Rabbit β-glucuronidase. Purification and properties, and the existence of multiple forms

1. beta-Glucuronidase (EC 3.2.1.31) was purified from rabbit liver by a procedure involving autolysis, (NH(4))(2)SO(4) fractionation, chromatography on DEAE-cellulose and hydroxyapatite, gel filtration, sedimentation in a sucrose gradient, and isoelectric focusing. 2. Electron microscopy revealed ferritin as the major contaminant in later stages of purification and also showed aggregates of enzyme molecules. Particular attention was paid to the removal of ferritin. 3. The purified enzyme was homogeneous in polyacrylamide-gel electrophoresis both in non-dissociating conditions and in the presence of sodium dodecyl sulphate, and in Ouchterlony gel diffusion and immunoelectrophoresis against polyspecific antisera. 4. Sedimentation in sucrose gradients gave a molecular weight of 300000, whereas gel filtration indicated 440000. 5. Subunits of 75000 molecular weight were observed in gel electrophoresis in the presence of sodium dodecyl sulphate and in gel filtration in the presence of urea. 6. The K(m) value for p-nitrophenyl beta-d-glucuronide was 0.6mm, and the enzyme was extremely sensitive to lactone inhibitors. It was also inhibited by Hg(2+) ions. 7. Multiple forms were observed in the pure enzyme by isoelectric focusing, with pI values of 4.5-5.8. Subunits showed similar heterogeneity. The origin of the multiple forms was investigated in detail, and the possibility of artifact generation largely excluded. Some of the forms of lowest pI disappeared after neuraminidase digestion. The nature of the residual heterogeneity remains to be elucidated.     

Science,philosophy, and politics in the work of J. B. S. Haldane, 1922–1937

This paper analyzes the interaction between science, philosophy and politics (including ideology) in the early work of J. B. S. Haldane (from 1922 to 1937). This period is particularly important, not only because it is the period of Haldane's most significant biological work (both in biochemistry and genetics), but also because it is during this period that his philosophical and political views underwent their most significant transformation. His philosophical stance first changed from a radical organicism to a position far more compatible with mechanical materialism. The primary intellectual influence that was responsible for this shift was that of F. G. Hopkins. Later, Haldane came to accept Marxism and its official metaphysics, dialectical materialism, a move that let him accept the materialist conception of the world while still maintaining a resolute distance from mechanism. Throughout all these changes, what is most obvious is the influence of science on Haldane's philosophical views. An influence in the opposite direction is far less apparent.Parts of this paper are extracted from a longer work which concerns the interactions between philosophy and science throughout Haldane's scientific career (Sarkar forthcoming). The general conclusions reached here, from a consideration of Haldane's work only from 1922 to 1937 (see Section 6), remain the same for the rest of his life, as is detailed in the longer work. Thanks are due to R. S. Cohen, J. F. Crow, A. R. Fersht, J. Maynard Smith, R. C. Olby, D. Paul, M. Ruse, J. Stachel and S. Sturdy for helpful discussions and comments and criticism of the positions outlined in this paper. This is Contribution No. BTBG-92-4 from the Theoretical Biology Group, Boston University.     

Ontogenesis of the α-MSH, β-MSH and ACTH cells in the foetal hypophysis of the rat. Correlation with the growth of the adrenals and adrenocortical activity

Pituitary sections from 15 to 21 day-old rat foetuses have been studied with the immunofluorescence technique, using antibodies anti alpha-MSH, anti beta-MSH and anti beta (1-24) ACTH. The first ACTH cells appear on day 17 of pregnancy in the pars distalis of the hypophysis and only on day 18 in the pars intermedia. beta-msh cells have been observed on day 16 in the pars anterior and on day 17 in the pars intermedia, while alpha-MSH cells appear only on day 18 and exclusively in the pars intermedia. The cytodifferentiation of the beta-MSH and ACTH cells occurs in the pars intermedia with about a 24 hours delay in comparison to that of the pars distalis. The first revealed cells are always located in the posterior half of the pituitary gland. The corticostimulating activity of the hypophysis has been tested with the fluorescence intensity of the corticotrophs, the adrenal weight, the adrenal content in corticosterone and the plasma corticosterone level. The fluorescence of the corticotrophs increases on day 18, shows a maximum on day 19 and decreases until term. The adrenal weight rises regularly between day 16 to day 20, thereafer the increase subsides. Adrenal and plasma corticosterone concentrations reach a peak on day 19 of pregnancy. These data suggest that hypophyseal corticostimulating activity is very high between days 18 and 19 and decreases between days 19 and 21.     

Reconstruction of the Allerød vegetation of the Neuwied Basin, western Germany, and its surroundings at 12,900 cal b.p.

During the late-glacial eruption of the Laacher See volcano 12,900 b.p. the whole area of the Neuwied Basin, middle Rhine region, was covered by several metres of the Laacher See Tephra (LST) damaging but also partly conserving the former vegetation. This gave the unique opportunity to study plant remains in situ and to estimate the spatial biodiversity of plant taxa within the area investigated at the end of the last glacial as if in a snapshot of 12,900 years ago. Different kinds of plant material can be found because of different preservation conditions namely imprints, charcoal and waterlogged remains. Investigations of this material from several places within the Neuwied Basin and the Brohl valley north of it allowed the reconstruction of the former vegetation at different scales from local to regional. This resulted in a large scale draft of a vegetation map of the studied area also based on the distribution and the stage of soil development at that time. As to whether thermophilous trees were already present is discussed but largely rejected. Electronic Supplementary Material Supplementary material is available for this article at     

Review of the fauna of Rotatoria,Cladocera, and Copepoda of the basin of the Anadyr’ River

The zooplankton composition is studied in the thermokarst, glacial and meteorite lakes, channels, former riverbeds, and hollows in the basin of Anadyr’. We found 174 taxa: 78, Rotatoria, 55, Cladocera, and 41, Copepoda. The most diverse is the lake fauna: 51 taxa of Rotatoria, 48, Cladocera, and 37, Copepoda. The thermokarst Lake Maiorskoe hosts 68 taxa: 31, Rotatoria, 14, Cladocera, and 23, Copepoda, wheras the cold ultraoligotrophic Lake El’gygytgyn features only one species of Cyclop of the group scutifer Cyclops neymanae Strel., and Rotatoria and Cladocera are present as allochtonous forms. The Copepoda illustrate the relations of the Anadyr’ fauna with those of Europe, North America, and Japan.     

Biochemical studies of the chromaffin granule. III. Redistribution of lipid phosphate,dopamine-β-hydroxylase and chromogranin a after freezing and thawing of the isolated granule membranes

Freezing and thawing a dialysed suspension of lysed chromaffin granules and sedimented membrane preparations resulted in redistribution of lipid phosphate and protein. By this treatment the high ratios of lipid phosphate/protein in the membrane fragments, isolated on sucrose density gradient from the dialysed suspensions and the sedimented membrane preparation, were reduced from 1.56 to 1.03 μmoles/mg and from 1.97 to 0.83 μmoles/mg, respectively.Multilamellar, liposomal structures could be isolated from the frozen and thawed membrane preparations and were found to sediment in the 0.4 M sucrose layer by density gradient centrigugation. This fraction was without morphological resemblance to the intact chromaffin granules or their membranes and was found to account for 53% of the lipid phosphate, 35% of chromogranin A, 21% of dopamine-β-hydroxylase activity and 12% of the protein of the total preparation. The specific activities of chromogranin A and dopamine-β-hydroxylase in the artificially formed liposomal structures closely resembled that of the solubilized protein and was significantly higher than in the lipid phosphate-depleted membrane fragments recovered in the 1.1 M sucrose layers.It is concluded that freezing and thawing as a means of purifying the isolated granule membranes lead not only to the solubilization of chromogranin A, but also to removal of dopamine-β-hydroxylase activity and lipid phosphate from the labile membrane fragments.     

Behaviour of selected PCBs,PAHs and γ-HCH in the Scheldt estuary,S.W. Netherlands

The behaviour of some organic micropollutants in the abiotic compartments water, particulate matter and bottom sediments of the Scheldt estuary, S.W. Netherlands, was studied between 1986 and 1989. Special attention was given to two individual PolyChlorinated Biphenyl congeners (PCB 52 and PCB 153), two Polycyclic Aromatic Hydrocarbons (PAHs) fluoranthene and benzo(a)pyrene and γ-HCH. Dissolved PCBs and γ-HCH behave conservatively during estuarine mixing. The concentration of γ-HCH in the fresh-water is extremely high in spring compared to the other seasons. Dissolved fluoranthene does not behave conservatively. The concentration of dissolved fluoranthene in the freshwater endmember shows a seasonal dependence with highest concentrations in winter and lowest in summer. Benzo(a)pyrene could hardly be detected in the dissolved phase with the methods used. PCBs and PAHs in particulate matter and sediments behave essentially conservatively as a result of the mixing of riverine particulates with high, and marine particulates with low organic micropollutant contents. Particulate fluoranthene is removed at low salinities under (nearly) anoxic conditions probably by microbial degradation. The rivers Scheldt and Rupel appear to be the major sources for the compounds studied. However, in particular the organic micropollutant contents of the sediments indicate that important emissions along the estuary were or are still present. Measurements of individual PCBs and PAHs in the <63 μm fraction of a dated sediment core from a salt marsh in the eastern part of the Scheldt estuary show that the recent input of PCBs and PAHs into the estuaryis probably 2–3 times lower than the maximum input in the mid-1960s. The PCB profile shows postdepositional congener-selective mobilization caused by advective transport. The PAH assemblage is remarkably uniform in the sediment core. The PAH ratios indicate that combustion of coal is the main source of PAHs in the Scheldt estuary.     

Spontaneous activity of foregut and hindgut visceral muscle of the locust,locusta migratoria—I. Normal activity and the effect of KCl depolarization and glutamate

1. Locust foregut and hindgut differed in their spontaneous contractility. The foregut was constantly active with regular contractions or with a fast rhythm incorporating larger irregular contractions.2. The hindgut showed bursts of large regular contractions interrupted by periods of quiescence.3. Foregut fluid, of unknown content, inhibited rhythmic contractions but induced large foregut tonic contractures.4. High KCl salines induced contractures in both gut sections, but the tension/[K]₀ curve showed no clear mechanical threshold.5. Glutamate (10⁻⁵ M) increased contraction frequency in the hindgut and stimulated quiescent preparations. At 10⁻⁴M, glutamate usually increased foregut contraction amplitude and frequency, this action being inhibited by 10⁻⁵ M tvramine.6. Considerable differences exist in natural rhythm, contractile properties and drug responses between these two divisions of locust gut.     

β-Mannoside mannohydrolase and the mobilization of the endosperm cell wall of lettuce seeds,cv. Grand Rapids

Mobilization of the endosperm cell-wall reserves of Lactuca sativa L. cv. Grand Rapids requires endo--mannanase and -galactosidase activity. A third enzyme, -mannoside mannohydrolase (EC 3.2.1.25) is also involved. We have determined the optimum extraction and assay conditions for this enzyme, which is soluble only in high-salt (1 M NaCl) buffer. It is located exclusively within the cotyledons, in association with a cellulosic cell-wall fraction. Its substrates are the products of endosperm cell-wall mobilization, mannobiose and mannotriose, which diffuse to the cotyledons and are hydrolyzed extracellularly by the -mannoside mannohydrolase.Abbreviation PNPM p-nitrophenyl--mannopyranoside We dedicate this paper to the memory of our friend and colleague Dr. Jacob D. Duerksen     

Ecological Partitioning of Cercopithecus campbelli, C. petaurista, and C. diana in the Taï Forest

I determined the degree of ecological partitioning among 3 species of guenons (Cercopithecus campbelli, C. petaurista, and C. diana) in the Taï Forest, Côte d’Ivoire and used the partitioning data to understand competitive relationships among them. Over a 13-mo period, I measured ecological partitioning in terms of food and canopy stratum use for 2 habituated groups of each guenon species and also collected data on food availability. I found that the study species diverged primarily in food items consumed and vertical strata occupied. Cercopithecus petaurista ate much more foliage than the other species did and used mostly the middle strata (5–20 m). Cercopithecus diana ate primarily fruit and used mostly the upper strata (>20 m). Cercopithecus campbelli ate mostly fruit together with large amounts of animal matter and primarily occupied the ground and low strata (<5 m). Of the specific pairs, the diets of Cercopithecus campbelli/C. diana overlapped the most overall and decreased during the season of low fruit availability. Cercopithecus campbelli and C. diana age/sex classes also overlapped more than the age/sex classes of other species pairs. The results suggest that the potential for competition was more intense for Cercopithecus campbelli/C.diana relations than it is for other species pairs. I compare my results from Taï with those from other primate and guenon communities and demonstrate that dietary overlaps and seasonal dietary divergence are lower in Taï than in most other guenon communities.     

Exploratory multiscale analysis of the fish assemblages and habitats of the lower St. Lawrence River,Québec,Canada

This work presents a multiscale analysis of the fish diversity of the lower St. Lawrence River which flows from the Laurentian Great Lakes of North America. A database of about 14,000 fish sampling stations from the lower St. Lawrence is linked to five different habitat maps of the study area: hydrographical units, water masses, depth, sediment type and wetland type. We hypothesize that species turnover (beta diversity) will be significant among these habitat maps. For each map, we calculate a UPGMA dendrogram based on the Jaccard coefficient of similarity in species occurrences as a first assemblage–habitat model. A randomization test is then used to identify the significant dendrograms from which we infer fish assemblages. We then show that many species are actually selecting or avoiding habitats. Finally, species selecting particular habitats are described by some selected ecological traits that are expected to occur frequently in those habitats. We found assemblages for the hydrographical units but not for the water masses. The fluvial section is selected by many stenohaline and some euryhaline species, while the freshwater estuary and the brackish estuary are only selected by euryhaline species. In the fluvial section at the hydrographical unit scale, many species associated with lentic habitats are limnophilic and more vegetation-dependent while many species selecting lotic habitats are rheophilic insectivorous species. Significant assemblages are defined for depth, sediment type and wetland type. Taken together, we found strong empirical evidence of a diverse actively selecting littoral assemblage of small, low mobility fishes opposing a channel assemblage of larger, more mobile fishes.