Colonization and emergence of midges (Chironomidae: Diptera) in slow sand filter beds

Colonization by midges, and temporal changes in their community structure, were examined in slow sand filter beds. The replicated beds allow the development of communities to be traced from a known starting point.The filter beds (rectangular concrete containers filled with water) have a substratum of sand on which a rich coating of organic particles develops during passage of the water through the bed. The containers (ponds) are drained from time to time and the organic layer is then scraped off the sand surface. This occurs on average, once a month. The length of time the ponds were filled with water (bed run) during the present study ranged from 16 to 77 days.In long bed runs small midges with a short aquatic phase (Cricotopus sylvestris, Psectrocladius limbatellus, Tanytarsus fimbriatus) produced adults after 16–20 days; other, larger midges,e.g. Psectrocladius barbimanus and the Tanypodinae required a longer aquatic phase. Of the Tanypodinae, the smallAblabesmyia phatta, had the shortest duration of the four species found, and was much the most numerous member of this subfamily. Some Chironomini only appeared when the organic coating had developed over the sand surface. Midges of this tribe frequently failed to complete their larval development within the duration of bed runs and were thus trapped on the substratum at the time of cleaning. When ponds were drained after short bed runs the succession in community structure observed in long runs was arrested.Three small midgesC. sylvestris, P. limbatellus andT. fimbriatus, were collected in high numbers throughout the life of all beds, except towards the end of the longest runs in the study. This suggests that small size, short life cycles, and the ability to colonize clean substrata, are important characteristics for the development of primary chironomid communities in short-lived temporary habitats.     

1H NMR studies on bovine cyclophilin: preliminary structural characterization of its complex with cyclosporin A

Cyclophilin (163 residues, Mr 17737), a peptidyl prolyl cis-trans isomerase, is a cytosolic protein that specifically binds the potent immunosuppressant cyclosporin A (CsA). The native form of the major bovine thymus isoform has been analyzed by 2D NMR methods, COSY, HOHAHA, and NOESY, in aqueous media. The 156 main-chain amides in CyP yield 126 observable NH/alpha CH couplings (81%, Gly pairs counted as 1). Following exhaustive D2O exchange, 44 amide resonances remain visible. Further analysis of the NH/NH, NH/alpha CH, and alpha CH/alpha CH regions of the COSY and NOESY data sets indicates that the residual amides in D2O form a coherent hydrophobic domain which yields 2D NMR features suggestive of a beta-sheet. Many (43/126) of the amide resonances have been classified according to amino acid type. In the aromatic region of the spectra, the assignment of the ring spin systems is nearly complete (12/15 Phe, 2/2 Tyr, 1/1 Trp, and 3/4 His). This has successfully lead to the complete assignment of all of their beta CH's, main-chain alpha CH resonances, and many of the backbone amide resonances (8/12 Phe, 2/2 Tyr, 1/1 Trp, and 2/3 His). In other regions of the spectrum, the side-chain and main-chain resonances for 10/23 Gly, 9/9 Ala, 5/11 Thr, 5/9 Val, and 1/6 Leu have been completely assigned. The drug-free cyclophilin and CsA-bound cyclophilin form two discrete protein structures that are in slow exchange on the NMR time scale. Comparison of the fingerprint regions from the COSY spectra obtained from the two forms of the protein reveals a minimum of 16 cross-peaks which are clearly shifted upon complexation. In fact, on the basis of chemical shift changes observed in assigned side-chain and main-chain resonances, only a relatively few of the amino acid residues identified to date are perturbed by complex formation. These include 3 Phe (8, 12, and 14) and the Trp in the aromatic region and 2 Ala (7 and 8) in the Ala/Thr region. In the upfield-shifted methyl region, an assigned Leu and Val spin system and a spin system labeled X10 (an Ile or Leu) are affected by complex formation. In addition, a new aliphatic spin system, labeled X11, which shows a close spatial relationship to the perturbed Phe12, is observed in this region of the spectrum. In summary, the regions of the protein altered by complex formation can be divided into two categories: a hydrophobic and a H2O-accessible domain.(ABSTRACT TRUNCATED AT 400 WORDS)     

Monoclonal antibodies to rabbit liver cytochrome P-448

Cytochrome P-448 (mol wt 55,000 Daltons) from rabbit liver was purified to a specific content of 16.6 nmol/mg. Mice were immunised with this preparation, their spleens removed and dissociated lymphocytes hybridised with myeloma cells. Four monoclonal antibodies against cytochrome P-448 were raised and partially characterised. All four antibodies interacted with cytochrome P-448 in intact microsomal fractions and selectively immunoadsorbed cytochrome P-448 from solubilised microsomal preparations. One of the antibodies inhibited benzo[a] pyrene hydroxylase activity in a reconstituted system, one had no effect on activity and two increased activity. The possible applications of such antibodies are discussed.     

Sociality as a life-history tactic of ground squirrels

Summary Multi-variate analysis of life-history traits of 18 species of burrowing sciurids indicates that reproductive effort is determined by body-size energetics. Other traits, such as age adult weight reached, age of dispersal, length of time of gestation, were significantly correlated with body size. A principal component analysis suggested that the complex of life-history traits could be reduced to four components: body size (=weight), seasonality, specific reproductive effort, and maturity. The variation in the sociality index was best explained by age of first reproduction and age adult weight reached. Generally, species are more social when large body size combined with a relatively short growing season is associated with delayed dispersal and occurs in those species typically breeding for the first time at age two or older. Sociality in these species may have evolved through retention of daughters within the maternal home range as a means of continuing reproductive investment beyond weaning.     

113Cd nmr study of the metal cluster structure of human liver metallothionein

Cadmium-113 nuclear magnetic resonance (¹¹³Cd nmr) was used to elucidate the structural properties of the cadmium binding sites in human liver metallothionein. The isotopically labeled ¹¹³Cd-metallothionein was prepared by the in vitro exchange of the native metals (greater than 94% zinc) for ¹¹³CdCl₂ during isolation. The two isoproteins, MT-1 and MT-2, showed ¹¹³Cd nmr resonances in the chemical shift range 610–670 ppm. The multiplet structure of the resonances is due to two bond scalar interactions between adjacent ¹¹³Cd ions linked by cysteine thiolate ligands. Homonuclear ¹¹³Cd decoupling experiments allowed the determination of the metal cluster structure, which, similar to the rabbit liver metallothionein, consists of a four- and a three-metal cluster designated cluster A and cluster B, respectively. Chemical shift similarities in the ¹¹³Cd nmr spectra of the human, rabbit and calf liver MT-1 and MT-2 are observed, especially for cluster A. Small variations in chemical shifts are explained in terms of differences in the primary structure between the two human isoproteins.     

The effects of mine drainage and organic enrichment on benthos in the river nent system,Northern Pennines

A preliminary survey of the benthic fauna of sites on the Nent system affected by elevated zinc levels, acid water and organic enrichment is described. Data on faunal composition are presented for 17 sites. A clustering technique was applied to the taxa/site matrix to demonstrate the degree of association between sites varying in zinc concentration. Chandler biotic scores were found to be lowest at sites affected by high zinc levels and/or dense growths of the alga Stigeoclonium tenue. The role of elevated zinc levels and algal growth in determining the diversity and abundance of benthos is discussed.     

Absorption and metabolism of nicotine from cigarettes.

Eight men volunteers each smoked a single cirgarette containing 14C-nicotine and gave arterial blood samples during and for 50 minutes after smoking. The maximum concentration of nicotine in the arterial blood ranged from 31 to 41 mug/l in four regular cigarette smokers who inhaled. Two non-smokers achieved maximum levels of 2 and 4 mug/l. On a separate occasion two of the inhalers received 1 mg. 14C-nicotine in 10 divided doses injected intravenously. In both cases the peak arterial nicotine concentrations bore a similar relationship to the intravenous dose, as did the peak nicotine concentrations to the retained doses during smoking.     

1H NMR studies on bovine cyclophilin: preliminary structural characterization of this specific cyclosporin A binding protein

High-field 1H NMR spectroscopy has been used to study the conformation of the cytosolic cyclosporin A binding protein cyclophilin. For the drug-free form of cyclophilin, spectral editing methods in conjunction with a pH titration were used to identify all four His residues present in the protein, and two-dimensional COSY and RELAY spectroscopy was used to elucidate the scalar connectivities in the aromatic and upfield methyl regions of the spectrum. From these scalar connectivities, it was possible to distinguish between inter- and intraresidue dipolar interactions within the aromatic and upfield methyl regions of cyclophilin in the NOESY spectrum. The results of this analysis showed extensive interresidue cross-relaxation among and between these latter spectral regions indicative of the proximal relationships of several of these residues and the presence of a hydrophobic core within cyclophilin.