Increased aberrance of cytokine expression in plasma of patients with more advanced squamous cell carcinoma of the head and neck

Patients with head and neck squamous cell carcinoma (HNSCC) are biased toward the Th2 phenotype as they have increased levels of the Th2 cytokines IL-4, IL-6 and IL-10 and diminished levels of the Th1 cytokine IFN-gamma. However, this bias is incomplete since levels of the Th1 cytokines IL-2 and GM-CSF are increased. This study examined the interrelationship among the plasma levels of Th1 and Th2 cytokines in 101 HNSCC patients and 40 age-matched controls without a known malignancy. Control subjects showed extensive interrelationships among levels of IL-1, IL-2, IL-4, and IFN-gamma, as well as between GM-CSF and TGF-beta. HNSCC patients showed an interrelationship in levels of IL-2, IL-4, IFN-gamma and GM-CSF, but to a less significant degree. What was prominent was a decline in correlation among cytokine levels with increased disease burden, such that there were no relationships among any of the cytokines in stage T4 patients. Nodal involvement also was associated with cytokine levels being more independent of each other, although the impact of nodal disease was less prominent than tumor burden. These results show a partial Th2 cytokine bias in HNSCC patients and a progressively more aberrant expression of cytokines with more advanced disease.     

Characterization of a spontaneously polarizing HT-29 cell line,HT-29/cl.f8

Summary A cloned cell line that spontaneously polarizes in standard glucose-containing media was derived from a single cell of the adenocarcinoma cell line HT-29. The cloned line, designated HT-29/cl.f8, has remained stable over 2 yr in culture, maintained high transepithelial resistance (300 ohm cm² or higher), and correctly sorted influenza virus and vesicular stomatitis virus to apical or basolateral domains, respectively. The newly cloned cells also displayed apical microvilli, tight junctions, and desmosomes, the morphological characteristics of mature epithelia. The cloned HT-29/cl.f8 cells function as epithelial enterocytes as shown by the apical expression of intestinal alkaline phosphatase, the expression of vimentin and cytokeratin, and lack of expression of mucin. We propose that the newly cloned HT-29/cl.f8 cells offer a viable alternative for studies of enterocyte function that will readily yield interpretable data not complicated by cell alterations due to the presence of drugs or chemicals that induce differentiation.     

Development and characterization of a reconstituted yeast translation initiation system

To provide a bridge between in vivo and in vitro studies of eukaryotic translation initiation, we have developed a reconstituted translation initiation system using components from the yeast Saccharomyces cerevisiae. We have purified a minimal set of initiation factors (elFs) that, together with yeast 80S ribosomes, GTP, and initiator methionyl-tRNA, are sufficient to assemble active initiation complexes on a minimal mRNA template. The kinetics of various steps in the pathway of initiation complex assembly and the formation of the first peptide bond in vitro have been explored. The formation of active initiation complexes in this system is dependent on ribosomes, mRNA, Met-tRNAi, GTP hydrolysis, elF1, elF1A, elF2, elF5, and elF5B. Our data indicate that elF1 and elF1A both facilitate the binding of the elF2 x GTP x Met-tRNAi complex to the 40S ribosomal subunit to form the 43S complex. elF5 stimulates a step after 43S complex formation, consistent with its proposed role in activating GTP hydrolysis by elF2 upon initiation codon recognition. The presence of elF5B is required for the joining of the 40S and 60S subunits to form the 80S initiation complex. The step at which each of these factors acts in this reconstituted system is in agreement with previous data from in vivo studies and work using reconstituted mammalian systems, indicating that the system recapitulates fundamental events in translation initiation in eukaryotic cells. This system should allow us to couple powerful yeast genetic and molecular biological experiments with in vitro kinetic and biophysical experiments, yielding a better understanding of the molecular mechanics of this central, complex process.     

Development in Japanese macaques (Macaca fuscata): Sexually dimorphic behavior during the first year of life

We have documented several sexually dimorphic patterns of behavior that develop during the first year of life in infant Japanese macaques and their mothers. Mothers treated their infants differently by sex—mothers of males broke contact with them and retrieved them more frequently than did mothers of females. And mothers of male infants moved more frequently than did mothers of female infants. Male infants played more, played in larger groups, and mounted more frequently; female infants groomed and spent more time close to other monkeys in larger social groups than did males. Female infants were also punished by other group members more frequently than were male infants. We conclude that male and female Japanese macaque infants receive differential treatment early in life by both their own mothers and other animals, and males and females in turn treat their mothers and other animals differently. There appears to be a reciprocal relationship between the behavior of infants, mothers and other social partners that contributes to the development of sexually dimorphic patterns of behavior.     

Fine-root growth in a forested bog is seasonally dynamic,but shallowly distributed in nutrient-poor peat

Background and aims

Fine roots contribute to ecosystem carbon, water, and nutrient fluxes through resource acquisition, respiration, exudation, and turnover, but are understudied in peatlands. We aimed to determine how the amount and timing of fine-root growth in a forested, ombrotrophic bog varied across gradients of vegetation density, peat microtopography, and changes in environmental conditions across the growing season and throughout the peat profile.

Methods

We quantified fine-root peak standing crop and growth using non-destructive minirhizotron technology over a two-year period, focusing on the dominant woody species in the bog: Picea mariana, Larix laricina, Rhododendron groenlandicum, and Chamaedaphne calyculata.

Results

The fine roots of trees and shrubs were concentrated in raised hummock microtopography, with more tree roots associated with greater tree densities and a unimodal peak in shrub roots at intermediate tree densities. Fine-root growth tended to be seasonally dynamic, but shallowly distributed, in a thin layer of nutrient-poor, aerobic peat above the growing season water table level.

Conclusions

The dynamics and distribution of fine roots in this forested ombrotrophic bog varied across space and time in response to biological, edaphic, and climatic conditions, and we expect these relationships to be sensitive to projected environmental changes in northern peatlands.

     

Managing biodiversity under climate change: challenges,frameworks, and tools for adaptation

The myriad challenges facing biodiversity under climate change are reflected in the challenges facing managers planning for these impacts. An ever-expanding number of recommendations and tools for climate change adaptation exist to aid managers in these efforts, yet navigating these various resources can lead to information overload and paralysis in decision-making. Here we provide a synthesis of the key considerations, approaches, and available tools for integrating climate change adaptation into management plans. We discuss principal elements in climate change adaptation—incorporating uncertainty through scenario planning and adaptive management—and review three leading frameworks for incorporating climate change adaptation into place-based biodiversity conservation planning. Finally, we identify the following key questions needed for long-term conservation planning and review the associated tools and techniques needed to address them: (1) How is the climate projected to change in my study area?; (2) How are non-climatic stressors projected to change?; (3) How vulnerable are species to climate change impacts?; (4) How are species ranges likely to respond?; and (5) How are management strategies expected to affect outcomes? In doing so, we aim to aid natural resource managers in navigating the burgeoning field of climate change adaptation planning and provide managers a roadmap for managing biodiversity under climate change.     

Combining field experiments and predictive models to assess potential for increased plant diversity to climate‐proof intensive agriculture

Agricultural production systems face increasing threats from more frequent and extreme weather fluctuations associated with global climate change. While there is mounting evidence that increased plant community diversity can reduce the variability of ecosystem functions (such as primary productivity) in the face of environmental fluctuation, there has been little work testing whether this is true for intensively managed agricultural systems. Using statistical modeling techniques to fit environment–productivity relationships offers an efficient means of leveraging hard‐won experimental data to compare the potential variability of different mixtures across a wide range of environmental contexts. We used data from two multiyear field experiments to fit climate–soil–productivity models for two pasture mixtures under intensive grazing—one composed of two drought‐sensitive species (standard), and an eight‐species mixture including several drought‐resistant species (complex). We then used these models to undertake a scoping study estimating the mean and coefficient of variation (CV) of annual productivity for long‐term climate data covering all New Zealand on soils with low, medium, or high water‐holding capacity. Our results suggest that the complex mixture is likely to have consistently lower CV in productivity, irrespective of soil type or climate regime. Predicted differences in mean annual productivity between mixtures were strongly influenced by soil type and were closely linked to mean annual soil water availability across all soil types. Differences in the CV of productivity were only strongly related to interannual variance in water availability for the lowest water‐holding capacity soil. Our results show that there is considerable scope for mixtures including drought‐tolerant species to enhance certainty in intensive pastoral systems. This provides justification for investing resources in a large‐scale distributed experiment involving many sites under different environmental contexts to confirm these findings.     

A major cathepsin B protease from the liver fluke Fasciola hepatica has atypical active site features and a potential role in the digestive tract of newly excysted juvenile parasites

The newly excysted juvenile (NEJ) stage of the Fasciola hepatica lifecycle occurs just prior to invasion into the wall of the gut of the host, rendering it an important target for drug development. The cathepsin B enzymes from NEJ flukes have recently been demonstrated to be crucial to invasion and migration by the parasite. Here we characterize one of the cathepsin B enzymes (recombinant FhcatB1) from NEJ flukes. FhcatB1 has biochemical properties distinct from mammalian cathepsin B enzymes, with an atypical preference for Ile over Leu or Arg residues at the P₂ substrate position and an inability to act as an exopeptidase. FhcatB1 was active across a broad pH range (optimal activity at pH 5.5–7.0) and resistant to inhibition by cystatin family inhibitors from sheep and humans, suggesting that this enzyme would be able to function in extracellular environments in its mammalian hosts. It appears, however, that the FhcatB1 protease functions largely as a digestive enzyme in the gut of the parasite, due to the localization of a specific, fluorescently labeled inhibitor with an Ile at the P₂ position. Molecular modelling and dynamics were used to predict the basis for the unusual substrate specificity: a P₂ Ile residue positions the substrate optimally for interaction with catalytic residues of the enzyme, and the enzyme lacks an occluding loop His residue crucial for exopeptidase activity. The unique features of the enzyme, particularly with regard to its specificity and likely importance to a vital stage of the parasite's life cycle, make it an excellent target for therapeutic inhibitors or vaccination.