Aspects of cAMP Signaling in Epileptogenesis and Seizures and Its Potential as Drug Target

Neurochemical Research - Epilepsy is one of the most common chronic neurological conditions. Today, close to 30 different medications to prevent epileptic seizures are in use; yet, far from all...     

Mercury Levels in Barbadian Fisher Folk and Commonly Caught Fish Species

Ocean fish are an important staple food for many Caribbean persons. Some species, however, have high Hg concentration levels and represent a significant source of Hg exposure. This study assessed Hg concentrations in fish species commonly caught and consumed in Barbados as well as Hg exposure of those who work in the fishing industry. Total Hg concentrations were measured in 100 fish samples. Human Hg concentrations were determined from hair snippets taken from 46 fisher folk and fish consumption habits evaluated using interview-administered questionnaires. Mercury levels varied by fish species with predatory fish species having the highest concentrations. Low Hg concentrations were found in dolphinfish (Coryphaena hippurus) and flyingfish (Hirundichthys affinis). Reported fish consumption rates were high and varied significantly between men and women, 2675 ± 1600 and 1275 ± 600 (g/week), respectively (p = 0.01). The median hair Hg concentration was 3.1 ± 2.3 μg/g (range: 0.01–15.6 μg/g) and significantly associated with consumption of “high-risk” fish species (p = .02). These results show that it is possible to make informed choices concerning fish consumption habits in order to maximize benefits while minimizing the effect of Hg exposures, especially for sensitive groups such as pregnant women and infants.     

Disulfide-bonded multimers of proteoglycan 4 (PRG4) are present in normal synovial fluids

Background

The proteoglycan 4 (PRG4) gene encodes for a mucin-like O-linked glycosylated protein with several names, including lubricin and superficial zone protein. The objective of this study was to analyze PRG4 in normal bovine calf and steer synovial fluids for evidence of native multimers formed by intermolecular disulfide bonds.

Methods

A combination of mucin biochemical techniques, with antibodies to both terminal domains and the mucin-like domain of PRG4, were used for analyses.

Results

Multimers were present in both calf and steer fluids, and reduction and alkylation converts the multimeric complex (likely dimeric) into monomeric subunits. Tandem mass spectrometry analyses supported the Western blot data and identified PRG4 in the reduced ∼ 345 kDa monomeric form. Interestingly, ∼ 70 kDa fragments released upon reduction contained peptides from both the N and C terminal regions, which most likely represent fragments of a sparsely glycosylated PRG4 population that are disulfide-linked to extensively glycosylated, intact monomers.

Conclusions

The analyses described here have demonstrated the presence of native disulfide-bonded multimers of PRG4 in normal bovine synovial fluids.

General significance

These structures are similar to those described for multimerization of mucins in general. Such multimerization and proteolytic cleavage of PRG4 may have functional significance in joint health and disease.     

Isolation of nuclei from label-retaining cells and measurement of their turnover rates in rat colon

We describe here a new technique for isolating nuclei from long-term label-retaining cells (LRCs), a subpopulation enriched with stem cells from colon, and for measuring their proliferation rates in vivo. A double-label approach was developed, combining the use of bromodeoxyuridine (BrdU) and ²H₂O. Male Fisher 344 rats were administered BrdU in drinking water continuously for 2–8 wk. BrdU was then discontinued (BrdU washout), and animals (n = 33) were switched to ²H₂O in drinking water and killed after 2, 4, and 8 wk. Nuclei from BrdU-positive cells (LRCs) were collected by flow cytometry. The percentages of LRCs were 7 and 3.8% after 4 and 8 wk of BrdU washout, respectively. Turnover rates of LRCs were measured on the basis of deuterium incorporation from ²H₂O into DNA of LRC nuclei, as determined by mass spectrometry. The proliferation rate of the LRCs collected was 0.33–0.90% per day (half-life of 77–210 days). Significant contamination from other potentially long-lived colon cells was excluded. In conclusion, this double-labeling method allows both physical isolation of nuclei from colon epithelial LRCs and measurement of their in vivo proliferation rates. Use of this approach may allow better understanding of mechanisms by which agents induce or protect against colon carcinogenesis. carcinogenesis; deuterated water; long-term label-retaining cells; stable isotopes     

Aplysamine-1 and related analogs as histamine H3 receptor antagonists

Aplysamine-1 (1), a marine natural product, was synthesized and screened for in vitro activity at the human and rat histamine H3 receptors. Aplysamine-1 (1) was found to possess a high binding affinity for the human H3 receptor (Ki = 30+/-4 nM). Synthetic analogs of 1, including des-bromoaplysamine-1 (10) and dimethyl-{2-[4-(3-piperidin-1-yl-propoxy)-phenyl]-ethyl}-amine (13), were potent H3 antagonists.