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Animal sterilization is suggested to promote food overconsumption, although it is unknown whether this effect is mediated by variations in satiety-related hormones, which are released in response to food intake. The aim of this study was to evaluate the effect of sterilization and of the main energy-delivery nutrients, fat and nonstructural carbohydrates, on food intake, blood concentration of satiety-related hormones, and activity level in dogs. In a 2-phase experiment (phase I [Ph.I], 74 d, and Ph.II, 84 d), 12 female Beagle dogs were assigned to a control group (intact in both phases) and a sterilization group (spayed 20 d before Ph.II). In each phase, dogs received a high-carbohydrate (HC) diet (313 and 105 g/kg DM starch and fat, respectively) and a high-fat (HF) diet (191 and 213 g/kg DM starch and fat, respectively), both high in total dietary fiber (>200 g/kg DM) and providing 27% ME as protein, in 2 consecutive periods following a crossover arrangement. During each period, dogs' voluntary DMI and activity level were recorded during 5 d. Then, energy allowance was restricted to 0.7 maintenance and the level of intake of a common challenge food offered 4 h after feeding the experimental diets (challenge food intake [ChFI]) was used as an index of the satiety state of dogs. Blood concentration of active ghrelin, cholecystokinin (CCK), total peptide YY (PYY), and insulin were determined before and 15, 60, 120, 240, and 360 min after feeding. Voluntary DMI was greater ( < 0.05) in HF-fed dogs, but ChFI did not differ between diets ( > 0.10). Dogs fed the HF diet showed a lower increase of CCK at 120 ( < 0.01) and 240 min ( < 0.05), resulting in a lower ( < 0.001) total area under the curve from 0 to 240 min (tAUC). A lower PYY elevation ( < 0.05) was also found in HF-fed dogs at 120 min. Only active ghrelin concentration at 240 min and insulin tAUC correlated ( < 0.05) with ChFI (r = 0.357 and r = -0.364, respectively), suggesting a role of these hormones in appetite. Dog sterilization did not affect voluntary DMI, ChFI, or blood hormones ( > 0.10) but led to a reduced activity level compared with control dogs ( < 0.05). In summary, dog sterilization was not associated with an impaired appetite control. Feeding dogs the HF diet led to energy overconsumption and to a lower blood elevation of CCK and PYY but was not associated with a weaker satiating effect 4 h later compared with the HC diet.

Ubiquitination plays a major role in many aspects of hematopoiesis. Alterations in ubiquitination have been implicated in hematological cancer. The ubiquitin ligase Triad1 controls the proliferation of myeloid cells. Here, we show that two RING (really interesting new gene) domains in Triad1 differentially bind ubiquitin-conjugating enzymes, UbcH7 and Ubc13. UbcH7 and Ubc13 are known to catalyze the formation of different poly-ubiquitin chains. These chains mark proteins for proteasomal degradation or serve crucial non-proteolytic functions, respectively. In line with the dual Ubc interactions, we observed that Triad1 catalyzes the formation of both types of ubiquitin chains. The biological relevance of this finding was studied by testing Triad1 mutants in myeloid clonogenic assays. Full-length Triad1 and three mutants lacking conserved domains inhibited myeloid colony formation by over 50%. Strikingly, deletion of either RING finger completely abrogated the inhibitory effect of Triad1 in clonogenic growth. We conclude that Triad1 exhibits dual ubiquitin ligase activity and that both of its RING domains are crucial to inhibit myeloid cell proliferation. The differential interaction of the RINGs with Ubcs strongly suggests that the ubiquitination mediated through UbcH7 as well as Ubc13 plays a major role in myelopoiesis.

Limonene-1,2-epoxide hydrolase (LEH) from Rhodococcus erythropolis DCL14, an enzyme involved in the limonene degradation pathway of this microlorganism, has a narrow substrate specificity. Of the compounds tested, the natural substrate, limonene-1,2-epoxide, and several alicyclic and 2-methyl-1,2-epoxides (e.g. 1-methylcyclohexene oxide and indene oxide), were substrates for the enzyme. When LEH was incubated with a diastereomeric mixture of limonene-1,2-epoxide, the sequential hydrolysis of first the (1R,2S)- and then the (1S,2R)-isomer was observed. The hydrolysis of (4R)- and (4S)-limonene-1,2-epoxide resulted in, respectively, (1S,2S,4R)- and (1R,2R,4S)-limonene-1,2-diol as the sole product with a diastereomeric excess of over 98%. With all other substrates, LEH showed moderate to low enantioselectivities (E ratios between 34 and 3).

In batch toxicity assays, azo dye compounds were found to be many times more toxic than their cleavage products (aromatic amines) towards methanogenic activity in anaerobic granular sludge. Considering the ability of anaerobic microorganisms to reduce azo groups, detoxification of azo compounds towards methanogens can be expected to occur during anaerobic wastewater treatment. In order to test this hypothesis, the anaerobic degradation of one azo dye compound, Mordant orange 1 (MO1), by granular sludge was investigated in three separate continuous upflow anaerobic sludge-blanket reactors. One reactor, receiving no cosubstrate, failed after 50 days presumably because of a lack of reducing equivalents. However, the two reactors receiving either glucose or a volatile fatty acids (acetate, propionate, butyrate) mixture, could eliminate the dye during operation for 217 days. The azo dye was reductively cleaved to less toxic aromatic amines (1,4-phenylenediamine and 5-aminosalicylic acid) making the treatment of MO1 feasible at influent concentrations that were over 25 times higher than their 50% inhibitory concentrations. In the reactor receiving glucose as cosubstrate, 5-aminosalicylic acid could only be detected at trace levels in the effluent after day 189 of operation. Batch biodegradability assays with the sludge sampled from this reactor confirmed the mineralization of 5-aminosalicylic acid to methane.

Animal sterilization is suggested to promote food overconsumption, although it is unknown whether this effect is mediated by variations in satiety-related hormones, which are released in response to food intake. The aim of this study was to evaluate the effect of sterilization and of the main energy-delivery nutrients, fat and nonstructural carbohydrates, on food intake, blood concentration of satiety-related hormones, and activity level in dogs. In a 2-phase experiment (phase I [Ph.I], 74 d, and Ph.II, 84 d), 12 female Beagle dogs were assigned to a control group (intact in both phases) and a sterilization group (spayed 20 d before Ph.II). In each phase, dogs received a high-carbohydrate (HC) diet (313 and 105 g/kg DM starch and fat, respectively) and a highfat (HF) diet (191 and 213 g/kg DM starch and fat, respectively), both high in total dietary fiber (>200 g/ kg DM) and providing 27% ME as protein, in 2 consecutive periods following a crossover arrangement. During each period, dogs' voluntary DMI and activity level were recorded during 5 d. Then, energy allowance was restricted to 0.7 maintenance and the level of intake of a common challenge food offered 4 h after feeding the experimental diets (challenge food intake [ChFI]) was used as an index of the satiety state of dogs. Blood concentration of active ghrelin, cholecystokinin (CCK), total peptide YY (PYY), and insulin were determined before and 15, 60, 120, 240, and 360 min after feeding. Voluntary DMI was greater (P < 0.05) in HF-fed dogs, but ChFI did not differ between diets (P > 0.10). Dogs fed the HF diet showed a lower increase of CCK at 120 (P < 0.01) and 240 min (P < 0.05), resulting in a lower (P < 0.001) total area under the curve from 0 to 240 min (tAUC^sub 0-240^). A lower PYY elevation (P < 0.05) was also found in HF-fed dogs at 120 min. Only active ghrelin concentration at 240 min and insulin tAUC^sub 0-240^ correlated (P < 0.05) with ChFI (r = 0.357 and r = ...0.364, respectively), suggesting a role of these hormones in appetite. Dog sterilization did not affect voluntary DMI, ChFI, or blood hormones (P > 0.10) but led to a reduced activity level compared with control dogs (P < 0.05). In summary, dog sterilization was not associated with an impaired appetite control. Feeding dogs the HF diet led to energy overconsumption and to a lower blood elevation of CCK and PYY but was not associated with a weaker satiating effect 4 h later compared with the HC diet.

Ligninolytic basidiomycetes were screened for their ability to produce the tetrachlorinated hydroquinone metabolites drosophilin A (DA, tetrachloro-4-methoxyphenol) and drosophilin A methyl ether (DAME, tetrachloro-1,4-dimethoxybenzene). Five fungal strains produced these metabolites in detectable amounts, including strains from Bjerkandera and Peniophora, which are genera not previously known for DA or DAME production. Phellinus fastuosus ATCC26.125 had the highest and most reliable production of DA and DAME in peptone medium, respectively 15-60 micromolar and 4-40 micromolar. This fungus was used to study culture conditions that could increase DAME production. A fourfold increase in DAME production was found after the addition of hydroquinone to growing cultures of P. fastuosus. Therefore, hydroquinone is postulated to be a possible biosynthetic precursor of DAME in the fungus. Antagonising P. fastuosus by adding filter-sterilised culture fluid of a competing fungus. Phlebia radiata, increased DAME production significantly by tenfold. This result suggests that DAME production is elicited by compounds present in the culture fluid of P. radiata indicating that DAME has an antibiotic function in P. fastuosus.

The ecologically important white rot basidiomycete Hypholoma fasciculare was previously shown to produce large amounts of adsorbable organic halogens (AOX). The purposes of this study were to identify the time period of AOX production in relation to the primary and secondary metabolic phases of the growth cycle of the fungus, to determine the maximal specific AOX production rates and final AOX yields on the different substrates and to account for the measured AOX in identifiable compounds. The AOX production was observed to take place during the transition between the primary and secondary metabolic phases of the growth cycle of the fungus. The maximum AOX production rates ranged from 0.63 to 3.23 mg AOX per gram of dry mycelium per day and the final AOX yields ranged from 0.88 and 1.50 percent of dry weight of mycelium on five different substrates including natural woody substrates. The AOX produced by the fungus was stable in all five substrates, even after prolonged incubation periods. However, the composition of the AOX changed drastically. Initially most of the AOX was accounted for by the compound 3,5-dichloro-p-anisyl alcohol; however, after prolonged incubation this compound was largely converted into 3,5-dichloro-p-anisic acid in N-rich medium and into unidentified organohalogens in N-limited medium.

The present study investigated the role of hypothalamic VIP in the regulation of the LH and PRL surge using immunoneutralization of endogenous VIP in mature ovariectomized (OVX), estradiol benzoate (EB)-treated female Wistar rats. We compared the effect of intracerebroventricular (i.c.v.) injections of a VIP antiserum (VIP-Ab) with that of saline (Ctr) on LH and PRL profiles in two separate groups of rats following two subcutaneous EB injections on days 8 and 9 after OVX. VIP-Ab or Ctr injections were given during the second half of the dark period, i.e. at 22:00 h (day 9), and, in addition, the following morning, i.e. at 08:00 h (day 10), just before the expected onset of the LH surge. Hourly blood samples were collected between 09:00 and 18:00 h on day 10. In addition, we studied the reproducibility of EB-induced LH and PRL surges and compared the effect of Ctr and VIP-Ab treatment on sequential surges in individual OVX females, i.e. 10 and 23 days after OVX, using each animal as its own control. Although we observeda large variation in the height and timing of LH and PRL peak levels between EB-treated females, the characteristics of successive surges of individual rats were highly reproducible. This reproducibility suggests that differences in functioning of the suprachiasmatic nucleus as well as in the response of the hypothalamus to steroid feedback largely explain the normal variation in hormone responses between rats. The VIP-Ab treatment resulted in a significant delay in the time course and a strong reduction of the magnitude of the afternoon LH and PRL surge. When analyzed within individual females, the effect of VIP-Ab treatment was even more pronounced due to a reduction in variability when each animal was used as its own control. These results suggest that hypothalamic VIP is an important regulator of both the timing and the magnitude of the EB-induced LH and PRL surge in the OVX rat, and suggest that its role may be stimulatory in this respect.

To evaluate whether the general trophic effect of gastrin on the oxyntic mucosa is an indirect effect mediated by histamine H2 receptors, sustained 24 hour hypergastrinaemia was induced in Sprague-Dawley rats by treatment with the long acting and potent histamine H2 antagonist loxtidine for five months. The trophic effect was assessed by weight, enumeration of total mucosal cells, parietal cells, and enterochromaffin like cells in smears stained for the actual cells after enzymatic dispersion of the mucosa, and by biochemical analysis of oxyntic mucosal homogenates. The weight of the whole stomach and the oxyntic mucosa increased by 12.7% (p = 0.016) and 27.5% (p = 0.006), respectively. Total oxyntic mucosal protein content increased by 28.7% (p = 0.058). Total numbers of mucosal cells and parietal cells increased by 11.9% (NS) and 24.1% (NS), respectively. The amount of the parietal cell specific enzyme H+,K(+)-ATPase was unchanged. On the other hand, the number of enterochromaffin like cells and related parameters, histidine decarboxylase activity and histamine content of the oxyntic mucosa, showed a pronounced and significant increase. It is concluded that the general trophic effect of gastrin on the oxyntic mucosa is not mediated by the histamine H2 receptor. The tropic effect of gastrin on the parietal cell seems, in contrast with that on the enterochromaffin like cell, not to be specific but only reflecting the general trophic effect on the oxyntic mucosa.