Cell proliferation of the rat gastrointestinal mucosa after treatment with E2 prostaglandins and indomethacin

The frequency of arrested mitoses after vincristine injection was studied in the gastrointestinal mucosa of rats treated with either natural prostaglandin E2 (0.2-5.0 mg X kg-1, b.d.), 15-R-15 methyl prostaglandin E2 (2 mg X kg-1, b.d.) or indomethacin (1.0-3.0 mg X kg-1, b.d.). In addition to the mitotic index, morphometric measurements including the mucosal thickness and the thickness of the proliferative and functional zones of the gastric corpus, antrum and jejunum were performed. Natural prostaglandin E2, at the highest dose range, reduced significantly the mitotic index in the gastric antrum. Normal values were found in the gastric corpus and jejunum and in the antrum with the lower doses. The mitotic index was unaffected by treatment with 15-R-15 methyl prostaglandin E2. Natural prostaglandin E2 produced trophic changes (i.e. increased thickness and/or hyperplasia) in the antrum, functional epithelial zone of the gastric corpus and in the jejunum. More pronounced trophic changes were observed in the mucosa of rats treated with the analogue. Indomethacin reduced the mucosal thickness in all examined epithelia and lowered the mitotic index in the jejunum. It is concluded that the trophic effects of E2 prostaglandins on gastrointestinal epithelia are not caused by increased production of new cells. The reduced mitotic index observed in the antral mucosa of prostaglandin-treated rats could be secondary to a negative feedback from the hyperplastic epithelium. The antitrophic effects of the prostaglandin-synthesis blocker (indomethacin) indicates that endogenous prostaglandins may participate in the epithelial cell regulation of the gastrointestinal tract.     

Cell cycle distribution of proliferative and functional cells of the rat jejunum after treatment with oral E2 prostaglandins

Proliferative and functional epithelial cells were isolated from jejunal specimens of the rat by means of vibrational treatment combined with differential air insufflation. This method gave a good separation between superficial cells of the villi and the crypt cells, as evaluated by flow cytometry, morphology, cytology, and incorporation of radioactive thymidine into DNA. Groups of Sprague-Dawley rats (260 g) were treated twice daily for 11 days with oral placebo, 15(R)15-methyl-prostaglandin E2 in the range of 0.125-2 mg X kg-1, or 5 mg X kg-1 natural PGE2. Isolated crypt cells and superficial cells of the jejunal villi were then analysed by flow cytometry. Morphometric measurements were performed on sections of some jejunal specimens not submitted to vibrational treatment. The cell cycle distribution of crypt cells was unaffected by treatment with the prostaglandin analogue despite the presence of trophic changes. The proportion of crypt cells in G2/M phase was slightly but significantly reduced in rats given natural PGE2 compared with controls. The cell cycle distribution of villus cells was not affected by prostaglandin treatment. Trophic changes in the absence of increased DNA synthesis (S phase) or increased mitotic activity suggests that the hyperplasia observed after prostaglandin treatment is due to a reduced cell loss and/or slower migration time of epithelial cells.     

Initial kinetic changes of prostaglandin E2-induced hyperplasia of the rat small intestinal epithelium occur in the villous compartments

This study was performed to further identify the sequence of cell kinetics that occurs in the development of gastric and intestinal epithelial hyperplasia after orally administered prostaglandins of the E series. A high-dose, short-treatment schedule was used to examine the initial effects on kinetic parameters in the rat small intestinal epithelium. Groups of rats were killed after a single dose of oral prostaglandin E2 at 1 h after in vivo labeling with [methyl-3H]thymidine and during continued treatment at 6, 12, 24, 48, 72, and 96 h. As evidenced by autoradiography, the earliest change produced by prostaglandin E2 was an increased cellularity of the villous compartment (p less than 0.05 after 24 h). There was no change of labeling index of the villous compartment or of the leading edge of labeled cells within 24 h. At 48 h, the increased cellularity was accompanied by a significantly elevated labeling index of the villi. Throughout the study period no significant differences were observed between groups in the number of cells or labeling indices in the jejunal crypts, or in cellular input from the crypts to the villi. Epithelial turnover time in the placebo and treatment groups was 69 and 71 h, respectively. To exclude the possibility that prostaglandin E2 initially affects cell birth rate and mean cell cycle time, a metaphase blocker was given after 4 days of treatment in a second study. Animals were killed after 0, 0.5, 1.0, 1.5, and 2.5 h. The rate of entry into mitoses was 8.1% cells/h in controls compared with 8.2% cells/h in treated rats. The distribution of mitoses within crypts was identical in the two groups and the mean cell cycle time was 13.6 and 13.2 h, respectively. Also in this study there were trophic changes of the villi. It is concluded that the hyperplasia produced by oral prostaglandin E2 starts in the villi of the small intestine and is initiated by reduced cell exfoliation from the villous tips. Previously recorded retention of cellular elements in villi and crypts, increased cellularity of the proliferative compartments, and reduced mitotic index are secondary events.     

Cell proliferation and DNA dependent DNA polymerase estimation in acute lymphoblastic leukaemia during treatment with prednisone and vincristine.

The presence of DNA polymerase and primer-template DNA in lymphoblast nuclei by measuring the in vitro incorporation of 3H-thymidine-5'-triphosphate (3H-TTP) was studied in 10 patients with acute lymphoblastic leukemia. Protein synthesis and various other cytokinetic parameters were also studied. After prednisone (P) administration a marked decrease in 3H-TTP labelling index (3H-TTP LI) was apparent together with an inhibition of 3H-leucine incorporation (3H-LEU LI) into lymphoblasts. A moderate decrease in 3H-TDR labelling index (3H-TDR LI) and a later decrease in mitotic index (MI) were seen. Single cell DNA measurements showed a depletion of 3H-TDR labelled lymphoblasts in early part of S-phase apparent at 24 h lasting up to 54 h after P administration. Vncristine given as a flash injection later in the study period caused an immediate rise of the MI, at the same time the P induced decline in 3H-TTP LI, 3H-TDR LI and 3H-LEU LI were continued in most patients. P is thought to damage the cells both in and outside the cell cycle. In the cell cycle the effect of P is an arresting effect in G1.     

Indomethacin accelerates clearance of labeled cells and increases DNA synthesis in gastrointestinal mucosa of the rat

Sprague-Dawley rats treated with placebo, parenteral indomethacin, or oral prostaglandin E₂ for six days were given an intraperitoneal injection of [³H]methyl-thymidine and killed at 45 min and 96 hr after labeling. Treatments were continued, until death. The dpm/DNA index was determined in mucosal scrapings of the stomach, duodenum, and jejunum and used to estimate DNA synthesis (45 min) and the clearance of labeled cells (96 hr). Indomethacin increased the DNA synthesis in both the duodenal and jejunal mucosa (P<0.05). In comparison to the controls, the clearance of labeled cells from the antral, duodenal, and jejunal mucosa was accelerated by indomethacin treatment, whereas the elimination of labeled cells from the antral and jejunal mucosa was slowed by PGE₂ treatment (P<0.05). DNA synthesis of the antral mucosa was significantly reduced by PGE₂ (P<0.05). The cyclooxygenase blocker did not affect the cell kinetic parameters of the oxyntic mucosa. The plasma levels of somatostatin were significantly higher both in PGE₂- and indomethacin-treated rats than in controls (P<0.05). It is concluded that indomethacin treatment increases the cell losses from the epithelial surface, which in turn trigger a compensatory trophic reaction. It is suggested that an important physiological action of endogenous prostaglandins is to regulate the outflow of cells from the superficial zones of the epithelium. Finally, this study disclosed the presence of hitherto unknown regulatory mechanism that promote cell proliferation in the gastrointestinal, mucosa despite inhibition of the synthesis of endogenous prostaglandins.     

16,16-Dimethyl prostaglandin E2 suppresses the increases in the proliferative activity of rat colonic epithelium induced by indomethacin and aspirin

Treatment of rats with indomethacin rapidly increased ornithine decarboxylase (4 h) of colonic mucosa and [3H]thymidine incorporation into colonic mucosal deoxyribonucleic acid (DNA) (1 or 5 days) when this parameter was examined in vivo and ex vivo. The changes in colonic mucosal ornithine decarboxylase and DNA synthesis induced by indomethacin were correlated temporally with suppression of colonic prostaglandin synthesis, as assessed from ex vivo colonic production of prostaglandin E, the dominant prostaglandin product of colon. Autoradiographic studies indicated that the enhancement of proliferative activity of colonic epithelium after treatment with indomethacin for 1 day was confined to the lower third of the colonic crypt (normal proliferative zone). After 5 days of indomethacin treatment, however, there was an extension of the proliferative zone to the upper third of the colonic crypts. Concurrent treatment of rats with the stable prostaglandin E2 analogue, 16,16-dimethyl prostaglandin E2, suppressed indomethacin-induced increases in colonic mucosal ornithine decarboxylase and DNA synthesis. Concurrent administration of 16,16-dimethyl prostaglandin E2 also prevented the extension of the proliferative zone of colonic epithelium induced by 5 days of indomethacin administration. 16,16-Dimethyl prostaglandin E2 alone for 1-5 days had no detectable effects on colonic mucosal ornithine decarboxylase and DNA synthesis compared with corresponding control values. Increases in colonic mucosal DNA synthesis were also induced by treatment of rats for 5 days with aspirin (ASA). The stimulation of colonic mucosal DNA synthesis induced by ASA was significantly suppressed by concurrent administration of 16,16-dimethyl prostaglandin E2 and was also correlated with the inhibition of colonic prostaglandin synthesis by ASA. The colons of rats treated with indomethacin for 1 day or ASA for 5 days appeared normal by light microscopy. However, treatment of rats for 5 days with indomethacin resulted in mild to moderate inflammation of the lamina propria and some goblet cell depletion at the mucosal surface, but no loss of surface epithelium. The ultrastructure of the surface epithelium of the colons of rats treated with indomethacin or ASA was normal as assessed by electron microscopy. The results thus demonstrate that inhibition of local colonic prostaglandin synthesis is associated with increases in the proliferative activity of colonic epithelium, and that these increases are suppressed by administration of 16,16-dimethyl prostaglandin E2.(ABSTRACT TRUNCATED AT 400 WORDS)     

Mucosal cell proliferation in duodenal ulcer and duodenitis.

Mucosal cell proliferation in the first part of the duodenum was studied in 24 patients using a tissue culture technique in which endoscopic biopsies were subjected to autoradiography after exposure to tritiated thymidine. Eight patients had a normal duodenum, eight had duodenal ulcer, and eight had symptomatic chronic non-specific duodenitis. The mean crypt labelling index (LI) in normal duodenum was 8.8 0.4% (SEM). Increased labelling indices of 15.6 +/- 1.7% were found near the edge of duodenal ulcers and 17.8 1.8% in duodenitis. Treatment with cimetidine reduced both the severity of duodenitis and the mean crypt LI. The LI of histologically normal duodenal mucosa distal to ulcer of duodenitis was similar to that of the control subjects' mucosa. The increased mucosal cell proliferation seen in severe duodenitis, either alone or associated with duodenal ulceration, suggested that erosions and ulcers arose when the crypts passed into 'high output failure' and were unable to compensate for further epithelial cell loss. There was no evidence in out study for a generalised failure of mucosal cell proliferation in duodenal ulcer or duodenitis.     

E2 prostaglandins modulate cell proliferation in the small intestinal epithelium of the rat.

Groups of Sprague-Dawley rats were administered 1 mg/kg indomethacin subcutaneously, indomethacin subcutaneously plus 200 micrograms/kg oral 15-R-15 methyl-prostaglandin E2 (MePGE2) or oral MePGE2 twice daily for 10 days. The animals were treated with antibiotics to prevent mortality. Two control groups were used: control 1 was given placebo and control 2 was treated with antibiotics. All rats were killed 4 h after injection of a metaphase blocker, and the proliferative activity of the distal small intestine was examined in histological sections by means of the cumulative mitotic index (MI). A reduction in the number of villous cells was observed in the rats given antibiotics (p < 0.05 vs. control 1). The small intestinal villi of the rats treated with indomethacin had fewer cells than those of both control groups (p < 0.05) whereas the crypts contained more cells (p < 0.05) and had a higher MI than those of the controls (p < 0.05 vs. controls 1 and 2). These changes were reverted by the prostaglandin analogue. The number of cells of the small intestinal crypts and the cumulative MI in the rats who received indomethacin and the prostaglandin analogue were similar to controls, and they were significantly lower than the values observed in the animals treated with indomethacin (p < 0.05). The animals treated with the prostaglandin analogue and placebo developed a marked hyperplasia of the small intestinal villi (p < 0.05 vs. both control groups), but the atrophy of the villi induced by indomethacin was not prevented by simultaneous administration of the analogue.(ABSTRACT TRUNCATED AT 250 WORDS)     

Trophic actions of E2 prostaglandins in the rat gastrointestinal mucosa. A quantitative morphologic study

Adult, male Sprague-Dawley rats in groups of 10 received one of the following treatments orally twice daily for 3 wk: prostaglandin E2 (PGE2) 7.5 mg X kg-1, 15(R)-15-methyl prostaglandin E2 (MePGE2) at 0.2 or 2.0 mg X kg-1, or vehicle. After 18 h of fasting and 10-12 h after the last dose, the rats were anesthetized, and the gastrointestinal tract was fixed and processed for macroscopic and microscopic investigations. Trophic changes were more pronounced in the gastric antrum than in the gastric corpus or small intestine. The thickness of the antral mucosa was significantly increased by PGE2 and in a dose-related way by MePGE2. The mucosa of the gastric corpus became significantly thicker only with the higher dose of MePGE2. In all the prostaglandin-treated groups, the proportion of endocrine cells was reduced. Small--but sometimes significant--changes were registered in the proportions of the various exocrine cells. The parietal cells became significantly larger (+88%) in the rats treated with high doses of MePGE2. The secretory surface of the parietal cells was markedly increased by PGE2 and MePGE2. The enlargement of the secretory surface in animals treated with prostaglandins corresponded to a marked elevation of the basal gastric acid secretion and an increase in plasma gastrin levels. Hypergastrinemia can explain some, but not all, of the trophic changes observed in this study. Light microscopic examination of the duodenal and jejunal mucosa showed dose-related increases in villus heights and crypt lengths after treatment with MePGE2. Only the duodenal villus heights were increased by PGE2.     

Beta-cell proliferation in normal and streptozotocin-treated newborn rats: site,dynamics and capacity

Summary Regeneration of neonatal beta cells after streptozotocin (STZ)-induced destruction may be due to either replication from pre-existing intra-islet beta cells or extra-islet precursor cells. To further investigate this issue, beta-cell growth was analysed in normal and streptozotocin-treated newborn rats (100 g/g body weight) at several time points during the first 20 days of life. Beta cells were identified by insulin immunostaining, non-isotopic in situ hybridization for rat preproinsulin mRNA, and electron microscopy. Their proliferative activity was recorded by bromodeoxyuridine-pulse labelling. Beta-cell size and total volume were determined by computerized morphometry. In normal rats, there was a threefold increase in total beta-cell volume during the first 5 days of life, with no further expansion till day 20. The bromodeoxyuridine labelling index of the intraislet beta cells was smaller than that of the extra-islet beta cells (2–3% vs 15–20%). Comparison of the cell birth rate, calculated from the beta-cell labelling index, with the observed increase in beta-cell volume suggested that in normal neonatal rats proliferation of the intra-islet beta-cell population could account for only 10% of the observed expansion. Administration of streptozotocin at birth resulted in more than 90% reduction of the total beta-cell volume at day 2, which then increased to 39% of the normal value by day 20. During this period of partial regeneration, which restored normoglycaemia, the labelling index of intra-islet beta cells was higher than in normal rats (9% vs 2%, p<0.001), whereas no change was seen in the extra-islet beta-cell labelling index. Comparison of cell birth rate with the increase in beta-cell volume indicated that 50–60% of the observed beta-cell growth could result from the intra-islet beta-cell proliferation. These results suggest that replication from pre-existing, surviving beta cells plays an important role in regeneration of neonatal beta cells after destruction by streptozotocin.Abbreviations STZ Streptozotocin - BrdU bromodeoxyuridine - DIG digoxigenin - SSC sodium chloride, sodium citrate - LI labelling index     

Alternating proliferative capacity in the rat gastrointestinal mucosa. Effects of E2 prostaglandins and indomethacin

Having previously observed an apparent uneven distribution of proliferating cells in the gastric corporic mucosa of the rat, we examined the mitotic distribution along 8-mm sections of gastric and jejunal epithelia. Metaphases were arrested with vincristine to facilitate mitotic count, and the effects of treatment with a prostaglandin E2 analogue and a cyclooxygenase blocker were examined. Clusters of mitotic figures alternating with non-proliferating areas were observed in the gastric corporic epithelium of control rats. During 4 h mitotic activity was absent over 21% of the corporic mucosa. Extending the examined area to about 240 glands reduced substantially the error of mitotic counts. An uneven distribution of mitoses was found in the antral and jejunal epithelium, but areas without proliferating cells were uncommon. Treatment with the prostaglandin E2 analogue reduced the number of mitosis-free areas in the gastric corpus to 13%, and clusters were less easily identified. The total mitotic count was unaffected by treatment. In the jejunum prostaglandin increased the absolute number of mitoses. The mitotic span was also increased, reflecting the uneven distribution. Indomethacin produced the opposite effects to the prostaglandin analogue, including reduction of epithelial height. Of the gastric corporic mucosa 35% was non-proliferating during the observation period, but the clustering phenomenon was still apparent. Absence of dose relationship was attributed to ulcerogenic actions of high doses of indomethacin. It is concluded that mitoses are unevenly distributed in the upper gastrointestinal epithelium of the rat and that safe estimates of mitotic count require examination of large corporic areas.(ABSTRACT TRUNCATED AT 250 WORDS)     

EGFR LI and Ki-67 LI are independent prognostic parameters influencing survivals of surgically treated squamous cell lung cancer patients

In literature there are still opinion differences concerning the prognostic significance of epidermal growth factor receptor (EGFR) expression and proliferative potential in patients with non small cell lung cancer (NSCLC). This prompted us to study those parameters. The Ki-67 labeling index (Ki-67 LI), EGFR labeling index (EGFR LI), and mitotic index (MI) were analyzed in the group of 78 consecutive, surgically treated squamous cell lung cancer (SqCLC) patients. The expression of Ki-67 and EGFR protein was visualized on formalin fixed, paraffin embedded sections using immunohistochemistry (IHC). Mitotic index was assessed on formalin fixed, paraffin embedded sections, stained with hematoxylin and eosin using morphological criteria. Mean values of Ki-67 LI and MI were higher for G2+G3 tumors than for G1 tumors. EGFR LI was higher for G1+G2 than for G3 tumors, and for pT3 than for pT1+pT2 tumors. Patients having tumors with Ki-67 < or =28% or (EGFR LI < or =13% or EGFR LI >80%) survived significantly shorter than those having tumors with Ki-67 LI >28% or 13%< EGFR LI < or =80%. In multivariate analysis, 13%> or = EGFR LI <80% and Ki-67 LI < or =28% were independent negative prognostic parameters influencing survivals of SqCLC patients.     

Response of rat alveolar macrophages to ozone: quantitative assessment of population size, morphology, and proliferation following acute exposure

The purpose of this study was to evaluate the in vivo effects of an acute exposure to low levels of ozone on rat pulmonary alveolar macrophages (PAM). Fisher 344 rats exposed to 0.0, 0.12, 0.8, or 1.5 ppm O3 for 6 h were killed immediately after and 3, 18, 42, or 66 h after ozone exposure and their lungs were lavaged. Compared to sham-exposed (control) rats, exposure to 0.12 ppm O3 had no measurable effect on the total number, labeling index (LI), mitotic index (MI), or morphology of rat alveolar macrophages. The number of neutrophils was significantly (p less than or equal to 0.001) greater than in controls at 3, 18, and 42 h after exposure to 1.5 ppm O3 and 42 h after exposure to 0.8 ppm O3. The number of PAM was approximately twice that of controls 42 and 66 h after exposure to 0.8 and 1.5 ppm O3. There was a significant (p less than or equal to 0.001) increase in PAM MI 42 and 66 h after exposure to 1.5 ppm O3 and 42 h after 0.8 ppm O3. The increase in the number of PAM in mitosis was preceded by an increase in PAM LI. The PAM LI was significantly (p less than or equal to 0.001) greater than controls 18 and 42 h after exposure but returned to near normal levels by 66 h after exposure. There was a transient decrease in the mean nuclear/cytoplasmic ratio of PAM from rats exposed to 1.5 ppm O3 18 and 42 h after exposure due to an increase in the mean PAM cytoplasmic area. Comparison of the PAM population doubling time (Dt) and cell cycle time (Ct) suggest that PAM proliferation played a significant role in the observed increase in PAM following exposure to 0.8 and 1.5 ppm O3. These results highlight the dynamic response of PAM to an acute exposure to ozone and suggest that the proliferative response of pulmonary alveolar macrophages may be a useful indicator of pulmonary damage following inhalation of an irritant oxidant.     

Effects of progestins and glucocorticoids on deoxyribonucleic acid synthesis in the uterus of the neonatal mouse

The effects of progestins and glucocorticoids on cellular proliferation were examined in the uterus of 5-day-old mice by monitoring either the labeling index (LI) after exposure to [methyl-3H]thymidine ([3H]TdR) in vivo or the mitotic index (MI) after colchicine-induced arrest of cells in metaphase. In untreated 5-day-old mice, epithelial LI was 31%, and stromal LI was 15%. Eighteen hours after a single ip injection of 40 mg/kg progesterone, epithelial LI was reduced to 2.3% and remained low for 48 h. Stromal LI increased transiently, reaching a zenith (40%) 18 h after administration of progesterone and returning to control levels by 24 h. When mitotic activity was assessed 24 h after progesterone treatment, epithelial MI was decreased (control, 3.1%; progesterone, 0.23%) and stromal MI was increased (control, 0.60%; progesterone, 2.1%). Thus, the measured effects on LI were indicative of altered proliferative activity of the tissues. Glucocorticoids also inhibited epithelial LI, but had no effect on stromal LI. Eighteen hours after a single ip injection, dexamethasone inhibited epithelial LI to the same extent as progesterone treatment. Corticosterone did not significantly decrease epithelial LI, while cortisol produced an intermediate inhibitory response. To determine whether the high baseline LI in uterine epithelium of neonatal mice was estrogen dependent, uteri of 1-day-old mice were grafted under the kidney capsule of ovariectomized adult mice. Eight days later, the hosts were treated with either progesterone or vehicle and then killed 18 h later. After labeling the tissue with [methyl-3H]thymidine in vitro, the mean LI of the epithelium of the grafted uteri was 11.5%, while that of the vehicle-treated hosts was 0.10%. Progesterone treatment reduced the LI of the grafted uterine epithelium to 1.0%. These data demonstrate that uterine tissues of the neonatal mouse proliferate rapidly in the absence of gonadal steroids. Progestins and glucocorticoids specifically inhibit this estrogen-independent DNA synthesis of uterine epithelium.     

The Labelling Index of Primitive Plasma Cells Determines the Clinical Behaviour of Patients with Myelomatosis

For patients with multiple myeloma the most important laboratory correlate of prognosis and disease activity is the bromodeoxyuridine (BrdUrd) plasma cell labelling index (LI). However, the traditional immunofluorescent microscope LI technique, like other manual enumeration assays, can suffer from poor precision and accuracy. In this study the LI of different subpopulations of plasma cells (CD38⁺⁺) as determined by flow cytometry was correlated with disease state. The mean LI of the total CD38⁺⁺ population was significantly higher (2.7±0.4%) than the LI determined by the traditional slide technique (0.6±0.1%) for 65 samples tested. Primitive plasma cells (CD38⁺⁺, CD45⁺⁺) had a higher labelling index than mature plasma cells (CD38⁺⁺, CD45⁻) (7.0±1.3% v 1.8%±0.3%) and in one patient the LI of the primitive plasma cells was 46%. In addition, the LI of the mature plasma cells was lower than the total plasma cell population. As expected, there was a significant difference between the LI of patients in plateau phase and progressive disease but this difference was greatest when the LI of the primitive plasma cells was studied (9.2±2.9% v 2.2±0.7%; z =19.9, P <0.001). This study has raised some concerns about the sensitivity and accuracy of the traditional labelling index and has shown that the increased LI associated with progressive disease is almost entirely attributable to an increase in the LI of the primitive plasma cell subpopulation and that the LI of primitive plasma cells provides a more clinically significant correlation with disease status than the traditional assay.     

Low-energy laser irradiation reduces formation of scar tissue after myocardial infarction in rats and dogs

BACKGROUND: Low-energy laser irradiation (LELI) has been found to attenuate various biological processes in tissue culture and experimental animal models. The aim of the present study was to investigate the effect of LELI on the formation of scar tissue in experimentally induced chronic infarct in rats and dogs. METHODS AND RESULTS: Myocardial infarction (MI) was induced in 50 dogs and 26 rats by ligation of the left anterior descending coronary artery. After induction of MI, the laser-irradiated (LI) group received laser irradiation (infrared laser, 803-nm wavelength) epicardially. Control MI-induced non-laser irradiated (NLI) dogs were sham-operated, and laser was not applied. All dogs were euthanized at 5 to 6 weeks after MI. Infarct size was determined by TTC staining and histology. The laser treatment (P:<0.05) lowered mortality significantly, from 30% to 6.5%, after induction of MI. The infarct size in the LI dogs was reduced significantly (P:<0.0001) (52%) compared with NLI dogs. Histological observation of the infarct revealed a typical scar tissue in NLI dogs and cellularity in most of the LI dogs. Only 14+/-3% of the mitochondria in the cardiomyocytes in the ischemic zone (4 hours after MI) of LI MI-induced rats were severely damaged, compared with 36+/-1% in NLI rats. Accordingly, ATP content in that zone was 7.6-fold (significantly) higher in LI than in NLI rats. CONCLUSIONS: Our observations indicate that epicardial LELI of rat and dog hearts after chronic MI caused a marked reduction in infarct size, probably due to a cardioprotective effect of the LELI.     

Variability of cell proliferation in the proximal and distal colon of normal rats and rats with dimethylhydrazine induced carcinogenesis

AIM: To investigate the patterns of cell proliferation inproximal and distal colons in normal rats and rats with 1,2-dimethylhydrazine (DMH) induced carcinogenesis using thethymidine analogue bromodeoxyuridine.METHODS: Colonic crypt cell proliferation wasimmunohistochemically detected using the anti-bromodeoxyuridine Bu20a monoclonal antibody.RESULTS: Marked regional differences were found in bothgroups. Total labelling index (LI) and proliferative zone sizein both normal (8.65±0.34 vs 7.2±0.45, 27.74±1.07 vs16.75±1.45) and DMH groups (13.13±0.46 vs 11.55±0.45,39.60±1.32 vs35.52±1.58) were significantly higher in distalthan in proximal colon (P＜0.05), although the number ofcells per proximal crypt was greater (31.45±0.20 vs34.45±0.39, 42.68±0.53 vs49.09±0.65, P＜0.0001). Crypt length,total LI and proliferative zone size all increased in both proximaland distal regions of DMH rats compared to normal controls(P＜0.0001). In DMH-treated rat colon a shift of labelled cellsto higher crypt cell positions was demonstrated distally whilsta bi-directional shift was evident proximally (P＜0.05).CONCLUSION: Our results show that changes in cellproliferation patterns, as assessed by bromodeoxyuridineuptake, can act as a reliable intermediate marker of coloniccancer formation. Observed differences between proliferationpatterns in distal and proximal colon may be associated withthe higher incidence of tumors in the distal colon.     

Effects of regional ischaemia, with or without reperfusion, on endothelium dependent coronary relaxation in the dog.

OBJECTIVE: The aim was to establish whether the duration of coronary ischaemia and coronary ischaemia with reperfusion selectively reduced the magnitude of relaxation mediated by endothelium dependent relaxing factor (EDRF) in response to thrombin, compared with relaxation produced by acetylcholine and calcimycin. METHODS: Adult male dogs, anaesthetised with sodium pentobarbitone (30 mg.kg-1 intravenously) were used. Coronary artery occlusions were maintained for either 15 or 45 min; in half the dogs from each timepoint, occlusion was followed by 60 min reperfusion. At the end of each in situ period, coronary arteries were removed from both ischaemic and non-ischaemic regions, cut into rings, and hung in isolated organ baths. Dose-response relationships to the EDRF dependent vasodilators thrombin, acetylcholine, and calcimycin, and to the EDRF independent vasodilator isoprenaline, were then established. RESULTS: Thrombin (0.003-0.3 units.ml-1) caused dose dependent relaxation in all tissues. Relaxant responses (E(max)) in the non-ischaemic vessels from both 15 and 45 min treatment groups were used as control data for the responses in ischaemic vessels. Maximum responses were not different in the non-ischaemic groups from either 15 or 45 min studies, at 82.7 (SEM 3.7)% after 15 min, and 82.1(2.4)% after 45 min. There was a small but significant reduction in E(max) after 15 min and 45 min ischaemia, to 74.4(3.2)% and 74.4(3.0)% respectively. Sixty minutes reperfusion provoked a further reduction in E(max) to 64.9(3.8)% after 45 min ischaemia, but not after 15 min ischaemia [70.3(4.2)%]. Neither 15 nor 45 min interventions altered E(max) of relaxation to acetylcholine or calcimycin (greater than 88.0% in each group). Similarly there were no significant differences between groups to the relaxation stimulated by isoprenaline (E(max) greater than 90.0%). CONCLUSIONS: The data suggest that loss of EDRF dependent relaxation to thrombin is more sensitive to ischaemia than the relaxation produced by either acetylcholine or calcimycin, and appears to be manifested early in the onset of ischaemic injury.     

Are Markers of Proliferation Valuable in the Histological Assessment of Pituitary Tumours?

The prediction of tumour behaviour and response to treatment has led to interest in the assessment of the proliferative potential of tumours. Pituitary tumours are usually histologically benign but are capable of aggressive growth and local invasion, although distant metastasis is limited to the very rare pituitary carcinoma. These differences in tumour behaviour may determine both the prognosis and also the effectiveness of treatment whether it be surgery, drugs or radiotheraphy. Immunohistochemistry using antibodies to Ki-67 and proliferating cell nuclear antigen (PCNA) which are expressed in cells that have entered the cell cycle, can be used to assess the proportion of the cells from a tumour that are proliferating. The percentage of positively stained nuclei (labelling index (LI)) may be helpful in predicting appropriate management, as there is a relationship in many tumours between labelling index, invasiveness and tumour recurrence. This has been shown to be true for pituitary tumours, although there may be significant overlap such that low LI may be seen in the rare, aggressive, metastatic pituitary carcinomas, and high LI in indolent tumours. Thus although assessment of proliferation may be helpful in arousing suspicion as to subsequent tumour recurrence or invasiveness, this technique also demonstrates that there are other important and as yet unidentified processes that determine pituitary tumour behaviour.     

Variability of cell proliferation in the proximal and distal colon of normal rats and rats with dimethylhydrazine induced carcinogenesis

AIM：To investigate the patterns of cell proliferation in proximal and distal colons in normal rats and rats with1,2-dimethylhydrazine(DMH)induced carcinogenesis using the thymidine analogue bromodeoxyuridine.METHODS:Colonic crypt cell proliferation was immunohistochemically detected using the anti-bromodeoxyuridine Bu20a monoclonal antibody.RESULTS:Marked regional differences were found in both groups.Total labelling index(LI)and proliferative zone size in both normal(8.65&#177;0.34vs7.2&#177;0.45,27.74&#177;1.07vs16.75&#177;1.45)andDMH groups(13.13&#177;0.46vs11.55&#177;0.45,39.60&#177;1.32vs35.52&#177;1.58)were significantly higher in distal than in proximal colon(P&lt;0.05).although the number of cells per proxmal crypt was greater(31.45&#177;0.20vs34.45&#177;0.39,42.68&#177;0.53vs49.09&#177;0.65,P&lt;0.001).Crypt length,total LT and proliferative zone size all increased in both proximal and distal regions of DMH rats compared to normal controls(P&lt;0.0001).In DMH-treated rat colon a shift of labelled cells to higher crypt cell positions was demonstrated distally whist a bi-directional shift was evident proximally(P&lt;0.05).CONCLUSION:Our results show that changes in cell proliferation patterns,as assessed by bromodeoxyuridine uptake,can act as a reliable intermediate marker of colonic cancer formation.Observed differences between proliferation patterns in distal and proximal colon may be associated with the higher incidence of tumors in t he distal colon.