A comparison of clomiphene citrate and human menopausal gonadotropin for use in conjunction with intrauterine insemination.

OBJECTIVE: To compare the outcome of superovulation using clomiphene citrate (CC) versus hMG in conjunction with IUI. DESIGN: Sequentially assigned, observational study. Couples initially were assigned to receive either CC or hMG for three cycles. SETTING: The Clinical Outpatient Department of the Dartmouth-Hitchcock Medical Center. PATIENT(S): Eighty-three infertile couples. INTERVENTION(S): IUI with hMG use. MAIN OUTCOME MEASURE(S): Conception rate, term pregnancy rate (PR), and pregnancy complications, such as spontaneous miscarriage and multiple gestation. RESULT(S): Of 83 couples who underwent at least one treatment cycle, 29 (35%) conceived during the study period. The relative rate of conception for hMG versus CC was 2.08 (95% confidence interval [CI], 0.93 to 4.68). The relative term PR was 2.10 (95% CI, 0.77 to 5.73) for hMG versus CC. There was no difference in the miscarriage rate for hMG versus CC. CONCLUSION(S): Both the conception rate and the term PR were higher using hMG, compared with CC, in combination with IUI, and showed a trend toward statistical significance.

Formation of immunochemical advanced glycosylation end products precedes and correlates with early manifestations of renal and retinal disease in diabetes.

Elevated levels of advanced glycosylation end products (AGEs) have been found in multiple tissues in association with diabetic vascular complications and during the microalbuminuric phase of diabetic nephropathy. In this study, we have used an AGE-specific enzyme-linked immunosorbent assay (ELISA) to measure skin AGEs to determine whether elevated levels can be detected before the onset of overt microangiopathy. Subjects with type I diabetes (n = 48) were graded for the degree of nephropathy (normal [23], microalbuminuria [12], or macroalbuminuria [12]) and retinopathy (none [13], background [20], or proliferative [15]). Subgroups with a premicroalbuminuric phase of albumin excretion (< or = 28 mg/24 h, n = 27) or with the earliest stages of retinopathy (n = 27) were identified. A significant increase in tissue AGEs was found as urinary albumin increased during the premicroalbuminuric phase of nephropathy even when the data were adjusted for age and duration of diabetes (P = 0.005). Immunoreactive AGEs also increased as normal renal status advanced to microalbuminuria and macroalbuminuria (P = 0.0001 across groups). Significant elevation of AGEs was also found in association with the earliest stages of clinically evident retinopathy (early background versus minimal grades). In addition, higher AGE levels were found in subjects with proliferative retinopathy when compared with those with less severe retinopathy (P < 0.004 across groups). In contrast, no significant differences were found in tissue AGE levels between groups with or without early retinopathy based on pentosidine or fluorescent AGE measurements, although fluorescent AGEs correlated with albumin excretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Anabolic-androgenic steroid and adrenal steroid effects on hippocampal plasticity.

Anabolic-androgenic steroids (AAS) are synthetic androgen-like compounds which are taken in high doses by athletes with the intention of enhancing muscular appearance, strength and/or athletic performance. Recent research indicates that high doses of AAS may influence the functions of the hippocampus. This evidence led us to explore the extent to which chronic AAS treatments influence spatial memory and the integrity of the hippocampus in the rat. Gonadally intact adult male Long-Evans rats were treated with either the AAS methandrostenolone, a steroid 'cocktail' (TNB; testosterone cypionate, boldenone undecylenate and nandrolone decanoate), or the oil vehicle daily for 12 weeks. A group of male rats treated with corticosterone (CORT; 10 mg/day) was also examined. Spatial memory was assessed in the Morris water maze after 10 weeks of hormone treatment. At 12 weeks, the animals were sacrificed, blood collected and the brain sectioned to assess hippocampal cell number. There were no impairments in the acquisition or retention of the Morris water maze in any hormone treatment group. Although serum testosterone levels were elevated in rats treated with TNB relative to the oil controls, neither the TNB or methandrostenolone treatments produced changes in hippocampal cell number. Serum CORT levels were significantly elevated in the rats treated with CORT and cell loss (15%) was detectable in the CA3b subfield in this group of animals. These results indicate that the AAS administered in the present study were not detrimental to hippocampal spatial memory or cell survival and that, while chronic CORT may produce mild hippocampal cell loss, this loss is not accompanied by deficits on a spatial memory task.

The effect of cigarette smoking on adrenal cortical hormones.

We assessed the association between cigarette smoking and basal levels of adrenal cortical hormones in 11 postmenopausal smokers and 11 postmenopausal nonsmokers and measured the acute adrenal effects of cigarettes in the smokers. After an overnight food, alcohol and tobacco fast, participants smoked or sham-smoked every hr for 8 hr and provided serum samples for hormone assay before and after every other cigarette/sham, as well as before and after a corticotropin stimulation test. The postmenopausal smokers had substantially higher basal levels of androstenedione (4.60 +/- 0.42 vs. 2.70 +/- 0.36 nmol/l, P < .05) and dihydroepiandrosterone sulfate (2.88 +/- 0.36 vs. 1.91 +/- 0.16 mumol/l, P < .05) and higher average levels of cortisol and androstenedione from 0800 to 1300 hr (351.0 +/- 17.5 vs. 295.5 +/- 17.1, nmol/l and 3.58 +/- 0.42 vs. 2.51 +/- 0.19 nmol/l, P = .03, and P < .05, respectively). There were small acute effects of individual cigarettes on the hormones, but the response to corticotropin was similar in smokers and nonsmokers. Our results indicate that cigarette smoking causes a generalized disturbance in adrenal cortical hormone levels. There is no evidence for acute tolerance to the adrenocortical affects of the hourly smoking of medium-nicotine cigarettes, but these acute effects do not explain the higher hormone levels in smokers. There is no evidence for a partial block in the cortisol synthesis pathway to explain the increased adrenal androgen levels in smokers.

Role of sex hormones in development of chronic mountain sickness in rats.

After chronic exposure to hypoxia, Hilltop Sprague-Dawley rats developed excessive polycythemia and severe pulmonary hypertension and right ventricular (RV) hypertrophy, signs consistent with human chronic mountain sickness; however, there were gender differences in the magnitude of the polycythemia and susceptibility to the fatal consequence of chronic mountain sickness. Orchiectomy and ovariectomy were performed to evaluate the role of sex hormones in the gender differences in these hypoxic responses. After 40 days of exposure to simulated high altitude (5,500 m; barometric pressure of 370 Torr and inspired Po2 of 73 Torr), both sham-gonadectomized male and female rats developed polycythemia and had increased RV peak systolic pressure and RV hypertrophy. The hematocrit was slightly but significantly higher in males than in females. Orchiectomy did not affect these hypoxic responses, although total ventricular weight was less in the castrated high-altitude rats. At high altitude, the mortality rates were 67% in the sham-operated male rats and 50% in the castrated animals. In contrast, ovariectomy aggravated the high-altitude-associated polycythemia and increased RV peak systolic pressure and RV weight compared with the sham-operated high-altitude female rats. Both sham-operated control and ovariectomized females suffered negligible mortality at high altitude. The present study demonstrated that 1) the male sex hormones play no role in the development of the excessive polycythemia, pulmonary hypertension, and RV hypertrophy during chronic hypoxic exposure or in the associated high mortality and 2) the female sex hormones suppressed both the polycythemic and cardiopulmonary responses in vivo during chronic hypoxic exposure.

Effects of 17 beta-estradiol and progesterone on pressor responses in conscious ovariectomized rats.

Attenuation of pressor responsiveness to several administered vasoconstrictors is a constant feature of normal gestation in humans and other species, such as the rat. However, the mechanism of this physiological adaptation remains uncertain. Because plasma levels of 17 beta-estradiol (E2) and progesterone (P) increase markedly during pregnancy, we tested the hypothesis that these hormones may mediate the reduced pressor responses. Seven days after bilateral ovariectomy and chronic instrumentation of rats, the pressor responses of arginine vasopressin, angiotensin II, and norepinephrine were tested on two occasions > or = 48 h apart. Then E2, P, or a combination of E2 and P was administered by subcutaneous implantation of 21-day-release steroid pellets. Pressor responses were again tested at various times throughout the period of steroid treatment. The plasma concentrations of the steroids were assessed by radio-immunoassay, and doses of the hormones were given that both approximated and exceeded circulating levels found in our laboratory for gravid rats. Despite chronic elevation of plasma E2 and/or P, we did not observe consistent attenuation of pressor responsiveness in any of the steroid-treatment regimens, nor was a decline in mean arterial pressure observed, which is typically found in rats during late gestation. In conclusion, we are unable to support the hypothesis that E2 and/or P contributes to the diminished pressor responsiveness of rat pregnancy.

Cell lines derived from pancreatic tumors of Tg(Ela-1-SV40E)Bri18 transgenic mice express somatostatin and T antigen.

Continuous cell lines have been isolated from islet cell, small cell anaplastic and acinar cell carcinomas arising in the pancreas of transgenic mice, line Tg(Ela-1-SV40E)Bri18. These mice carry the pseudogene construct composed of elastase-1 promoter linked to the SV40 T antigen. Cells derived from islet cell or small cell anaplastic tumors secreted insulin and somatostatin during the early period of culture. Phenotypic alterations occurred during culture, whereby insulin secretion ceased and cells instead secreted somatostatin, indicating a change from beta-cell to delta-cell phenotype. Acinar cell lines did not secrete amylase or lipase.

Increased collagen-linked pentosidine levels and advanced glycosylation end products in early diabetic nephropathy.

RATIONALE: Advanced glycosylation end products (AGEs) may play an important role in the development of diabetic vascular sequelae. An AGE cross-link, pentosidine, is a sensitive and specific marker for tissue levels of AGEs. OBJECTIVES: To evaluate the role of AGEs in the development of diabetic nephropathy and retinopathy, we studied pentosidine levels and the clinical characteristics of 48 subjects with insulin-dependent diabetes mellitus. Diabetic nephropathy was classified as normal, microalbuminuria, or gross proteinuria, and retinopathy was graded as none, background, or proliferative. NEWLY OBSERVED FINDINGS: Significant elevation of pentosidine (P = 0.025) was found in subjects with microalbuminuria or gross proteinuria (73.03 +/- 9.47 vs 76.46 +/- 6.37 pmol/mg col) when compared with normal (56.96 +/- 3.26 pmol/mg col). Multivariate analysis to correct for age, duration of diabetes, and gender did not modify the results. Elevated pentosidine levels were also found in those with proliferative when compared with those with background retinopathy (75.86 +/- 5.66 vs 60.42 +/- 5.98 pmol/mg col) (P < 0.05). CONCLUSIONS: Microalbuminuria is associated with elevated levels of pentosidine similar to those found in overt diabetic nephropathy suggesting that elevated AGE levels are already present during the earliest detectable phase of diabetic nephropathy.

Antisperm antibodies in women: variability in antibody levels in serum, mucus, and peritoneal fluid.

OBJECTIVE: To look for patterns of antisperm antibody expression in women by exploring the levels of antisperm antibodies in different body fluids. This was achieved by studying sequential serum samples from individual patients and by comparing the levels of antisperm antibodies in serum from a number of patients with the levels of antisperm antibodies in cervical mucus or peritoneal fluid (PF). DESIGN: Prospective studies were performed on sequential serum samples within a menstrual cycle. Retrospective studies were done to compare antisperm antibodies in serum and mucus or PF. The immunobead assay was used to measure antisperm antibodies in these fluids. SETTING: Dartmouth-Hitchcock Medical Center, Lebanon, New Hampshire. PATIENTS: A random sample of patients undergoing evaluation for infertility. RESULTS: The levels of antisperm antibodies in sera drawn from patients at different points in a menstrual cycle stimulated by the presence of exogenous hormones did not change during the follicular phase of the menstrual cycle. Also, in many samples, the antisperm antibody level in serum did not correlate with the antisperm antibody levels in mucus or PF. CONCLUSIONS: The data suggest that measurement of antisperm antibodies at a single point in time or from a single fluid is not sufficient when evaluating a woman for immunological infertility. The data also suggest that numerous and complex factors contribute to the expression of antisperm antibodies in women.

Ventilatory and hematopoietic responses to chronic hypoxia in two rat strains.

Hilltop (H) and Madison (M) strains of Sprague-Dawley rats exhibit strikingly different susceptibilities to the effects of chronic altitude exposure. The H rats develop greater polycythemia, hypoxemia, and pulmonary hypertension. We studied ventilation, pulmonary gas exchange, tissue oxygenation, and hematologic adaptations in the two rat strains during a 50-day exposure to a simulated altitude (HA) of 5,500 m (18,000 ft). There were no strain differences among the variables we studied under sea level (SL) conditions. Within the first 14 days of hypoxic exposure, the only significant strain differences were that erythropoietin (EPO) rose much higher and erythroid activity was greater in the H rats, even though arterial Po2 and PCo2 (Pao2 and PaCo2, respectively), renal venous PO2 (Prvo2), and ventilation (VE) were equivalent in the two strains during this time. By day 14 at HA, the H rats had significantly higher erythroid activity, hematocrit (Hct), and EPO levels, significantly lower PaO2 and PrvO2, but equivalent VE and PaCO2. These changes persisted for the remainder of the exposure, except that the Hct continued to rise and the increase was greater in H rats. Despite the greater O2-carrying capacity of H rats in the later stages of hypoxic exposure, PaO2 and PrvO2 were significantly lower in H rats. There were no strain differences at either SL or HA in ventilatory responses to hypercapnia or hypoxia, in blood O2 affinity or 2,3-diphosphoglycerate, in extrarenal production of EPO, or in EPO clearance. We conclude that early in the hypoxic exposure the H rats produce more EPO at apparently equivalent levels of hypoxia, and this is the first step in the pathogenesis of the maladaptation to HA manifest by H rats. We find no consistent evidence that differences in VE contribute to the variable susceptibility to hypoxia in the two rat strains.

Inhibitory effect of streptozotocin on tumor development in transgenic mice bearing an elastase I-SV40 T-antigen fusion gene.

Transgenic mice, bearing a fusion gene of rat elastase I promoter and SV40 T-antigen, developed acinar cell tumors of the pancreas, as predicted by the model. In addition, they developed insulinomas and somatostatin (delta)-cell hyperplasia of the pancreatic islets. The insulinomas and the delta-cell hyperplasia appeared to be functional, as evidenced by changes in plasma glucose, insulin, and somatostatin. Streptozotocin, which has been shown to inhibit pancreatic carcinogenesis in the hamster model, significantly reduced the numbers of insulinomas and delta-cell hyperplasias. Streptozotocin did not cause a statistically significant reduction in exocrine tumors.

Hypodiploid, pseudodiploid, and normal karyotypes prevail in cytogenetic studies of medullary carcinomas of the thyroid and metastatic tissues.

Medullary carcinoma of the thyroid (MCT), often a dominantly inherited neoplasm, derived from intrathyroid C-cells of neural crest origin, is one of the solid tumors least studied cytogenetically. The cells are difficult to grow in culture, only two cell lines having ever been established. Cytogenetic studies of only 5 tumors have been reported previously. In this paper we report on the cytogenetic analyses of 8 specimens of primary and/or metastatic MCT tumor tissue from 6 patients with familial disease, including more recent metastatic tumors in lymph node and femur of a patient whose thyroid and earlier lymph node metastases were described previously. Some of these specimens were harvested sequentially over time. Hypodiploid or diploid modal numbers prevailed with normal, pseudodiploid, or hypodiploid karyotypes.

Inhibition of a transplantable pancreatic carcinoma by castration and estradiol administration in rats.

Influence of sex steroids on the growth of an azaserine-induced transplantable rat pancreatic carcinoma, DSL-2, was studied. This established transplantable tumor has been maintained in syngeneic rats. Inbred male Lewis rats were pretreated with castration and s.c. implantation of 1.0-mg 17 beta-estradiol (CAS: 50-28-2; estradiol) pellets at 7 weeks of age. Tumor cells were inoculated s.c. on the back of intact male, castrated male, or 17 beta-estradiol-treated castrated male rats. Additional male rats served as non-tumor-bearing controls. There was no difference in the body weight between tumor-bearing and non-tumor-bearing male rats. A distinct difference in the tumor growth was observed in variously conditioned recipients. In castrated male hosts, the serum testosterone levels and the epididymis weights were significantly decreased, and the tumor weights were significantly less as compared to intact control hosts. Additional pretreatment with 17 beta-estradiol caused a markedly slower growth of tumors and increases of the serum 17 beta-estradiol levels and the pituitary weights in castrated male recipients. The remarkable response of tumor growth to castration was also observed in a fast-growing tumor derived from DSL-2. Moreover, close positive relationships between tumor weights and the activities of both serum amylase and lipase were observed. Results showed that the pretreatment with castration alone or in combination with 17 beta-estradiol treatment was able to inhibit the growth of the transplantable tumor. In addition, tumor cells had an ability to produce amylase and lipase, and the amount of enzymic activity was related to the tumor volume. Thus, these data indicate that the transplantable rat pancreatic carcinoma retains physiological function. Our previous study has shown the modulation by sex steroids of azaserine-induced preneoplastic lesions of pancreas in rats. Therefore, androgens and estrogens may play key roles as promoters and inhibitors during the process of pancreatic carcinogenesis.

The renin-angiotensin system during pregnancy in chronically instrumented, conscious rats.

Whether the renin-angiotensin system is activated during rat gestation is controversial. Therefore we serially assessed plasma renin activity in unrestrained, chronically instrumented conscious rats during pregnancy and the postpartum period. Plasma renin activity was 3.26 +/- 0.30, 2.80 +/- 0.31, and 2.70 +/- 0.26 ng.ml-1.hr-1 on gestational days 6, 12, and 20, respectively. When the same rats were studied after delivery, plasma renin activity was 1.87 +/- 0.29, 1.81 +/- 0.09, and 2.31 +/- 0.35 ng.ml-1.hr-1 on postpartum days 3, 6, and 11, respectively. Levels measured during pregnancy were significantly greater than in the postpartum period (p less than 0.05 or less than 0.01). We then evaluated potential functional consequences of the renin-angiotensin system in gravid rats. Near term, renal hemodynamics fall from the peak levels of midgestation; we tested whether angiotensin II mediates this apparent vasoconstriction. Captopril (1.5 mg/kg, 1.5 mg.kg-1.hr-1) was acutely administered to lower circulating angiotensin II. The drug produced an 80% inhibition of angiotensin I pressor response, a tenfold elevation in plasma renin activity, but caused the same degree of mild renal vasodilation in rats whether they were virgin or pregnant. We also tested whether prior occupancy of receptors by endogenous hormone or receptor downregulation mediates the attenuated pressor response to angiotensin II observed during late pregnancy. Acute administration of captopril failed to augment refractory pressor responsiveness. Chronic treatment with enalaprilat (2.0 mg.kg-1.day-1 by osmotic minipump) also did not restore pressor responsiveness. But, in our hands, chronic administration of enalaprilat most likely failed to lower plasma angiotensin II. In summary, we suggest that during rat gestation (1) the renin-angiotensin system is activated, (2) angiotensin II does not mediate the apparent renal vasoconstriction observed near term, (3) prior receptor occupancy by endogenous hormone is not responsible for the attenuated pressor response to angiotensin II, and (4) long-term treatment with enalaprilat can produce hypotension without reducing plasma concentrations of angiotensin II.

Inhibitory effects of estrogen and castration on the early stage of pancreatic carcinogenesis in Fischer rats treated with azaserine.

Effects of sex steroids on pancreatic carcinogenesis during the early stage were studied in azaserine-treated rats of both sexes. Fischer rats were given weekly i.p. injections of azaserine (30 mg/kg) [CAS:115-02; diazoacetate serine(ester)] at 2 and 3 weeks of age and were divided into six groups. Castration, ovariectomy, and s.c. implantations of either a 0.3-mg or a 1.0-mg 17 beta-estradiol (CAS:50-28.2; estradiol) pellet were performed at 7 weeks of age. The groups were as follows: group 1, intact male; group 2, castrated; group 3, castrated plus 0.3 mg estradiol; group 4, castrated plus 1.0 mg estradiol; group 5, ovariectomized; and group 6, intact female. Rats were killed 4 months after the last injection of azaserine. Azaserine treatment induced atypical acinar cell foci and nodules (AACN) in both sexes. The acidophilic AACN are considered preneoplastic lesions. An apparent sex difference was observed; the number of acidophilic AACN was greater in male rats than in female rats. Castration caused a significant decrease in both the serum testosterone levels and the number of acidophilic AACN, which were comparable to those in ovariectomized female rats. Furthermore, when estradiol treatment was administered to the castrated male rats, a linear decrease in the number of acidophilic AACN and an elevation in the serum estradiol levels were observed and were dose dependent. There were also positive relationships between estradiol treatments and the mean pituitary and pancreas weights. These results showed that estradiol treatment and the drop in testosterone levels caused by castration were highly effective in inhibiting the development and growth of preneoplastic lesions of the pancreas of the rats treated with azaserine. This estradiol effect was dose dependent. The present study, therefore, provides evidence that estrogen may act as an inhibitor and androgen as a promoter in the early stage of pancreatic carcinogenesis in rats.

Cardiovascular and hormonal response to hemorrhage in conscious rats.

The effect of arginine vasopressin (AVP) pressor blockade on the response to graded hemorrhage was investigated on conscious, unstressed, freely moving rats. The parameters studied were mean arterial pressure (MAP), heart rate (HR), blood velocity in the ascending aorta (ABV), and plasma concentrations of AVP (pAVP), renin (PRC), corticosterone (pCS), and catecholamines. After the first hemorrhage (0.75% of body wt over 5 min), MAP remained unchanged while ABV declined and HR increased. The two subsequent hemorrhages brought about significant reduction in MAP, HR, and ABV. pAVP, pCS, and PRC increased gradually during the experiment, while plasma catecholamine levels remained unchanged except for epinephrine, which increased after the third hemorrhage. After pretreatment with the AVP-pressor antagonist, [d(CH2)5Tyr(Me)]AVP, the hemorrhage-induced cardiovascular changes were practically identical to those seen in control animals. Results with AVP blockade performed after the third hemorrhage were also negative. A pressor role of AVP after hypotensive hemorrhage could only be revealed in the presence of converting-enzyme inhibition and alpha-adrenergic blockade. In addition, [d(CH2)5Tyr(Me)]AVP did not modify the effect of hemorrhage on pCS and catecholamines and caused only a slight enhancement of the increase in PRC. It is concluded that conscious, nonstressed rats, if all compensatory mechanisms are allowed full expression, exhibit a normal cardiovascular compensatory response to hemorrhage in the absence of functional AVP pressor receptors.

Epidural anesthesia and analgesia in high-risk surgical patients.

The authors conducted a randomized controlled clinical trial to evaluate the effect of epidural anesthesia and postoperative analgesia (EAA) on postoperative morbidity in a group of high-risk surgical patients. A total of 53 patients were admitted to the study, 28 received EAA, and 25 received standard anesthetic and analgesic techniques without EAA. Surgical "risk" was evaluated preoperatively and found to be comparable in the two groups. When compared to control patients, patients who received EAA had a reduction in the overall postoperative complication rate (P = 0.002) and in the incidence of cardiovascular failure (P = 0.007) and major infectious complications (P = 0.007). Urinary cortisol excretion, a marker of the stress response, was significantly diminished during the first 24 postoperative hours in the group receiving EAA (P = 0.025). Finally, hospital costs were significantly reduced for patients who received EAA (P = 0.02). The authors conclude that EAA exerted a significant beneficial effect on operative outcome in a group of high risk surgical patients.

Effect of castration and hormone replacement on azaserine-induced pancreatic carcinogenesis in male and female Fischer rats.

Previous reports have shown that pancreatic cancer was induced preferentially in male versus female azaserine-treated rats. This study was designed to determine the importance of estrogen and testosterone in this phenomenon. Fischer (F344) rats received a single injection of azaserine (30 mg/kg) at 21 days of age. At 28 days of age, they were weaned and divided into 12 groups of 9-10 rats as shown below. Surgery (castration or sham operation) was performed at 4 weeks of age. All drugs (estradiol, the antiestrogen tamoxifen, testosterone propionate and/or the antiandrogen flutamide) were administered, starting at weaning, in 3-week timed-release pellets until autopsy. Rats were killed 4 months after the administration of azaserine. The pancreas was weighed and prepared for quantitative histologic analysis of atypical acinar cell nodules (AACNs) which are putative preneoplastic lesions. Both number and size of AACNs were analyzed. In intact female rats, AACN burden was smaller than in intact males (P less than 0.05). Ovariectomy increased the AACN burden (P less than 0.05), while estradiol or tamoxifen treatments to ovariectomized females resorted the burden to control levels (P less than 0.05). Testosterone with tamoxifen treatment to ovariectomized females led to a significant increase in AACN burden over control values. In intact male rats, orchiectomy decreased the AACN burden (P less than 0.05). In orchiectomized rats, testosterone treatment slightly increased the AACN burden, flutamide treatment alone increased this parameter (P less than 0.05) but flutamide with estradiol decreased the AACN burden (P less than 0.01). These data strongly support the hypothesis that sex steroids play a major role in the higher incidence of pancreatic cancer in male versus female rats.