Can Surgery Be Avoided? Exclusive Antibiotic Treatment for Pelvic Actinomycosis.

Pelvic actinomycosis is an uncommon, slowly progressing granulomatous infection that has been associated with the presence of intrauterine devices. Due to its unspecific clinical and radiologic findings, it can mimic pelvic or intra-abdominal malignancy leading to mutilating surgery of high morbidity. Rarely, diagnosis is made preoperatively and in most cases surgical intervention is necessary. The patient in our case is a 42-year-old female with an IUD for 15 years diagnosed with pelvic actinomycosis. Patient was uniquely diagnosed preoperatively through paracentesis and treated conservatively with prolonged antibiotic therapy and without any type of surgical intervention. Follow-up at 1 year showed almost complete radiologic resolution of the inflammatory mass, nutritional recovery, and absence of symptoms. Pelvic actinomycosis can be successfully diagnosed and treated medically without surgical interventions.

In utero exposure to bisphenol A alters the development and tissue organization of the mouse mammary gland.

Exposure to estrogens throughout a woman's life, including the period of intrauterine development, is a risk factor for the development of breast cancer. The increased incidence of breast cancer noted during the last 50 years may have been caused, in part, by exposure of women to estrogen-mimicking chemicals that are released into the environment. Here, we investigated the effects of fetal exposure to one such chemical, bisphenol A (BPA), on development of the mammary gland. CD-1 mice were exposed in utero to low, presumably environmentally relevant doses of BPA (25 and 250 microg/kg body weight), and their mammary glands were assessed at 10 days, 1 mo, and 6 mo of age. Mammary glands of BPA-exposed mice showed differences in the rate of ductal migration into the stroma at 1 mo of age and a significant increase in the percentage of ducts, terminal ducts, terminal end buds, and alveolar buds at 6 mo of age. The percentage of cells that incorporated BrdU was significantly decreased within the epithelium at 10 days of age and increased within the stroma at 6 mo of age. These changes in histoarchitecture, coupled with an increased presence of secretory product within alveoli, resemble those of early pregnancy, and they suggest a disruption of the hypothalamic-pituitary-ovarian axis and/or misexpression of developmental genes. The altered relationship in DNA synthesis between the epithelium and stroma and the increase in terminal ducts and terminal end buds are striking, because these changes are associated with carcinogenesis in both rodents and humans.

The mouse uterotrophic assay: a reevaluation of its validity in assessing the estrogenicity of bisphenol A.

The prevalence of synthetic chemicals in our environment that are capable of mimicking the female hormone estrogen is a growing concern. One such chemical, bisphenol A (BPA), has been shown to leach from a variety of resin-based and plastic products, including dental sealants and food and beverage containers, in concentrations that are sufficient to induce cell proliferation in vitro. The response to BPA in vivo has been varied; thus the aims of this study were to investigate a) whether BPA has an estrogenic effect in CD-1 mice, a strain that is useful for developmental studies; and b) whether the uterotrophic assay is a valid means of determining the estrogenicity of BPA by comparing it with other end points measured in the uterus. Immature female CD-1 mice were exposed to BPA in concentrations ranging from 0.1 to 100 mg/kg body weight for 3 days. Results showed that BPA induced a significant increase in the height of luminal epithelial cells within the uterus at concentrations of 5, 75, and 100 mg/kg and that BPA induced lactoferrin at concentrations of 75 and 100 mg/kg. A uterotrophic response (increase in uterine wet weight) was induced by 100 mg/kg BPA only. Further, the proportion of mice showing vaginal opening was greater after exposure to 0.1 and 100 mg/kg BPA, relative to the control animals and those receiving intermediate doses of BPA. These results demonstrate that BPA induces changes in the mouse uterus that differ depending on the exposure dose and the end point measured, and reveal that certain tissue effects show a nonmonotonic relationship with dose. These data also demonstrate that BPA induces estrogenic changes in the uterus of the CD-1 mouse, and highlight the need to reevaluate the validity of the mouse uterotrophic assay as an end point for determining the estrogenicity of suspected environmental estrogens.

Morphometric and endocrine analyses of the effects of nutrition on the testis of mature Merino rams.

The effects of nutrition on the testis were investigated in groups of five mature Merino rams that were fed either a sub-maintenance (low) diet or a supra-maintenance (high) diet for 69 days. Testosterone, oestradiol and inhibin were measured in blood plasma sampled simultaneously from jugular and testicular veins after an i.v. injection of 200 ng ovine LH kg-1. Plasma concentrations of testosterone, inhibin and oestradiol were higher in testicular than in jugular vein plasma for both diets (P < 0.01). After the LH injection, jugular plasma testosterone increased more rapidly (P < 0.01) in rams fed the high diet than in rams fed the low diet. This was not seen in the testicular vein. Oestradiol concentrations were higher in rams on the high diet than in those on the low diet, in both the testicular (P < 0.0001) and the jugular vein (P < 0.02). Diet did not affect inhibin concentrations. Testes were surgically removed and processed for light microscopy. Testicular mass and seminiferous tubule length and diameter were higher with the high diet than the low diet (P < 0.01). The number of Sertoli cell nuclei per testis was also affected (high diet: 120 +/- 6 x 10(8); low diet: 77 +/- 7 x 10(8); P < 0.001), whereas the proportion of testis occupied by Sertoli cell nuclei was not affected. The number of Leydig cells per testis was not affected by diet, but Leydig cells occupied a greater volume of testis in rams on the high diet than in those on the low diet (P < 0.001). The effects of nutrition on Leydig and Sertoli cells are consistent with changes in the endocrine and exocrine functions of the testis. The finding that Sertoli cell population was altered in adult rams may be explained by the GnRH-independent effects of nutrition.

Oxidative stress differentially regulates the expression of gamma-glutamyl transpeptidase mRNAs in the initial segment of the rat epididymis.

Reactive oxygen species (ROS) have a powerful cytotoxic effect on spermatozoa and have been implicated in spermatozoal dysfunction and male infertility. gamma-Glutamyl transpeptidase (GGT) is essential to the metabolism of the antioxidant glutathione and, as such, is believed to be important in protecting spermatozoa against oxidative stress. The aims of this study were 1) to establish in vitro conditions in which ROS were generated and 2) to determine whether oxidative stress regulated the expression of GGT mRNAs I-IV in the initial segment of the epididymis. Initial segments were collected from adult male rats and incubated in culture media to which ROS-generating compounds, hypoxanthine and xanthine oxidase, were added. By 6.5 hours, incubation of tissue in high-oxidative stress conditions caused a 56% decrease in reduced glutathione concentration, a concomitant 240% increase in oxidized glutathione concentration, and a 25% decrease in adenosine triphosphate concentration. RNase protection analyses demonstrated an approximate 70% up-regulation of GGT mRNAs II-IV in a differential manner, depending on the concentration of oxidizing agents and the type of ROS generated. gamma-Glutamyl transpeptidase mRNA I was not expressed. These results support the hypothesis that expression of GGT mRNAs is regulated by oxidative stress in the initial segment of the rat epididymis.

Effects of ischaemia on the caput epididymis and its relationship to higher epididymal obstruction: a qualitative study in the ram.

The objective of this study was to determine if localized ischaemia of the caput epididymis in the ram causes morphological changes similar to those characteristic of higher epididymal obstruction in humans. This was tested by performing unilateral occlusion of the superior epididymal artery in 10 rams. At the end of 4 (n = 5) or 28 weeks (n = 4), rams were castrated and the testes and epididymides were weighed. Analysis of histological sections at the light microscope level provided quantitative data on lumen diameter and epithelial cell height of the efferent ducts. Qualitative analysis and specific histochemical stains for identification of lipofuscin pigment provided further information on tissue changes. Electron microscopy was performed on the efferent ducts to assess ultrastructural changes. The results revealed that localized ischaemia of the proximal epididymis caused a dramatic change in tubule calibre of the efferent ducts and initial segment resulting in obstruction of the distal lumina. These changes were more severe following 28 weeks of arterial occlusion. The epithelial cells of the proximal region showed an increase in the number of lysosomes and they became active in phagocytosis of spermatozoa. Lipofuscin pigment accumulated within the epithelial cells and also in macrophage-like cells that had invaded the lumina, interstitium and intra-epithelial regions of the ducts. On the basis of these observations we conclude that the tissue changes which occur in the ram epididymis as a result of localized ischaemia show a striking similarity to those seen in men exhibiting higher epididymal obstruction. This suggests the possible implication of vascular disorders in the aetiology of obstructive azoospermia in men.

Relationship between testicular morphology and sperm production following ischaemia in the ram.

Arteriosclerosis was induced in the internal spermatic artery of rams to determine if this condition is implicated in the aetiology of testicular pathology which causes male infertility. Data were collected on sperm concentration and motility for 56 days following surgery to provide an index of testicular function. Testes were then weighed and a testicular biopsy score count was performed on histological sections to assess spermatogenic potential of seminiferous tubules. Vascular disturbance caused focal damage of the seminiferous epithelium, similar to that seen among infertile men, and a reduction in ejaculate volume, sperm concentration and sperm motility. Sperm concentration decreased following ischaemia yet was maintained to some degree by a germ-cell depleted spermatogenic epithelium. Normal testicular morphology was maintained above a testis weight of about 120 g (for an individual testis), but below this threshold spermatogenesis was severely impaired. In conclusion, these data have provided information on the relationship between testicular morphology and function following ischaemia in the ram. Furthermore, the morphological changes induced in the testis were similar to those seen among infertile men and, by their focal nature, could explain the distinction between oligozoospermia and azoospermia in men exhibiting spermatogenic arrest.

Testicular morphology and androgen profiles following testicular ischaemia in rams.

Arteriosclerotic changes were induced in the internal spermatic artery of rams to determine whether there is a link between this condition and some pathological conditions of the testes, similar to those that cause infertility in men. Eight weeks after the induction of testicular ischaemia, blood plasma was collected simultaneously from the jugular and spermatic veins after an LH injection (10 micrograms) and assayed for testosterone. The rams were then castrated and sections of the testis, ductuli efferentes and spermatic cord were examined quantitatively and qualitatively. Vascular disturbance decreased the percentage of normal spermatogenic epithelium (P < 0.01) and the diameter of the seminiferous tubules (P < 0.001). These effects were accompanied by an increase in the percentage of the interstitial region within the testis (P < 0.05). Macrophages, lymphocytes and other inflammatory cells became numerous in the interstitium as damage to the seminiferous epithelium progressed. The most striking feature of the ischaemic testis was the focal damage of the spermatogenic epithelium, that is, sections of the same testis exhibited both normal and germ cell-depleted seminiferous tubules. Concentrations of testosterone in peripheral plasma were not significantly altered by either unilateral or bilateral testicular ischaemia; however, the concentration of testosterone was higher in the experimental spermatic vein than in the contralateral spermatic vein (P < 0.05) as was the ratio of LH:testosterone (P < 0.05). Unilateral vascular disturbance of the testis did not cause damage in the contralateral testis. The ductuli efferentes of these rams also showed structural changes as a result of vascular disturbance.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of dietary zinc deficiency on the reproductive system of young male sheep: testicular growth and the secretion of inhibin and testosterone.

The effects of dietary zinc deficiency on testicular development in young Merino rams (initial live mass, 22 kg) were tested. Four groups of five rams were fed ad libitum with diets containing 4, 10, 17 or 27 micrograms Zn g-1. To control the effects of loss of appetite caused by zinc deficiency, a fifth group (pair-fed control) was fed the diet containing 27 micrograms Zn g-1, but the amount of feed offered was restricted to that eaten voluntarily by the zinc deficient (4 micrograms Zn g-1) rams they were paired with. After 96 days on the diets, epididymal and testicular masses did not differ significantly between the animals fed 10, 17 or 27 micrograms Zn g-1 ad libitum, but were significantly lower in pair-fed controls, and lowest in the zinc-deficient animals. Testicular responsiveness to LH, as measured by testosterone production, increased substantially in most rams as the experiment progressed, the only exception being the zinc-deficient group, in which the response to LH was lower than in any of the other groups. Testicular concentrations of zinc and testosterone were lower in the zinc-deficient animals than in all the other groups. Plasma inhibin concentrations fell as the experiment progressed in rams fed 17 and 27 micrograms Zn g-1 ad libitum, but not in the other groups. The pair-fed control rams had smaller seminiferous tubules and less lumen development than did the controls fed ad libitum (27 micrograms Zn g-1), which were similar to the animals fed 10 or 17 micrograms Zn g-1. In zinc-deficient rams, the tubule development was further retarded and the interstitial regions were more extensive than in the other groups. We conclude that the overall effect of zinc deficiency on testicular development is due to a combination of a non-specific effect (low gonadotrophin concentrations caused by the low feed intake) and a specific effect due to the lack of zinc. The zinc-specific effect is localized within the testis where it reduces the development of the capacity to produce testosterone, leading to low intratesticular concentrations of testosterone, a critical factor for the growth, development and function of the seminiferous tubules.

A quantitative description of the epididymis and its microvasculature: an age-related study in the rat.

The morphology of the epididymal duct and, in particular, the epididymal microvasculature was examined at the light microscope level in young sexually-mature rats (3-5 months) and aged rats (18 months) to investigate the structural changes that may occur within the organ as a result of ageing, and which may predispose the organ to pathological changes. Quantitative data on the microvascular network of the epididymis (percentage of capillaries in the interstitial region, average area and surface density of the capillary lumen) were collected in 4 regions of the epididymis: the initial segment, caput, corpus and cauda. Epithelial cell height, epididymal lumen diameter, number of smooth muscle cells and percentage of smooth muscle surrounding the duct were also assessed within the same 4 regions. The data for both young and aged groups revealed a trend of decreasing capillary size from the initial segment of the epididymis to the cauda by 23%. Further, the percentage of capillaries within the interstitial region of the epididymis decreases dramatically (52%) in the same direction. The possible contribution of lymphatic capillaries to the data is discussed. The data revealed that none of the parameters assessed changed significantly up to 18 months of age. The quantitative data on the microvascular morphology of the epididymis presented in this study provide the basis for subsequent studies directed at the blood flow dynamics of the organ.

Quantitative studies of hydroperoxide reduction by prostaglandin H synthase. Reducing substrate specificity and the relationship of peroxidase to cyclooxygenase activities.

The peroxidase activity of prostaglandin H (PGH) synthase catalyzes reduction of 5-phenyl-4-pentenyl hydroperoxide to 5-phenyl-4-pentenyl alcohol with a turnover number of approximately 8000 mol of 5-phenyl-4-pentenyl hydroperoxide/mol of enzyme/min. The kinetics and products of reaction establish PGH synthase as a classical heme peroxidase with catalytic efficiency similar to horseradish peroxidase. This suggests that the protein of PGH synthase evolved to facilitate peroxide heterolysis by the heme prosthetic group. Comparison of an extensive series of phenols, aromatic amines, beta-dicarbonyls, naturally occurring compounds, and nonsteroidal anti-inflammatory drugs indicates that considerable differences exist in their ability to act as reducing substrates. No correlation is observed between the ability of compounds to support peroxidatic hydroperoxide reduction and to inhibit cyclooxygenase. In addition, the resolved enantiomers of MK-410 and etodolac exhibit dramatic enantiospecific differences in their ability to inhibit cyclooxygenase but are equally potent as peroxidase-reducing substrates. This suggests that there are significant differences in the orientation of compounds at cyclooxygenase inhibitory sites and the peroxidase oxidation site(s). Comparison of 5-phenyl-4-pentenyl hydroperoxide reduction by PGH synthase and horseradish peroxidase reveals considerable differences in reducing substrate specificity. Both the cyclooxygenase and peroxidase activities of PGH synthase inactivate in the presence of low micromolar amounts of hydroperoxides and arachidonic acid. PGH synthase was most sensitive to arachidonic acid, which exhibited an I50 of 0.6 microM in the absence of all protective agents. Inactivation by hydroperoxides requires peroxidase turnover and can be prevented by reducing substrates. The I50 values for inactivation by 15-hydroperoxy-5,8,11,13-eicosatetraenoic acid are 4.0 and 92 microM, respectively, in the absence and presence of 500 microM phenol, a moderately good reducing substrate. The ability of compounds to protect against hydroperoxide-induced inactivation correlates directly with their ability to act as reducing substrates. Hydroquinone, an excellent reducing substrate, protected against hydroperoxide-induced inactivation when present in less than 3-fold molar excess over hydroperoxide. The presence of a highly efficient hydroperoxide-reducing activity appears absolutely essential for protection of the cyclooxygenase capacity of PGH synthase. The peroxidase activity is, therefore, a twin-edged sword, responsible for and protective against hydroperoxide-dependent inactivation of PGH synthase.(ABSTRACT TRUNCATED AT 400 WORDS)

Spectral properties of the higher oxidation states of prostaglandin H synthase.

Prostaglandin H (PGH) synthase reacts with organic hydroperoxides and fatty acid hydroperoxides on a millisecond time scale to generate an intermediate that is spectrally similar to compound I of horseradish peroxidase. Compound I of PGH synthase is converted to compound II within 170 ms. Compound II decays to resting enzyme in a few seconds. Thus, the peroxidase reaction of PGH synthase appears to involve a cycle of native enzyme, compound I, and compound II, typical of heme-containing peroxidases. The Soret absorption maximum of compound I appears to occur at 412 nm but a small amount of compound II may be present. Soret maxima occur at 420, 433, and 419 for compound II, the ferrous enzyme, and the oxyferrous enzyme (compound III), respectively. Rapid scan analysis of the reaction of PGH synthase with arachidonic acid reveals the absorbance of compound II but no evidence for ferrous or oxyferrous enzyme.

Enzymatic reduction of 5-phenyl-4-pentenyl-hydroperoxide: detection of peroxidases and identification of peroxidase reducing substrates.

5-Phenyl-4-pentenyl-hydroperoxide (PPHP) is reduced to 5-phenyl-4-pentenyl-alcohol (PPA) by plant and animal peroxidases in the presence of reducing substrates. PPHP and PPA are rapidly isolated with solid phase extraction, separated by isocratic reverse-phase high-performance liquid chromatography, and quantitated with a fixed-wave-length ultraviolet detector. The procedure described is suitable for detecting peroxide-reducing enzymes, determining the kinetic properties of heme- and non-heme-containing peroxidases, and evaluating oxidizable compounds as reducing substrates for peroxidases. Horseradish peroxidase (HRP) and phenol reduce PPHP with a Km for phenol of 252 microM and a turnover number of 1.05 X 10(4) min-1. Under similar conditions, the Km of HRP for PPHP is 18 microM in the oxidation of guaiacol. A series of 21 compounds was evaluated for the ability to serve as reducing substrates for HRP. The results indicate that the procedure described can not only identify compounds that are reducing substrates but also rank them for relative activity. This may provide a new method with which to identify novel antithrombotic, antimetastatic, or anti-inflammatory drugs as well as to detect and characterize mammalian peroxidases.