Prenatal vitamin C and E supplementation in smokers is associated with reduced placental abruption and preterm birth: a secondary analysis.

OBJECTIVE: Smoking and pre-eclampsia (PE) are associated with increases in preterm birth, placental abruption and low birthweight. We evaluated the relationship between prenatal vitamin C and E (C/E) supplementation and perinatal outcomes by maternal self-reported smoking status focusing on outcomes known to be impacted by maternal smoking. DESIGN/SETTING/POPULATION: A secondary analysis of a multi-centre trial of vitamin C/E supplementation starting at 9-16 weeks in low-risk nulliparous women with singleton gestations. METHODS: We examined the effect of vitamin C/E by smoking status at randomisation using the Breslow-Day test for interaction. MAIN OUTCOME MEASURES: The trial's primary outcomes were PE and a composite outcome of pregnancy-associated hypertension (PAH) with serious adverse outcomes. Perinatal outcomes included preterm birth and abruption. RESULTS: There were no differences in baseline characteristics within subgroups (smokers versus nonsmokers) by vitamin supplementation status. The effect of prenatal vitamin C/E on the risk of PE (P = 0.66) or PAH composite outcome (P = 0.86) did not differ by smoking status. Vitamin C/E was protective for placental abruption in smokers (relative risk [RR] 0.09; 95% CI 0.00-0.87], but not in nonsmokers (RR 0.92; 95% CI 0.52-1.62) (P = 0.01), and for preterm birth in smokers (RR 0.76; 95% CI 0.58-0.99) but not in nonsmokers (RR 1.03; 95% CI 0.90-1.17) (P = 0.046). CONCLUSION: In this cohort of women, smoking was not associated with a reduction in PE or the composite outcome of PAH. Vitamin C/E supplementation appears to be associated with a reduction in placental abruption and preterm birth among smokers.

PP170. Prenatal vitamin C and E supplementation is associated with a reduction in placental abruption and preterm birth in smokers.

INTRODUCTION: Maternal tobacco use increases the incidence of numerous adverse pregnancy outcomes including miscarriage, small for gestational age infants, spontaneous preterm birth and placental abruption. OBJECTIVES: We evaluated the relationship between prenatal vitamin C/E supplementation and perinatal outcomes by maternal smoking status. METHODS: Secondary analysis of a multicenter trial of vitamin C/E starting at 9-16 weeks in low-risk nulliparous women with singletons. We examined the effect of vitamin supplementation by reported smoker or non-smoker at time of randomization. The primary outcomes were preeclampsia (new onset hypertension and proteinuria) and a composite outcome of severe pregnancy associated hypertension (severe hypertension OR mild or severe hypertension with elevated liver enzymes, elevated serum creatinine, thrombocytopenia, eclamptic seizure, fetal growth restriction, medically indicated preterm birth or perinatal death). Perinatal outcomes included preterm birth and abruption. The Breslow-Day test was used to ascertain whether there was an interaction between smoking status and vitamin supplementation. RESULTS: Of 9969 women, 4993 received vitamins C/E and 4976 received placebo. The prevalence of smoking (15.6% overall; 788 vitamin, 763 placebo) was similar in treatment groups. The analysis of vitamin C/E by smoking status for perinatal outcomes is given (Table 1). The effect of prenatal vitamin C/E on the risk preeclampsia or pregnancy associated hypertension composite outcome did not differ by smoking status. Vitamin C/E was protective for placental abruption and preterm birth among smokers. CONCLUSION: The effect of vitamin C/E supplementation on preeclampsia/pregnancy associated hypertension did not differ by smoking status. However, vitamin C/E supplementation was associated with a >40% reduction in placental abruption and >30% reduction in preterm birth among smokers warranting further study. ACKNOWLEDGEMENT: This project supported in part by National Institutes of Health Grant PO1-HD30367.

Phenylephrine-induced neurogenic prostatitis facilitates the promotion of PIN-like lesions in rats.

BACKGROUND: Atypical prostate hyperplasia, resembling PIN lesions in men, together with chronic inflammatory exudates, have been reported after chronic administration of phenylephrine (PE). The present study aims to characterize the evolution of the expression of typical leukocyte markers immunohistochemically as correlated to the promotion of PIN lesions. METHODS: Adolescent rats received injections of PE (subcutaneously) (10 mg/kg BW/per day) and were euthanized 3, 8 hr, 1, 3, 7, 14 days thereafter. The dissected prostates were fixed in formalin, and paraffin embedded sections were cut and immunoreacted. RESULTS: PE exerted a time related bi-phasic effect on the rat prostate. A first inflammatory reaction phase took place at 3-8 hr post injection characterized by vascular dilatation, congestion, edema, and massive leukocytic infiltrate, mainly of ED1+ cells. At 24 hr, the amount of ED1+ cells decreased approaching the equivalent values of ED2+, CD8+ cells, and mastocytes. Their values remained relatively high for the rest of the experimental period. A second phase of proliferative changes, consisting of healing fibrosis as well as dysplastic epithelial lesions, similar to human PIN lesions, appeared on the 7th day. CONCLUSIONS: Neuro-immune interactions promote prostatic fibrosis and dysplastic changes; these being preceded by an acute and transient inflammatory reaction.

Effects of senescence and citral on neuronal vacuolar degeneration in rat pelvic ganglia.

A significant part of the morbidity in elderly men involves pelvic organs and their autonomic neural regulation. Environmental stimuli also impair the structure and function of pelvic organs. One of these factors is citral, a widely-used cosmetic fragrance constituent, which causes severe prostatic hyperplasia in rats. In this study, we assessed the effect of topical administration of citral (30 days) on the morphology of pelvic ganglia (PG) in young adult and old Wistar rats. Neuronal vacuolar degeneration with preserved nuclei of PG neurons was observed in untreated senescent, but not young rats. Citral significantly increased the rate of vacuolated neurons in old rats (from 3 to 14%), but only slightly in young ones (from 0 to 0.5-0.3%). Similar lesions were not found in inferior cervical or celiac ganglia, in either group. This shows that environmental stimuli enhance age-related processes of vacuolar neuronal degeneration in PG, and may contribute to the dysfunction of pelvic organs in the elderly.

Dominant negative Met reduces tumorigenicity-metastasis and increases tubule formation in mammary cells.

Activation of the Met tyrosine kinase growth factor receptor by its ligand HGF/SF has been shown to increase in vitro invasiveness in epithelial cell lines. To study the effect of Met-HGF/SF signaling in breast cancer cells, we transfected met, hgf/sf and dominant negative (DN) forms of met into the poorly differentiated metastatic murine mammary adenocarcinoma cell line DA3. These cells express moderate levels of endogenous Met, which is rapidly phosphorylated in response to HGF/SF treatment. Met+hgf/sf transfection results in significantly increased tumorigenic and metastatic activity in vivo accompanied by reduced tubule formation. DA3 cells transfected with DN forms of Met (DN-DA3) exhibit reduced Met phosphorylation following exposure to HGF/SF. Furthermore, as compared to the parental cells, the DN-DA3 cells exhibit diminished in vitro scattering and invasiveness, while in vivo they display greatly reduced tumorigenicity and spontaneous metastasis. Tumors emanating from DN-DA3 cells injected to BALB/C mice are highly differentiated and display extensive tubule formation. These results suggest that Met-HGF/SF signaling is a determining factor in the delicate balance between differentiation/tubule formation and tumorigenicity-metastasis. Oncogene (2000) 19, 2386 - 2397

Spontaneous hyperplasia of the ventral lobe of the prostate in aging genetically hypertensive rats.

Recent studies have shown that the prostatic autonomic innervation takes part in its homeostasis and growth. Other works showed that spontaneously hypertensive rats (SHR) show excessive sympathetic activity, accompanied by lower urinary tract symptoms, increased growth capacity of prostatic stromal cells, and increased levels of androgens and their receptors. Furthermore, young SHR were reported to present incipient stages of benign prostatic hyperplasia (BPH). The aim of the present study was to examine whether this strain indeed develops spontaneous BPH with age, and can thus serve as a genuine natural model for this disorder. For this purpose, ventral lobes of prostates of one-year-old, male SHR and their normotensive counterparts, Wistar Kyoto (WKY) rats, were examined histopathologically, and the degree of hyperplasia was evaluated according to a score-chart protocol (histoscore). SHR exhibited severe adenomatous spontaneous BPH, characterized by piling-up of epithelial cells, with papillary formations, accompanied by a mild increase in the amount of fibrocytes and smooth muscle cells in the stroma. This was reflected by histoscore values of 38 +/-2. Thickening of prostatic arterioles also was noted, as well as mild chronic inflammatory exudate. WKY rats did not show any of these features of BPH despite their age (histoscore 17 +/- 3, significantly different from that of SHR). We conclude that SHR can serve as a rodent model for the spontaneous development of BPH with age, most probably due to the excessive neuroendocrine activity characteristic of this rat strain.

Induction of atypical prostatic hyperplasia in rats by sympathomimetic stimulation.

BACKGROUND: Prostatic innervation may participate in its homeostasis and growth. alpha-Adrenergic inhibition alleviates clinical symptoms in benign prostatic hyperplasia. However, the prostatic effect of adrenergic agonists has not been investigated. This study deals with the prostatic effect of subchronic sympathomimetic stimulation. METHODS: Male rats received daily subcutaneous injections of the alpha-adrenergic agonist phenylephrine, 1, 10, or 20 mg/kg per day, the beta-adrenergic agonist isoproterenol, 1, 2.5 or 5 mg/kg per day, or saline, for 30 days, and the prostates were removed for histopathological examination. RESULTS: Phenylephrine induced atypical prostatic hyperplasia, characterized by piling-up with papillary and cribriform patterns, and budding-out of epithelial cells. It decreased prostatic secretions and total weight. Similar results were observed in orchidectomized rats receiving exogenous testosterone supplementation. Isoproterenol had no prostatic morphological effect. CONCLUSIONS: These results raise the possibility that sympathetic stimuli play a role in normal and aberrant growth and differentiation of prostatic epithelium, and suggests neurostimulants-treated animals as a model to study the etiology and development of prostatic hyperplasia.

Role of chronic inflammation in the promotion of prostatic hyperplasia in rats.

PURPOSE: Chronic prostatitis is a common histopathological finding in prostatectomized patients. A possible interrelationship between the presence of leukocyte exudate and the extent of the hyperplastic lesions has been suggested. The aim of the present study was to analyze the effect of the immunomodulator compounds, Complete Freund Adjuvant (CFA) and cyclosporin A (CsA), administered alone or together with citral on the induction and extent of rat prostatic hyperplasia. MATERIALS AND METHODS: Adolescent Wistar rats (42 days old) were given citral alone or combined with CFA or CsA for one month. Semiquantitative analysis of the extent of the hyperplastic lesions was made with the histoscore protocol. RESULTS: CsA did not induce hyperplastic changes or abolish the ability of citral to promote hyperplastic changes or to affect the extent of the lymphocytic exudate in the stroma. CFA itself, however, had a proliferative action on the prostatic epithelium, and it augmented the hyperplastic changes induced by citral and even induced atypical transformations of the acinar epithelium. CONCLUSIONS: Immunoinflammatory stimulators might play a role in the prostatic epithelial cell growth and proliferation processes, most probably by modulation of the cytokine system.

Human prostatic tissue constituents respond differently to thermal injury induced by Nd-YAG laser ITT fiber applied ex-situ.

BACKGROUND AND OBJECTIVE: The human prostate is a glandular organ that has intervening fibromuscular elements. The objective of this study was to evaluate the thermocoagulative effect on the different components of the organ exposed to Nd-YAG laser irradiation using a new ITT optic fiber design (Interstitial Thermal Therapy). STUDY DESIGN/PATIENTS AND METHODS: Twelve fresh transvesical prostatectomy specimens were irradiated ex-situ at 4 to 6 sites using an ITT fiber with laser energies ranging from 900 to 4500 joules (J) applied at times ranging from 60 to 300 seconds corresponding to 10 to 15 watts. The specimens were serially sectioned to include the proximal, mid and distal regions of the centrally coagulated channel. An ocular grid was utilized to measure the radius (r) of thermal injury from the margin of the centrally coagulated channel to the furthermost detectable area of thermally induced changes. RESULTS: The maximal extent of thermal injury was seen at the mid-length of the irradiated channel at energy of 2700 J (15w/180 sec). The fibromuscular compartment of the prostate was more affected (r = 855 microm) and showed more interstitial vacuolization and charring than the adenomatous one (r = 495 microm). Utilization of higher energy doses did not significantly alter the depth of injury but did reduce the extent of interstitial vacuolization of both components. CONCLUSION: The study indicates that the extent of laser induced coagulative necrosis depends on the histologic architecture of the prostate and also varies in extent along the length of the channel surrounding the fiber.

Citral and testosterone interactions in inducing benign and atypical prostatic hyperplasia in rats.

Citral is a monoterpene in wide use as an aromatic supplement in the cosmetics and food industries. Previous studies in our laboratory have shown that cutaneous application of citral on adolescent rats may induce benign prostatic hyperplasia (BPH)-like and even atypical hyperplastic changes in the ventral lobes. In the present study we investigate the possible interactions between citral and serum testosterone levels on the induction of hyperplastic changes in the ventral prostate of adolescent rats. In addition, the study includes a comparative analysis of normal intact rats showing circadian variations of serum testosterone levels and rats in whom this rhythmic pattern was abolished either by excessive supplementation of exogenous androgen or by castration. Our results demonstrate an induction of benign as well as atypical prostatic hyperplasia following citral application. The most severe atypical changes were noted in the citral-treated rats with high serum testosterone levels. Although the mechanism of action of citral is yet unknown, the present results suggest a synergism between citral and testosterone resulting in hyperplastic changes in the rat ventral prostate. In addition, the results reconfirm that serum testosterone levels fluctuate according to a circadian rhythm in intact young and adolescent male rats. The application of citral tends to lower the morning circadian peaks, and the circadian pattern was abolished in orchiectomized rats and in those treated with testosterone implants.

Intraarticular minocycline injection in experimental synovitis.

Minocycline, a semi-synthetic tetracycline, was injected into one hind joint of twenty-two rabbits with zymosan-induced arthritis, while the contralateral joint served as a control. A local inflammatory reaction was observed a few days after the zymosan injection. Most clinical parameters such as knee diameter, systemic temperature, sedimentation rate and blood cell count did not change throughout the experiment both in control and minocycline treated rabbits. However, the zymosan platelet counts rose from 3.4x10(5)/microL to 5x10(5)/microL, as well as the level of serum fibrinogen (from 99 mg% to 370 mg%). Microscopically, a perivascular infiltrate consisting of lymphocytes and polymorphonuclear cells was seen. Lymphoid follicles as well as plasma cells epitheloid and giant cells were also observed. A mild tendency to fibrosis and lesser inflammatory reaction in the minocycline treated knees was noted. Our data suggest that intraarticular minocycline treatment did not alleviate the course of the rheumatoid-like inflammatory reaction of the knee joint.

Measured limits to contamination of optical surfaces by elastomers in vacuum.

We have monitored the reflectivity of mirrors that were exposed to a fluoroelastomer (3M-Fluorel 2176) and a room-temperature vulcanizing silicone rubber (RTV-615) in vacuum. The 95% confidence limit on the decrease of mirror reflectivities was less than 0.35 ppm/week for Fluorel and <0.29 ppm/week for RTV-615.

Immediate vasoactive effect of citral on the adolescent rat ventral prostate.

The vasoactive effect of a fragrance compound, citral, on rat ventral prostate is presented. A combined method of Indian-ink perfusion and postfixation transparency was used. One single dose of citral applied on the back skin of the rat induced an immediate triple response-like effect on the prostatic vascular bed. A slight vasodilatation appeared already after 2 1/2 min, followed by a marked vasoconstriction process after 5-min postcitral administration. Toward the period of 10 min, a second vasodilatation process was noted which persisted for the following 24 hr of observation. This study has demonstrated that changes in the microvascularization of the rat prostate can be estimated by counting the number of carbon-stained blood vessels. The results obtained by this technique are closely related with the findings in the vascular network profile of the macroangiographic observations. The mechanism of action of citral on the microvascularization of the rat ventral prostate is yet unknown. Based on the present findings, we suggest that the neoplastic capacity of citral upon the prostatic epithelia is activated via a nonspecific inflammatory reaction modulated either by local release of neurotransmitters or throughout a direct effect of citral on the endothelial cells.

Comparative study of experimentally induced benign and atypical hyperplasia in the ventral prostate of different rat strains.

The induction of prostatic lesions in the rat by hormonal manipulation and/or chemical carcinogens in different strains is well documented in literature. However, the selection of a strain for such studies was merely based on the laboratory husbandry facilities rather than on the animal genotype. It is well accepted that the morphology and function of the prostate exhibit a marked species-specific differences that makes extrapolation to other animals and man difficult. Our group has developed an experimental model for the study of rat prostatic hyperplasia and/or dysplasia induced by citral--a flavor constituent. Previous observations have revealed that the Wistar rat ventral prostate is reactive to citral, especially during its adolescent growth period as well as during the redifferentiation stage among postcastrated rats treated with exogenous testosterone. The present study presents data on the ventral prostate susceptibility of three additional strains selected by their interrelated endocrine morbidity. In order to facilitate an objective comparative analysis of the prostatic lesions a histopathological score chart was elaborated. The present findings showed that the Wistar and Sprague-Dawley strains were the most susceptible to developing benign and atypical prostatic hyperplasia both in intact and postcastrated rats. The F344 and ACl/Ztm rat strains remained refractory toward all of these treatments. Our data provide evidence that the strain genotype and, more precisely, their endocrine background, play a determinative role in the prostatic responsiveness towards citral. It is suggested that additional consideration be given to the selection of an adequate animal strain when studying experimental neoplastic transformation, especially when hormonal interactions might be involved.

Translocation of bacteria due to direct mucosal damage caused by Gastrografin. An experimental study in newborn rats.

A study was carried out on 40 newborn rats to determine the effect of Gastrografin (a hyperosmolar solution, 1700 mosm/liter) on the gastrointestinal tract. All the newborns received an inoculum of Klebsiella bacteria to the gastrointestinal tract. Thirty received, in addition to maternal milk, a feeding of Gastrografin twice daily. The Gastrografin was found to cause severe and irreversible damage to the mucosa of the small intestine, causing the death of 24 rats less than a week after start of the experiment. The 10 rats who received no Gastrografin and served as controls showed no signs of disease or damage to the intestinal tract. Cultures taken from the peritoneal cavity after sacrifice were all positive for Klebsiella in the Gastrografin group (30 rats) and negative in the controls (10 rats). This study has therefore demonstrated that severe damage to the small intestine mucosa will lower the intestinal barrier and lead to transmural translocation of bacteria into the peritoneal cavity.

Morphometric studies of pancreatic acinar granule formation in NCTR-Balb/c mice.

NCTR-Balb/c mice are afflicted with a cholesterol lysosomal storage disorder stemming from a defect in intracellular cholesterol processing. The clinical and biochemical abnormalities expressed in the mice resemble Niemann-Pick type C and D disorders in humans. One of the proposed mechanisms to explain the pathophysiology of the disorder implies a defect in the process of membrane transport that normally takes place in the vesicular movement of cholesterol to specific target sites in the cell. Secretory granule formation in pancreatic acinar cells is one of the biological processes known to involve massive membrane flow. Thus, we have undertaken a morphometric study of the regranulation mechanism in the pancreatic acinar cells of the mutant mice, as a way of studying cellular membrane movement. Electron micrographs of pancreatic acinar cells from mutant and normal mice were taken at several time points after extensive degranulation induced by pilocarpine injection. Two hours after stimulation the pancreatic cells demonstrated a complete loss of granules, and at later time points newly formed granules appeared. Identical unit granule volumes were observed in both groups, indicating that the progranules were of normal size. However, the rate of granule formation and maturation was reduced in the mutant mice, which might be the result of a defect in membrane function.