Preliminary observation on carboxyl-methyl Poria cocos polysaccharide (CMPCP) in treating chronic viral hepatitis.
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Basidiomycota↗
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A plasmid from S. citri strain M14 hybridizes with extrachromosomal DNAs from other spiroplasmas, including corn stunt spiroplasma E275, tick spiroplasma 277F, and coco spiroplasma N525.
A plasmid, pM41, has been isolated from the Spiroplasma citri strain M4 (group I-1) and characterized by restriction mapping. Using a 32P-labeled probe specific of the plasmid, we have shown by DNA-DNA hybridization that a plasmid identical to pM41 or a closely related plasmid, is present in several, but not all, S. citri strains. DNA sequences that hybridize to pM41 were also identified in three other spiroplasmas not belonging to the S. citri species. Protein patterns of several S. citri strains have been compared in order to investigate the effect of pM41 on the spiroplasma protein profiles or maps. In fact, the presence of pM41 does not appear to modify the protein pattern.
Bacterial Proteins↗
Intrauterine programming of hypertension in the rat: nutrient interactions.
The feeding of low protein diets to pregnant rats is known to programme hypertension in their offspring though a glucocorticoid-dependent mechanism. To establish whether diets of differing fatty acid composition could produce the same effects, or modulate the effects of a low protein diet, pregnant rats were fed 1 of 4 different diets. Control animals were fed an 18% casein:corn oil (18 + Corn) diet, while experimental groups were fed 18% casein:coconut oil (18 + Coco), 9% casein:corn oil (9 + Corn) or 9% casein:coconut oil (9 + Coco). Feeding of coconut oil diets significantly reduced weight gain of the rats, an effect attributable to a lower food intake. Despite this, reproductive performance of the rats was not compromised. While pups of the 9 + Corn group were of normal birthweight, the dams in the 18 + Coco and 9 + Coco groups gave birth to pups of significantly lower weight. At 7 weeks of age male and female pups exposed to the 9 + Corn diet in utero had significantly elevated systolic blood pressure. Males exposed to the 9 + Coco and 18 + Coco diets also had higher blood pressures than control animals, but the effect was not observed in females. Female rats exposed to the three experimental diets in utero had enlarged hearts, relative to controls and the same effect of maternal diet was observed in male rats exposed to the 9 + Coco and 18 + Coco diets. Effects of maternal diet upon liver size were also noted in males exposed to 18 + Coco and 9 + Corn diets (smaller livers). Males exposed to 9 + Corn had smaller lungs than control rats, whilst the rats from the 9 + Coco group had significantly enlarged lungs. In rats exposed to the 9 + Corn diet in utero there was evidence of increased glucocorticoid action in both liver and hypothalamus, where activities of corticosteroid inducible enzymes were elevated. There was no evidence of a role for glucocorticoids in programming the hypertension of male rats exposed to high saturated fat diets in utero. The data would suggest that high saturated fatty acid, or low linoleic acid intake in pregnancy may programme hypertension in the fetal rat. This effect of maternal diet occurs through a mechanism different to that initiated by protein restriction and is sex specific.
Animal Nutritional Physiological Phenomena↗
Dietary fat saturation produces lipid modifications in peritoneal macrophages of mouse.
We investigated the effects of a saturated fat diet on mice lipid metabolism in resident peritoneal macrophages. Male C57BL/6 mice were weaned at 21 days of age and assigned to either the experimental diet, containing coconut oil (COCO diet), or the control diet, containing soybean oil as fat source. Fat content of each diet was 15% (w/w). Mice were fed for 6 weeks until sacrifice. In plasma of mice fed the COCO diet, the concentration of triglyceride, total cholesterol, HLD- and (LDL+VLDL)-cholesterol, and thiobarbituric acid-reactive substances (TBARS) increased, without changes in phospholipid concentration, compared with the controls. In macrophages of COCO-fed mice, the concentration of total (TC), free and esterified cholesterol, triglyceride, phospholipid (P) and TBARS increased, while the TC/P ratio did not change. The phospholipid compositions showed an increase of phosphatidylcholine and phosphatidylserine + phosphadytilinositol, a decrease of phosphatidylethanolamine, and no change in phosphatidylglycerol. (3)H(2)O incorporation into triglyceride and phospholipid fractions of macrophages increased, while its incorporation into free cholesterol decreased. Incorporation of [(3)H]cholesterol into macrophages of COCO-fed mice and the fraction of [(3)H]cholesterol ester increased. COCO diet produced an increase in myrystic, palmitic and palmitoleic acids proportion, a decrease in linoleic and arachidonic acids and no changes in stearic and oleic acids, compared with the control. Also, a higher relative percentage of saturated fatty acid and a decrease in unsaturation index (p <0.001) were observed in macrophages of COCO-fed mice. These results indicate that the COCO-diet, high in saturated fatty acids, alters the lipid metabolism and fatty acid composition of macrophages and produces a significant degree of oxidative stress.
Animals↗
Effect of dietary fat saturation on lipid metabolism, arachidonic acid turnover and peritoneal macrophage oxidative stress in mice.
We investigated the effects of a saturated fat diet on lipid metabolism and arachidonic acid (AA) turnover in mouse resident peritoneal macrophages. The pro-oxidative effect of this diet was also studied. Female C57BL/6 mice were weaned at 21 days of age and assigned to either the experimental diet containing coconut oil (COCO diet), or the control diet containing soybean oil as fat source (10 mice per group). The fat content of each diet was 15% (w/w). Mice were fed for 6 weeks and then sacrificed. The concentration of total lipids, triglycerides, (LDL+VLDL)-cholesterol, thiobarbituric acid-reactive substances (TBARS) and reduced glutathione were increased in the plasma of mice fed the COCO diet, without changes in phospholipid or total cholesterol concentrations compared to control. The concentrations of total cholesterol, free and esterified cholesterol, triglycerides, and TBARS were increased in the macrophages of COCO-fed mice, while the content of total phospholipids did not change. The phospholipid composition showed an increase of phosphatidylcholine and a decrease of phosphatidylethanolamine. The [3H]-AA distribution in the phospholipid classes showed an increase in phosphatidylcholine and phosphatidylethanolamine. Incorporation of [3H]-cholesterol into the macrophages of COCO-fed mice and into the cholesterol ester fraction was increased. The COCO diet did not affect [3H]-AA uptake but induced an increase in [3H]-AA release. The COCO diet also enhanced AA mobilization induced by lipopolysaccharide. These results indicate that the COCO diet, high in saturated fatty acids, alters the lipid metabolism and AA turnover of peritoneal macrophages in female mice and also produces a significant degree of oxidative stress.
Animals↗
Classification of emergency department chief complaints into 7 syndromes: a retrospective analysis of 527,228 patients.
STUDY OBJECTIVE: Electronic surveillance systems often monitor triage chief complaints in hopes of detecting an outbreak earlier than can be accomplished with traditional reporting methods. We measured the accuracy of a Bayesian chief complaint classifier called CoCo that assigns patients 1 of 7 syndromic categories (respiratory, botulinic, gastrointestinal, neurologic, rash, constitutional, or hemorrhagic) based on free-text triage chief complaints. METHODS: We compared CoCo's classifications with criterion syndromic classification based on International Classification of Diseases, Ninth Revision (ICD-9) discharge diagnoses. We assigned the criterion classification to a patient based on whether the patient's primary diagnosis was a member of a set of ICD-9 codes associated with CoCo's 7 syndromes. We tested CoCo's performance on a set of 527,228 chief complaints from patients registered at the University of Pittsburgh Medical Center emergency department (ED) between 1990 and 2003. We performed a sensitivity analysis by varying the ICD-9 codes in the criterion standard. We also tested CoCo on chief complaints from EDs in a second location (Utah). RESULTS: Approximately 16% (85,569/527,228) of the patients were classified according to the criterion standard into 1 of the 7 syndromes. CoCo's classification performance (number of cases by criterion standard, sensitivity [95% confidence interval (CI)], and specificity [95% CI]) was respiratory (34,916, 63.1 [62.6 to 63.6], 94.3 [94.3 to 94.4]); botulinic (1,961, 30.1 [28.2 to 32.2], 99.3 [99.3 to 99.3]); gastrointestinal (20,431, 69.0 [68.4 to 69.6], 95.6 [95.6 to 95.7]); neurologic (7,393, 67.6 [66.6 to 68.7], 92.7 [92.6 to 92.8]); rash (2,232, 46.8 [44.8 to 48.9], 99.3 [99.3 to 99.3]); constitutional (10,603, 45.8 [44.9 to 46.8], 96.6 [96.6 to 96.7]); and hemorrhagic (8,033, 75.2 [74.3 to 76.2], 98.5 [98.4 to 98.5]). The sensitivity analysis showed that the results were not affected by the choice of ICD-9 codes in the criterion standard. Classification accuracy did not differ on chief complaints from the second location. CONCLUSION: Our results suggest that, for most syndromes, our chief complaint classification system can identify about half of the patients with relevant syndromic presentations, with specificities higher than 90% and positive predictive values ranging from 12% to 44%.
Bayes Theorem↗
Hypocholesterolemic effect of physically refined rice bran oil: studies of cholesterol metabolism and early atherosclerosis in hypercholesterolemic hamsters.
Physically refined rice bran oil containing 2-4% nontriglyceride components as compared to other vegetable oils appears to be associated with lipid lowering and antiinflammatory properties in several rodent, primate and human models. These experiments were designed to investigate possible mechanisms for the hypocholesterolemic effect of the physically refined rice bran oil and to examine its effect on aortic fatty streak formation. In the first experiment, 30 hamsters were fed, for 8 weeks, chow-based diets plus 0.03% added cholesterol and 5% (wt/wt) coconut, canola, or physically refined rice bran oil (COCO, CANOLA or PRBO animal groups, respectively). Both plasma total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) were significantly reduced in PRBO but not in CANOLA relative to COCO. PRBO also showed a significant 15-17% reduction in cholesterol absorption and significant 30% increase in neutral sterol (NS) excretion with no effect on bile acid (BA) excretion. Both CANOLA and PRBO showed a significant 300-500% increase in intestinal 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and significant (>25%) decrease in hepatic HMG-CoA reductase activities with respect to COCO. In a second experiment, 36 hamsters were fed chow-based diets with 0.05% added cholesterol, 10% coconut oil and 4% additional COCO, CANOLA or PRBO. Relative to COCO and CANOLA, plasma TC and LDL-C were significantly reduced in PRBO. Early atherosclerosis (fatty streak formation) was significantly reduced (48%) only in PRBO, relative to the other two. These results suggest that the lipid lowering found in PRBO is associated with decreased cholesterol absorption, but not hepatic cholesterol synthesis, and that the decrease in fatty streak formation with this oil may be associated with its nontriglyceride components not present in the other two diets.
Animals↗
Influence of host cultivars and Rhizobium species on the growth and symbiotic performance of Phaseolus vulgaris under salt stress.
In order to study the effect of salt stress on the Rhizobium-common bean symbiosis, we investigated the response of both partners, separately and in symbiosis. The comparison of the behaviour of five cultivars of Phaseolus vulgaris differing in seed colour, growing on nitrates and different concentrations of NaCl, showed genotypic variation with respect to salt tolerance. Coco Blanc was the most sensitive cultivar, whereas SMV 29-21 was the most tolerant one. At the Rhizobium level, two strains previously selected for their salt tolerance were used: Rhizobium tropici strain RP163 and Rhizobium giardinii strain RP161. Their relative growth was moderately decreased at 250mM NaCl, but they were able to grow at a low rate in the presence of 342 mM NaCl. Their viability at the minimal inhibitory concentration was slightly affected. The effect of salinity on Rhizobium-plant association was studied by using the tolerant variety SMV 29-21 and the sensitive one Coco Blanc inoculated separately with both strains. In the absence of salinity, the strains induced a significantly higher number of nodules on the roots of the cultivar SMV 29-21 compared to those of Coco Blanc. Concerning effectiveness, both strains were similarly effective with SMV 29-21, but not with Coco Blanc. In the presence of salinity, Coco Blanc was more severely affected when associated with RP163 than with RP161. Salinity affected the nodulation development more than it affected the infection steps. Neither of the two strains was able to nodulate SMV 29-21 under saline conditions, in spite of the fact that this was considered the most salt-tolerant variety. The unsuccessful nodulation of SMV 29-21 could be related to the inhibition by salt of one or more steps of the early events of the infection process. In conclusion, N-fixing plants were found to be more sensitive to salt stress than those depending on mineral nitrogen. Evidence presented here suggests that a best symbiotic N2 fixation under salinity conditions could be achieved if both symbiotic partners, as well as the different steps of their interaction (early events, nodule formation, activity, etc.), are all tolerant to this stress.
Adaptation, Physiological↗
Independent effects of dietary saturated fat and cholesterol on plasma lipids, lipoproteins, and apolipoprotein E.
Nine normolipidemic males (18-37 years) were fed formula diets containing (as % of calories) egg white protein (15%), glucose polymer:sucrose, 3:1 (54%), and fats (31%) as one of the following: corn oil (corn), corn oil plus 1 gram/day cholesterol (corn+), coconut oil (coco), coconut oil plus 1 gram/day cholesterol (coco+). Two dietary periods of 18 days each were separated by 1 month during which plasma lipid levels returned to prestudy values. A given dietary period consisted of 9 days of either corn or coco feeding allowed by 9 days of corn+ or coco+, respectively. Fasting plasma samples were taken the last 3 days of each 9-day interval. Lipids were determined by standard procedures and the apoE levels in lipoprotein fractions isolated by discontinuous density gradient ultracentrifugation were determined by radioimmunoassay. The biochemical variables measured were: total plasma, VLDL, IDL + LDL, and HDL, cholesterol, triglyceride, and apoE levels, as well as the apoE of plasma d greater than 1.17 g/ml. The effects of apoE phenotype, the type of dietary oil (corn versus coco), the presence or absence of dietary cholesterol, and the day of sampling within triplicates on the above variables were assessed statistically. The type of oil had the only significant effect on any variable. At P less than 0.01, the coconut oil diets were associated with significant elevations (as compared to corn oil) of the following nine variables: total, VLDL, IDL + LDL, and HDL cholesterol; total, VLDL, and IDL + LDL apoE; total and VLDL triglycerides.
Adolescent↗
Vibrational frequencies of the homoleptic cobalt carbonyls: Co4(CO)12 and Co6(CO)16.
The homoleptic cobalt carbonyls Co4(CO)12 and Co6(CO)16 are characterized by their equilibrium geometries, thermochemistry, and vibrational frequencies using density functional theory (DFT) methods with the B3LYP, BLYP, and BP86 functionals. The B3LYP predicted CoCo distances are 2.51 and 2.47 A for the C3v and Td structures, respectively, of Co4(CO)12. The global minimum for Co4(CO)12 has C3v symmetry with three bridging and nine terminal carbonyls. The 2.51 and 2.52 A CoCo distances suggest the single bond required for an 18-electron configuration for the metal atoms. This structure is close to an experimentally realized structure. A more symmetrical Co4(CO)12 structure with Td symmetry, analogous to that observed in the valence isoelectronic Ir4(CO)12 molecule, lies approximately 28 kcal/mol higher in energy and exhibits a small imaginary vibrational frequency ( approximately 40i). It has a slightly shorter CoCo distance of 2.47 A. Both Co4(CO)12 structures satisfy the 18-electron rule. The Co6(CO)16 structure has Td symmetry and satisfies the Wade-Mingos rules for an octahedral cluster. The nu(CO) carbonyl frequencies for both Co4(CO)12 and Co6(CO)16 computed with the BP86 functional are closer to the experimental values than those computed with the B3LYP and BLYP functionals. The structure of Co6(CO)16 is not known experimentally, but the BP86 functional predicts 2.56 A (CoCo), 1.77 and 2.02 A (CoC), and 1.66 and 1.20 A (CO) for the bond distances.
Carbon↗
Dietary fibers from mushroom sclerotia: 3. In vitro fermentability using human fecal microflora.
The in vitro fermentability of three novel dietary fibers (DFs) prepared from mushroom sclerotia, namely, Pleurotus tuber-regium, Polyporous rhinocerus, and Wolfiporia cocos, was investigated and compared with that of the cellulose control. All DF samples (0.5 g each) were fermented in vitro with a human fecal homogenate (10 mL) in a batch system (total volume, 50 mL) under strictly anaerobic conditions (using oxygen reducing enzyme and under argon atmosphere) at 37 degrees C for 24 h. All three novel sclerotial DFs exhibited notably higher dry matter disappearance (P. tuber-regium, 8.56%; P. rhinocerus, 13.5%; and W. cocos, 53.4%) and organic matter disappearance (P. tuber-regium, 9.82%; P. rhinocerus, 14.6%; and W. cocos, 57.4%) when compared with those of the cellulose control. Nevertheless, only the W. cocos DF was remarkably degraded to produce considerable amounts of total short chain fatty acids (SCFAs) (5.23 mmol/g DF on organic matter basis, with a relatively higher molar ratio of propionate) that lowered the pH of its nonfermented residue to a slightly acidic level (5.89). Variations on the in vitro fermentability among the three sclerotial DFs might mainly be attributed to their different amounts of interwoven hyphae present (different amounts of enzyme inaccessible cell wall components) as well as the possible different structural arrangement (linkage and degree of branching) of their beta-glucans.
Basidiomycota↗
Dietary fibers from mushroom sclerotia. 4. In vivo mineral absorption using ovariectomized rat model.
The effect of three novel dietary fibers (DFs) prepared from mushroom sclerotia, namely, Pleurotus tuber-regium, Polyporus rhinocerus, and Wolfiporia cocos, on calcium and magnesium absorption was evaluated in ovariectomized (OVX) rats fed with sclerotial DF based and low Ca (0.3%) diets for 14 days. The animals in the W. cocos DF diet group possessed significantly (p < 0.05) higher levels of cecal total short-chain fatty acids (204 mumol/g of cecal content) and had an acidic pH (5.88) in their cecum when compared with those of the cellulose control group. Such an acidic environment was found to promote the ionization of the unabsorbed Ca and Mg in their cecum, which in turn significantly (p < 0.05) increased the concentrations of cecal soluble Ca (2.56-fold) and Mg (1.22-fold). Besides, the apparent Ca and Mg absorptions of the W. cocos DF group were also significantly (p < 0.05) enhanced (Ca, 16.5%; Mg, 15.3%) together with significantly (p < 0.05) higher serum Ca (3.61 mmol/L) and Mg (1.07 mmol/L) levels when compared with those of the cellulose control group. These data suggest that ingestion of W. cocos DF could improve the overall Ca and Mg absorptions of the OVX rats fed a low Ca diet. The potential use of sclerotial DFs as a functional food ingredient for enhancing mineral absorption is also discussed.
Absorption↗
Influence of traditional Chinese anti-inflammatory medicinal plants on leukocyte and platelet functions.
The enzymes 5-lipoxygenase and elastase are therapeutic targets in dermatological disorders such as psoriasis. Fifteen extracts from traditional Chinese medicinal plants used to treat topical inflammations were screened for their inhibitory effect on lipoxygenase, cyclooxygenase and elastase activity in intact leukocytes and platelets. Astragalus membranaceus, Forsythia suspensa and Poria cocos inhibited 5-lipoxygenase, with IC50 values of 141, 80 and 141 microg mL(-1), respectively. The latter two species, along with Angelica dahurica and Angelica pubescens, also inhibited elastase (IC50 values of 80, 123, 68 and 93 microg mL(-1), respectively), while A. pubescens, Atractylodes macrocephala, Lentinus edodes, Rehmannia glutinosa and Paeonia lactiflora selectively inhibited 12-(S)-HHTrE production, a valid marker of cyclooxygenase activity. The inhibition of phospholipase A(2) activity by P. cocos is discussed. Dehydrotumulosic and pachymic acids, which have been isolated from P. cocos, were shown to inhibit leukotriene B(4) release. The results indicate that both P. cocos and F. suspensa are potentially valuable species in the management of skin pathologies involving chronic inflammation.
Animals↗
Comparison of two major emergency department-based free-text chief-complaint coding systems.
INTRODUCTION: Emergency departments (EDs) using free-text chief-complaint data for syndromic surveillance face a unique challenge because a complaint might be described and coded in multiple ways. OBJECTIVE: Two major ED-based free-text chief-complaint coding systems were compared for agreement between free-text interpretation and syndrome coding. METHODS: Chief-complaint data from 21,736 patients at an urban ED were processed through both the New York City Department of Health and Mental Hygiene (DOHMH) syndrome coding system as modified by the Chicago Department of Public Health and the Real-Time Outbreak Detection System Complaint Coder (CoCo, version 2.1, University of Pittsburgh). To account for differences in each system's specified syndromes, relevant syndromes from the DOHMH system were collapsed into the corresponding CoCo categories so that a descriptive comparison could be made. DOHMH classifications were combined to match existing CoCo categories as follows: 1) vomit+diarrhea = Gastrointestinal; 2) cold+respiratory+asthma = Respiratory; 3) fevflu = Constitutional; 4) rash = Rash; 5) sepsis+other = Other, 6) unknown = Unknown. RESULTS: Overall agreement between DOHMH and CoCo syndrome coding was optimal (0.614 kappa). However, agreement between individual syndromes varied substantially. Rash and Respiratory had the highest agreement (0.711 and 0.594 kappa, respectively). Other and Constitutional had an intermediate level of agreement (0.453 and 0.419 kappa, respectively), but less than optimal agreement was identified for Gastrointestinal and Unknown (0.270 and 0.002 kappa, respectively). CONCLUSIONS: Although this analysis revealed optimal overall agreement between the two systems evaluated, substantial differences in classification schemes existed, highlighting the need for a consensus regarding chief-complaint classification.
Algorithms↗
Characterisation of copia-like retrotransposons in oil palm ( Elaeis guineensis Jacq.).
The work aimed to isolate and characterise copia-like sequences from Elaeis guineensis. Thirty-two different RT (reverse transcriptase) sequences were isolated from a single oil palm genome using degenerate primers. Extreme sequence heterogeneity was observed. The DNA and protein sequences were assigned to three different classes (A, B, C) on the basis of bootstrapping. We estimated the copy number of the three different classes by using a dot-blot analysis. The comparative results suggest that class-B RT sequences occur at a higher copy number in Cocos nucifera than in Elaeis guineensis and E. oleifera. Class-C RT sequences, which comprise the bulk of isolated sequences, occur in much higher copy number in Elaeis guineensis than in Cocos nucifera, and E. oleifera. Class-A sequences, which have low copy number in Elaeis guineensis and Cocos nucifera, are not present in E. oleifera. Our preliminary results suggest that class-C sequences represent about 5% of the E. guineensis genome, class B, 1% and class A, 0.1%. The methylation status of genomic domains specified by the RT probes was analysed using two pairs of restriction enzymes, each pair having the same recognition sites but different methylation sensitivities ( MspI, HpaII and Sau3AI, MboI). Results from these experiments showed clearly that the genomic domains specified by the RT probes are methylated. These also suggest that a higher copy number appears to correlate to a higher degree of methylation. Our preliminary results suggest that copia-like retrotransposons, because of their ubiquity and diversity, have great potential as genetic markers for plant genome and biodiversity analysis in E. guineensis.
Journal Article↗
Inhibition of the aminopeptidase from Aeromonas proteolytica by L-leucinethiol: kinetic and spectroscopic characterization of a slow, tight-binding inhibitor-enzyme complex.
The peptide inhibitor L-leucinethiol (LeuSH) was found to be a potent, slow-binding inhibitor of the aminopeptidase from Aeromonas proteolytica (AAP). The overall potency (K(I)*) of LeuSH was 7 nM while the corresponding alcohol L-leucinol (LeuOH) was a simple competitive inhibitor of much lower potency (K(I) = 17 microM). These data suggest that the free thiol is likely involved in the formation of the E x I and E x I* complexes, presumably providing a metal ligand. In order to probe the nature of the interaction of LeuSH and LeuOH with the dinuclear active site of AAP, we have recorded both the electronic absorption and EPR spectra of [CoCo(AAP)], [CoZn(AAP)], and [ZnCo(AAP)] in the presence of both inhibitors. In the presence of LeuSH, all three Co(II)-substituted AAP enzymes exhibited an absorption band centered at 295 nm, characteristic of a S --> Co(II) ligand-metal charge-transfer band. In addition, absorption spectra recorded in the 450 to 700 nm region all showed changes characteristic of LeuSH and LeuOH interacting with both metal ions. EPR spectra recorded at high temperature (19 K) and low power (2.5 mW) indicated that, in a given enzyme molecule, LeuSH interacts weakly with one of the metal ions in the dinuclear site and that the crystallographically identified mu-OH(H) bridge, which has been shown to mediate electronic interaction of the Co(II) ions, is likely broken upon binding LeuSH. EPR spectra of [CoCo(AAP)]-LeuSH, [ZnCo(AAP)]-LeuSH, and [Co_(AAP)]-LeuSH were also recorded at lower temperature (3.5-4.0 K) and high microwave power (50-553 mW). These signals were unusual and appeared to contain, in addition to the incompletely saturated contributions from the signals characterized at 19 K, a very sharp feature at g(eff) approximately 6.5 that is characteristic of thiolate-Co(II) interactions. Combination of the electronic absorption and EPR data indicates that LeuSH perturbs the electronic structure of both metal ions in the dinuclear active site of AAP. Since the spin-spin interaction seen in resting [CoCo(AAP)] is abolished upon the addition of LeuSH, it is unlikely that a mu-S(R) bridge is established.
Aeromonas↗
Paramagnetic cobalt(II) as a probe for kinetic and NMR relaxation studies of phosphate binding and the catalytic mechanism of Streptomyces dinuclear aminopeptidase.
Phosphate was proposed to be a bridging ligand in the structure 1xjo.pdb of Streptomyces dizinc aminopeptidase (sAP), which prompted further studies of phosphate binding to this enzyme. Phosphate inhibits sAP and its Co(2+)-substituted derivatives in a noncompetitive manner from pH 6.0 to 9.0, with strongest inhibition observed at lower pHs (K(i) = 0.6, 8.2, and 9.1 mM for ZnZn-, CoCo-, and CoZn-sAP, respectively, at pH 6.0), which indicates that phosphate does not compete with substrate binding to the dinuclear active site and that monobasic phosphate has a higher binding affinity. The inhibition K(i)-pH profiles for phosphate inhibition of both the native and the Co(2+)-substituted derivatives reveal a similar pK(a) around 7.0, reflecting that phosphate binding is not affected by the metal centers of different Lewis acidities. Modification of ZnZn- and CoCo-sAP with the arginine-specific reagent phenylglyoxal reveals a significant weakening in phosphate and substrate binding by showing approximately a 10-fold increase in the dissociation constant K(i) for phosphate binding and approximately 4-8-fold increase in K(m). The catalysis is also influenced by the modification as reflected by a significant decrease in k(cat) in both cases. Furthermore, phosphate and the transition-state inhibitor 1-aminobutyl phosphonate can protect arginine from the modification, strongly suggesting that Arg202 near the active site is involved in phosphate binding and in stabilizing the transition state. The effect on (31)P NMR relaxation of phosphate caused by the paramagnetic metal center in Co(2+)-substituted derivatives of sAP has been measured, which reveals that only one phosphate is bound to sAP with the Co(2+)-(31)P distance in the range of 4.1-4.3 A. The (1)H NMR relaxation of the bulk water signal in the CoCo-sAP sample remains unchanged in the presence of phosphate, further indicating that phosphate may not bind to the active-site metals to displace any metal-bound water/hydroxide. These results strongly support that the phosphate binding site is Arg202 and that this residue plays an important role in the action of sAP.
Aminopeptidases↗
The importation of the black rhinoceros (Diceros bicornis) from Zimbabwe into Australia.
The paper describes a program to import and breed black rhinoceros ex situ at Western Plains Zoo in Australia. Nine rhinoceros (7 females and 2 males) captured in 1992 in Chete National Park, Zimbabwe, were transported to Australia via Cocos Island. The veterinary treatment of the animals before and during quarantine in Zimbabwe and on Cocos (Keeling) Islands is described. Three animals died; an adult male on Cocos Islands and a juvenile male and an adult female at Western Plains Zoo, Dubbo, New South Wales. The juvenile male died as a result of trauma sustained shortly after arrival and the two adults after developing a severe hepatopathy. The group of 6 females and an additional 4 males imported from the USA in 1994 have adapted well to captivity and to the climate and environment of central west New South Wales.
Adaptation, Physiological↗