AND METHODS: 40 patients with severe idiopathic constipation were operated on between 1981 and 1993. Patients were accepted for a colectomy and an ileo-rectal anatomosis after a thorough gastro-intestinal investigation. Pre-operative bowel frequency was less than 2 movements per week, and slow transit was documented. Postoperative complications occurred in eight patients. Early re-operation was performed in 2 patients for small bowel obstruction. RESULTS: Mean follow-up was 11 (range 5-16) years. The defaecation frequency at follow-up was 3.0 +/- 1.9 per day. Twenty-nine patients stated that they were satisfied and 11 were dissatisfied with the procedure. The outcome did not correlate with observed signs of outlet obstruction, blunted rectal sensation or presence of a psychiatric diagnosis. At 5-16 years after the procedure 33 patients still retain the ileo-rectal anastomosis. Seven patients have had further procedures: Five patients have an ileo-anal pouch, one has a continent ileostomy and one has a conventional ileostomy. Small bowel obstruction was encountered in 17 patients, in 10 of these surgical treatment was necessary. CONCLUSION: It is concluded that colectomy and ileorectal anastomosis for the treatment of severe idiopathic constipation causes an increase in the number of bowel motions, but is deemed successful only by 3 out of 4 patients. Secondary morbidity is considerable.

The trephine stoma: formation of a stoma without laparotomy.

A trephine stoma may be an attractive alternative for those patients who require a stoma but not a laparotomy. Twenty-seven consecutive patients were candidates for formation of a trephine stoma. A loop ileostomy was successfully constructed in seven patients and an end sigmoid colostomy in 15, while conversion to a formal laparotomy was necessary in five patients mainly because the sigmoid colon was immobile or there were extensive adhesions. The absence of a major abdominal wound made the postoperative course simple with little pain, a quick recovery, and early discharge from hospital.