Two new chemical constituents from Lonicera japonica.

Two new chemical constituents, japopenoid D (1), and japopenoid E (2), were isolated and identified from the flower buds of Lonicera japonica Thunb. The structures of these compounds were elucidated based on spectroscopic analysis (HR-ESI-MS, NMR), and the absolute configurations of 1 and 2 were determined by comparison of their electronic circular dichroism (ECD) spectra with literature and theoretical calculation. The anti-inflammatory activities of the isolates were evaluated by measuring their inhibitory effects on PGE2 and IL-6 production in LPS stimulated RAW 264.7 macrophages. As a result, compound 1 could reduce PGE2 and IL-6 levels in LPS-activated RAW 264.7 macrophages with IC50 values of 6.78 and 42.07 µM, respectively.[Formula: see text].

Prostaglandins in semen compromise the immune response against sexually transmitted pathogens.

Seminal plasma is not just a spermatozoa carrier. It induces the expression of inflammatory cytokines and chemokines and a massive infiltration of neutrophils, monocytes and dendritic cells in the female genital mucosa after coitus, enabling the innate immune system to fight against sexually transmitted pathogens. However, exposure to seminal plasma not only turns on an inflammatory response but also induces regulatory mechanisms that allow the fetus (a semiallograft) to grow and develop in the uterus. In mouse models it has been shown that seminal plasma induces the expansion of regulatory T cells specific to seminal Ags in the receptive partner, thus promoting tolerance to paternal alloantigens and avoiding allogeneic fetal rejection. These mechanisms appear to be mainly induced by prostaglandins of the E series (PGE) and TGF-ß, which are present at huge concentrations in the seminal plasma. Moreover, we have recently shown that exposure to seminal plasma induces the differentiation of dendritic cells into a tolerogenic profile through a mechanism dependent on the activation of the prostanoid receptors EP2 and EP4 by seminal PGE. Our hypothesis proposes that this tolerogenic response induced by seminal PGE, while promoting fertility by inducing tolerance toward paternal alloantigens, might also compromise the development of the adaptive immune response against sexually transmitted pathogens in the receptive partner.

Phosphatidylserine inhibits inflammatory responses in interleukin-1ß-stimulated fibroblast-like synoviocytes and alleviates carrageenan-induced arthritis in rat.

Recently, phosphatidylserine (PS) has received attention for its anti-inflammatory effect; however, the molecular mechanisms of its action have not been fully understood. Thus, we hypothesized that PS might have antiarthritic and anti-inflammatory effects. To test this hypothesis, the in vitro anti-inflammatory effect of soybean-derived PS was tested on interleukin (IL)-1ß-stimulated fibroblast-like synoviocytes from rheumatoid arthritis patients (RA-FLS) by measuring the levels of IL-6, IL-8, prostaglandin E(2), and vascular endothelial growth factor by enzyme-linked immunosorbent assay. The analgesic and antiarthritic activities of PS were investigated in rat models of carrageenan-induced acute paw pain and arthritis. The former was evaluated with a paw pressure test; the latter, by measuring paw volume and weight distribution ratio. In addition, the participation of mitogen-activated protein kinase signaling in the anti-inflammatory and antiarthritic effects of PS was investigated in RA-FLS. Phosphatidylserine inhibited the production of inflammatory mediators IL-6; IL-8; vascular endothelial growth factor; and, in particular, prostaglandin E(2) in IL-1ß-stimulated RA-FLS. These effects were associated with abrogation of inhibitor of nuclear factor-κBα phosphorylation and suppression of p38 and c-jun amino terminal kinase but not extracellular signal-regulated kinase 1/2 phosphorylation. In rats, PS also showed a significant inhibitory effect on arthritic and nociceptive symptoms induced by carrageenan. These findings suggest that PS has anti-inflammatory and antiarthritic effects in vitro and in in vivo animal models; thus, PS should be further studied to determine its potential use as either a pharmaceutical or dietary supplement for alleviating arthritic symptoms.

Molecular mechanisms of a novel selenium-based complementary medicine which confers protection against hyperandrogenism-induced polycystic ovary.

The objective was to evaluate ovarian functionality and oxidative response in hyperandrogenism-induced polycystic ovary (PCO) and the protective effects of immunomodulator drug (IMOD), an electromagnetically-treated, selenium-based, herbal medicine. Daily oral administration of letrozole (1 mg/kg) for 21 consecutive days induced ovarian cysts in female rats. An effective dose of IMOD (30 mg/kg per day) was given intraperitoneally for 21 days. Biomarkers of ovarian function, serum concentrations of estradiol, progesterone, testosterone, and ovarian prostaglandin-E (PGE), were analyzed. To determine the role of oxidative stress (OS) in hyperandrogenism-induced PCO, concentrations of cellular lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), peroxynitrite (ONOO), and tumor necrosis factor (TNF)-α as a marker of inflammation and apoptosis were measured in serum and ovaries. Letrozole-induced PCO resulted in significant increases in concentrations of lipid peroxidation and peroxynitrite in serum and ovary, but significantly decreased superoxide dismutase, catalase, and glutathione peroxidase. Serum concentrations of testosterone and TNF-α, and ovarian prostaglandin-E were increased (P < 0.001) in animals with cysts versus control, whereas estradiol and progesterone were decreased (P < 0.01 and P < 0.001, respectively). When compared with controls, letrozole induced irregular cycles and PCO characterized by a high incidence of subcapsular ovarian cysts with a diminished granulosa cell layer, luteinized granulosa cells in the cyst wall, significantly more atretic preantral and antral follicles, and absence of CL. There were almost no intact primary, secondary, and tertiary follicles in PCO rats. All end points assessed were significantly improved by IMOD and reached close to normal levels. In conclusion, the present study provided evidence that toxic free radicals and TNF-α were involved in the pathogenesis of PCO; furthermore, IMOD prevented ovarian histopathologic, endocrine, and biochemical alterations induced by hyperandrogenism.

Endometrial gene expression of acute phase extracellular matrix components following estrogen disruption of pregnancy in pigs.

In pigs, administration of estrogen to gilts on Days 9 and 10 of pregnancy causes conceptus fragmentation and death between Days 15 and 18 of gestation. Conceptus degeneration is associated with breakdown of the microvilli surface glycocalyx on the lumenal epithelium (LE). We previously identified endometrial expression of inter-α-trypsin inhibitor (ITI) and hyaluronic acid (HA), which are key components of extracellular matrix (ECM), during the period of conceptus attachment to the uterine surface in the pig. Tumor necrosis factor-α-inducible protein-6 (TNFAIP6) serves as a linker for ECM expansion and is stimulated by prostaglandin E (PGE). We hypothesized that early estrogen administration alters the normal ECM components forming glycocalyx on the LE. Bred gilts (4 gilts/trt/day) were treated with either 5mg estradiol cypionate (E) or corn oil (CO) on Days 9 and 10 of gestation. The uterus was surgically removed on either Days 10, 12, 13, 15 and 17 of gestation and endometrial tissue snap frozen in liquid nitrogen. Endometrial tumor necrosis factor-α (TNF), TNFAIP6, interleukin 6 (IL6), and inter-α-trypsin inhibitor heavy chains (ITIH) were detected during early pregnancy thereby indicating all components for maintenance of the extracellular glycocalyx are present in the endometrium of pigs. However, only gene expression of ITIH2 was suppressed by E-treatment. TNFAIP6 protein was detected across all days of gestation but was not affected by E-treatment. The present study demonstrates that while the pig endometrium expresses key components of ECM only ITIH2 gene expression was altered by E-treatment. A decrease in ITIH2 could lead to the possible loss of the uterine glycocalyx leading to conceptus degeneration; however, other factors may be involved with the loss of glycocalyx during implantation in the pig following E-treatment.

Effect of DHEA and metformin on corpus luteum in mice.

We evaluated the effect of hyperandrogenism in ovaries with functional and regressing corpora lutea (CL) and the action of metformin in preventing these possible alterations using a mouse model. To obtain a CL functional for 9+/-1 days, immature female mice of the BALB/c strain were injected i.p. with 10 IU/mouse of pregnant mare's serum gonadotropin (PMSG). DHEA (60 mg/kg body weight s.c., 24 and 48 h prior to kill) decreased both serum progesterone (P) and estradiol (E(2)) levels and increased the activity of superoxide dismutase (SOD) from ovaries with functional CL (on day 5 after PMSG). It increased P and E(2) and the activities of SOD and catalase (CAT) and decreased lipoperoxidation of ovaries with regressing CL (on day 9 after PMSG). Treatment with DHEA did not affect the production of prostaglandin F(2alpha) (PGF(2alpha)) or PGE by ovaries with functional CL, whereas DHEA decreased PGF(2alpha) and increased PGE production by ovaries with regressing CL. Metformin (50 mg/kg body weight, orally) given together with DHEA restored E(2) levels from mice with ovaries with functional CL and serum P, PGF(2alpha) and PGE levels, and oxidative balance in mice with ovaries with regressing CL. Metformin alone was able to modulate serum P and E(2) levels, lipoperoxidation, SOD and CAT, and the 5,5-dimethyl-1-pyrroline N-oxide/(*)OH signal. These findings suggest that hyperandrogenism is able to induce or to rescue CL from luteolysis and metformin treatment is able to prevent these effects.

Ibuprofen use during extreme exercise: effects on oxidative stress and PGE2.

PURPOSE: Oxidative stress was examined with use (N = 29) or nonuse (N = 25) of ibuprofen in ultramarathoners after the Western States Endurance Run. METHODS: Oxidative stress was assessed by measuring plasma and urinary F2-isoprostanes, plasma nitrite, and plasma urate. A urinary prostaglandin E2 metabolite (PGE-M) was used as an end point to assess ibuprofen use. Ibuprofen users consumed 600 and 1200 mg of ibuprofen the day before and on race day, respectively, and nonusers avoided all antiinflammatory medications. Blood and urine were collected in the morning before the race and immediately after the race. RESULTS: Use compared with nonuse of ibuprofen significantly increased plasma (P

Effect of epidermal growth factor and prostaglandin on the expression of aromatase (CYP19) in human adrenocortical carcinoma cell line NCI-H295R cells.

We investigated the effects of epidermal growth factor (EGF) and prostaglandins (PG) on the expression of aromatase (CYP19) in human adrenocortical carcinoma cell line NCI-H295R cells. EGF significantly increased aromatase activity and CYP19 gene transcript in NCI-H295R cells. Exon PII was selected from among several tissue-specific exon I regions. Promoter II that abuts on exon PII was activated by EGF. PGE(2) also significantly increased aromatase activity, CYP19 gene transcript, and promoter II activity. The results of experiments using protein kinase (PK) inhibitors suggest that the cAMP-PKA signaling pathway is involved in the up-regulation of aromatase expression by EGF. PGE(2) activated promoter II activity in 4 h, while 12 h was required for its activation by EGF. In addition, PGE(2) was secreted from NCI-H295R cells in response to EGF. Selective agonists for prostaglandin receptors EP(1) and EP(2) significantly increased aromatase activity, which was decreased by the corresponding antagonists. Finally, antagonists for EP(1) and EP(2) inhibited the up-regulation of aromatase expression following EGF. These results suggest that PGE(2) secondarily acts as an autocrine signal in the up-regulation of aromatase expression by EGF in NCI-H295R cells.

Respuesta peritoneal inmediata a la contaminación durante la cirugía laparoscópica en un modelo experimental en el ratón / Immediate peritoneal response to contamination during laparoscopic surgery in an experimental rat model

Introducción. En diversos estudios se ha demostrado que la cirugía laparoscópica (CL) minimiza el traumatismo quirúrgico y preserva la repuesta inmunológica. Otra ventaja es la observación de una menor incidencia de complicaciones infecciosas. Sin embargo, en diferentes estudios in vitro se ha demostrado que la atmósfera con CO2 afecta a la fisiología del macrófago, lo que incidiría en la respuesta a la contaminación peritoneal. Sin embargo, éste es un aspecto controvertido ante la evidencia experimental de una respuesta mejor conservada a la contaminación peritoneal. Este estudio se planteó para investigar la repuesta inmediata del peritoneo a la contaminación en una atmósfera con CO2. Material y método. Se distribuyeron 192 ratones CD-1 en 3 grupos: grupo I, LP, n = 64 (laparotomía); grupo II, LC-CO2, n = 64 (laparoscopia-CO2), y grupo III, LC-T, n = 64, (laparoscopia-tracción). Los ratones fueron aleatorizados para recibir 1 ml de una suspensión de Escherichia coli (1 x 104 UFC/ml) (contaminación [C]) o suero salino (sin contaminación [SC]). Se obtuvo fluido peritoneal a las 1,5, 3, 6 y 12 h tras la cirugía. Se determinaron MCP-1, IL-6 y PGE-2. Resultados. Los valores MCP-1 fueron significativamente superiores y de forma más precoz en el grupo II (LC-CO2-SC) que en el grupo I (LP-SC) (p < 0,007). De manera simultánea, el incremento en el grupo tracción (LC-T-SC, grupo III) fue significativamente mayor (p < 0,002) que tras la laparotomía, sin diferencias respecto al grupo II (LC-C02-SC). Cuando se añadió la contaminación hubo un incremento significativo en los 3 grupos (p < 0,5). Las modificaciones de MCP-1 en el grupo LP-C fueron estadísticamente superiores y aparecieron de forma más tardía que en los grupos con tracción LC-T-C (p < 0,002) y LC-CO2-C (p < 0,02). Interlieucine (IL)-6: los 3 modelos presentaron un incremento significativo, que fue más tardío en el grupo LP-SC. Simultáneamente, el incremento de IL-6 fue más precoz y significativamente superior en el grupo LC-T-SC que en el grupo LP (p < 0,003), sin diferencias entre LC-CO2-SC y LC-T-SC. Se observó una diferencia significativa entre los grupos contaminados y no contaminados en el modelo LC-CO2. El modelo de tracción (grupo LC-T-C) presentó un incremento superior respecto a los grupos LP-C y LC-CO2-C (p < 0,001). Prostaglandine E2 (PGE-2): en los 3 modelos sin contaminación se observó un incremento significativo. Sin embargo, no se encontraron diferencias cuando se añadió la contaminación. Conclusión. El neumoperitoneo con CO2 induce una respuesta peritoneal cualitativamente diferente de la cirugía abierta y modifica la respuesta a la contaminación, con una menor elevación de MCP-1 e IL-6 (AU)

Introduction. Several studies have shown that laparoscopic surgery (LS) minimizes surgical trauma and preserves immune response. Another advantage is the lower incidence of infectious complications. However, several in vitro studies have shown that an atmosphere with CO2 affects macrophage physiology, which would affect the response to peritoneal contamination. This observation is controversial, given the experimental evidence of a better conserved response to peritoneal contamination. The aim of the present study was to investigate the immediate response of the peritoneum to contamination in an atmosphere with CO2. Material and method. A total of 192 CD-1 rats were distributed into three groups: group I, LP, n=64, (laparotomy); group II, LC-CO2, n=64, (laparoscopy-C02), group III, LC-T, n= 64, (laparoscopy-traction). The rats were randomized to receive 1 ml of a suspension of Escherichia coli (1x104 CFU/ml) (contamination [C]) or saline serum (no contamination [NC]). Peritoneal fluid was obtained at 1.5, 3, 6, and 12 h after surgery. Monocyte chemotactic protein-1 (MCP-1), interleukin (IL)-6 and prostaglandin•E2 (PGE2) were determined. Results. MCP-1 levels were significantly higher and increased earlier in group II (LC-C02-NC) than in group I (LP-NC) (p<.007). Simultaneously, the increase in the traction group was significantly higher (p<.002) than after laparotomy, without differences with respect to group II (LC-C02-NC). When contamination was added, there was a significant increase in the three groups (p<.5). The modifications in MCP-1 in the LP-C group were statistically significantly greater and appeared earlier than those in the traction groups, LC-T-C (p<.002) and LC-C02-C (p<.02). Interleukin 6: the three models showed a significant increase, which appeared later in the LP-NC group. Simultaneously, the increase in IL-6 appeared earlier and was significantly greater in the LC-T-NC group than in the LP group (p<.003), with no differences between the LC-C02-NC and LC-T-NC groups. There was a significant difference between contaminated and uncontaminated groups in the LC-CO2 model. The traction model (LC-T-C group) showed a greater increase than the LP-C and LC-C02-C groups (p<.001). PGE2: a significant increase was observed in the three models without contamination. However, when contamination was added, no differences were observed. Conclusion. Pneumoperitoneum with CO2 provokes a peritoneal response that is qualitatively different from open surgery and modifies the response to contamination with a greater increase in MCP-1 and IL-6 (AU)

[Role of prostaglandins in the pathogenesis of stomach ulcer and gastropathy caused by non-steroid anti-inflammatory drugs].

GOAL OF THE STUDY: To assess the pathogenetic significance of the prostaglandin level in the mucous coat of the stomach in patients with osteoarthritis who take several types of non-steroid anti-inflammatory drugs. MATERIALS AND METHODS: The study of the E2 and F2alpha (prostaglandins levels in the blood and mucous coat of the stomach was conducted in 20 patients with stomach ulcer, 15 patients with osteoarthritis taking diclofenac and 16 patients taking celecoxib. RESULTS: It has been shown that the E2 prostaglandin level decreases both in the blood and in the mucous coat of the stomach in patients with stomach ulcer. If non-steroid anti-inflammatory drugs are taken, the E2 prostaglandin level in the mucous coat of the stomach decreases even if there are no erosive-ulcerous lesions. If celecoxib is taken, the changes in the prostaglandin E2 and F2beta level in the mucous coat of the stomach are less marked. Patients with the low content of E2 prostaglandin in the mucous coat of the stomach develop gastropathies more frequently even before non-steroid anti-inflammatory drugs are taken. CONCLUSION: Reduction of the basal prostaglandin secretion in the mucous coat of the stomach is a risk factor of non-steroid anti-inflammatory drugs gastropathy appearance. The use of selective COX-2 inhibitors makes it possible to reduce the risk of gastropathy appearance in this group of patients.

Influence of vedaprofen (Quadrisol) on quality and freezability of stallion semen.

The objective of this study was to evaluate the effect of the non-steroidal anti-inflammatory drug (NSAID) vedaprofen (Quadrisol) on quality and freezability of stallion semen. Experiments were performed using 22 Franches Montagnes stallions from the National Stud in Avenches (Switzerland) randomly divided into a control and test group. Vedaprofen was given orally to all stallions of the test group at the recommended therapeutic dose (initial dose of 2mg/kg followed by 1mg/kg body weight every 12h) for 14 days. Control animals received the same amount of carrier substance. During treatment, blood samples of five stallions in both test and control group were collected for PGF(2 alpha)-metabolite (PG-metabolite) determination. Ejaculates from all stallions were collected and cryopreserved weekly for 14 weeks from September to December. Concentrations of PG-metabolite, PGF and PGE were measured in the seminal plasma of ejaculates collected 2 weeks before, during and 2 weeks after treatment. In fresh semen the volume, concentration, motility and number of normal sperm and sperm with major defects (acrosome defects, abnormal heads, nuclear vacuoles, proximal droplets, midpiece defects) were evaluated. In frozen-thawed semen samples motility as well as viability (SYBR-14/PI) were tested and the hypoosmotic swelling test (HOS) was performed. Results demonstrate that vedaprofen had no effect on blood plasma concentration of PG-metabolite but significantly inhibited both, PGF and PGE concentrations in seminal plasma. Furthermore, all quality parameters in fresh and frozen-thawed semen were not affected by vedaprofen treatment but the time of semen collection had a significant (P<0.05) effect on motility, normal sperm and sperm with nuclear vacuoles in fresh semen.

Effects of dehydroepiandrosterone on ovarian cystogenesis and immune function.

The purpose of the present report was to study the possible relationship between ovarian functionality and the immune response during cystogenesis induced by androgenization with dehydroepiandrosterone (DHEA). Daily injection of DHEA (6 mg/kg body weight) for 20 consecutive days induced ovarian cysts in BALB/c mice. As markers of ovarian function, serum estradiol (E) and progesterone (P) and the ovarian inmunomodulator prostaglandin E (PGE) were analyzed. In order to know how the integrity of the tissue was altered after induction of cystogenesis, the oxidative status was also evaluated. Serum E and P levels, and ovarian PGE concentration, were increased in animals with cysts compared with healthy controls. The oxidant status (quantified by malondialdehyde (MDA) formed after the breakdown of the cellular membrane by free radical mechanisms) was augmented, meanwhile the antioxidant (evaluated by the glutathione (GSH) content) diminished during the induction of cystogenesis. Both immunohistochemical and flow cytometry assays demonstrated that DHEA treatment increased the number of T lymphocytes infiltrating ovarian tissue. Therefore, while ovarian controls showed equivalent expression of CD4+ and CD8+ T cell subsets, injection of DHEA yielded a selective ovarian T cell infiltration as demonstrated by enhanced CD8+ and diminished CD4+ T lymphocyte expression. These results show that the development of cysts involves changes in ovarian function and an imbalance in the oxidant-antioxidant equilibrium. We observed also both an increased and selective T lymphocyte infiltration.

Antithrombin reduces ischemia/reperfusion-induced liver injury in rats by activation of cyclooxygenase-1.

This study was conducted to determine which isoform of cyclooxygenase (COX) is more significantly involved in the anti-thrombin (AT)-induced increase in prostaglandin production in the liver of rats, subjected to hepatic ischemia/reperfusion (I/R). Hepatic tissue levels of 6-keto-PGF(1alpha), a stable metabolite of prostacyclin (PGI(2)), and PGE(2) were transiently increased 1 hour after reperfusion. Thereafter, hepatic PGE2 levels were gradually increased until 6 hours after reperfusion, while hepatic 6-keto-PGF(1alpha) levels were decreased to the pre-ischemia levels at 6 hours after reperfusion. AT significantly enhanced increases in hepatic tissue levels of 6-keto-PGF(1alpha) and PGE(2) seen 1 hour after reperfusion, while it inhibited increases in hepatic PGE(2) levels seen 6 h after reperfusion. Neither dansyl-Glu-Gly-Arg-chloromethyl ketone-treated factor Xa (DEGR-Xa), a selective inhibitor of thrombin generation, nor Trp(49)-modified AT which lacks affinity for heparin, showed any effects on these changes. Pretreatment with indomethacin (IM), a non-selective inhibitor of COX, inhibited AT-induced increases in hepatic tissue levels of 6-keto-PGF(1alpha) and PGE(2) seen 1 hour after reperfusion, whereas pretreatment with NS-398, a selective inhibitor of COX-2, did not. The increase in hepatic tissue blood flow and inhibition of hepatic inflammatory responses seen in animals given AT were reversed by pretreatment with IM, but were not affected by pretreatment with NS-398. Administration of ilo-prost, a stable analog of PGI(2), and PGE(2) produced effects similar to those induced by AT. Increases in hepatic tissue levels of PGE(2) 6 hours after reperfusion were inhibited by pretreatment with NS-398. Although AT did not affect COX-1 mRNA levels 1 hour after reperfusion, it inhibited the I/R-induced increases in hepatic tissue levels of both PGE(2) and COX-2 mRNA 6 hours after reperfusion. These observations strongly suggested that AT might reduce the I/R-induced liver injury by increasing the production of PGI2 and PGE2 through activation of COX-1. Furthermore, since TNF-alpha is capable of inducing COX-2, inhibition of TNF-alpha production by AT might inhibit COX-2-mediated PGE(2) production. These effects induced by AT might contribute to its anti-inflammatory activity.

Localization of isoketal adducts in vivo using a single-chain antibody.

Isoketals are highly reactive gamma-ketoaldehydes formed by the oxidation of arachidonic acid that rapidly adduct to proteins. To investigate the formation of isoketal adducts in vivo, we isolated and characterized a single-chain antibody from a phage displayed recombinant ScFv library that bound a model peptide adducted with synthetic 15-E2-isoketal. Recognition of isoketal adduct by this anti-isoketal adduct single-chain antibody was essentially independent of the amino acid sequence of adducted peptides or proteins. The antibody did not cross-react with 4-hydroxynonenal or 4-oxononanal adducts or with 15-F2t-isoprostane (8-iso-prostaglandin F2alpha). We investigated the formation of isoketal adducts in a well-established model of oxidative injury, hyperoxia. Exposure to >98% oxygen for 7 h dramatically increased both the number of immunoreactive airway epithelial cells and the intensity of immunoreactivity compared with animals exposed to normal room air (21% oxygen). We conclude that isoketal adducts form in epithelial cells as a result of high oxygen exposure and that this single-chain antibody provides a valuable tool to localize the formation of isoketal adducts in tissues in vivo.

The impact of induced stress during days 13 and 14 of pregnancy on the composition of allantoic fluid and conceptus development in sows.

Stress due to regrouping of breeding females is difficult to avoid completely in loose-housing systems. The effects of stress during the maternal recognition of pregnancy on fetal development and survival at Day 30 of pregnancy was, therefore, studied in 17 sows allocated into one control (C-) group, one group deprived of food during Days 13 and 14 (FD-), and one group (A-), which was treated with ACTH (0.01 mg/kg body weight of Synacthen Depot) every sixth hour during the same period. Total number of fetuses, fetal survival rate, volume of allantoic fluid, and the weight and length of total fetal unit, placentas, allantochorion and fetuses were determined. The concentrations of progesterone (P4), PGFM, PGF2, PGE, estrone-sulfate, and estradiol-17beta in the allantoic fluid were analyzed. No significant differences between groups were found for any parameter measured except for P4. Food deprivation increased P4 concentration in the allantoic fluid, and there was a positive correlation between the P4 concentration and the weight of the placenta. It is, therefore, suggested that P4 influences the placenta size among food-deprived sows.

Possible association between amniotic fluid micro-organism infection and microflora in the mouth.

OBJECTIVE: To determine whether oral bacteria are found in the amniotic cavity. DESIGN: Laboratory based analysis of clinical samples. SETTING: Royal London Hospital, Whitechapel. POPULATION: Forty-eight women attending for elective caesarean section. METHODS: Dental plaque, a high vaginal swab, amniotic fluid and chorioamnion tissue were taken from women with intact membranes. MAIN OUTCOME MEASURES: Samples were investigated using culture and microscopy for the presence of microorganisms. Amniotic fluid was analysed by polymerase chain reaction (PCR) for the presence of the ubiquitous 16S rRNA gene specific to most eubacteria. Samples were analysed using PCR genus and species specific primers directed to bacterial taxa found as part of the normal oral microflora (Streptococcus spp. and Fusobacterium nucleatum). Levels of prostaglandin E2 and cytokines were measured in amniotic fluid. RESULTS: Amniotic fluid was positive for universal bacteria PCR, Streptococcus spp. PCR and F. nucleatum PCR in 34/48, 20/48 and 7/48 of cases, respectively. Streptococcus spp. and F. nucleatum were cultured from the dental plaque, vagina and amniotic fluid of 48/48, 14/48, 0/48 and 29/48, 6/48, 0/48 subjects, respectively. A significant association was found between detection of microbial DNA (universal and F. nucletum) and complications in previous pregnancies including miscarriage, intrauterine death, neonatal death, preterm delivery and premature rupture of membranes (P < 0.05 and P < 0.01, respectively). Prostaglandin E2 and cytokine levels, with the exception of IL-1alpha, were not significantly different between women with and without evidence of infection. CONCLUSIONS: The results indicate that Streptococcus spp. and F. nucleatum in the amniotic fluid may have an oral origin.

Optimum intensities of ultrasound for PGE(2) secretion and growth of osteoblasts.

This study compared the effects of different intensity ultrasound (US) on osteoblasts in the far-field model with effects of the near-field model from the literature, to understand the relations between prostaglandin E(2) (PGE(2)) and osteoblast growth. We used an in vitro model to investigate the effects of 1-MHz, pulsed 1:4, and five different spatial-average temporal-peak intensity (150, 300, 600, 1200 and 2400 mW/cm(2)) US stimulations in far-field exposure (240 mm) on osteoblasts for 15 min. Optimum intensity in this study was 600 mW/cm(2), and cell density and PGE(2) secretion could be significantly stimulated at this intensity. This research may indicate that the growth of osteoblasts by US stimulation was, at least partly, due to increases in the synthesis and secretion of PGE(2). This well-controlled model can lead to further research on the biologic mechanisms for US.

[Inhibitory effect of alveolar macrophages on the proliferation of pulmonary fibroblasts].

The effect of alveolar macrophages (AM) harvested from Wistar rats by lung lavage on proliferation of human embryo pulmonary fibroblasts in culture was investigated. It was observed that supernatants of AM decreased the uptake of (3)H TdR by the pulmonary fibroblasts. The AM activated with opsonized zymosan (OPZ) showed a stronger inhibitory effect on fibroblast proliferation compared with inactivated AM. Following pretreatment with indomethacin, the inhibitory effect of AM was abolished and reversed to stimulatory effect on pulmonary fibroblast proliferation. The PGE content in AM supernatant was measured with radioimmunoassay. It was observed that the inhibitory effect of AM was highly correlated to prostaglandin (PGE) content in the supernatant of AM. The results suggest that AM has both inhibitory and stimulatory effects on the proliferation of pulmonary fibroblast; the inhibitory effect is primary under normal conditions. This inhibitory action is mainly due to PGE secreted from AM. It is, therefore, suggested that AM plays an important role in suppressing pulmonary fibrosis under normal conditions.