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The natural anti-cancer agent bromelain is found to be beneficial for either single or multi-targeted therapy in gastric and skin carcinoma, by inhibiting cancer cell growth. Importantly, the presence of peroxidase enhances its biological efficiency. We have now evaluated a panel of cancer cell lines with bromelain in presence or absence of peroxidase to identify that the combination has higher apoptosis inducing potential in all those cell lines. Bromelain plus peroxidase (BM-PR) inhibited acute myeloid (K562) cell proliferation and altered the morphological features. Incidence of apoptosis was established by using annexin V exposure and this was confirmed that the cell cycle was arrested at G0/G1 phase in a concentration-dependent manner. BM-PR increased the intracellular ROS level and altered the mitochondrial membrane potential, as detected using dichlorofluores cin diacetate (DCFDA). It also regulated the expression of apoptosis-related proteins like Bax, Bcl2, caspase-3 and cytochrome besides causing up-regulation of p53 as determined by western blot analysis. These results suggest that BM-PR from pineapple induces apoptosis better than only bromelain in acute myeloid leukemia cells possibly via mitochondria dependent pathway.

The present study records the occurrence of major lipid fractions and their fatty acids in a digenetic trematode parasiteParamphistomum cervi, and the rumen fluid and liver of the goat (Capra hircus). The amount of neutral lipids (NL), glycolipids (GL) and phospholipids (PL) of goat liver, rumen fluid and of the parasite shows that the rumen fluid is rich in NL, which is also in maximum quantity in the parasite, while the liver is rich in PL followed by NL. The number of fatty acids of total lipids (TL), NL and PL is greater in the parasite than those of the liver and rumen fluid. The number of fatty acids of GL is higher in the liver than in the parasite and the rumen fluid. Comparison of unsaturated fatty acid (UFA), C18total and C18UFA of TL, NL, GL and PL of the liver, rumen fluid and the parasite shows that the amount of C18UFA is higher inP. cerviin all the lipid fractions, except for GL, than in the rumen fluid and the liver. The results reveal thatP. cerviabsorbs a greater number of fatty acids than its host.

Manganese (Mn) is an essential mineral that is found in varying amounts in aerosols or dust. Exposure to atmospheric Mn at high concentration is a risk factor in humans that can manifest as neuronal degeneration resembling Parkinson's disease (PD). Since the underlying mechanism of Mn and dopamine (DA) interaction-induced cell death remains unclear, here, we showed that Mn exposure alone to mesencephalic cells for 24h induced minimal apoptotic cell death. However, cells pre-exposed to DA for 2h accelerated Mn-induced apoptosis. The vulnerability of Mn-induced apoptotic cell death to DA was determined by measuring lactate dehydrogenase (LDH) and Apoptag TUNEL staining (terminaldeoxynucleotidyl transferase DNA labeling). This was further confirmed by the cell viability assay to support our hypothesis that DA at the cellular level interacts with Mn and causes cells to be more susceptible. Pretreatment with nitric oxide blocker (7-nitroindazole, 7-NI), vitamin E or NF-kappaB inhibitor (SN50) significantly protected the cells from Mn and DA interaction-induced reactive oxygen species (ROS) and apoptosis. Western blot analysis showed that Mn in the presence of DA markedly induced induction of NOS (iNOS) expression. Pretreatment with 7-NI, SN50 or vitamin E significantly attenuated increased iNOS expression indicating that iNOS expression is regulated by ROS and the transcription factor NF-kappaB. Further, the generation of ROS as an early event in Mn and DA interaction is not controlled by NF-kappaB as SN50 pretreatment did not prevent ROS. These findings suggest that NF-kappaB induction and the activation of nitric oxide synthase through ROS represent a proximate mechanism for Mn-induced neurotoxicity.

Summary The coexistence of brain parenchymal cysts at various stages of evolution, both intraventricular and subarachnoid racemose, is reported in a patient with neurocysticercosis. The condition has a variety of presentations, depending on the location of the cyst. This case is of particular interest because of the rarity of this condition in India.

Time-synchronous development of endometrium and embryo under adequate progesterone dominance is considered integral to the process of blastocyst implantation in the human. It now appears that hypothesis-driven and candidate-based deductive approach fails to explain this complex control process. We propose a systems biology approach to elucidate the control process underlying the physiological basis of successful interfacing between embryo and endometrium towards blastocyst implantation. Elucidation of the time course pattern of transcriptomics involved in the process of blastocyst implantation in mid-luteal phase endometrium with and without progesterone dominance, as well as, with and without viable embryo shall elaborate upon the polygenic and multifactorial nature of the process of blastocyst implantation. Accordingly, a large scale homeodynamic model of hierarchical arrangement of functional networks of regulatory genomic expressional elements at the level of endometrial receptivity shall emerge. It is anticipated that such a systems biology approach shall provide an integrated picture of the process and shall also open up novel areas of basic, strategic and translational research in the biology of blastocyst implantation.

Japanese encephalitis virus (JEV) induces an acute infection of the central nervous system, the pathogenic mechanism of which is not fully understood. To investigate host response to JEV infection, 14-day-old mice were infected via the extraneural route, which resulted in encephalitis and death. Mice that received JEV immune splenocyte transfer were protected from extraneural JEV infection. Pathology and gene expression profiles were then compared in brains of mice that either succumbed to JEV infection or were protected from infection by JEV immune cell transfer. Mice undergoing progressive JEV infection had increased expression of proinflammatory cytokines, chemokines, and signal transducers associated with the interferon (IFN) pathway. In contrast, mice receiving immune cell transfer had increased production of the Th2 cytokine IL-4, and of IL-10, with subdued expression of IFN-gamma. We observed IL-10 to be an important factor in determining clinical outcome in JEV infection. Data obtained by microarray analysis were further confirmed by quantitative RT-PCR. Together, these data suggest that JEV infection causes an unregulated inflammatory response that can be countered by the expression of immunomodulatory cytokines in mice that survive lethal infection.

A polyclonal antibody based coproantigen detection enzyme linked immunosorbent assay (cAg-ELISA) for diagnosis of experimental and natural Oesophagostomum columbianum infection in goats was developed and evaluated. Adult O. columbianum worms, collected from the caecum and colon of slaughtered goats, were triturated and cultured for obtaining infective third stage larvae (L 3 ) and also used for preparation of excretory–secretory antigen (ESAg). Experimental goats were orally infected each with 600 L 3 /kg of the body weight. Filter sterilized faecal supernatant, i.e. the coproantigen (cAg) was harvested from the rectal faeces of all the infected goats, on alternate days from day-5 till day-31 after the infection. Hyperimmune serum (HIS) against ESAg of O. columbianum was raised in rabbits. Molecular and antigenic characterization of ES products of O. columbianum by HIS revealed that 50 and 39 kDa polypeptides were immuno-dominant. Coproantigen detection ELISA was standardized by using the cAg as coating antigen and its subsequent binding with the HIS against ESAg of O. columbianum . The sensitivity, specificity and accuracy of the standardized assay were determined by evaluating the assay on the faecal supernatant of 96 slaughtered goats taking into consideration their recorded parasitological status in respect of the abomasal and the intestinal parasites. The cAg-ELISA detected the prepatent oesophagostomosis on 20–24-day-post-infection with a sensitivity, specificity and accuracy of 88, 89.13 and 88.54%, respectively. The assay is relatively easy to perform and would serve as a reliable tool for detection of caprine nodular oesophagostomosis.

Pathogenic Aeromonas hydrophila (strain VB21), a multiple-drug resistance strain contains a plasmid of about 21 kb. After curing of plasmid, the isolates became sensitive to antimicrobials, to which they were earlier resistant. The cured bacteria exhibited significant alterations in their surface structure, growth profile and virulence properties, and failed to cause ulcerative disease syndrome (UDS) when injected into the Indian catfish Clarias batrachus. Routine biochemical studies revealed that the plasmid curing did not alter the biochemical properties of the bacteria. After transformation of the plasmid into cured A. hydrophila the bacterium regained its virulence properties and induced all the characteristic symptoms of UDS when injected into fish. Thus, the plasmid plays a pivotal role in the phenotype, growth and virulence of A. hydrophila and pathogenesis of aeromonad UDS.

Successful blastocyst implantation depends upon the synchronous dialogue between age- and stage-matched embryo and adequately primed maternal endometrium. Endometrial signals present in the uterine lumen influence the growth and the viability of preimplantation stage embryo. Thus, uterine secretion of embryotoxic cytokines may affect the preimplantation stage embryo. Our previous study in the rhesus monkey has indicated that tumor necrosis factor-alpha (TNF-alpha) is one such candidate present in the uterine lumen, which may act as an embryotoxic agent. In the present study, the effect of TNF-alpha on de novo protein synthesis by mouse morulae (n = 100) and blastocysts (n = 100) in vitro was investigated by 2D-polyacrylamide gel electrophoresis. A total of 35 and 40 protein spots were detected in lysates of control morulae and blastocysts, respectively. Exposure of embryos to TNF-alpha (50 ng/ml) reduced the number of protein spots to 15 and 17 compared to that of control morulae and blastocysts. Seven spots in morula and 13 protein spots in blastocyst flourograms showed quantitative changes in their expressions with exposure to TNF-alpha. Morulae and blastocysts exposed to TNF-alpha expressed 8 and 17 protein spots, respectively, that were not seen in control embryos. It appears from the present study that exposure of preimplantation stage embryos to TNF-alpha affects their protein synthesis both quantitatively and qualitatively.

Synchronous attainment of maternal endometrial receptivity allows implantation-stage adhesive blastocyst to undertake apposition, attachment and invasion. In the present essay, we propose a model according to which luteal phase progesterone induces a basic drive in endometrium toward receptivity and as a result, adequately primed endometrium differentiates through certain steps in a fixed action pattern. The implantation-stage embryo senses such endometrial responsiveness circumstantially by the factors secreted by maternal endometrium and undertakes differentiation to implant by secreting factors which act on maternal endometrial cells to further potentiate them to implantation stage-specific changes. Such a dynamic temporo-spatial manner of interaction involving a set of specific factors acting synchronously leads to the activation of innate releasing process in both compartments towards embryo attachment followed by successful intrusion and controlled invasion of trophoblast cells into maternal endometrium. In the present review we discuss the potential role of various endocrine and paracrine factors in the process of blastocyst implantation in the human.

A single low dose administration of a high affinity anti-progestin agent like mifepristone during the early luteal phase inhibits blastocyst implantation in human and non-human primates. Though it has been observed that luteal phase serum concentrations of estradiol and progesterone were not affected by the application of anti-nidatory dose of early luteal phase mifepristone suggesting that ovarian steroidogenic function is not compromised, it is nevertheless possible that ovarian physiology at the local tissue level is affected in this treatment schedule. In the present study, healthy, mature, proven fertile female rhesus monkeys were divided into two groups. Group 2 animals were treated with a single dose of mifepristone (2 mg/kg body weight), while group 1 animals were injected with vehicle (1:4 benzoyl benzoate: olive oil, v/v, s.c.) on day 2 post-ovulation. The morphological examination including that of vascularity, as well as, histometric determination of profiles of immunopositivity for IL-1alpha and TGF-beta1 in stromal, follicular and luteal compartments of mid-luteal phase ovaries from animals with or without a single, anti-nidatory dose of mifepristone applied on day 2 after ovulation failed to reveal any significant change between the two groups. Thus, it appears that early luteal phase administration of a single antinidatory dose of mifepristone does not affect the ovarian physiology in the treatment cycle.

Ischaemia induced germ cell apoptosis in rat testis was studied in detail to find out (i) spermatogenic stage or seminiferous epithelium region specific involvement of germ cells in apoptosis, (ii) preferential specificity of a particular germ cell type to become apoptotic and (iii) the ratio of live and dead testicular cells isolated in vitro after various period of ischaemic induction. Cell apoptosis, as observed in histological sections increased from 1 to 24 h of ischaemia. Apoptosis was not restricted to any specific germ cell type but was observed simultaneously in all the cell types in the initial hours (1-6 h) of ischaemia. No spermatogenic stage specific preference in apoptotic induction was also observed. However, as the duration of ischaemia progressed, the cell types observed to be most affected in number and morphology were the spermatids followed by spermatocytes. Centrally located tubules of testis were affected first than those located in the periphery. Overexpression of Bax staining was limited to few germ cell nuclei only. More than 95% of the germ cells in the control testis that earlier showed trypan blue dye exclusion were found stained after 12 h of ischaemia. Starting from early hours (1 h), lipid peroxidation rose proportionally with the duration of ischaemia while superoxide dismutase (SOD) and catalase activities were found decreased. Significant (p0.05) increase in the activities of glutathion-s-transferase and levels of hydrogen peroxide were observed after 6 h of ischaemia. These findings indicate that the physiological processes of oxidative stress have a direct linkage to the extent of germ cell apoptosis in the seminiferous epithelium.

The testicular inhibitory effect of the aqueous fraction of methanol extract of Stephania hernandifolia leaf was studied in male Wistar rats.The supernatent and the precipitate part of aqueous fractions of the methanol extract of the leaf were gavaged separately to rat at a similar dose of 200 mg/mL per 100 g body weight per day for 28 days. After cessation of treatment, various observations were conducted.In both treated groups, there were significant decreases in the relative weights of the sex organs, the testicular key androgenic enzymes activities, the plasma level of testosterone, the number of different germ cells at stage VII of seminiferous epithelial cell cycle and the seminiferous tubular diameter in comparison to the controls. Neither of the parts had somatic, renal and hepatic toxicity. This study suggested that the active molecules present in the aqueous fraction of methanol extract of Stephania hernandifolia leaves might be steroids as indicated by thin layer chromatography using specific staining substance for steroid molecules.In rats, the aqueous fraction of methanol extract of the S. hernandifolia leaves possesses certain testis-inhibitory substances, which may be steroid-like agents.

This study concerned the minimum and optimum effective doses of calcium chloride needed for induction of chemosterilization in male albino rats, 30 days after a single intratesticular injection of calcium chloride (CaCl2.2H2O) solution at 2.5, 5, 10 or 20 mg per 100 g body weight per testis. There was a significant diminution in the relative wet weight of the sex organs (p0.01), epididymal sperm count (p0.001), plasma concentration of testosterone (p0.01), testicular activities of delta5,3beta-hydroxy steroid dehydrogenase (delta5,3beta-HSD), 17beta-hydroxy steroid dehydrogenase (17beta-HSD) (p0.01), glutathione S-transferase (GST) (p0.01), superoxide dismutase (SOD) (p0.01), and peroxidase (p0.01), significant elevations in testicular content of malondialdehyde (MDA) and conjugated dienes (p0.01), along with derangement of seminiferous tubular architecture and degeneration of the Leydig cells in the testis and elevations in the concentrations in the plasma of LH and FSH (p0.01), commencing at a dose of 5 mg, with the greatest effects at a dose of 20 mg. No significant alterations in these factors occurred at the dose of 2.5 mg in comparison to the control that received only the vehicle. There was no significant alteration in the plasma concentrations of prolactin (p0.05), corticosterone (p0.05) or fasting blood glucose or in the rectal temperature (p0.05) at any of the doses relative to the control group, suggesting that this chemosterilizing procedure did not exert any chronic stress on the experimental animals. From these observations, it may be suggested that 5 mg should be considered as the minimum dose, and 10 mg or 20 mg as the optimum dose, whereas 2.5 mg was ineffective for induction of chemosterilization. There would seem to be little point in using more than 20 mg of calcium chloride for this purpose. Intratesticular injection of calcium chloride at an effective dose may be considered as an alternative to surgical castration.

Intravaginal administration of an anti-angiogenic agent, fumagillin, during blastocyst implantation inhibits pregnancy establishment in a dose-related manner in the rhesus monkey. In the present study, mated female rhesus monkeys were vaginally inserted with tampons containing vehicle (group 1; n = 5) and test agent (fumagillin, 4 mg/animal; group 2; n = 6) on cycle day 20, and endometrial tissue samples were collected on cycle day 24 from all monkeys and processed for histological examination and immunohistochemical localization for LIF, IL-6, TGF-beta and VEGF. Concentrations of estradiol-17 beta, progesterone and chorionic gonadotrophin in peripheral circulation were determined. From the serum profiles of the hormones, 2 monkeys in group 1, and 1 monkey in group 2 appeared pregnant. However, endometrial morphology revealed histological evidence of pregnancy in 3 out of 6 fumagillin-treated animals. Histometric analysis of immunohistochemical staining in epithelial, stromal and vascular compartments revealed that per cent areas occupied by immunoprecipitate for the cytokines studies did not change in epithelial and stromal compartments, except that for TGF-beta which was higher (P0.05) in epithelial compartment in group 2. No change was observed in immunoprecipitation areas for IL-6 in epithelial, stromal and vascular compartments. On the other hand, changes (P0.05) for LIF, TGF-beta and VEGF were evident in the vascular compartment. It is possible that disparate responses observed in glandular, stromal and vascular compartments in implantation stage endometrium following fumagillin treatment actually caused from associated decline in progesterone concentration in peripheral circulation. It is also possible that fumagillin, an angiostatic agent, affects the synthesis and secretion of cytokines primarily in the vascular compartment of implantation stage endometrium, and thereby manifests differential responses in epithelial, stromal and vascular compartments.

In the present study isolated uterine epithelial cells from normal rabbits were maintained in culture on free floating rat-tail collagen matrix, and the morphological characteristics of these cells were examined. Additionally, the pattern of protein synthesis and secretion by rabbit uterine epithelial cells grown on free floating collagen gels following estradiol and/or progesterone treatment in vitro was examined. Isolated epithelial cells cultured on collagen gels in complete medium containing serum attached to form monlayers, and eventually the gels became free floating and contracted giving rise to luminal arrangements. These cells were cytokeratin positive epithelial cells and were ultrastructurally polarized. These cells also exhibited differential upregulation and down regulation in the synthesis and secretion of proteins in response to estradiol, progesterone, and estradiol plus progesterone. Additionally, a permissive action between progesterone and estradiol in the synthesis of two species of secretory proteins was observed. It however remains to be examined whether different species of proteins produced in vitro in response to estradiol and progesterone bear any association with physiological states in reproductive cycle in this species.

Porcine testicular carbonyl reductase (PTCR) belongs to the short chain dehydrogenases/reductases (SDR) superfamily and catalyzes the NADPH-dependent reduction of ketones on steroids and prostaglandins. The enzyme shares nearly 85% sequence identity with the NADPH-dependent human 15-hydroxyprostaglandin dehydrogenase/carbonyl reductase. The tertiary structure of the enzyme at 2.3 A reveals a fold characteristic of the SDR superfamily that uses a Tyr-Lys-Ser triad as catalytic residues, but exhibits neither the functional homotetramer nor the homodimer that distinguish all SDRs. It is the first known monomeric structure in the SDR superfamily. In PTCR, which is also active as a monomer, a 41-residue insertion immediately before the catalytic Tyr describes an all-helix subdomain that packs against interfacial helices, eliminating the four-helix bundle interface conserved in the superfamily. An additional anti-parallel strand in the PTCR structure also blocks the other strand-mediated interface. These novel structural features provide the basis for the scaffolding of one catalytic site within a single molecule of the enzyme.

Steroidogenic key enzymes, i.e. delta 5-3 beta and 17 beta-hydroxysteroid dehydrogenase (delta 5-3 beta and 17 beta-HSD) activities, in the testis and Bidder's organ of the toad were inhibited and ascorbic acid synthesis in these organs was decreased by a wide range of lithium concentration in in vitro study. A significant inhibition was noted at a concentration of 2.0 mM, which is easily achieved in the blood during the treatment of manic patients by lithium chloride. This experiment reflected that lithium exerts a direct inhibitory effect on hydroxysteroid dehydrogenase activities in the testis and Bidder's organ--a rudimentary ovary in Bufo.

In this study, effects of timed sequential exposure to various concentrations of chloroquine and its two metabolites (SA100387/050 B, SA 100487/053 B), quinine, hydroquinine and quinidine were monitored by morphological analyses, using synchronous cultures of Plasmodium falciparum. The trophozoites and schizonts were sensitive to chloroquine and its metabolites. However, quinine and two of its derivatives were lethal without particularly being stage-specific.

Steroid dehydrogenase enzymes influence mammalian reproduction, hypertension, neoplasia, and digestion. The three-dimensional structures of steroid dehydrogenase enzymes reveal the position of the catalytic triad, a possible mechanism of keto-hydroxyl interconversion, a molecular mechanism of inhibition, and the basis for selectivity. Glycyrrhizic acid, the active ingredient in licorice, and its metabolite carbenoxolone are potent inhibitors of human 11 beta-hydroxysteroid dehydrogenase and bacterial 3 alpha, 20 beta-hydroxysteroid dehydrogenase (3 alpha, 20 beta-HSD). The three-dimensional structure of the 3 alpha, 20 beta-HSD carbenoxolone complex unequivocally verifies the postulated active site of the enzyme, shows that inhibition is a result of direct competition with the substrate for binding, and provides a plausible model for the mechanism of inhibition of 11 beta-hydroxysteroid dehydrogenase by carbenoxolone. The structure of the ternary complex of human 17 beta-hydroxysteroid dehydrogenase type 1 (17 beta-HSD) with the cofactor NADP+ and the antiestrogen equilin reveals the details of binding of an inhibitor in the active site of the enzyme and the possible roles of various amino acids in the catalytic cleft. The short-chain dehydrogenase reductase (SDR) family includes these steroid dehydrogenase enzymes and more than 60 other proteins from human, mammalian, insect, and bacterial sources. Most members of the family contain the tyrosine and lysine of the catalytic triad in a YxxxK sequence. X-ray crystal structures of 13 members of the family have been completed. When the alpha-carbon backbone of the cofactor binding domains of the structures are superimposed, the conserved residues are at the core of the structure and in the cofactor binding domain, but not in the substrate binding pocket.

There is a growing awareness of a need for developing novel methods of contraceptive technology which should not only be effective in providing protection against conception but also take into consideration the reproductive health issues confronting men and women. This paper considers the process of embryo implantation as one such potential target. The hormonal basis of embryo implantation in primates has been discussed to indicate that progesterone, and not estrogen, from ovarian source is the primary determinant of embryo-endometrial maturation and their synchronization for implantation. Thus, low dose administration of the anti-progesterone, mifepristone, during early luteal phase has been shown to be an effective anti-implantation approach to for fertility control. Furthermore, the dissociation of endometrial-hormonal synchrony at the time of blastocyst implantation following the post-ovulatory mifepristone administration has been shown to be the physiological basis of its anti-implantation effect with undisturbed circulatory hormone profiles and ovarian functions. Further studies are required to appreciate the full potential and to mollify the limitations of this approach.

(1) A significant seasonal variation in the membrane fluidity (as sensed by DPH-fluorescence polarization), membrane lipid components (phospholipid and neutral lipid), fatty acid composition of membrane phospholipid (phosphatidylcholine, phosphatidylethanolamine and sphingomyelin), positional distribution of fatty acids at Sn-1 and Sn-2 position of phosphatidyl-choline and -ethanolamine is noticed in the brain membranes (myelin, synaptosomes, and mitochondria) of a tropical air breathing teleost, Clarias batrachus. (2) A `partial compensation' of membrane fluidity during seasonal adaptation is observed in myelin and mitochondria membrane fractions. Synaptosomes membrane fraction exhibits a different response. Depletion (about 15–70%) of membrane lipid components (phospholipid, cholesterol, diacylglycerol and triacylglycerol) per unit of membrane protein is the characteristic feature of summer adaptation. An increase (about 20–100%) in the level of oleic acid and decrease (about 20–60%) in the level of stearic acid are almost common features in membrane phospholipid fractions of winter-adapted Clarias. (3) From the tissue slice experiment it is evident that there is an activation of cellular phopholipase A2 at lower growth temperature and of cellular phosphalipase A1 at higher growth temperature and this suggests the reorganization of molecular architecture of the membrane during seasonal adaptation. (4) Accumulation of oleic acid in Sn-1 position and polyunsaturated fatty acids in Sn-2 position of phophatidyl-choline and -ethanolamine during winter indicates an increase in the concentration of 1-monoenoic, 2-polyenoic molecular species of phospholipid in order to maintain the stability of membrane lipid bilayer.

The characteristics of implantation stage endometrium following a single-dose, early luteal phase application of mifepristone (RU486) in proven conception cycles has been examined in the rhesus monkey in an attempt to understand the physiological basis of the anti-implantation activity of the drug. Endometrial samples were collected from monkeys subjected to vehicle (group 1, n = 14) and RU486 (2 mg/kg body weight; group 2, n = 12) on day 2 after the presumed day of ovulation of successfully mated cycles. The average diameter of glands (P0.05), number of vacuolated cells (P0.01), number of supranuclear vacuolated cells (P0.05) in glandular epithelium and amount of glandular secretion (P0.05) were significantly lower in RU486-treated endometrium compared with control tissue samples. Additionally, 18% of glandular epithelial cells showed apoptotic and degenerative features in RU486-treated tissue samples. These data, together with the observed significant decreases in precipitate area (P0.02) and in the optical absorbance of alkaline phosphatase reaction end-product (P0.05), confirm that retardation in glandular differentiation in the upper functionalis is a likely target of antiprogestin action in implantation stage endometrium. An increased frequency of mitosis in stromal cells (P0.05) and a greater degree of extravasation (P0.05) were also observed after RU486 exposure. Despite an apparent indication of constriction and regression in few RU486-exposed endometria compared with controls, morphometric analyses did not show any changes in capillary structure. Whether endometrial vasculature in progesterone-exposed uterus is a target of antiprogestin action during the peri-implantation stage remains to be determined. Further studies are required to explain the observed increase (P0.02) in the area of precipitate of von Willebrand (vW) factor with no change in vW factor-positive vessels, and the apparent increase in collagen IV immunostain in subepithelial and perivascular basement membrane in implantation stage endometrium after early luteal phase RU486 treatment in monkeys.

The present work reports the synthesis and preliminary pharmacological characterization of 8,9-dihydroxy-2,3,7,11b-tetrahydro-1H-naph[1,2,3-de] isoquinoline (4, dinapsoline). This molecule was designed to conserve the essential elements contained in our D1 agonist pharmacophore model (i.e., position and orientation of the nitrogen, hydroxyls, and phenyl rings). It involved taking the backbone of dihydrexidine [3; (+/-)-trans-10, 11-dihydroxy-5,6,6a,7,8,12b-hexahydrobenzo[a] phenanthridine], the first high-affinity full D1 agonist, and tethering the two phenyl rings of dihydrexidine through a methylene bridge and removing the C(7)-C(8) ethano bridge. Preliminary molecular modeling studies demonstrated that these modifications conserved the essential elements of the hypothesized pharmacopore. Dinapsoline 4 had almost identical affinity (KI = 5.9 nM) to 3 at rat striatal D1 receptors and had a shallow competition curve (nH = 0.66) that suggested agonist properties. Consistent with this, in both rat striatum and C-6-mD1 cells, dinapsoline 4 was a full agonist with an EC50 of ca. 30 nM in stimulating synthesis of cAMP via D1 receptors. The design and synthesis of dinapsoline 4 provide a powerful test of the model of the D1 pharmacophore we have developed and provide another chemical series that can be useful probes for the study of D1 receptors. An interesting property of 3 is that it also has relatively high D2 affinity (K0.5 = 50 nM) despite having an accessory phenyl ring usually though to convey D1 selectivity. Dinapsoline 4 was found to have even higher affinity for the D2 receptor (K0.5 = 31 nM) than 3. Because of the high affinity of 4 for D2 receptors, it and its analogs can be powerful tools for exploring the mechanisms of "functional selectivity" (i.e., that 3 is an agonist at some D2 receptors, but an antagonist at others). Together, these data suggest that 4 and its derivatives may be powerful tools in the study of dopamine receptor function and also have potential clinical utility in Parkinson's disease and other conditions where perturbation of dopamine receptors is useful.

Injection of lithium chloride at the dose of 200 micrograms/toad/alternate day for 7, 14 and 21 days caused a significant reduction in the activities of testicular delta 5-3 beta, 17 beta-hydroxysteroid dehydrogenase and acid phosphatase enzymes. There was a marked inhibition in spermatogenesis in lithium chloride treated toad for 14 and 21 days of treatment, but 7 days of treatment has no effect.

Mifepristone (RU 486), an antiprogesterone, is a promising luteal phase contraceptive agent for human use. However, at present its use is limited by the practical constraint of determining the day of ovulation for an LH + 2 day administration of the drug as indicated from experimental and clinical studies. The aim of the present study was to identify the effective period of luteal phase (luteal phase window) when a single administration of mifepristone would induce antinidatory activity without disturbing menstrual cyclicity and ovulatory pattern in the rhesus monkey. RU 486 (2 mg/kg body weight in benzyl benzoate/olive oil, 1:3) was given to mated monkeys (n = 9) on cycle day 16 in the first treatment cycle (treatment group T1, n = 9), and in the following cycle on cycle day 20 (treatment group T2, n = 8). A single s.c. injection of this antiprogestin during early to midluteal phase (days 1-10 after ovulation, as determined from retrospective analysis of serum concentrations of estrogen and progesterone) provided a one hundred per cent protection against pregnancy, with no apparent side effects. There were no changes in cycle lengths (F = 3.5; P0.3), day of ovulation (F = 1.8; P0.7) and duration of menses (F = 3.5; P0.3) compared with the pre-treatment and post-treatment cycles. Pooled analyses of serum concentrations of estrogen and progesterone during luteal phases of T1 and T2 cycles also showed no variations with those in pre- and post-treatment cycles.(ABSTRACT TRUNCATED AT 250 WORDS)

The morphological characteristics of endometrium on day 6 after ovulation of conception (group 1) and non-fecund, menstrual (group 2) cycles have been studied in the rhesus monkey (n = 30). A conception cycle was distinguished by the presence of a developmentally normal, age-stage-synchronized embryo. Thus, 78% of the mated cycles (n = 18) yielding synchronous embryos (12 zona-encased and two zona-free blastocysts) were used for this study. On day 6 after ovulation, no significant changes in the serum concentrations of oestrogen and progesterone were seen in conception cycles (n = 14) compared with the non-mated, normal ovulatory cycles (n = 12). Morphometric analyses revealed that on day 6 of gestation (n = 8), endometrium differed from the corresponding non-mated luteal phase (n = 7) with significant increases in epithelial mitosis (P0.01), height of glandular epithelium (P0.05), volume ratio of gland cell to gland (P0.03), degree of pseudostratification (P0.02), and higher frequency of supranuclear, adluminal accumulation of vacuoles in gland cells (P0.05). The degree of stromal oedema was higher (P0.02) in fecund cycles but there was no change in venular diameter. In a separate set of experiments, estimates of tissue vascular response revealed a higher (P0.02) endometrial extravascular albumin space on the same day of gestation; there were no differences, however, in endometrial blood volume, or in the number of von Willebrand antigen-positive capillaries and small vessels between the two groups (group 1, n = 6; group 2, n = 5).(ABSTRACT TRUNCATED AT 250 WORDS)

The contranidatory action of mifepristone (RU 486) given as a single application at different dosages to mated rhesus monkeys (Macaca mulatta) on second day after ovulation has been examined in the present study. In group 1, monkeys (n = 3) received only vehicle (benzyl benzoate: olive oil, 1:4, v/v) and were treated as controls. In group 2 monkeys (n = 4), RU 486 was given by gavage at 10 mg/kg in group 4 (n = 5). The patterns of cycles and profiles of serum estrogen and progesterone were monitored for assessing the occurrence of implantation and pregnancy. At a single dose of 10 mg/kg, RU 486 was found to be ineffective in preventing nidation, resulting pregnancy in three females out of four treated monkeys. Similarly, an s.c. administration of 1 mg/kg could provide pregnancy protection in two of the four treated monkeys. In these monkeys, however, the menstrual cycle characteristics were not affected as compared to pretreatment cycles. Interestingly, the administration of 2 mg/kg, s.c., RU 486 could provide a hundred percent pregnancy protection in mated monkeys, and there was no significant changes in the pattern of menstrual cycle characteristics. It appears that an early post-ovulatory administration of RU 486 may be successfully used in human as an effective once-a-month, early luteal phase contranidatory agent.

Effect of a single subcutaneous injection of cadmium chloride at the dose of 0.5 mg/toad on adrenal delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSD) was observed after 7 days. The activity of delta 5-3 beta-HSD was measured histochemically. The experiments indicate that cadmium chloride resulted in a significant decrease in the activity of adrenal delta 5-3 beta-HSD in toad during breeding season (June-July).

The purpose of this investigation was to make a correlative study between spermatogenesis and testicular acid and alkaline phosphatase activities in mature prostatectomized animals at different post-operative periods. The results demonstrate that there was a significant augmentation in the activity of testicular acid and alkaline phosphatases subsequent to 14 and 21 days of prostatectomy. A parallel quantitative study of spermatogenesis at stage VII of the seminiferous cycle, namely, type A spermatogonia (ASg), preleptotene spermatocytes (pLSc), mid-pachytene spermatocytes (mPSc) and step 7 spermatids (7sd), revealed that there was a significant reduction in the number of step 7 spermatids after 14 and 21 days. No change was observed in the above testicular enzymes and spermatogenesis after 7 days of prostatectomy. Therefore, it is concluded that prostatectomy can alter the above testicular enzyme activities and spermatogenesis in chronic prostatectomized state.

Changes in the adrenal weight, adrenal 5-ene-3 beta-hydroxysteroid dehydrogenase (5-ene-3 beta-HSD) activity and serum levels of corticosterone were observed in male wistar rats after the treatment of lithium chloride in the doses of 100, 200 and 400 micrograms/100 g of body weight/day for 7, 14 and 21 days. The experiments indicate that 200 and 400 micrograms/100 g.b.w. administered for 14 and 21 days caused a significant stimulation in the activities of adrenal 5-ene-3 beta-HSD along with elevation of adrenal weights and serum levels of corticosterone. 100 micrograms of lithium chloride was not able to modulate the adrenal activity. Moreover, plasma levels of lithium remain in therapeutic range in this experiment at the doses of 200 and 400 micrograms/100 g body weight. Therefore, our data suggest that lithium can alter the adrenal activity within its therapeutic range according to the duration of treatment.

Activities of delta 5-3 beta- and 17 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSD and 17 beta-HSD), Leydig cell nuclear area (LCNA) and spermatogenesis in the testis were observed after injection of lithium chloride in the 'antiserum to luteinizing hormone (LH)' treated toad. A significant decrease in the activities of steroidogenic enzymes, LCNA and spermatogenesis were noticed after the injections of 'antiserum to LH' to toads. Further decrease in the activities of the above parameters was observed in the lithium chloride--'antiserum to LH' treated toad. It is suggested that lithium chloride may inhibits testicular function without modulating the pituitary activity.

Weight, number of follicles and corpora lutea of Bidder's organ increased significantly after castration of adult toad in breeding season. Removal of pituitary along with testes decreased both weight and number of follicles and corpora lutea. Administration of luteinizing hormone (LH) in castrated and hypophysectomized toad showed more or less similar result as that of control indicating that LH has an effect on the histology of Bidder's organ in the toad.

Changes in the activities of acid and alkaline phosphatase were observed in the testis, prostate and seminal vesicle after the injection of lithium chloride at the doses of 100, 200 and 400 micrograms/100 g body weight/day for 7, 14 and 21 days. The studies indicate that 200 and 400 micrograms/100 g body weight for 14 days and 21 days showed a significant inhibition in the activity of acid phosphatase in all the above reproductive organs. There is a significant stimulation of alkaline phosphatase activity at the doses of 200 and 400 micrograms of lithium after 21 days of treatment in testis, prostate and seminal vesicle along with significant decrease in accessory sex organs weight in comparison to control animal. Therefore, it is evident that the effect of lithium on male reproductive organs mainly depends on the amount of the drug being injected and the duration of treatment to it.

Prostaglandin synthetase was immobilized by adsorption of goat vesicular microsomes on silica gel containing CaSO4 (silica gel G). Repeated cycles of enzymatic conversion of arachidonic acid to prostaglandin by the immobilized microsomes increased the product yield by 1.5 fold, in comparison to the same by free microsomal particles. The presence of Ca2+ in silica gel is responsible for this improved yield of prostaglandin as the divalent metal ion stabilized prostaglandin synthetase activity in a remarkable way. Microsomal particles immobilized on solid supports like alumina G and controlled pore glass were not very effective.

Testicular delta 5-3 beta- and 17 beta-hydroxysteroid dehydrogenase (delta 5-3 beta- and 17 beta-HSD) activities of rat are inhibited in vitro by a wide range of lithium concentration. The inhibitory effects of lithium are evident at a concentration of 2.5 mM which is easily achieved during the treatment of acute manic patients with lithium. This suggests that lithium exerts a direct inhibitory effect on testicular hydroxysteroid dehydrogenase activities.

Lithium chloride at a dose of 200 micrograms/100 g body weight/day given for 21 days caused a significant increase in adrenal weight, adrenal 5-ene-3 beta-hydroxysteroid dehydrogenase (5-ene-3 beta-HSD) activity along with elevation in serum level of corticosterone on the 22nd day in the rat. Administration of testosterone for the last 14 days to lithium treated rats caused a significant decrease in adrenal weight, adrenal 5-ene-3 beta-HSD activity and serum level of corticosterone in comparison to lithium treated animals.

The present work continues our aim of establishing an experimental model to study the decidual cell reaction to an artificial deciduogenic stimulus in the long-term ovariectomized rhesus monkey treated with oestrogen followed by progesterone. The fine structural details of decidual, granular and plaque cells, which constituted the endometrial cellular response to the deciduogenic stimulation in the present study, revealed striking similarities with those reportedly present in an endometrial response to blastocyst implantation in the rhesus monkey. Plaque epithelia showed a significant degree of hypertrophy, hyperplasia and differentiation followed by a steady degeneration by day 32 (equivalent to day 16 after trauma) of treatment. The plaque cells were shown to contain numerous regular-shaped mitochondria, polyribosomes and large amounts of rough endoplasmic reticulum (RER) in their cytoplasm and were characteristically arranged in clusters or acini formation surrounded by discrete basal laminae. As early as day 28 of treatment, the initiation of stromal decidual cell transformation was noted and, by day 48, a sizeable pool of decidual cells was found. The decidual cells had rounded nuclei and elaborate arrangements of interconnected cisternae of RER which were often moderately dilated and filled with amorphous, electron-dense material. Granular cells were characterized by eccentrically located nuclei and numerous membrane-bound, electron-dense granules in their cytoplasm and were found in increasing numbers in the stroma around decidual cells, blood vessels and glandular epithelia. Based on the ultrastructural and temporal characteristics of the endometrial cells studied it has been suggested that the cells have secretory functions, and that the differential maturation profiles of the cell types might be caused by their differences in sensitivities and by their integral response to progesterone. Journal of Endocrinology (1990) 124, 53–57

Summary The hepatic enzyme tryptophan pyrrolase was assayed from male and female Swiss/ICR mice of different ages, for example, the neonate, 5-day, 10-day, 25-day, 30-day, 35-day, 40-day and 45-day olds. These data show an increase through the 15–20th day followed by a decline in enzymic activity, which drop is succeeded by a rise in activities in both sexes by the 30th day of development. These enhanced activities are steeper in the males compared to the females. Males show a steady level of enzymic activity by day 35, whereas females tend to exhibit, around this time period, an oscillation in enzymic activity. Possible explanations of these results are offered.

Pretreatment with triamterene (2,4,7-triamino-6-phenylpteridine) results in significant increase in the level of endogenous holotryptophan pyrrolase over the induction caused by hydrocortisone alone. The pteridine perhaps results in a favorable stoichiometric ratio in vivo of apotryptophan pyrrolase and the heme coenzyme.

Triamterene (2,4,7-triamino-6-phenylpteridine), a diuretic drug, affects the hepatic endogenous holotryptophan pyrrolase (EC 1.13.1.12) in the mouse, with stress from sham-operation as a preconditioning factor; under these conditions, maximum levels of enzymatic activities are obtained. These values range from 6.87 to 9.84 units with a mean value of 8.252 mumoles of kynurenine per gram tissue per hour. Employing similar experimental designs, the effects of cortisol were studied for comparative evaluation; these activities range from 4.154 to 4.999 units with a mean value of 4.49. The significance and the possible mechanism of action of triamterene are discussed.

The purpose of the present study was to show the effects of lithium on testicular activities in the toad. Adult male toads were injected with lithium chloride (200 micrograms/toad) of alternate days for 21 days. At the moment of sacrifice on 22nd day, lithium treated animal showed decreases in testicular weight and Leydig cell nuclear area along with inhibition of spermatogenesis and testicular delta 5-3 beta as well as 17 beta-hydroxysteroid dehydrogenase activities. The results of our present experiment suggest that lithium administration might be associated with significant adverse effects on testicular activity in toad.

Acid phosphatase activity in both total and subcellular fractions of endometrium of rhesus monkey on day 5 of gestation was significantly (P

A variation of temporal recursive filtering for intravenous and intra-arterial digital subtraction angiography (DSA) is described. It results in a dynamic tomographic DSA display that may improve intravenous DSA performance by removing the confusion introduced by vessel overlap and reducing the severity of motion artifacts. A conventional circular tomographic apparatus was modified by the addition of a 23-cm image intensifier-television chain and made to run continuously at one revolution each 1.5 seconds. The video output was processed by the recursive filter in a two-step process resulting in a dynamic tomographic display of arterial opacification. Theory and results of initial animal studies are presented.