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1.
The conformational properties of the oligosaccharide chain of GM1 ganglioside containingN-glycolyl-neuraminic acid, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer, were studied through NMR nuclear Overhauser effect investigations on the monomeric ganglioside in dimethylsulfoxide, and on mixed micelles of ganglioside and dodecylphosphocholine in water. Several interresidual contacts for the trisaccharide core--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-were found to fix the relative orientitation of the three saccharides, while the glycosidic linkage of the terminal -Gal-was found to be quite mobile as the -Gal-(1-3)--GalNAc-disaccharide exists in different conformations. These results are similar to those found for two GM1 gangliosides containingN-acetyl-neuraminic acid and neuraminic acid [1].Abbreviations Ganglioside nomenclature is in accordance with Svennerholm [23] and the IUPAC-IUB Recommendations [24] GM3(Neu5Ac) II3Neu5AcLacCer, -Neu5Ac-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM3(Neu5Gc) II3Neu5GcLacCer, -Neu5Gc-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Ac) II3Neu5AcGgOse4Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Gc) II3Neu5GcGgOse4Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu) II3NeuGgOse4Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu-(2-3)]--Glc-(1-1)-Cer - GD1a IV3Neu5AcII3Neu5AcGgOse4Cer, -Neu5Ac-(2-3)--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GalNAc-GD1a IV4GalNAcIV3Neu5AcII3Neu5AcGgOse4Cer, -GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - Neu neuraminic acid - Neu5Ac N-acetyl-neuraminic acid - Neu5Gc N-glycolyl-neuraminic acid - Cer ceramide  相似文献   

2.
Pseudomonas maltophilia is an uncommon cause of hospital-acquired infection and is resistant to most of the antimicrobial agents used in the treatment of gram-negative infections. Susceptibility of 108 isolates ofP. maltophilia to ceftazidime, aztreonam, defpirome, gentamicin, imipenem, enoxacin, and ticarcillin plus clavulanic acid was determined by an agar dilution method. The isolates were in general resistant to the antibiotics. Imipenem and cefpirome were not active at clinically achievable levels. Of the isolates, 20% were susceptible to 16 g/ml ceftazidime, 53% were susceptible to 4 g/ml enoxacin, 10% were susceptible to 4 g/ml gentamicin, and 25% were susceptible to 64 g/ml ticarcillin plus 2 g/ml clavulanic acid.  相似文献   

3.
We have measured the levels of interferon (IFN), tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-1, and IL-2 in the whole blood cell culture supernatants of 43 tumor patients undergoing a treatment with biological response modifiers or a conventional therapy with 5-fluorouracil and leucovorin. In the blood cell cultures of the 16 patients who received 5-fluorouracil and leucovorin IFN levels decreased (P0.01) and TNF levels rose (P0.05) during each therapy cycle. However, in the blood samples a declining number of total leukocytes and lymphocytes was measured (P0.05). Progressive disease could be correlated to a tendency towards lower IFN levels in the pretherapeutic cultures of these patients. The second group analyzed consisted of 8 patients receiving a low-dose IL-1 therapy. In this group we found either an unchanged or an augmented IFN production of the blood cells during treatment. In the group of 13 patients receiving low-dose recombinant IL-2 (4.5×106IU m–2 day–1) significantly increasing IFN levels were seen in the blood cell cultures during the therapy (P0.05), although total leukocyte counts decreased. In this group, 4 had stable disease for at least 2 months and 9 patients had tumor progression under therapy. In the cultures of the latter a tendency towards lower IFN values was found. Finally, the cytokine production in the blood cell cultures of 6 patients receiving a combination therapy of IFN and high-dose IL-2 was studied. During this therapy a dramatically reduced production not only of IFN but also of all other measured cytokines was found. In this group all patients had tumor progression under therapy. It is concluded that the measurements of cytokine production in a reproducible whole blood culture system may be useful for monitoring immunological therapies and may help us to find out which doses of biological response modifiers have enhancing or suppressive effects on the functions of the immune cells.  相似文献   

4.
Summary Cholic acid (3,7,12-trihydroxy-5-cholanoic acid) was completely and selectively transformed into 12-ketoursodeoxycholic acid (3,7-dihydroxy-12-oxo-5-cholanoic acid) by means of two consecutive enzymatic steps catalyzed, the first, by 7- and 12-hydroxysteroid dehydrogenase and, the second, by 7-hydroxysteroid dehydrogenase. Coenzyme regeneration was carried out with -ketoglutarate-glutamate dehydrogenase and glucose-glucose dehydrogenase, respectively.  相似文献   

5.
Barley anthers from cold pretreated spikes produced no or few calluses when plated with both loculi in contact with the medium (flat). When anthers were plated with only one loculus in contact with the medium (up), a high proportion of the anthers produced calluses. The top loculus of the up anthers was most productive. Flat anthers, when compared with up anthers, were not only slower to produce multicellular pollen grains (MCPs) and microcalluses, but also produced fewer of them and ceased production earlier. The MCPs and microcalluses in flat anthers grew more slowly and few developed beyond the 30 cell stage. These results establish the importance of anther orientation for barley anther culture.  相似文献   

6.
Summary Stationary phase cells of strain phr/MC 2 ofE. coli are not photoreactivable but the frequency of UV-induced mutations to low Streptomycine-resistance (S 3, 3/ml) is decreased strongly by illumination with light of fluorescence tubes (310 to 500 nm) after UV-irradiation. Also dark-reversion (DRM) of these mutations due to keeping UV-irradiated cells in saline is observed. Illumination before UV-irradiation decreases the frequency of the mutations (photoprotection against mutation=PPM) to the same extent as the combined action of photoreversion (PRM) and DRM. The lag-phase of cell division is prolonged strongly by illumination from 80 min without light to 150 min by the light-dose of highest activity. The additional lag is nearly the same if the illumination is done before, after or without UV-irradiation; this lag is about additive to the small lag caused by UV. Pre-illumination of the stationary-phase cells does not cause photoprotection against killing (PP), it even decreases the survival after high UV-doses. The observations support the hypothesis that PRM in this strain may be indirect, i.e. caused by the light-induced additional division lag which enhances the dark repair of UV-premutations. Also spontaneous premutations which are apparently present in the stationary-phase cells seem to be influenced by the light in this way.  相似文献   

7.
Chong  D.K.X.  Roberts  W.  Arakawa  T.  Illes  K.  Bagi  G.  Slattery  C.W.  Langridge  W.H.R. 《Transgenic research》1997,6(4):289-296
A 1177 bp cDNA fragment encoding the human milk protein -casein was introduced into Solanum tuberosum cells under control of the auxin-inducible, bidirectional mannopine synthase mas12) promoters using Agrobacterium tumefaciens-mediated leaf disc transformation methods. Antibiotic-resistant plants were regenerated and transformants selected based on luciferase activity carried by the expression vector containing the human -casein cDNA. The presence of human -casein cDNA in the plant genome was detected by PCR and DNA hybridization experiments. Human -casein mRNA was identified in leaf tissues of transgenic plants by RT-PCR analysis. Human - casein was identified in auxin-induced leaf and tuber tissues of transformed potato plants by immunoprecipitation and immunoblot analysis. Human -casein produced in transgenic plants migrated in polyacrylamide gels as a single band with an approximate molecular mass of 30 kDa. Immunoblot experiments identified approximately 0.01% of the total soluble protein of transgenic potato leaf tissue as -casein. The above experiments demonstrate the expression of human milk - casein as part of an edible food plant. These findings open the way for reconstitution of human milk inedible plants for replacement of bovine milk in baby foods for general improvement of infant nutrition, and for prevention of gastric and intestinal diseases in children  相似文献   

8.
Summary Theoretical studies on the necessary number of components in mixtures (for example multiclonal varieties or mixtures of lines) have been performed according to yielding ability. All theoretical investigations are based upon a Gram-Charlier frequency distribution of the component means with skewness 1 and kurtosis 2. The selected fraction p of the best components constitutes the mixture under consideration. The same selection differential S = S (p, 1, 2) can be realized by different parameter values of p, 1 and 2. Therefore, equal yield levels of the mixture can be achieved by different selected fractions p which implies different numbers of components in the mixture. Numerical results of S = S(p) for different values of 1 and 2 are presented and discussed. Of particular interest are the selected fractions p which lead to a maximal selection differential S. These results on S for large populations must be reduced in the case of finite population size. For this correction term we used an approximation B = B (p, n, 1, 2) given by Burrows (1972) where n = number of selected components. For given parameter values of 1, 2 and p, the necessary number n of components can be calculated by using the condition: Burrows-correction less than a certain percentage g of S — for example with g = 0.05 or g = 0.01. For given 1 and 2, the number n leading to a maximal selection differential S can be regarded as necessary number of components (necessary = maximum gain of selection under the given conditions). Numerical results are given for 2 = 0 and for eight situations which are defined by linear relations 2 = c 1 between skewness and kurtosis. These cases will contain all possible numerical situations for 1 and 2, which may be relevant for practical applications. The necessary number of components turns out to be nearly independent of the numerical value of the kurtosis 2. The n-intervals leading to selected fractions p from 0.01 to 0.20 approximately are: 2 n 4 for g = 0.05, 6 n 20 for g = 0.01 and 11 n 40 for g = 0.005, respectively. However, percentages g less than 0.01 would be unrealistically excessive. Therefore, following the assumptions and restrictions given in this paper one may conclude that n = 20 seems to be an appropriate upper bound for the necessary number of components in mixtures.  相似文献   

9.
Purification and properties of gammagamma-enolase from pig brain   总被引:1,自引:0,他引:1  
Isoelectric focusing revealed three enolase isoforms in pig brain, which were designated as - (pI = 6.5), - (pI = 5.6), and -enolase (pI = 5.2). The pI of purified -enolase was also 5.2. The -enolase isoform of enolase was purified from pig brain by a purification protocol involving heating to 55°C for 3 min, acetone precipitation, ammonium sulfate precipitation (40%–80%), DEAE Sephadex ion-exchange chromatography (pH 6.2), and Sephadex G200 gel filtration. The final specific activity was 82 units/mg protein. As with other vertebrate enolases, -enolase from pig proved to be a dimer with a native mass of 85 kDa and a subunit mass of 45 kDa. The pH optimum for the reaction in the glycolytic direction is 7.2. The K m values for 2-PGA, PEP, and Mg2+ were determined to be 0.05, 0.25, and 0.50 mM, respectively, similar to K m values of other vertebrate enolases. The amino acid composition of pig -enolase, as determined by amino acid analysis, shows strong similarity to the compositions of -enolases from rat, human, and mouse, as determined from their amino acid sequences. Despite the differences seen with some residues, and considering the ways that the compositions were obtained, it is assumed that pig -enolase is more similar than the composition data would indicate. Moreover, it is likely that the sequences of pig -enolase and the other -enolases are almost identical. Li+ proved to be a noncompetitive inhibitor with either 2-PGA or Mg2+ as the variable substrate. This enolase crystallized in the monoclinic space group P2, or P21. An R symm <5% was obtained for data between 50 and 3.65 Å, but was a disappointing 30% for data between 3.65 and 3.10 Å, indicating crystal disorder.  相似文献   

10.
The plant growth substance N6-(2-isopentenyl) adenine (i6Ade) significantly inhibits the rates of ethanol oxidation and acetaldehyde reduction in vitro by cell-free extracts of Zymomonas mobilis and of an Escherichia coli recombinant strain, containing the Z. mobilis adhB gene. The two-substrate kinetics of ethanol oxidation (forward) is affected by increasing values of dissociation constants for coenzyme and coenzyme —enzyme complexes in the presence of i6Ade, whereas the reaction maximum velocity (V m) remains unchanged and reflects the competitive type of inhibition. Changes of the kinetic constants of acetaldehyde reduction (back) are similar, except the increasing value of V m and correspond to the CIS (competitive inhibition and stimulation) type of inhibition. The estimated values of inhibition constants of the forward and back reactions are 0.39 ± 0.05 mM and 0.19 ± 0.06 mM, respectively.  相似文献   

11.
The binding of benzo()pyrene, ellipticine, and cis-parinaric acid to native, esterified, and alkylated -lactoglobulin was followed by enhancement of the ligand fluorescence. Three studied ligands bind to native or modified -lactoglobulin in apparent molar ratios varying between 1/8 and 2/1, with apparent dissociation constants in the range of 10–8 M for ligand/-lactoglobulin complexes. The studied, chemically modified -lactoglobulin derivatives display higher binding affinities for all studied ligands, cis-parinaric acid excluded. The reductive alkylation of -NH2 lysyl residues of -lactoglobulin increases the apparent molar ratios of benzo()pyrene and cis-parinaric acid, and decreases it for ellipticine. The esterified and native -lactoglobulin complexed to the investigated ligands display similar stoichiometries. Dynamic light scattering study of ligand--lactoglobulin complexes in solution shows the formation of aggregates: the apparent hydrodynamic radius value of -lactoglobulin dimer (3.4 nm) reaches 49, 46, and 74 nm upon addition and binding of benzo()pyrene, ellipticine, and cis-parinaric acid, respectively.  相似文献   

12.
A comparison of different habitats, biogas plant, rumen fluid and sewage sludge, for cellulolytic organisms indicated sewage studge was the best source. Enrichment cultura gave a mixed culture which exhibited CMCase activity as well as extracellular Avicelase, xylanase, -glucosidase, -xylosidase activities and cell-bound -glucosidase, and -xylosidase production in a synthetic medium with eleven different cellulosic and lignocellulosic substrates. The activity of extracellular -glucosidase and -xylosidase production was significantly higher than endogenous activities. Hemicellulases were induced better than cellulases. The anzyme system was stable under aerobic conditions. Of the different lignocellulosic substrates, kallar grass was the best inducer of extracellular enzymes.
Résumé La comparaison de différents habitats: digesteur méthanique, fluide du rumen ou boue de station d'épuration, pour leur contenu en organismes cellulolytiques, indiquent que la boue de station d'épuration est la meilleure source. Une culture par enrichissement a produit une culture mixte qui a exhibé aussi bien une activité CMCase que des activitiés extracellulaires avicelasique, xylanasique, -glucosidasique et -xylosidasique et qu'une production de -glucosidase et de -xylosidase liées à la cellule, dans un milieu synthétique et pour onze substrats cellulosiques et lignocellulosiques différents. L'activité de la -glucosidase extracellulaire et la production de -xylosidase sont significativement plus élevées que les activitiés endogènes. Les hemicellulases sont mieux induites que les cellulases. Le système enzymatique est stable dans des conditions aérobies. Parmi les divers substrats lignocellulosiques, l'herbe Kallar est le meilleur inducteur d'enzymes extracellufaires.
  相似文献   

13.
Cytological investigations combined with cytophotometric DNA determinations on Feulgen stained squash preparations of ovarioles from third and fourth instar nymphs of Carausius morosus revealed that during meiotic prophase a largely despiralized stage follows pachytene and that in this stage an extra reduplication of the nuclear DNA takes place (pachyreduplication phase). Meiotic prophase then proceeds with tetrapachytene, a pachytene-like stage with twice the amount of DNA as compared to oocyte nuclei in earlier meiotic stages, and with more than half the somatic number of elements, being probably autobivalents.Supported by Deutsche Forschungsgemeinschaft.  相似文献   

14.
The effect of carbonnitrogenphosphorus (CNP) ratio of organic substrates on the regeneration of ammonium and phosphate was investigated by growing natural assemblages of freshwater bacteria in mineral media supplemented with the simple organic C, N, and P sources (glucose, asparagine, and sodium glycerophosphate, respectively) to give 25 different substrate CNP ratios. Both ammonium and phosphate were regenerated when CN and NP atomic ratios of organic substrates were 101 and 161, respectively. Only ammonium was regenerated when CN and NP ratios were 101 and 10–201, respectively. On the other hand, neither ammonium nor phosphate was regenerated when CN and NP ratios were 151 and 51, respectively. In no case was phosphate alone regenerated. As bacteria were able to alter widely the CNP ratio of their biomass, the growth yield of bacteria appeared primarily dependent on the substrate carbon concentration, irrespective of a wide variation in the substrate CNP ratio.  相似文献   

15.
Summary The distribution of different laminin polypeptides, type VII collagen and tenascin has been studied in adult and foetal colorectal mucosa by using the indirect immunofluorescence technique. Immunoreactivity for laminin 1 chain was located to basement membranes of epithelia, muscularis mucosae, and blood vessels, respectively in different segments of adult colon and rectum. Laminin 1 and 1 chains were additionally expressed in lamina propria. Laminin 2 chain was also found in lamina propria around the pericyptal fibrollasts. Immunoreactivity for laminin 2 chain was restricted to basement membranes in the muscularis mucosae and arteries. Laminin 3 and 3 chains, suggestive for laminin-5, were confined especially to surface epithelial basement membranes. Immunoreactivity for type VII collagen was confined to basement membrane of surface epithelium in a punctate manner, while that for tenascin was seen slightly more broadly in the basement membrane zone and also in the muscular layer. The distribution of laminin chains in 16-week-foetal colon mostly resembled that of corresponding adult tissue, although immunoreactivities for laminin 2 and 2 chains were lacking. Type VII collagen and the high molecular weight isoform of tenascin also absent from the foetal colon. The results show that the basement membrane of the surface epithelium of colon and rectum express the components of epithelial adhesion complex, laminin-5 (3-3-2) and type VII collagen, resembling in this respect small intestine and stomach while laminin-2 (2-1-1) appears to be associated with pericryptal fibroblasts, and laminin-1 (1-1-1) widely in most basement membranes.  相似文献   

16.
The antimicrobial activity of amphotericin B, 5-fluorocytosine, nystatin, clotrimazole and miconazole were compared in vitro against 244 strains of yeasts that had been isolated from clinical specimens. The yeasts used in this study included 20 species of Candida, Cryptococcus, Saccharomyces Geotrichum, Rhodotorula, Torulopsis and Trichosporon. The majority of the strains (78%) had an MIC of 0.5 g/ml for amphotericin B, 81% an MIC of 1 g/ml for 5-fluorocytosine, 99% 8 g/ml for nystatin, 91%, 8.0 g/ml for clotrimazole and 98% had an MIC of 4.0 for miconazole. Of the anti-fungal agents tested, 5-fluorocytosine and nystatin were found to have the greatest antifungal activity.  相似文献   

17.
Aegilops tauschii (2n=2 x=14, DD) is a rich source of genetic variability for hexaploid wheat (Triticum aestivum, 2n=6 x=42, AABBDD) improvement. This variability can be accessed through utilizing synthetic hexaploid wheat lines, which contain genomes from Ae. tauschii and T. turgidum (2n=4 x=28, AABB). Numerous desirable characteristics can and have been introgressed into common hexaploid wheat with this germplasm. In this work, the genetic variability in the two puroindoline genes (a and b) contained on the D genome, and the relationship that sequence polymorphisms in these genes have on endosperm texture among a population of 75 CIMMYT synthetic hexaploid accessions is described. Kernel texture was evaluated using the single kernel characterization system (SKCS). Kernel texture differed significantly (P0.0001) among the synthetic hexaploid accessions (range 2.6–40.9) and the parent types, durum or Ae. tauschii. The interaction term between parent types was also a significant effect (P0.0001). In addition to the wild-type protein sequences of the puroindoline genes (those present in Chinese Spring and all other soft wheats), three other translated sequences were identified in puroindoline a and two others in puroindoline b. These protein sequences were associated with significantly (P0.0001) softer endosperm textures than the wild-type protein sequences. As the softer alleles are expressed in a hexaploid background, they are immediately available to wheat breeding programs.  相似文献   

18.
Summary The circular DNA decamer 5-dpCGC-TT-GCG-TT-3 was studied in solution by means of NMR spectroscopy and molecular dynamics in H2O. At a temperature of 269 K, a 50/50 mixture of two dumbbell structures (denoted L2L2 and L2L4) is present. The L2L2 form contains three Watson-Crick C-G base pairs and two two-residue loops in opposite parts of the molecule. On raising the temperature from 269 K to 314 K, the L2L4 conformer becomes increasingly dominant (95% at 314 K). This conformer has a partially disrupted G(anti)-C(syn) closing base pair in the 5-GTTC-3 loop with only one remaining (solvent-accessible) hydrogen bond between NH of the cytosine dC(1) and O6 of the guanine dG(8). The opposite 5-CTTG-3 loop remains stable. The two conformers occur in slow equilibrium (rate constant 2–20 s–1). Structure determination of the L2L2 and L2L4 forms was performed with the aid of a full relaxation matrix approach (IRMA) in combination with restrained MD. Torsional information was obtained from coupling constants. Coupling constant analysis (3JHH, 3JHP, 3JCP) gave detailed information about the local geometry around backbone torsion angles , , and , revealing a relatively high flexibility of the 5-GTTC-3 loop. The values of the coupling constants are virtually temperature-independent. Weakly constrained molecular dynamics in solvent was used to sample the conformational space of the dumbbell. The relaxation matrices from the MD simulation were averaged over r–3 to predict dynamic NOE volumes. In order to account for the 1:1 conformational mixture of L2L2 and L2L4 present at 271 K, we also included S2 factors and r–6 averaging of the r–3-averaged relaxation matrices. On matrix averaging, the agreement of NOE volumes with experiment improved significantly for protons located in the thermodynamically less stable 5-GTTC-3 loop. The difference in stability of the 5-CTTG-3 and 5-GTTC-3 loops is mainly caused by differences in the number of potential hydrogen bonds in the minor groove and differences in stacking overlap of the base pairs closing the minihairpin loops. The syn conformation for dC(1), favored at high temperature, is stabilized by solvation in the major groove. However, the conformational properties of the dC(1) base, as deduced from R-factor analysis and MD simulations, include a large flexibility about torsion angle .  相似文献   

19.
Part of a -amylase genomic DNA sequence from the oomycete, Achlya bisexualis was cloned by polymerase chain reaction (PCR) using degenerate oligonucleotide primers derived from the conserved regions of other known -amylase sequences. The 5- and 3-regions of the -amylase gene were amplified by genome walking method. The Ach. bisexualis -amylase gene consisted of a 1338bp open reading frame, encoding a protein of 446 amino acids with a molecular weight of 49 381Da, and was not interrupted by any intron. The deduced amino acid sequence of the -amylase gene had 67% similarity to the -amylase of Saprolegnia ferax, followed by 40% similarity to that ofArabidopsis thaliana. The -amylase gene was expressed in Saccharomyces cerevisiae placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.  相似文献   

20.
Hydrological variables have influenced fish recruitment in lakes, reservoirs and rivers. We evaluated how annual and seasonal hydrological variables were related to year-class strength (i.e., residuals from catch curves) of sportfish across eight Florida waterbodies (four rivers and four lakes). Multiple regression equations computed for black bass Micropterus spp. were combined across rivers and year-class strength was negatively related to spring median flow rates and in some cases positively related to winter median flow rates (all p0.10). Conversely, Lepomis spp. residuals combined from the rivers indicated that year-class strength was positively related to median flow rates in the fall prior to spawning and negatively related to post-spawn fall median flow rates (all p0.10). Fish recruitment combined across lakes were not related to water levels in this study, although within lake relationships did occur in some instances. Ecological implications of this work include regulations such as minimum flows and levels (MFLs) regarding sportfish species. Impacts of hydrology on year-class strength of sportfish were stronger in rivers than in lakes for these Florida systems. High flows at least once every 3years in the fall may allow inundation of floodplain habitat, providing favorable environmental conditions for Lepomis spp. reproduction. Setting MFLs during periods of drought (i.e., 3years or more) should consider impacts to short-lived species such as Lepomis spp.  相似文献   

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