Effect of Metal-Rich Sewage Sludge Application on the Bacterial Communities of Grasslands

The effect of long-term application of heavy metal-laden sewage sludge on the total heterotrophic aerobic and the cadmium-resistant soil bacterial communities was studied. Gram-positive bacteria were completely absent from resistant communities. These findings suggest that this group is highly susceptible to Cd. Shannon's diversity indices estimated for total communities did not reveal negative effects on the communities that developed in the presence of sludge. However, Cd-resistant communities isolated from long-term sludge-amended soils were more diverse than the resistant communities from a control sample, suggesting that adaptation to Cd as a stressor had occurred in the presence of sludge constituents. This higher diversity was attributed to Cd resistance in pseudomonads and gram-negative fermenters. Resistance did not develop by dissemination of Cd resistance plasmids, because these were rarely detected in the genomes of resistant strains.     

Phylogenetic Survey of Proteins Related to Synapsin I and Biochemical Analysis of Four Such Proteins from Fish Brain

A phylogenetic survey of proteins immunologically related to Synapsin I, a major synaptic vesicle-associated phosphoprotein in mammals was carried out. Proteins antigenically related to Synapsin I were found by use of radioimmunoassay and other radioimmunochemical techniques in the nervous systems of several vertebrate and invertebrate species, which included birds, reptiles, amphibians, fish, echinoderms, arthropods, and mollusks. Four proteins present in fish brain, antigenically related to Synapsin I, were further studied and found to resemble mammalian Synapsin I in several respects. Like Synapsin I, the fish proteins were present in high amounts in nervous tissue, were enriched in synaptosomal fractions of brain where they were substrates for endogenous protein kinases, were acid extractable, and were sensitive to digestion by collagenase. In addition, two-dimensional peptide-mapping analysis revealed some homology between major phosphopeptide fragments of Synapsin I and the fish proteins. The results indicate that proteins related to Synapsin I are wide-spread in the animal kingdom.     

Class I and class II restriction pattern polymorphisms associated with independently derived RT1 haplotypes in inbred rats

This communication reports the DNA level identification of class I and class II sequences associated with 20 RT1 haplotypes which have been assigned previously to eight RT1 groups. Sixteen to 22 bands in genomic blots hybridized with the mouse pH-2III class I cDNA probe. Only the three RT1 ^khaplotypes associated with identical class I restriction fragment patterns. Differences in restriction bands between putatively identical RT1 haplotypes were either less than or equal to 6%, or greater than 50%, suggesting a relatively high level of recombination between serologically identified RT1.A genes and the majority of class I sequences. Restriction fragment patterns associated with three RT1 ^uhaplotypes differed by less than 6%. However, intra-RT1 ^a,intra-RT1 ^b,and intra-RT1 ^lrestriction fragment differences were between 50 and 64%. In specific cases, different RT1 haplotypes associated with identical class I restriction patterns, e.g., RT1 ^m(MNR) and RT1 ^d(MR); higher resolution confirmed the difference (two bands) between RT1 ^mand RT1 ^d.Results of hybridization with the human DC₁ probe confirmed that the AVN RT1 ^aand NSD RT1 ^bhaplotypes were generated by recombinations within the vicinity of the RT1.B : RT1.D regions. These results demonstrate that a previous classification of RT1 haplotypes was incomplete and did not include the majority of class I and class II sequences which distinguish RT1 haplotypes.     

A rapid naphthol yellows method for measuring the cellular protein content of anchorage cultures

Summary A rapid method has been developed for measuring the cellular protein content of mono- and multilayered anchorage cultures. Fixed or air dried cultures are stained for 30 min with 0.2% Naphthol Yellow S (NYS) dissolved in 1% acetic acid. Unbound dye is removed by a series of four 2.5 min washes in 1% acetic acid, and protein-bound dye extracted with 10 mM unbuffered Tris base for spectrophotometric optical density determination at 433 nm. The NYS method exhibited a least-squares correlation coefficient of 0.99997 with the Oyama-Eagle Lowry method.     

Reduction in headache patients' medical expenses associated with biofeedback and relaxation treatments

Comparisons are made of self-reported medical costs from a sample of headache patients who underwent various combinations of relaxation training and biofeedback training. The average costs for the 2 years prior to self-regulatory treatment were $955±480 (3 SEM) for 45 patients; for the 2 years after completing treatment the average costs were $52±28 (3 SEM) for patients. Within the limitations of the study, medical costs do seem to have been markedly reduced.This research was supported by a grant from NINCDS, NS-15235.     

A note on the characterization of an estuarine microbial community enriched with the herbicide Fenuron

M inney , S.F., P arkes , R.J. & B ull , A.T. 1985. A note on the characterization of an estuarine microbial community enriched with the herbicide Fenuron. Journal of Applied Bacteriology 59 , 17—22.
A stable, five-membered estuarine microbial community was enriched in the presence of 1,1 dimethyl-3-phenylurea (Fenuron), phenylurea and aniline. At a dilution rate of 0μD01/h in the chemostat, partial aniline utilization occurred and the rate increased with dilution rate. At the higher dilution rates the phenylurea component was also utilized. At a dilution rate of mD015/h in a fluidized bed system, complete removal of both aniline and phenylurea occurred. There was no evidence of Fenuron degradation under any of the conditions examined. The importance of the use of heterogeneous culture conditions in the laboratory is discussed.     

REPRODUCTIVE MATURITY AND SEASONALITY OF MALE SPOTTED DOLPHINS, STENELLA ATTENUATA, IN THE EASTERN TROPICAL PACIFIC

We estimated age at attainment of sexual maturity and examined reproductive seasonality for male spotted dolphins, Stenella attenuata , from the eastern tropical Pacific Ocean. Maturity was determined by histological examination of testes. Average age at sexual maturation was 14.7 yr (the mean of two readers' age estimates). Testis and epididymis weight and seminiferous tubule diameters were reliable indicators of maturity, whereas age, length and color phase were less reliable. Seasonality was determined by changes in testis and epididymis weight, relative quantity of spermatids and spermatozoa, and lumen diameter, as well as an index of testis development (weight of the right testis and epididymis divided by length of the right testis). Testis and epididymis weights and index values peaked in July and August, midway between two predicted mating seasons for the northern offshore stock, but spermatozoa levels were elevated during the predicted breeding seasons.     

Cell surface changes in preimplantation mouse embryos during compaction investigated using FITC conjugated lectins after proteolytic enzyme treatment

Summary Fluorescein-conjugated lectins were used to examine the reappearance of glycoproteins on the surface of 8-cell mouse embryos after treatment with proteolytic enzymes. Embryos were decompacted in calcium free medium, treated with various proteases and the process of recompaction monitored. The most effective enzymes in delaying recompaction were subtilopeptidase A and proteinase K at 1 mg/ml; the initiation of recompaction was delayed by about 5 h and 90% recompaction by 14–18 h. Papain and -chymotrypsin were only effective in the absence of calcium. The reappearance of receptors for fluorescein-conjugated Con-A, MPA, RCA-I, FBP, BSL-II and DBA was examined photometrically at 0,8–10 and 17–18 h after proteinase K treatment. There was an increase in binding of MPA, RCA-I, FBP and BSL-II in control embryos during the period of the experiment, between approx. 61 and 80 h post coitum in which embryos passed from the 8-cell stage to the 16–32 cell stage. Con-A binding remained the same and that of DBA decreased. By the time that 50% of enzyme treated embryos had recompacted (8–10 h) binding of Con-A was similar to control embryos. Binding of FBP had almost reached control levels while that of BSL-II, DBA, RCA-I and MPA had reached 60–85% of control levels. When embryos were fully compact (17–18 h) Con-A, FBP and DBA were bound in equal or slightly greater amounts to enzyme treated as to control embryos, and receptors for BSL-II, MPA and RCA-I had recovered almost to control levels. The results clearly show that the recovery of glycoproteins on the surface of 8–16 cell embryos parallels recompaction, providing further evidence for the role of these molecules in compaction.     

Repair inhibition of potential mutations of ultraviolet-irradiatedEscherichia coli by chloroquine and quinacrine

The frequency of ultraviolet(UV)-induced mutations drops rapidly whenEscherichia coli Hcr⁺ cells (strains WP-2 Hcr⁺; B/r) are incubated on phosphate-buffered agar (PBA), but is reduced only slightly if chloroquine or quinacrine are incorporated into the medium. The excision-deficient WP-2 Hcr^– strain shows little reduction in the number of mutants when incubated on PBA. During postirradiation incubation on PBA, cell viability was relatively unaffected by the presence of the chemicals in the PBA (25 g/ml quinacrine; 50 g/ml chloroquine). When cells were given optimal doses of photoreactivating light, no further decline in mutations was obtained during subsequent incubation on PBA. Approximately 64% of the mutants seen when cells are treated with UV-PBA-chloroquine and 90% seen with UV-PBA-quinacrine can be repaired if cells are incubated on PBA. When these chemicals were added to the PBA, both excision-proficient strains (WP-2 Hcr⁺; B/r) demonstrated a marked reduction in the repair of UV-induced mutations to streptomycin resistance. Our results indicate that these chemicals interfere with the excision of UV-induced pyrimidine dimers, a process that normally occurs during postirradiation incubation on PBA.