Novel phosphoserine phosphatase inhibitors

Phosphoserine phosphatase (EC 3.1.1.3) catalyzes the final step in the major pathway of L-serine biosynthesis in brain. This enzyme may also regulate the levels of glycine and D-serine, the known and putative co-agonists for the glycine site of the N-methyl-D-aspartate receptor in caudal and rostral brain regions, respectively. Using L-phosphoserine as substrate, the rank order potency for inhibition of phosphoserine phosphatase was p-chloromercuriphenylsulfonic acid (CMPSA) > glycerophosphorylcholine > hexadecylphosphocholine > or = phosphorylcholine > N-ethylmaleimide > or = L-serine > fluoride > D-2-amino-3-phosphonopropionic acid (D-AP3). Glycerylphosphorylcholine (IC50 18 microM) was found to be an uncompetitive inhibitor of phosphoserine phosphatase. Glycerylphosphorylcholine probably binds a novel site on the enzyme since the known allosteric inhibitor L-serine is highly selective for its feedback regulatory site, indicated by the inactivity of 25 L-serine analogs. Fluoride ion (IC50 770 microM) may bind the active site as has been shown for other Mg2+-dependent enzymes. The sulfhydryl reagent CMPSA is a potent, noncompetitive inhibitor of the enzyme using L-phosphoserine as substrate (IC50 9 microM) but is > 300-fold less potent using D-phosphoserine as substrate. Substrate-dependent differences are also observed with the sulfhydryl alkylator N-ethylmaleimide, which inhibits L-phosphoserine, but stimulates D-phosphoserine hydrolysis. These sulfhydryl reagents may dissociate multimeric forms of the enzyme to form monomers; the multimeric forms and monomers may preferentially cleave L- and D-phosphoserine, respectively. Phosphorylcholine esters and sulfhydryl reagents may prove useful in determining the contribution of phosphoserine phosphatase to the biosynthesis of glycine and D-serine in neuronal tissue in vitro.     

Prostate specific antigen in the expressed prostatic fluid of men with benign prostatic hyperplasia and prostate carcinoma

PURPOSE: We tested the hypothesis that the histochemically demonstrated prostate specific antigen (PSA) content of prostate carcinoma cells does not necessarily reflect PSA production and secretion by evaluating expressed prostatic fluid. MATERIALS AND METHODS: Expressed prostatic fluid and serum from 152 men with clinical benign prostatic hypertrophy (BPH), 132 with histologically proved BPH and 46 with prostate carcinoma were analyzed with the Hybritech PSA assay. RESULTS: Expressed prostatic fluid PSA levels from carcinoma patients (median 1.70 mg./ml., mean 2.25) were significantly higher than in the histologically proved BPH group (median 1.28 mg./ml., mean 1.42, p < 0.05). CONCLUSIONS: PSA concentration is increased in the expressed prostatic fluid of prostates of men with carcinoma compared to those with histological BPH. This finding may be a functional manifestation of a field change or paracrine effects within the prostate.     

A comparison of computational color constancy Algorithms. II. Experiments with image data

For pt.I see ibid., vol. 11, no.9, p.972-84 (2002). We test a number of the leading computational color constancy algorithms using a comprehensive set of images. These were of 33 different scenes under 11 different sources representative of common illumination conditions. The algorithms studied include two gray world methods, a version of the Retinex method, several variants of Forsyth's (1990) gamut-mapping method, Cardei et al.'s (2000) neural net method, and Finlayson et al.'s color by correlation method (Finlayson et al. 1997, 2001; Hubel and Finlayson 2000). We discuss a number of issues in applying color constancy ideas to image data, and study in depth the effect of different preprocessing strategies. We compare the performance of the algorithms on image data with their performance on synthesized data. All data used for this study are available online at http://www.cs.sfu.ca//spl sim/color/data, and implementations for most of the algorithms are also available (http://www.cs.sfu.ca//spl sim/color/code). Experiments with synthesized data (part one of this paper) suggested that the methods which emphasize the use of the input data statistics, specifically color by correlation and the neural net algorithm, are potentially the most effective at estimating the chromaticity of the scene illuminant. Unfortunately, we were unable to realize comparable performance on real images. Here exploiting pixel intensity proved to be more beneficial than exploiting the details of image chromaticity statistics, and the three-dimensional (3-D) gamut-mapping algorithms gave the best performance.     

A comparison of subcellular element concentrations in frozen-dried, plastic-embedded, dry-cut sections and frozen-dried cryosections

Biological X-ray microanalysis of diffusible elements within cellular and subcellular compartments requires preparation methods to retain electrolytes in the compartments they occupied in vivo. X-ray microanalysis of frozen-dried, plastic-embedded samples has been used to quantitate electrolytes at the cellular level. We have compared the subcellular elemental distribution in dry cut sections from such samples with that in ultrathin frozen-dried cryosections. Rat pancreases were quench-frozen onto a helium-vapor-cooled copper block. Cryosections were cut at 130-150 K, transferred using a Gatan cold stage, frozen-dried in the column and analysed at 190 K. Tissue fragments were frozen-dried at 190 K, and cut on a dry knife at 293 K. Both samples provided images permitting unambiguous identification of all major compartments except the Golgi complex. Intracellular potassium-to-sodium ratios obtained on frozen-dried plastic-embedded sections were lower than for cryosections (e.g. 1.77 in basal cytoplasm in plastic sections as compared to 4.34 for cryosections) and varied with the pre-embedding procedure (e.g. 1.77 in formaldehyde-fixed as compared to 2.87 in osmium-fixed plastic sections). Potassium gradients between adjacent organelles were large in cryosections and insignificant in plastic-embedded material. Higher cytoplasmic phosphorus, potassium and sulfur concentrations were observed in cryosections. Therefore, a redistribution of electrolytes and covalently bound elements occurred subcellularly in the plastic sections. Calcium was quantifiable in most organelles in cryosections but the plastic lowered sensitivity too much to permit routine calcium quantification. We conclude that in our hands frozen-dried, plastic-embedded samples were compromised and provided lower sensitivity than cryosections.(ABSTRACT TRUNCATED AT 250 WORDS)     

Housing quality and resilience in New Zealand

There is extensive research on the negative health impacts of poor housing quality. However, little is known about the potential health benefits of high-quality housing in poor neighbourhoods. Neighbourhoods with unexpectedly good health outcomes despite high levels of deprivation have been deemed resilient places and housing quality in these areas may be a contributor to this resilience. This study aimed to evaluate whether an indicator of neighbourhood housing quality was associated with a previously quantified resilience index (RINZ) in New Zealand. It was found that areas with high housing quality tended to have higher median income, greater proportions of partnered people and shorter-term residents, and very low proportions of Māori. A positive association was found between housing quality and resilience, after adjustment for deprivation. There was no indication of differences by heterogeneity in housing quality within the aggregate unit of analysis. These findings pose the hypothesis that improving housing quality in similarly deprived areas that have poor health outcomes could potentially boost health. To extend this understanding, further development of a more sophisticated housing quality indicator is recommended.     

Observations of wind turbine wakes and surface roughness effects on wind flow variability

Wind data collected at nine meteorological towers at the Goodnoe Hills MOD-2 wind turbine site were analyzed to characterize the wind flow over the site both in the absence and presence of wind turbine wakes. Free-flow characteristics examined were the variability of wind speed and turbulence intensity across the site as a function of wind direction and surface roughness. The nine towers' data revealed that scattered areas of trees upwind of the site caused pronounced variations in the wind flow over the site. At two towers that were frequently downwind of an extensive grove of trees, up to 30% reductions in wind speed and a factor of 2 to 3 increase in turbulence intensity were measured. A substantial increase in the magnitude of the wind gusts, as well as a considerable decrease in the mean wind speed, was observed when a tower was downwind of the trees.Wind turbine wake characteristics analyzed included the average velocity deficits, wake turbulence, wake width, wake trajectory, vertical profile of the wake, and the stratification of wake properties as a function of the ambient wind speed and turbulence intensity. The wind turbine rotor disk spanned a height of 15 m to 107 m. The nine towers' data permitted a detailed analysis of the wake behavior at a height of 32 m at various downwind distances from 2 to 10 rotor diameters (D). The relationship between velocity deficit and downwind distance was surprisingly linear, with average maximum deficits ranging from 34% at 2 D to 7% at 10 D. Largest deficits were at low wind speeds and low turbulence intensities. Average wake widths were 2.8 D at a downwind distance of 10 D. Implications for turbine spacing are that, for a wind farm with a 10-D row separation, array losses would be significantly greater for a 2-D than a 3-D spacing because of incremental effects caused by overlapping wakes. Other interesting wake properties observed were the wake turbulence (which was greatest along the flanks of the wake). the vertical variation of deficits (which were greater below hub height than above), and the trajectory of the wake (which was essentially straight).     

Truncated human endothelin receptor A produced by alternative splicing and its expression in melanoma

PURPOSE: Traumatic sphincter disruption frequently is associated with a rectovaginal fistula, but the effect of a persistent sphincter defect on the outcome of rectovaginal fistula repair is poorly documented. We analyzed the outcome of rectovaginal fistula repairs based on preoperative sphincter status. PATIENTS AND METHODS: We identified 52 women who underwent 62 repairs of simple obstetrical rectovaginal fistulas between 1992 and 1995. Fourteen patients (27 percent) had preoperative endoanal ultrasound studies and 25 (48 percent) had anal manometry studies. Follow-up was by mailed questionnaire in 36 patients (69 percent) and by telephone interview in 12 (23 percent), for a total response rate of 92 percent. Median age was 30.5 (range, 18-70) years, and median follow-up was 15 (range, 0.5-123) months. Twenty-five patients (48 percent) complained of varying degrees of fecal incontinence before surgery. There were 27 endorectal advancement flaps and 35 sphincteroplasties (28 with and 8 without levatoroplasty). RESULTS: Success rates were 41 percent with endorectal advancement flaps and 80 percent with sphincteroplasties (96 percent success with and 33 percent without levatoroplasty; P = 0.0001). Endorectal advancement flap was successful in 50 percent of patients with normal sphincter function but in only 33 percent of patients with abnormal sphincter function (P = not significant). For sphincteroplasties, success rates were 73 vs. 84 percent for normal and abnormal sphincter function, respectively (P = not significant). Results were better after sphincteroplasties vs. endorectal advancement flaps in patients with sphincter defects identified by endoanal ultrasound (88 vs. 33 percent; P = not significant) and by manometry (86 vs. 33 percent; P = not significant). Poor results correlated with prior surgery in patients undergoing endorectal advancement flaps (45 percent vs. 25 percent; P = not significant) but not sphincteroplasties (80 vs. 75 percent; P = not significant). CONCLUSIONS: All patients with rectovaginal fistula should undergo preoperative evaluation for occult sphincter defects by endoanal ultrasound or anal manometry or both procedures. Local tissues are inadequate for endorectal advancement flap repairs in patients with sphincter defects and a history of previous repairs. Patients with clinical or anatomic sphincter defects should be treated by sphincteroplasty with levatoroplasty.