Bioassay for determining 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEs) in human hepatoma Hepg2 cells

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), and benz[a]anthracene (BA) highly induce cytochrome P4501A1, determined by aryl hydrocarbon hydroxylase (AHH) activity, in human hepatoma HepG2 cells within 24 h. AHH activity induced by TCDD and TCDF persists for at least 48 h. In contrast, AHH activity induced by BA rapidly declines, although the amounts applied are 4-5 orders of magnitude higher than those of TCDD or TCDF. AHH induction in HepG2 cells differs from that in rat hepatoma cells H4IIEC3/T in two aspects: (1) HepG2 cells are 20 times less sensitive to the test compounds than H4IIEC3/T cells. (2) TCDF-induced AHH activity does not persist in the rat cells. The results suggest that human HepG2 cells, because of their low sensitivity, are inferior to rat H4IIEC3/T cells for determining TCDD equivalents in environmental samples. They may be useful for investigating species dependent differences in the toxicokinetics of individual polyhalogenated aromatic hydrocarbon congeners.     

Markedly altered colchicine kinetics in a fatal intoxication: examination of contributing factors

1. Colchicine poisoning, which is relatively rare, is associated with significant morbidity and mortality. Whilst a new treatment modality, in the form of colchicine-specific Fab fragments is on the horizon, currently available therapy is largely supportive. 2. The elimination of colchicine occurs primarily by hepatic metabolism, following a first-order process, with significant enterohepatic circulation. Renal extraction is responsible for approximately 20% of colchicine elimination. 3. We report a case of colchicine intoxication, complicated by the presence of co-ingestants, in which serum colchicine concentrations remained quasi-constant over the 3 days of the patient's survival, consistent with marked alterations both in metabolism and excretion. The initial presentation was relatively benign but the subsequent course was one of severe colchicine poisoning, resulting in death. 4. Severe colchicine toxicity appears to have resulted in a vicious cycle of progressive organ dysfunction and impaired elimination. 5. Josamycin, one of the co-ingestants and an inhibitor of P-glycoprotein, the membrane pump responsible for multidrug resistance, may have played a significant role in impeding the cellular and biliary elimination of colchicine. Co-ingested opioid and anticholinergic compounds may have altered colchicine absorption and gastrointestinal transit. 6. This case serves as a reminder of the need for attention to co-ingested drugs, to early aggressive therapy, and if available, to consideration of immunotherapy.     

Purification, characterization, and spasmogenic activity of neurotensin from the toad Bufo marinus

Neurotensin (NT) was isolated from an extract of the intestine of the cane toad, Bufo marinus and its primary structure established as: pGlu-Ala-Ile-Val-Ser-Lys-Ala-Arg-Arg-Pro-Tyr-Ile-Leu. This amino acid sequence shows five substitutions (Leu2 --> Ala, Tyr3 --> Ile, Glu4 --> Val, Asn5 --> Ser, and Pro7 --> Ala) compared with bovine NT. Synthetic Bufo NT (pD2 = 8.05 +/- 0.28) was equipotent and equally effective as bovine NT (pD2 = 8.24 +/- 0.38) in producing spasmogenic contraction of isolated segments of toad small intestine. However, the maximum response produced by Bufo NT was only 35 +/- 2% of that produced by substance P. The potencies, but not the maximum responses, to Bufo and bovine NT were significantly (p < 0.05) attenuated by pre-treatment with atropine but neither parameter was significantly diminished by tetrodotoxin and indomethacin. The data suggest that the action of NT involves interaction with receptors on toad intestinal smooth muscle that recognize the C-terminal region of NT (residues 8-13) that has been fully conserved during evolution of tetrapods. Contractile activity is mediated, at least in part, by release of acetylcholine.     

Cerebrospinal fluid selenium and chromium levels in patients with Parkinson's disease

We compared CSF and serum levels of selenium and chromium, measured by atomic absorption spectrophotometry, in 28 patients with Parkinson's disease (PD) and 43 matched controls. The CSF and serum levels of these trace metals did not differ significantly between PD patients and controls. CSF selenium and chromium levels were not correlated with age, age at onset, duration of the disease, scores of the Unified Parkinson Disease Rating Scale of the Hoehn and Yahr staging in the PD group. Although antiparkinsonian therapy did not influence significantly the CSF levels of selenium, PD patients not treated with levodopa had significantly higher CSF selenium levels than controls (p < 0.01). It is possible that increased CSF selenium levels could indicate an attempt of protection against oxidative stress. The normality of CSF and serum chromium levels suggest that these values are not related with the risk for PD.     

Marriage and the parenting alliance: longitudinal prediction of change in parenting perceptions and behaviors

The study evaluates how marriage and the parenting alliance affect parenting experiences over time. Couples (N = 79) with school-age children who have mental retardation completed self-report and observational measures of marriage, the parenting alliance, and parenting attitudes and behaviors at 2 periods, 18-24 months apart. Longitudinal structural equation modeling demonstrated significant effects of marital quality on changes over time in self-reports of perceived parenting competence for both the mothers and the fathers, and in observed negative mother-child interactions. Also, in all cases, the parenting alliance mediated the effects of marriage on parenting experiences. There was little evidence of reciprocal causation in which parenting variables predicted change in the quality of marriage and the parenting alliance. Interactions involving child age suggested that teenagers as opposed to younger children were more reactive to negative features of their parents' marital functioning and parenting alliance. Implications are discussed regarding stable but negative marital functioning and regarding possible differences in mothers' and fathers' parenting in the context of marital distress.     

Computerized tomographic assessment of the composition of gallstones

We present the results of an in vitro study of biliary lithiasis (n = 106) with computerized tomography (CT). We analyzed the correlation between composition of gallstones (crystallographic analysis of thin sections and atomic absorption spectrophotometric analysis of total calcium content) with CT patterns (densitometry) and their corresponding attenuations. Six main CT patterns were distinguished: hypodense (homogeneous and heterogeneous), dense, homogeneous hyperdense, ringed hyperdense and irregular hyperdense. Statistically significant differences (p < 0.05) were found between attenuations for cholesterol (mixed and pure) and pigmentary stones. We found a good inverse correlation between attenuations and cholesterol content; the contents of pigment, inorganic calcic salts and total calcium content also showed significant direct correlations (p < or = 0.01). Of the samples classified as pure cholesterol type stones, 86.4% showed hypodense patterns, and the remaining 13.6% showed irregular hyperdense patterns. Of the pigmentary stones, 80% showed homogeneous hyperdense images. All stones that contained more than 3% calcium produced hyperdense patterns, whereas 72.4% of the stones that contained less calcium produced hypodense images. Tomodensitometric measurements provided more information than simple radiography, and made it possible to distinguish a greater variety of gallstone types based on structural complexity.     

Evidence that the mediobasal hypothalamus is the primary site of action of estradiol in inducing the preovulatory gonadotropin releasing hormone surge in the ewe

Although a neural site of action for estradiol in inducing a LH surge via a surge of GnRH is now well established in sheep, the precise target(s) for estrogen within the brain is unknown. To address this issue, two experiments were conducted during the breeding season using an artificial model of the follicular phase. In the first experiment, bilateral 17beta-estradiol microimplants were positioned in either the medial preoptic area (MPOA) or the mediobasal hypothalamus (MBH), and LH secretion was monitored. An initial negative feedback inhibition of LH secretion was observed in ewes that had estradiol microimplants located in the MPOA (6 of 6 ewes) or caudal MBH in the vicinity of the arcuate nucleus (4 of 4). In contrast, a normal LH surge was only found in animals bearing estradiol microimplants in the MBH (5 of 10). Detailed analysis of estradiol microimplant location with respect to the estrogen receptor-alpha-immunoreactive cells of the hypothalamus revealed that 4 of the 5 ewes exhibiting a LH surge had microimplants located bilaterally within or adjacent to the area of estrogen receptor-expressing cells of the ventromedial nucleus. Two of these ewes exhibited a LH surge without showing any form of estrogen negative feedback. In the second experiment, we used the technique of hypophyseal portal blood collection to monitor GnRH secretion directly at the time of the LH surge induced by estradiol delivered either centrally or peripherally. Central estradiol implants induced the GnRH surge. The duration and mean plasma concentration of GnRH during the surge were not different between animals given peripheral or central MBH estradiol implants. Cholesterol-filled MBH microimplants did not evoke a GnRH surge. We conclude that the ventromedial nucleus is the primary site of action for estradiol in stimulating the preovulatory GnRH surge of the ewe, whereas the MPOA and possibly the caudal MBH are sites at which estrogen can act to inhibit LH secretion. These data provide evidence for the sites within the ovine hypothalamus responsible for mediating the bimodal influence of estradiol on GnRH secretion and suggest that different, and possibly independent, neuronal cell populations are responsible for the negative and positive feedback actions of estradiol.