Parent bisphenol A accumulation in the human maternal-fetal-placental unit

Bisphenol A (BPA), an endocrine disruptor, is employed in the manufacture of a wide range of consumer products. The suggestion that BPA, at amounts to which we are exposed, alters the reproductive organs of developing rodents has caused concern. At present, no information exists concerning the exposure of human pregnant women and their fetuses to BPA. We therefore investigated blood samples from mothers (n = 37) between weeks 32 and 41 of gestation. Afer the births, we also analyzed placental tissue and umbilical cord blood from the same subjects. We developed a novel chemical derivatization-gas chromatography/mass spectrometry method to analyze parent BPA at concentrations < 1 micro g/mL in plasma and tissues. Concentrations of BPA ranged from 0.3 to 18.9 ng/mL (median = 3.1 ng/mL) in maternal plasma, from 0.2 to 9.2 ng/mL (median = 2.3 ng/mL) in fetal plasma, and from 1.0 to 104.9 ng/g (median = 12.7 ng/g) in placental tissue. BPA blood concentrations were higher in male than in female fetuses. Here we demonstrate parent BPA in pregnant women and their fetuses. Exposure levels of parent BPA were found within a range typical of those used in recent animal studies and were shown to be toxic to reproductive organs of male and female offspring. We suggest that the range of BPA concentrations we measured may be related to sex differences in metabolization of parent BPA or variable maternal use of consumer products leaching BPA.     

A Pharmacokinetic/Pharmacodynamic Approach to Predict the Cumulative Cortisol Suppression of Inhaled Corticosteroids

The suppression of endogenous cortisol release is one of the major systemic side effects of inhaled corticosteroids in the treatment of asthma. The circadian rhythm of the endogenous cortisol release and the resulting plasma concentrations as well as the release suppression during corticosteroid therapy could previously be described with an integrated PK/PD model. Based on this model, a PK/PD approach was developed to quantify and predict the cumulative cortisol suppression (CCS) as a surrogate marker for the systemic activity of inhaled corticosteroid therapy. The presented method was applied to predict CCS after single doses and during short-term multiple dosing of the inhaled corticosteroids flunisolide (FLU), fluticasone propionate (FP), and triamcinolone acetonide (TCA), and after oral methylprednisolone as systemic reference therapy. Drug-specific PK and PD parameters were obtained from previous single-dose studies and extrapolated to the multiple-dose situation. For single dosing, a similar CCS within the range of 16–21% was predicted for FP 250 g, FLU 500 g, and TCA 1000 g. For multiple dosing, a respective CCS of 28–33% was calculated for FLU 500 g bid, FP 250 g, bid, and TCA 1000 g bid. Higher cortisol suppression compared to these single and multiple dosing regimens of the inhaled corticosteroids was predicted after oral doses of only 1 mg and 2 mg methylprednisolone, respectively. The predictive power of the approach was evaluated by comparing the PK/PD-based simulations with data reported previously in clinical studies. The predicted CCS values were in good correlation with the clinically observed results. Hence, the presented PK/PD approach allows valid predictions of CCS for single and short-term multiple dosing of inhaled corticosteroids and facilitates comparisons between different dosing regimens and steroids.     

Chromosomal Aberrations in Lymphomas of the Gastrointestinal Tract

B-cell lymphomas of the gastrointestinal (GI) tract have represented a field of extensive research ever since a close association was shown with chronic inflammatory processes such as Helibacter pylon infection. Much evidence has accumulated to suggest that the mucosa-associated lymphoid tissue (MALT) induced by inflammation and autoimmune processes is the environment which gives rise to the small cell lymphomas of the GI tract (e.g. extranodal marginal B-cell lymphoma according to REAL). The small B-cell lymphoma may then progress to the large cell variants. Hence, B-cell lymphomas of the GI tract may present a model for lymphomagenesis and progression. In this review, recent cytogenetic data are discussed which yield new insights into the biology of gastrointestinal lymphomas.     

Identification of 3{alpha}-hydroxy-cyproterone acetate as a metabolite of cyproterone acetate in the bile of female rats and the potential of this and other already known or putative metabolites to form DNA adducts in vitro

Cyproterone acetate (CPA) is a synthetic steroid hormone usedin the therapy of prostate cancer in men and different formsof acne and hirsutism in women. CPA has been shown by ³²P-postlabelinganalysis to bind covalently to hepatic DNA of rats in vivo andin vitro. A prerequisite for DNA adduct formation of CPA ismetabolic activation of the drug to a reactive intermediate.In the present study bile was collected from [³H]CPA-treatedfemale rats and, following chromatographic separation of bileextracts, fractions of the eluate were examined for the presenceof reactive metabolites which were able to form adducts withcalf thymus DNA in vitro. The formation of adducts was detectedby ³²P-postlabeling analysis. One major metabolite of CPA presentin the bile extracts was isolated and, following a thoroughstructural elucidation by mass spec-trometry and ¹H-NMR, thismetabolite was identified as 3     

Pathophysiological role of poly(ADP-ribose) polymerase (PARP) activation during acetaminophen-induced liver cell necrosis in mice.

DNA fragmentation in hepatocytes occurs early after acetaminophen (AAP) overdose in mice. DNA strandbreaks can induce excessive activation of poly(ADP-ribose) polymerases (PARP), which may lead to oncotic necrosis. Based on controversial findings with chemical PARP inhibitors, the role of PARP-1 activation in AAP hepatotoxicity remains unclear. To investigate PARP-1 activation and evaluate a pathophysiological role of PARP-1, we used both PARP inhibitors (3-aminobenzamide; 5-aminoisoquinolinone) and PARP gene knockout mice (PARP-/-). Treatment of C3Heb/FeJ mice with 300 mg/kg AAP resulted in DNA fragmentation and alanine aminotransferase (ALT) release as early as 3 h, with further increase of these parameters up to 12 h. Few nuclei of hepatocytes stained positive for poly-ADP-ribosylated nuclear proteins (PAR) as indicator for PARP-1 activation at 4.5 h. However, the number of PAR-positive cells and staining intensity increased substantially at 6 and 12 h. Pretreatment with 500 mg/kg 3-aminobenzamide before AAP attenuated hepatic glutathione depletion and completely eliminated DNA fragmentation and liver injury. Delayed treatment several hours after AAP was still partially protective. On the other hand, liver injury was not attenuated in PARP-/- mice compared to wild-type animals. Similarly, the specific PARP-1 inhibitor 5-aminoisoquinolinone (5 mg/kg) was not protective. However, 3-aminobenzamide attenuated liver injury in WT and PARP-/- mice. In summary, PARP-1 activation is a consequence of DNA fragmentation after AAP overdose. However, PARP-1 activation is not a relevant event for AAP-induced oncotic necrosis. The protection of 3-aminobenzamide against AAP-induced liver injury was due to reduced metabolic activation and potentially its antioxidant effect but independent of PARP-1 inhibition.     

Phase shift in the REM sleep rhythm

Carbohydrate depletion during exercise was measured in the liver, in the three different types of skeletal muscle, and in the blood of exercise-trained and untrained rats. The acute exercise test consisted of 45 min of treadmill running of progressively increasing intensity. The training program consisted of 6 hrs of swimming per day, 5 days per week for 14 weeks; the training induced an increase of approximately 35 percent in the respiratory capacity of gastrocnemius muscle, and a 14 percent incrase in heart weight. Glycogen stores in fast-twitch red, fast-twitch white, and slow-twitch red types of skeletal muscle, were depleted significantly more slowly in the trained than in the untrained animals during the treadmill exercise test. Resting glycogen stores in the liver were higher and were depleted more slowly during exercise in the trained than the untrained animals. Blood lactate concentration was significantly lower in the trained than in the untrained rats at the end of the exercise test. These results provide evidence that endurance exercise training induces adaptation which protect against the depletion of glycogen from the liver and from the tree types of skeletal muscle during prolonged exercise.     

Osteoconductive modifications of Ti-implants in a goat defect model: characterization of bone growth with SR muCT and histology

In this work the osteoconductive potential of coatings for titanium implants using different extracellular matrix components was evaluated. Cylindrical implants with two defined cavities A and B were coated with collagen type I, type III, or RGD peptide, and placed in the femur of goats together with an uncoated reference state. Bone contact and volume were determined after 5 and 12 weeks implantation, using both histomorphometry and synchrotron radiation micro computed tomography (SR muCT) as the methods complement each other: SR muCT allows for a high precision of bone detection due to the large number of analysed slices per sample, while histology offers a better lateral resolution and the possibility of additionally determining bone contact. Both methods revealed similar tendencies in bone formation for the differently bio-functionalized implants, with the SR muCT data resulting in significant differences. After 5 and 12 weeks, all three coatings showed a significant increase in bone volume over the uncoated reference, with the highest results for the collagen coatings. The coating consisting of just the RGD-sequence to improve cell adhesion showed only a slight improvement compared with the reference material. For uncoated titanium, RGD, and especially collagen type I, the response in cavity A, situated in denser bone, was stronger than in cavity B. Collagen type III, on the other hand, appeared to be the more effective coating in areas of lesser bone density as represented by cavity B. These results indicate that matrix molecules (or combinations thereof) are capable of generating the appropriate signals for the specific microenvironment around implants and can thus accelerate the bone formation process and increase the stability of implants.     

Synthesis and cell surface display of class II determinants by long-term propagated rat T line cells

We have investigated the capacity of the encephalitogenic BS rat T cell line bs 83 and its variant clone bs 83.III.C6 to synthesize and express RT1.B-specific class II molecule subsets defined by monoclonal antibodies (mAb) MRC-OX6 and MRC-OX3. Earlier studies had indicated that mAb MRC-OX6 recognizes three distinct molecular species: an immature oligomeric polypeptide chain complex comprised of the polymorphic subunits alpha, beta and the invariant proteins of the gamma group; a biosynthetic intermediate composed of post-translationally modified alpha, beta and gamma chain (denoted p35) and a fully glycosylated alpha, beta two-chain complex derived from the plasma membrane. MRC-OX3 was shown to recognize a serologically distinct alpha, beta two-chain complex that coexists with the MRC-OX6-specific heterodimer at the cell surface. Here we show that premutant bs 83 cells were unable to synthesize class II molecules of either set. In contrast endogeneous synthesis by mutant cells of MRC-OX6-specific molecules was demonstrated. Unlike control spleen cells variant cells failed to synthesize the mature MRC-OX3-reactive class II subset. Instead a three-polypeptide chain complex comprised of the terminally glycosylated subunits alpha, beta and invariant chain p35 was present at the cell surface. This complex appears to represent the preserved biosynthetic intermediate that failed to release invariant chain p35 upon its transit into the plasma membrane. These latter observations support our notion of gamma chain-induced epitope diversification during post-translational maturation of RT1.B-specific class II molecules.     

Astrocytes Upregulate Glial Fibrillary Acidic Protein (GFAP), but not Insulin-like Growth Factor-I (IGF-I) during Experimental Autoimmune Neuritis (EAN)

T cell-mediated autoimmune neuritis produces rapid activation of spinal cord microglia. To determine whether this microglial response upregulates astrocytic expression of IGF-related proteins, we induced EAN and used in situ hybridization and immunocytochemistry to examine the mRNAs and peptides for glial fibrillary acidic protein (GFAP), insulin-like growth factor-I (IGF-I), IGF-I receptor (IGFR-I) and IGF binding protein-2 (IGFBP-2). Relative levels of GFAP mRNA and peptide were highest in the lumbar spinal cord 4–10 d following T cell transfer and significant GFAP elevations were still present after three weeks. The astrocytes expressing GFAP mRNA and peptide were localized around motoneurons which were related topographically to axons in peripheral nerve inflammatory lesions. In the nucleus gracilis, where terminals of dorsal root ganglion neurons are located, astrocytic levels of GFAP mRNA and peptide rose later and did not reach their highest levels until 21 d after T cell transfer. Even though microglia were activated in both locations 2–4 d after transfer, astrocytic levels of IGF-I, IGFR-I and IGFBP-2 mRNA and peptide did not differ significantly from those observed in controls. The dissociation of GFAP and IGF-I expression in EAN suggests that these astrocytic responses may be independently regulated. We also suggest that the type and severity of remote neuronal injury are probably more important inducers and regulators of these astrocytic responses than microglial cell activation.