Nematode‐specific PCR primers for the 18S small subunit rRNA gene

A set of polymerase chain reaction primers were designed, which amplify a c. 1 kb fragment of the 18S ribosomal DNA gene, and are specific to the phylum Nematoda. These have proven useful in isolating nematode genes from samples mixed with other biological material, particularly with application to DNA barcoding. Optimal reaction conditions are described. These primers have successfully amplified the correct fragment from a wide phylogenetic range of nematodes, and have so far generated no sequences from any other organismal group.     

Spartina anglica C. E. Hubbard: a natural model system for analysing early evolutionary changes that affect allopolyploid genomes

Spartina anglica arose during the end of the 19th century in England by hybridization between the indigenous Spartina maritima and the introduced East American Spartina alterniflora and following genome duplication of the hybrid ( S.  ×  townsendii ). This system allows investigations of the early evolutionary changes that accompany stabilization of a new allopolyploid species in natural populations. Various molecular data indicate that S. anglica has resulted from a unique parental genotype. This young species contains two distinctly divergent homoeologous genomes that have not undergone extensive change since their reunion. No burst of retroelements has been encountered in the F₁ hybrid or in the allopolyploid, suggesting a 'structural genomic stasis' rather than 'rapid genomic changes'. However, modifications of the methylation patterns in the genomes of S.  ×  townsendii and S. anglica indicate that in this system, epigenetic changes have followed both hybridization and polyploidization.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 82 , 475–484.     

Integrating evidence-based treatments for common mental disorders in routine primary care: feasibility and acceptability of the MANAS intervention in Goa,India

Common mental disorders, such as depression and anxiety, pose a major public health burden in developing countries. Although these disorders are thought to be best managed in primary care settings, there is a dearth of evidence about how this can be achieved in low resource settings. The MANAS project is an attempt to integrate an evidence based package of treatments into routine public and private primary care settings in Goa, India. Before initiating the trial, we carried out extensive preparatory work, over a period of 15 months, to examine the feasibility and acceptability of the planned intervention. This paper describes the systematic development and evaluation of the intervention through this preparatory phase. The preparatory stage, which was implemented in three phases, utilized quantitative and qualitative methods to inform our understanding of the potential problems and possible solutions in implementing the trial and led to critical modifications of the original intervention plan. Investing in systematic formative work prior to conducting expensive trials of the effectiveness of complex interventions is a useful exercise which potentially improves the likelihood of a positive result of such trials.     

FOUNDING A NEW SCIENCE: MIND GENOMICS

We present in this article our vision for a new science, modeled on the emerging science of genomics and the technology of informatics. Our goal in this new science is to better understand how people react to ideas in a formal and structured way, using the principles of stimulus–response (from experimental psychology), conjoint analysis (from consumer research and statistics), Internet‐based testing (from marketing research) and multiple tests to identify patterns of mind‐sets (patterned after genomics). We show how this formal approach can then be used to construct new, innovative ideas in business. We demonstrate the approach using the development of new ideas for an electronic color palette for cosmetic products to be used by consumers.     

Water Content and the Responsivity of Lettuce Seeds to Light

Lettuce seed (Lactuca sativa L. cv. Grand Rapids) prevented from attaining maximum water content are still able to respond to irradiation if the seed moisture content is above 6%. When the limitation to water uptake is by immersion in a solution of either polyethylene glycol 400 (PEG 400), mannitol, or sucrose, the responsiveness of the seed is lessened. With PEG 400 the molarity which reduces germination by about 50% is 0.03 while mannitol and sucrose must be present at 0.07M and 0.05 M respectively to bring about the same reduction. While red light overcomes any additional effect 0.1 M mannitol and 0.025 M PEG 400 have on lettuce seed at 30°C (at which temperature the sample was positively photoblastic), irradiation did not overcome the effect of 0.1 M sucrose. Gibberellic acid did not have any effect on any of the treated seed. The solutes used cannot be considered as inert. Some chemical activity is associated with them and it is unwise to consider using these molecules to establish a particular level of water potential in the liquid bathing the seed.     

Local selection modifies phenotypic divergence among Rana temporaria populations in the presence of gene flow

In ectotherms, variation in life history traits among populations is common and suggests local adaptation. However, geographic variation itself is not a proof for local adaptation, as genetic drift and gene flow may also shape patterns of quantitative variation. We studied local and regional variation in means and phenotypic plasticity of larval life history traits in the common frog Rana temporaria using six populations from central Sweden, breeding in either open‐canopy or partially closed‐canopy ponds. To separate local adaptation from genetic drift, we compared differentiation in quantitative genetic traits (Q_ST) obtained from a common garden experiment with differentiation in presumably neutral microsatellite markers (F_ST). We found that R. temporaria populations differ in means and plasticities of life history traits in different temperatures at local, and in F_ST at regional scale. Comparisons of differentiation in quantitative traits and in molecular markers suggested that natural selection was responsible for the divergence in growth and development rates as well as in temperature‐induced plasticity, indicating local adaptation. However, at low temperature, the role of genetic drift could not be separated from selection. Phenotypes were correlated with forest canopy closure, but not with geographical or genetic distance. These results indicate that local adaptation can evolve in the presence of ongoing gene flow among the populations, and that natural selection is strong in this system.     

NO signalling in cytokinin-induced programmed cell death

Cell death can be induced by cytokinin 6-benzylaminopurine (BA) at high dosage in suspension-cultured Arabidopsis cells. Herein, we provide evidence that BA induces nitric oxide (NO) synthesis in a dose-dependent manner. A reduction in cell death can be observed when the cytokinin is supplemented with the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) or the nitric oxide synthase (NOS) inhibitors: 2-aminoethyl-isothiourea (AET) and N^G.-monomethyl- l -arginine ( l -NMMA), which suggests that NO is produced via a NOS and is a signalling component of this form of programmed cell death. In BA-treated cells, mitochondrial functionality is altered via inhibition of respiration. This inhibition can be prevented by addition of either cPTIO or AET implying that NO acts at the mitochondrial level.     

Dimeric DOTA-α-Melanocyte-Stimulating Hormone Analogs: Synthesis and In Vivo Characteristics of Radiopeptides with High In Vitro Activity

Dimeric analogs of α-melanocyte-stimulating hormone (α-MSH) labeled with radiometals are potential candidates for diagnosis and therapy of melanoma by receptor-mediated tumor targeting. Both melanotic and amelanotic melanomas (over-)express the melanocortin-1 receptor (MC1-R), the target for α-MSH. In the past, dimerized MSH analogs have been shown to display increased receptor affinity compared to monomeric MSH, offering the possibility of improving the ratio between specific uptake of radiolabeled α-MSH by melanoma and nonspecific uptake by the kidneys. We have designed three linear dimeric analogs containing a slightly modified MSH hexapeptide core sequence (Nle-Asp-His-d-Phe-Arg-Trp) in parallel or antiparallel orientation, a short spacer, and the DOTA chelator for incorporation of the radiometal. In vitro, all three peptides were more potent ligands of the mouse B16-F1 melanoma cell melanocortin-1 receptor (MC1-R) than DOTA-NAPamide, which served as standard. The binding activity of DOTA-diHexa(NC-NC)-amide was 1.75-fold higher, that of diHexa(NC-NC)-Gly-Lys(DOTA)-amide was 3.37-fold higher, and that of DOTA-diHexa(CN-NC)-amide was 2.34-fold higher. Using human HBL melanoma cells, the binding activity of diHexa(NC-NC)-Gly-Lys(DOTA)-amide was sixfold higher than that of DOTA-NAPamide. Uptake by cultured B16-F1 cells was rapid and almost quantitative. In vivo, however, the data were less promising: tumor-to-kidney ratios 4 hr postinjection were 0.11 for [¹¹¹In]DOTA-diHexa(NC-NC)-amide, 0.26 for diHexa(NC-NC)-Gly-Lys([¹¹¹In]DOTA)-amide, and 0.36 for [¹¹¹In]DOTA-diHexa(CN-NC)-amide, compared to 1.67 for [¹¹¹In]DOTA-NAPamide. It appears that despite the higher affinity to the MC1-R of the peptide dimers and their excellent internalization in vitro, the uptake by melanoma tumors in vivo was lower, possibly because of reduced tissue penetration. More striking, however, was the marked increase of kidney uptake of the dimers, explaining the unfavorable ratios. In conclusion, although radiolabeled α-MSH dimer peptides display excellent receptor affinity and internalization, they are no alternative to the monomeric DOTA-NAPamide for in vivo application.     

Abiotic effects predominate under prolonged livestock‐induced disturbance

Despite the widespread recognition that disturbance by livestock affects multiple indices of landscape health, few studies have examined their effects on both biotic and abiotic processes. We examined the effects of livestock disturbance on soil, vascular plants and reptiles across a disturbance gradient in a semi‐arid Australian woodland. Our gradient ranged from long‐ungrazed water remote sites, through intermediately grazed recovering sites, to currently grazed sites close to water. Our aim was to examine the nature of the effects of grazing‐induced disturbance on biotic and abiotic processes along the gradient. We detected small biotic effects, but no abiotic effects, at low levels of disturbance (intermediate sites). We could not detect a consistent biotic effect on plants or reptiles along the gradient, except between the extreme disturbances. In contrast, we recorded substantial reductions in abiotic structure and function at the most disturbed sites. Structural changes included reductions in the cover of shrub hummocks and increases in bare soil, and reductions in cryptogamic soil crusts. Structural changes were associated with declines in function (soil stability and nutrient indices). Our data are consistent with the notion that abiotic effects predominate at high levels of disturbance in rangelands. Given the extent of abiotic modification at currently grazed sites, the cover of abiotic elements such as hummocks and soil surfaces would seem a better indicator of the long‐term effect of grazing‐induced disturbance than biotic components. The extent of disturbance at currently grazed sites across large areas of rangeland suggests that autogenic recovery will be protracted.