N-isobutyryl-His-Trp-Ala-Val-D-Ala-His-Leu-NHMe (ICI 216140) a potent in vivo antaconist analogue of bombesin/gastrin releasing peptide (BN/GRP) derived from the C-terminal sequence lacking the final methionine residue

The GRP receptor mediated growth response in Swiss 3T3 cells has been used to identify BN/GRP antagonists. Analysis of bombesin antagonism by substance P analogues and by truncated GRP analogues revealed that deletion of the C-terminal methionine residue was important for antagonism. Des-Met analogues showing potent antagonist activity in the in vitro 3T3 system (IC50 approximately 2nM) were synthesized. Further structural modification of these peptides led to the identification of (CH3)2CHCO-His-Trp-Ala-Val-D-Ala-His-Leu-NHCH3 (ICI 216140) which reduced bombesin-stimulated rat pancreatic amylase secretion to basal levels when administered subcutaneously at 2.0 mg per kg.     

Down-regulation of phytochrome mRNA abundance by red light and benzyladenine in etiolated cucumber cotyledons

Northern blot analysis revealed that a single 4.2 kb phytochrome mRNA species was detectable in cotyledons excised from five-day-old etiolated cucumber seedlings. Intact etiolated five-day-old cucumber seedlings were given a red light or benzyladenine treatment, and cotyledons were harvested at various times following treatment. The abundance of phytochrome mRNA in the cotyledons was quantitated using ³²P-labeled RNA probes and slot blot analysis. By 2 h after irradiation the phytochrome mRNA level was reduced to 40% of the initial abundance and reaccumulation began by 3 h after irradiation. Reaccumulation of phytochrome mRNA to the time-zero dark control level was achieved by 10 h after treatment. A decrease in phytochrome mRNA abundance was evident by 2 h after benzyladenine treatment, and a maximal reduction to 45% of the time-zero dark control was attained by 4 h after treatment. No recovery of the phytochrome mRNA level was evident by 8 h after benzyladenine treatment. The abundance of actin mRNA was unaffected by benzyladenine treatment.     

Mechanism of cadmium uptake by activated sludge

Summary The significance of metabolic activity in cadmium uptake by unacclimated activated sludge was studied. Below 30 mg/l cadmium in solution, biosorption was found to follow the Freundlich isotherm, which is the most common pattern for physico-chemical adsorption. More than 95% of total cadmium uptake was achieved within 5 min metal-sludge contact time. Biosorption increased strongly when the initial cadmium concentration in solution was raised from 10 to 100 mg/l, whereas in the same concentration range the metabolic activity of the sludge, as measured by respiratory activity and extracellular protein production, was very significantly inhibited. The addition of nutrients at low but significant levels failed to increase cadmium uptake in 2 h contact time, while in 24 h the addition of nutrients caused the biosorption to increase by only 5–10% without any significant growth of the biomass. Biosorption was found to increase with temperature between 5° C and 40° C, in correlation with an increase in the metabolic activity of the sludge. Pretreatment of the sludge with metabolic inhibitors (NaN₃ and UV rays) appeared to cause only a very slight decrease (5–10%) of biosorption. These results suggest that metabolic uptake of cadmium was low and that adsorption to the surface of the cells was the major mechanism of uptake.Offprint requests to: S. S. Sofer     

Preservative-mediated changes of the surface properties ofEscherichia coli

The effectiveness of two commonly used preservatives, sodium benzoate and potassium disulfite, was evaluated in terms of their bactericidal activity and capacity to induce changes in the surface properties ofEscherichia coli isolated from commercial food preserves. Preservative treatment over a five-week test period resulted in controlling the multiplication of these organisms and causing a decline in cell-surface hydrophobicity, hemagglutinating ability and adherence capacity to rat intestinal cells ofE. coli isolates. A loss in motility was also exhibited.     

Intensive Linkage Mapping in a Wasp (Bracon Hebetor) and a Mosquito (Aedes Aegypti) with Single-Strand Conformation Polymorphism Analysis of Random Amplified Polymorphic DNA Markers

The use of random amplified polymorphic DNA from the polymerase chain reaction (RAPD-PCR) allows efficient construction of saturated linkage maps. However, when analyzed by agarose gel electrophoresis, most RAPD-PCR markers segregate as dominant alleles, reducing the amount of linkage information obtained. We describe the use of single strand conformation polymorphism (SSCP) analysis of RAPD markers to generate linkage maps in a haplodiploid parasitic wasp Bracon (Habrobracon) hebetor and a diploid mosquito, Aedes aegypti. RAPD-SSCP analysis revealed segregation of codominant alleles at markers that appeared to segregate as dominant (band presence/band absence) markers or appeared invariant on agarose gels. Our SSCP protocol uses silver staining to detect DNA fractionated on large thin polyacrylamide gels and reveals more polymorphic markers than agarose gel electrophoresis. In B. hebetor, 79 markers were mapped with 12 RAPD primers in six weeks; in A. aegypti, 94 markers were mapped with 10 RAPD primers in five weeks. Forty-five percent of markers segregated as codominant loci in B. hebetor, while 11% segregated as codominant loci in A. aegypti. SSCP analysis of RAPD-PCR markers offers a rapid and inexpensive means of constructing intensive linkage maps of many species.     

BRCA1 maps proximal to D17S579 on chromosome 17q21 by genetic analysis

Previous studies have demonstrated linkage between early-onset breast cancer and ovarian cancer and genetic markers on chromosome 17q21. These markers define the location of a gene (BRCA1) which appears to be inherited as an autosomal dominant susceptibility allele. We analyzed five families with multiple affected individuals for evidence of linkage to the BRCA1 region. Two of the five families appear to be linked to BRCA1. One apparently linked family contains critical recombinants, suggesting that the gene is proximal to the marker D17S579 (Mfd188). These findings are consistent with the maximum-likelihood position estimated by the Breast Cancer Linkage Consortium and with recombination events detected in other linked families. Linkage analysis was greatly aided by PCR-based analysis of paraffin-embedded normal breast tissue from deceased family members, demonstrating the feasibility and importance of this approach. One of the two families with evidence of linkage between breast cancer and genetic markers flanking BRCA1 represents the first such family of African-American descent to be reported in detail.     

Canadian outpatients and advance directives: poor knowledge and little experience but positive attitudes.

OBJECTIVE: To examine the knowledge of, previous experience with, attitudes toward and perceived barriers to completing advance directives among outpatients at two general medicine clinics. DESIGN: Cross-sectional questionnaire administered in face-to-face structured interviews. SETTING: General internal-medicine outpatient clinics at a university teaching hospital. PATIENTS: One hundred and five adult outpatients who could communicate in spoken English and who consented to be interviewed. RESULTS: Of 167 patients approached, 58 were excluded because they could not communicate in spoken English, and 4 refused to participate. Of the remaining 105 patients, 17 (16%) knew about living wills, 12 (11%) about durable powers of attorney for health care and 4 (4%) about advance directives. Twenty-three (22%) had thought about their preferences for life-sustaining treatment, 20 (19%) had discussed them, none had written them down, and 45 (43%) had thought about choosing a proxy. Sixty-one (58%) wanted to think about their preferences for treatment, 65 (62%) wanted to discuss them, 32 (30%) wanted to write them down, and 80 (76%) wanted to choose a proxy. The perceived barriers to completing an advance directive were inability to write, the belief that an advance directive was unnecessary, a fatalistic attitude, previous discussion of preferences, a desire to leave the decision to doctors, uncertainty about preferences, a desire to discuss preferences rather than document them, a desire to wait until the situation arose, a desire to write down preferences in the future and a desire to avoid thinking about preferences or advance directives. Respondents with more knowledge of life-sustaining treatments were more likely to want to complete an advance directive. CONCLUSIONS: Outpatients have positive attitudes toward advance directives, but their knowledge and experience are limited. These data underscore the need for patient education and for policies to eliminate the barriers to completing advance directives that patients face.     

First chromosome scale genomes of ithomiine butterflies (Nymphalidae: Ithomiini): Comparative models for mimicry genetic studies

The ithomiine butterflies (Nymphalidae: Danainae) represent the largest known radiation of Müllerian mimetic butterflies. They dominate by number the mimetic butterfly communities, which include species such as the iconic neotropical Heliconius genus. Recent studies on the ecology and genetics of speciation in Ithomiini have suggested that sexual pheromones, colour pattern and perhaps hostplant could drive reproductive isolation. However, no reference genome was available for Ithomiini, which has hindered further exploration on the genetic architecture of these candidate traits, and more generally on the genomic patterns of divergence. Here, we generated high-quality, chromosome-scale genome assemblies for two Melinaea species, M. marsaeus and M. menophilus, and a draft genome of the species Ithomia salapia. We obtained genomes with a size ranging from 396 to 503 Mb across the three species and scaffold N50 of 40.5 and 23.2 Mb for the two chromosome-scale assemblies. Using collinearity analyses we identified massive rearrangements between the two closely related Melinaea species. An annotation of transposable elements and gene content was performed, as well as a specialist annotation to target chemosensory genes, which is crucial for host plant detection and mate recognition in mimetic species. A comparative genomic approach revealed independent gene expansions in ithomiines and particularly in gustatory receptor genes. These first three genomes of ithomiine mimetic butterflies constitute a valuable addition and a welcome comparison to existing biological models such as Heliconius, and will enable further understanding of the mechanisms of adaptation in butterflies.     

Cytogenetics of modern sugar canes

Modern sugar cane varieties are derived from interspecific crosses involving as many as four species. Because a chromosome increase accompanies certain crosses and backcrosses, modern varieties have very high aneuploid chromosome numbers and complicated genetics. Despite this complexity, the chromosome behavior of some modern varieties approaches that of allopolyploids. In achieving homozygosity, therefore, such varieties should respond to inbreeding almost like diploids. The meiotic chromosome behavior of F₁ hybrids and modern varieties indicates little or no genetic exchange between chromosomes ofSaccharum officinarum andS. spontaneum. Irradiation may break linkages between desirable and undesirableS. spontaneum genes not ordinarily broken by crossing-over between the chromosomes of the two species. The quick success of nobilizingS. spontaneum (recurrently back-crossing to “noble canes”) depends on a peculiar increase of the chromosomes ofS. officinarum. Experience with nobilizingS. spontaneum should not make breeders impatient when they turn to interspecific crosses unaccompanied by chromosome increases.     

Three new Ecuadorian species of Axinaea (Melastomataceae)

Three new Ecuadorian species of Axinaea (Melastomataceae) are described: Axinaea flava, A. glauca, and A.lawessonii. All three species are endemic to southern Ecuador. Axinaea flava and A.glauca grow in the transition zone between montane forest and pbramo. They share characters such as shrubby habit, dense to moderate pubescence and coriaceous, rigid, erect leaves, which may be related to their high altitude habitat. Axinaea flava is the only species of the genus with a yellow corolla. Axinaea glauca, a shrub less than 1 m high, is the smallest Axinaea species known. Field observations show that these two species have a restricted habitat and their known populations are small, probably less than 100 individuals. The third species, A.lawessonii, grows in wet montane forests. It can grow as a shrub or a slender tree and its leaves are glabrous and less rigid than those of A.flava and A.glauca, characters which are probably a consequence of the more humid and benign habitat. Axinaea lawessonii is the only species of the genus in which the leaf margins have uncinate teeth. The species is rather frequent in southern Ecuador and has been collected in a dozen localities, most of which are within the Podocarpus National Park.