Timing of Maternal Exposure and Foetal Sex Determine the Effects of Low-level Chemical Mixture Exposure on the Foetal Neuroendocrine System in Sheep.

We have shown that continuous maternal exposure to the complex mixture of environmental chemicals (ECs) found in human biosolids (sewage sludge), disrupts mRNA expression of genes crucial for development and long-term regulation of hypothalamic-pituitary gonadal (HPG) function in sheep. The present study investigated whether exposure to ECs only during preconceptional period or only during pregnancy perturbed key regulatory genes within the hypothalamus and pituitary gland and whether these effects were different from chronic (life-long) exposure to biosolid ECs. The findings demonstrate that the timing and duration of maternal EC exposure influences the subsequent effects on the foetal neuroendocrine system in a sex-specific manner. Maternal exposure prior to conception, or during pregnancy only, altered the expression of key foetal neuroendocrine regulatory systems such as gonadotrophin-releasing hormone and kisspeptin to a greater extent than when maternal exposure was 'life-long'. Furthermore, hypothalamic gene expression was affected to a greater extent in males than in females and, following EC exposure, male foetuses expressed more 'female-like' mRNA levels for some key neuroendocrine genes. This is the first study to show that 'real-life' maternal exposure to low levels of a complex cocktail of chemicals prior to conception can subsequently affect the developing foetal neuroendocrine system. These findings demonstrate that the developing neuroendocrine system is sensitive to EC mixtures in a sex-dimorphic manner likely to predispose to reproductive dysfunction in later life.

The dynamics of microbial colonization of barrier membranes for guided tissue regeneration.

The microbial colonization of expanded polytetrafluoroethylene membrane by putative periodontopathogens at 3 minutes of intraoral manipulation was determined in 42 patients with 42 mandibular posterior two- to three-wall defects. Twenty patients exhibited no periodontal pockets of > or = 5 mm, other than the study site, and low levels of pathogens (group A). Twenty-two patients revealed multiple periodontal pockets of 5 mm or more and numerous pathogens (group B). Within the preceding 3 months of regenerative surgery, group A patients had received apically positioned flap surgery with osseous recontouring (except for the study site), and group B patients had been enrolled in a non-surgical maintenance program. The subgingival microbiota was examined prior to regenerative therapy, and the membrane microbiota was examined at 3 minutes and at the time of removal at 6 weeks by culture, DNA probes, and phase-contrast microscopy. The mean initial defect depth was 7.4 mm for group A and 7.2 mm for group B. At 6 months, the difference in mean clinical attachment gain was statistically significant (P < 0.001; group A: 3.4 mm; group B: 1.4 mm). At 3 minutes, putative pathogens were detected in seven (16.7%) membranes in group B (group Binfected), and the associated sites gained only 0.6 mm in clinical attachment at 6 months. Clinical attachment gain was modeled as a linear function of the explanatory variables (r2 = 86%). The presence of Porphyromonas gingivalis detected by DNA probe at 3 minutes was associated with 1.5 mm less expected gain (P = 0.0002). Total microbial counts and the percentage of Peptostreptococcus micros and Capnocytophaga species at baseline, and of motile rods on the membrane surface facing the gingiva at 6 weeks, were statistically significant negative predictors of clinical attachment. For each week the membrane remained covered, an additional 0.5 mm gain could be expected (P = 0.002); and for every 10 sites that exhibited bleeding on probing, the clinical attachment gain was 0.6 mm less at the site of regeneration (P < 0.0001). The present results showed that putative pathogens may colonize membranes within 3 minutes of intraoral manipulation. The patient group treated with periodontal osseous surgery revealed the lowest levels of periodontal pathogens in the membranes and exhibited the most gain in clinical attachment.

Ontogeny of photic-induced c-fos mRNA expression in rat suprachiasmatic nuclei.

The suprachiasmatic nucleus (SCN) of the hypothalamus contains a circadian pacemaker that controls a variety of physiological and behavioral rhythms. Photic induction of immediate early genes such as c-fos in the SCN occurs in a circadian-phase dependent manner, suggesting that c-fos may be part of the pathway for entrainment of circadian rhythms. The purpose of this study was to determine the point in development when photic stimuli can first activate c-fos mRNA expression in the rat SCN. The results indicate that photic stimulation can induce statistically significant c-fos mRNA expression at circadian time 22 (CT22) on postnatal day 1 (P1), although hybridization above background levels can be detected in the SCN of some rat pups earlier in the subjective night. We infer that a multi-step pathway between visual transduction by the retina and regulation of c-fos transcription in SCN cells must be mature by CT22 on P1.

c-fos mRNA in the suprachiasmatic nuclei in vitro shows a circadian rhythm and responds to a serotonergic agonist.

The mammalian suprachiasmatic nuclei (SCN) contain a circadian clock that produces approximately 24 h rhythms of physiology and behavior even during constant dark. Under such conditions, light stimuli applied during the subjective night induce phase shifts of circadian rhythms and increase immediate early gene expression (c-fos) in the SCN. In vitro preparations of the SCN continue to show circadian rhythms of metabolic rate and neuronal firing rates, which can be phase shifted by non-photic stimuli. This study was designed to investigate whether the SCN display a rhythm of c-fos mRNA levels in vitro and whether quipazine, which phase-shifts the SCN circadian clock, induces c-fos expression in vitro. Levels of c-fos mRNA were found to be significantly higher in the subjective day than subjective night in the SCN in vitro. This rhythm parallels other in vivo and in vitro rhythms in SCN metabolic and neuronal activity and is consistent with previous in vivo work showing higher daytime levels of Fos-like immunoreactivity in animals maintained under constant dark conditions. Quipazine treatment during the subjective day (which phase-advances the circadian rhythm of neuronal firing in the SCN) decreased c-fos mRNA levels in the dorsomedial but not ventrolateral SCN, but quipazine did not affect c-fos levels when administered at night. This effect is consistent with serotonergic agonists inhibiting SCN neuronal activity and is the first evidence that a non-photic phase-shifting stimulus alters c-fos in the SCN at a phase-appropriate time.