Proliferative lesions and reproductive tract tumors in male descendants of mice exposed developmentally to diethylstilbestrol.

Prenatal exposure to diethylstilbestrol (DES) is associated with reproductive tract abnormalities, subfertility and neoplasia in experimental animals and humans. Studies using experimental animals suggest that the carcinogenic effects of DES may be transmitted to succeeding generations. To further evaluate this possibility and to determine if there is a sensitive window of exposure, outbred CD-1 mice were treated with DES during three developmental stages: group 1 was treated on days 9-16 of gestation (2.5, 5 or 10 microg/kg maternal body weight) during major organogenesis; group II was treated once on day 18 of gestation (1000 microg/kg maternal body weight) just prior to birth; and group III was treated on days 1-5 of neonatal life (0.002 microg/pup/day). DES-exposed female mice (F(1)) were raised to maturity and bred to control males to generate DES-lineage (F(2)) descendants. The F(2) males obtained from these matings are the subjects of this report; results in F(2) females have been reported previously [Newbold et al. (1998) CARCINOGENESIS:, 19, 1655-1663]. Reproductive performance of F(2) males when bred to control females was not different from control males. However, in DES F(2) males killed at 17-24 months, an increased incidence of proliferative lesions of the rete testis and tumors of the reproductive tract was observed. Since these increases were seen in all DES treatment groups, all exposure periods were considered susceptible to perturbation by DES. These data suggest that, while fertility of the DES F(2) mice appeared unaltered, increased susceptibility for tumors is transmitted from the DES 'grandmothers' to subsequent generations.

The jejunal pouch as a rectal substitute after proctocolectomy.

Our hypothesis was that a jejunal pouch used as a rectal substitute after proctocolectomy would slow enteric transit, delay defecation, and decrease stool frequency compared to an ileal pouch so used. Twelve dogs underwent proctocolectomy; six had a jejunal pouch-distal rectal anastomosis and six had an ileal pouch-distal rectal anastomosis. After recovery, postprandial mouth-to-anus transit was slower in jejunal pouch dogs (253 +/- 18 minutes [mean +/- SEM]) than in ileal pouch dogs (112 +/- 7.9 minutes; P <0.05). Moreover, jejunal pouch dogs passed only 4.1 +/- 0.3 stools during the 12 hours after eating, whereas ileal pouch dogs passed 6.3 +/- 0. 9 stools (P <0.05). The mean frequency of proximal ileal pacesetter potentials after feeding was less in jejunal pouch dogs (12 +/- 0.4 cycles/min) than in ileal pouch dogs (16 +/- 0.3 counts/min; P = 0. 01), and jejunal pouches had more action potentials (jejunal = 82% +/- 4.3% of pacesetter potentials had action potentials, ileal = 61% +/- 3.0%; P <0.05). In contrast, gastric emptying and pouch motility, emptying, mucosal integrity, and bacteriologic and histologic properties were similar in the two groups of dogs. We concluded that the jejunal pouch operation slowed enteric transit, delayed defecation, and decreased postprandial stooling compared to the ileal pouch operation.

Restoration of myoelectrical propagation across a jejunal transection using microsurgical anastomosis.

The aim of this study was to determine whether microsurgical anastomosis can restore propagation of jejunal pacesetter potentials (PPs) across a site of canine jejunal transection and preserve motility and transit in bowel distal to the transection. A complete jejunal transection with exact microsurgical anastomosis was performed in five dogs, while five dogs with intact jejunum and five dogs with complete transection and end-to-end conventional macrosurgical anastomosis were used as controls. Long-term recording electrodes and intraluminal, open-tipped pressure catheters were implanted in all dogs. The mean frequency of PPs decreased distal to the transection in both groups of transected dogs. However, aborad propagation of PPs across the anastomosis occurred episodically by 3 months in each dog that had a microsurgical anastomosis, but never occurred in any dog with a conventional macroanastomosis. Moreover, the motility and transit in bowel distal to the transection were unaltered in the dogs with a microsurgical anastomosis, whereas they decreased in the dogs with a macroanastomosis. The conclusion was that microsurgical anastomosis of transected canine jejunum restored episodic propagation of PPs across the anastomosis, and preserved motility and maintained transit in bowel distal to the anastomosis. The conventional macroanastomosis did none of these.

Gastric accommodation in non-ulcer dyspepsia and the roles of Helicobacter pylori infection and vagal function.

BACKGROUND: The pathophysiological mechanisms in non-ulcer dyspepsia are incompletely understood. AIMS: To compare gastric motor and sensory functions in Helicobacter pylori positive or negative patients with non-ulcer dyspepsia. PATIENTS: Seventeen patients with non-ulcer dyspepsia and 16 asymptomatic controls. METHODS: The following were evaluated: gastrointestinal symptoms; gastric emptying and orocaecal transit of solids; abdominal vagal function; gastric compliance; fasting and postprandial gastric tone and phasic contractions; symptoms during ingestion of cold water and during the distension of an intragastric bag; and somatic sensitivity and personality profile (Minnesota Multiphasic Personality Inventory, MMPI). RESULTS: Gastric accommodation was reduced in H pylori negative dyspeptics relative to controls; the degree of accommodation was unrelated to H pylori status in dyspeptics. Increased postprandial gastric sensation was more frequent among H pylori positive patients (4/5 H pylori positive versus 4/12 H pylori negative patients). Intragastric meal distribution and orocaecal transit were normal; gastric emptying at four hours was abnormal in 4/17 patients. Vagal dysfunction was rare. Eight of 17 patients had somatisation or depression on MMPI. CONCLUSION: Impaired gastric accommodation is frequent in non-ulcer dyspepsia and seems to be unrelated to vagal efferent dysfunction. H pylori infection does not seem to influence gastric accommodation, but is associated with heightened sensitivity in dyspeptics. Therapeutic approaches that restore normal postprandial accommodation and gastric sensitivity should be tested in non-ulcer dyspepsia.

Increased tumors but uncompromised fertility in the female descendants of mice exposed developmentally to diethylstilbestrol.

Prenatal exposure to diethylstilbestrol (DES) has been associated with the subsequent development of reproductive tract abnormalities, including poor reproductive outcome and neoplasia, in experimental animals and humans. Experimental animal studies with chemical carcinogens have raised the possibility that adverse effects of DES may be transmitted to succeeding generations. To evaluate this possibility and to determine if there is a sensitive window of developmental exposure, outbred CD-1 mice were treated with DES during three stages of development: group 1 was treated on days 9-16 of gestation (2.5, 5 or 10 microg/kg maternal body wt), the time of major organogenesis; group II was treated once on day 18 of gestation (1000 microg/kg maternal body wt) just prior to birth; group III was treated on days 1-5 of neonatal life (0.002 microg/pup/day). Female mice (F1) in each group were raised to sexual maturity and bred to control males. As previously reported, fertility of the F1 DES-exposed females was decreased in all groups. Female offspring (DES lineage or F2) from these matings were raised to maturity and housed with control males for 20 weeks. The fertility of these DES lineage female mice was not affected by DES exposure of their 'grandmothers'. DES lineage mice were killed at 17-19 and 22-24 months of age. An increased incidence of malignant reproductive tract tumors, including uterine adenocarcinoma, was seen in DES lineage mice but not in corresponding controls; the range and prevalence of tumors increased with age. Because uterine adenocarcinomas were seen in all three DES groups, all developmental exposure periods were considered susceptible to the adverse effects of DES. These data suggest that the reduced fertility observed in the DES F1 female mice was not transmitted to their descendants; however, increased susceptibility to tumor formation is apparently transmitted to subsequent generations.

Gastric mechanosensory and lower esophageal sphincter function in rumination syndrome.

Our hypothesis was that rumination syndrome is associated with gastric sensory and motor dysfunction. We studied gastric and somatic sensitivity, reflex relaxation of the lower esophageal sphincter (LES), and gastric compliance and accommodation postprandially and postglucagon. A barostatically controlled gastric bag and esophageal manometry were used to compare gastric sensorimotor functions and LES relaxation to gastric distension in 12 patients with rumination syndrome and 12 controls. During bag distensions, patients had greater nausea, bloating, and aggregate score, but not pain, compared with controls (P < 0.05). At 4 and 8 mmHg gastric distension, LES tone reduction was greater in patients than in controls (P < 0.05). Gastric compliance, accommodation to a standard meal, and response to glucagon were not different in patients and controls; however, 6 of 12 patients had no gastric accommodation; the latter patients had significantly greater pain perception during distension (P < 0.05) but normal somatic sensitivity compared with healthy controls. Rumination syndrome is characterized by higher gastric sensitivity and LES relaxation during gastric distension. A subgroup of patients also had absent postprandial accommodation.

Canine small bowel motor patterns and contractions are not neurally regulated during enteric nutrient infusion.

The aims of this study were to determine the effects of duodenal and jejunoileal nutrient infusions on small intestinal motor patterns and intestinal contractions in neurally intact and neurally isolated small bowel. Fifteen dogs were prepared with duodenal and jejunal infusion and manometry catheters and a diverting jejunal cannula. Ten of the dogs underwent in situ neural isolation of the jejunoileum. A mixed nutrient meal (0.5 kcal/ml) was infused into the duodenum or jejunum at 3 ml/min for 5 h. Control experiments involved infusion of a balanced salt solution. Manometric data collected on-line to a microcomputer were analyzed for direction, distance, and velocity of spread of single pressure waves (SPW) and clustered contractions. Isolated duodenal and jejunoileal nutrient infusions inhibited the fasting motor pattern in neurally intact and neurally isolated small bowel. Motor activity (motility index) increased slightly during nutrient infusion within groups, but there were few differences between groups. Neither neural isolation nor nutrient infusion had a consistent effect on spread of SPW or migration of clustered contractions. Isolated duodenal and jejunoileal nutrient infusions in the dog inhibit fasting motor patterns and increase motor activity slightly but have little effect on characteristics of individual and clustered contractions. Extrinsic innervation to the jejunoileum or intrinsic neural continuity of the jejunum with the duodenum had little effect on single or grouped contractions. Although the changes in motor activity demonstrated in this study appear small, alterations in intestinal transit and absorption may still occur and may be of importance physiologically.

Rectal tone, distensibility, and perception: reproducibility and response to different distensions.

Increasing interest is focusing on the role of intestinal tone, distensibility, and mechanosensation in the genesis of abdominal symptoms. Experimental approaches usually feature balloon distension of the bowel with measurements of perception, tone, and compliance and/or elastance; however, the methodologies are standardized incompletely. We examined the reproducibility of repeated assessments of sensory perception, basal tone, and compliance and/or elastance of the rectum during distension. We also evaluated the response to inflations that varied in regard to control of pressure or volume, pattern of distension, and rate of inflation. Five healthy volunteers were studied under two separate protocols. The first featured a series of experiments on each of 5 days; the other consisted of 2 separate days of study. Repeated distensions evoked reproducible responses of sensation and compliance and/or elastance on a single day, providing a conditioning distension preceded them. Day-to-day variability was also sufficiently small to allow valid comparisons to be made on different days in healthy persons. The configuration of the distension profile (phasic, staircase, or ramp) and the rate of inflation (from 1 to 40 ml/s) had little effect on distensibility or perception. Perceptions were sometimes transient and sometimes constant, but no relationship was found between these temporal features and the magnitude of the stimulus. These observations help provide a basis as to how the responses to rectal distension can be best studied.

Influence of H. pylori infection on gastric motor and sensory function in asymptomatic volunteers.

The effect of H. pylori infection on gastric motility and sensation is unclear. Our hypothesis is that H. pylori infection increases gastric sensation and reduces gastric accommodation and emptying. In eight H. pylori-positive and eight H. pylori-negative asymptomatic subjects, infection was proven by antral histology or culture. We evaluated: (1) gastric emptying of solids, (2) proximal gastric compliance, (3) fasting and postprandial proximal gastric tone and phasic contractions, (4) gastric sensation during balloon inflations or ingestion of cold water, and (5) abdominal vagal function. H. pylori infection was associated with lower gastric accommodation (median 75% postprandial increase in barostat balloon volume compared to fasting) when compared to the accommodation in uninfected volunteers (median 211% change from fasting). One H. pylori-positive subject had an abnormal abdominal vagal function test and her gastric accommodation response was reduced. Other motor and sensory functions in the two groups were similar. In asymptomatic volunteers, H. pylori infection and gastritis result in reduced accommodation (diastolic dysfunction) but no change in overall sensation or motor functions of the stomach.

Adrenergic modulation of human colonic motor and sensory function.

The effects of pharmacological modulation of adrenergic receptors on colonic motor and sensory function are unclear. We studied 40 healthy volunteers in a single-blind design; 12 received saline, and the remaining 28 received either clonidine, yohimbine, phenylephrine, or ritodrine. A barostat-manometric assembly in the left colon recorded drug effects on fasting and postprandial motor function, compliance, and sensation in response to standardized phasic balloon distensions delivered in random order. Clonidine reduced and yohimbine increased fasting, but not postprandial tone, by 63.2 +/- 22.3% and 24.8 +/- 8.8% (SE), respectively. Clonidine tended to reduce fasting phasic activity in the descending and sigmoid colon. A power exponential model provided the best fit to the compliance curve. Clonidine significantly increased colonic compliance. Clonidine reduced and yohimbine increased colonic perception of pain but not gas sensation during distension. Phenylephrine and ritodrine did not influence colonic motor or sensory function in the present studies. Thus alpha 2-receptors modulate fasting colonic tone and compliance and alter perception of pain but not gas during mechanical stimulation of the colon.

Alpha 2-adrenergic modulation of colonic tone during hyperventilation.

Our aims were to assess the role of adrenergic modulation in the hyperventilation-induced increase in colonic tone. Of 40 healthy volunteers, 12 received placebo (saline) and the remaining 28 received either clonidine, yohimbine, phenylephrine, or ritodrine. Time-frequency mapping of heart rate based on Wigner distribution assessed variations in parasympathetic and sympathetic activity during hyperventilation. Tone in the descending colon was recorded by a barostat balloon before, during, and after 5 min of hyperventilation. Heart rate spectral analysis suggested diminished sympathetic and vagal activity during hyperventilation and increased sympathetic and vagal activity after hyperventilation. Adrenergic agents influenced (P = 0.01) the tonic response after, but not during, hyperventilation. Yohimbine reduced the increment in colonic tone after hyperventilation compared with saline (P < 0.05) and clonidine (P = 0.002); phenylephrine and ritodrine had no effects. Different mechanisms modulate the increase in colonic tone during and after hyperventilation. Yohimbine attenuates the increase in colonic tone after hyperventilation probably by enhancing inhibitory sympathetic input to the colon.

Pharmacological modulation of rectal tone alters perception of distention in humans.

OBJECTIVE: Drugs can alter perception of balloon distention of the GI tract. It has been proposed that the mechanism by which this occurs is through effects on visceral afferent pathways. Our hypothesis was that modulation of rectal tone will also influence the perception of rectal balloon distention. METHODS: Fasting and postprandial rectal tone, compliance, and perception of rectal distention were measured in 25 healthy subjects, using a five-armed, parallel, single-blinded study design. Each subject received either glucagon, nitroglycerin, clonidine, yohimbine, or saline. RESULTS: Rectal tone, but not compliance, influenced perception as measured by balloon distention of the rectum (r = 0.6, p = 0.002). Glucagon, nitroglycerin, and clonidine reduced and yohimbine increased fasting tone compared with saline. Compliance and postprandial tone were similar in all groups. Yohimbine increased rectal perception of distention. CONCLUSIONS: Tone is one of the factors that influences the sensory perception of balloon distention in the human rectum. Alpha2-adrenergic agents, a nitric oxide donor, and glucagon altered fasting rectal tone, but postprandial tone was similar after administration of each agent.

Accelerated onset of uterine tumors in transgenic mice with aberrant expression of the estrogen receptor after neonatal exposure to diethylstilbestrol.

The role of estrogen and the estrogen receptor (ER) in the induction and promotion of tumors was investigated by using transgenic MT-mER mice, which overexpress the ER. It was hypothesized that because of this abnormal expression of the ER, the reproductive-tract tissues of the MT-mER mice may be more susceptible to tumors after neonatal exposure to the potent synthetic estrogen diethylstilbestrol (DES). Normally non-estrogen responsive tissues that may have expressed ER as a result of the transgene were also studied for DES-induced tumors. Wild-type and MT-mER littermates were treated with 2 micrograms/pup/d DES 1-5 d after birth and then killed at 4, 8, 12, and 18 mo of age. The DES-treated MT-mER mice demonstrated a significantly higher incidence of uterine adenocarcinoma at 8 mo (73%) than the DES-treated wild-type mice (46%). The tumors of the MT-mER mice were often more aggressive than those in the wild-type animals. These tumors were also preceeded at 4 mo by a significantly higher incidence of the preneoplastic lesion atypical hyperplasia in the MT-mER mice (26% compared with 0% in the wild-type mice). Other DES-induced abnormalities were observed at equal rates in the wild-type and MT-mER mice. Although no tumors were observed in untreated wild-type females, a single untreated MT-mER female had uterine adenocarcinoma at 18 mo. These data indicate that the level of ER present in a tissue may also be a determining factor in development of estrogen-responsive tumors.

Ontogeny of lactoferrin in the developing mouse uterus: a marker of early hormone response.

Lactoferrin (LF) was mapped during organogenesis of the murine reproductive tract, starting on fetal Day 12, as a marker of estrogen responsiveness. To induce LF expression, pregnant outbred CD-1 mice were injected s.c. with diethylstilbestrol (DES; 100 microg/kg maternal body weight), and fetal genital tract tissues were removed; neonatal and immature mice received s.c. injections of DES (2 microg/pup per day). Corn oil-treated and untreated mice at corresponding ages provided the controls. Immunocytochemical techniques using a polyclonal antibody showed no detectable LF in control genital tract tissues until late gestation. However, after DES treatment, LF was localized in uterine epithelial cells as early as fetal Day 14; the intensity of LF staining increased with age and number of DES treatments. Control uterine tissues responded to the rise of circulating estrogens at parturition (fetal Day 19) by producing LF, although the magnitude of response was lower than that of DES-treated tissues. Uterine tissue homogenates from control and DES mice were analyzed by SDS-PAGE and Western blots, verifying the protein to be LF. Isolation of mRNA and Northern blot analysis further showed that LF mRNA was present in the developing Mullerian duct and that DES stimulated early induction of the LF gene. The early appearance of LF suggests that it may play an important role in the hormonal regulation of growth and differentiation of developing uterine tissues.

SDZ HTF 919 stimulates canine colonic motility and transit in vivo.

Effects of the nonbenzamide 5-hydroxytryptamine4 agonist SDZ HTF 919 on gastrointestinal motility are unclear. Our aim was to assess the in vivo effects on gastrointestinal and colonic transit of radiolabeled residue and on colonic phasic contractility. In six female dogs, transit was measured over a period of 2 days by radioscintigraphy and colonic motility was measured by pneumohydraulic perfusion manometry of the proximal and distal colon. SDZ HTF 919 was administered initially by bolus i.v. infusion, followed by s.c. injection 8 and 16 hr later. Doses tested were 0.03, 0.1 and 0.3 mg/kg, and isotonic saline and vehicle served as controls in each dog. Stomach and small bowel transit was not significantly altered by SDZ HTF 919. Overall, i.v. SDZ HTF 919 accelerated colonic transit during the first 1 hr, compared with controls. These effects were significant even with the lowest dose of SDZ HTF 919. Responses to higher infusion doses were more variable. SDZ HTF 919 did not cause significant changes in quantitative pressure indices, such as amplitude or motor index, in the small bowel or colon. Prolonged postprandial colonic contractions, each lasting >30 sec, were noted after each i.v. agent and were significantly more frequent with the 0.03 mg/kg dose than with control (vehicle) treatment. Thus, SDZ HTF 919 accelerates canine colonic transit in vivo during the first 1 hr after i.v. administration. SDZ HTF 919 appears to be a promising agent for stimulation of mammalian colonic transit.

A pilot study of motility and tone of the left colon in patients with diarrhea due to functional disorders and dysautonomia.

OBJECTIVE: Our aim was to identify qualitative or quantitative colonic motor patterns induced postprandially in a pilot study of patients with diarrhea due to functional disease or dysautonomia to identify objective endpoints for future studies. METHODS: In patients with functional diarrhea (n = 5) or dysautonomia (n = 4) in whom GI transit was documented by scintigraphy, we studied colonic motility by combined manometry and barostat measurements for 1 h fasting and 2 h postprandially (1000-kcal meal). Data were compared with those of healthy control subjects. RESULTS: There were no differences in compliance, overall phasic motility of the left colon, fasting tone, or maximal change in postprandial tone in the diarrhea group as compared with the control group. The diarrhea group showed more high amplitude propagated contractions 4.4 +/- 3.6 (SD)/2 h, p < 0.05) compared with the control group (0.7 +/- 1.4/2 h); the mean postprandial tonic response (12 +/- 14%, p < 0.05) and its duration were reduced in the diarrhea group compared with the control group (27 +/- 17%). Two dysautonomic patients showed a paradoxical relaxation of the colon postprandially. CONCLUSION: Reduced duration of increased colonic tone postprandially and increased number of high amplitude propagated contractions seem to be useful objective endpoints for future studies.

Effect of cholecystokinin octapeptide and atropine on human colonic motility, tone, and transit.

The role of cholecystokinin (CCK) in postprandial control of colonic motility is controversial. To test the hypothesis that CCK stimulates colonic tone, motility, and transit we measured these colonic functions in 16 healthy subjects using intraluminal manometry, barostatic balloon measurements, and radioscintigraphy. This was a randomized-order, double-blind, sequential study design in each subject of saline and either atropine (0.01 mg/kg stat and 0.01 mg/kg/hr by infusion) or CCK-octapeptide (OP, 30 ng/kg stat and 60 ng/kg/hr by infusion). Atropine was used as control to demonstrate responsiveness of selected parameters of colonic motility. Atropine significantly reduced whole colon (change from fasting = 52 +/- 11%) and left colon (change from fasting 61 +/- 8%) phasic pressure activity and transverse colon tone (change from fasting 159 +/- 40%); CCK-OP had no significant effects on phasic contractility, tone or transit. Thus, a CCK-OP infusion that maximally stimulates pancreatic exocrine secretion and gallbladder contraction has no effect on motor function or transit in prepared colon of healthy subjects.

Motility and tone of the left colon in constipation: a role in clinical practice?

OBJECTIVES: Colonic motor mechanisms deranged in constipation are not understood completely. Our aim was to measure left colonic motility and tone, during fasting and postprandially in patients with chronic constipation. METHODS: During 1 h fasting and 2 h postprandially, we measured pressures (multilumen manometry) and tone (barostat) in the left colon of 15 healthy controls and 40 patients with chronic constipation associated with slow (n = 15) or normal colonic transit (n = 12) or outlet obstruction (n = 13). RESULTS: Fasting tone was similar in all groups, and all demonstrated a significant increase in motor activity to food. There was lower postprandial tone (p < 0.05) in the slow transit and outlet obstruction groups. There were no differences in the timing of the tonic response or the number or amplitude of high-pressure propagated contractions. The slow transit group had lower postprandial phasic responses in the rectosigmoid (p < 0.05) and descending (p < 0.1) colon; the outlet obstruction group had lesser descending (p < 0.05) and rectosigmoid (p < 0.1) colon phasic motility. CONCLUSIONS: Colonic intraluminal measurements alone do not discriminate subgroups of chronic constipation more accurately than transit and pelvic floor tests, and currently have a limited role in clinical practice. However, manometry and tone measurements may be helpful in confirming a diagnosis of slow transit constipation (colonic inertia) in patients considered candidates for surgical treatment.