Standard metabolic rate models for Carmine Shiner (Notropis percobromus) and Common Shiner (Luxilus cornutus) across different temperature regimes

Standard metabolic rates (SMR) were measured empirically for carmine shiner Notropis percobromus and common shiner Luxilus cornutus to develop SMR models that predict metabolic responses of each species under thermal conditions observed in the wild. SMR increased significantly with body mass and rising water temperature, ranging from 0.05 mg O₂ h⁻¹ at 10°C to 0.89 mg O₂ h⁻¹ at 20°C for N. percobromus weighing 0.6–2.5 g and from 0.11 mg O₂ h⁻¹ at 10°C to 0.98 mg O₂ h⁻¹ at 20°C for L. cornutus weighing 0.8–6.6 g. SMR models significantly differed between sympatric species on account of differences in model intercepts (RA) and temperature coefficients (RQ), however, the allometric relationships between mass and SMR did not significantly differ between species. Known distribution of N. percobromus and L. cornutus includes the Birch River located in Manitoba, Canada, where N. percobromus is listed as Endangered. Little is known about the physiology of N. percobromus or the species' ability to acclimate or adapt to different environmental conditions. While size differences between species contributed, in part, to differences in SMR predictions for Birch River populations, SMR trends (< 2 mg O₂ h⁻¹) for individuals weighing 1 g were similar for both species across daily temperatures. Respirometry experiments contributed to developing species-specific SMR models and inform on the effect of natural and anthropogenic stressors, namely water temperature, on the conservation of N. percobromus in this ecosystem.     

Impact of demography on extinction/fixation events

In this article we consider diffusion processes modeling the dynamics of multiple allelic proportions (with fixed and varying population size). We are interested in the way alleles extinctions and fixations occur. We first prove that for the Wright–Fisher diffusion process with selection, alleles get extinct successively (and not simultaneously), until the fixation of one last allele. Then we introduce a very general model with selection, competition and Mendelian reproduction, derived from the rescaling of a discrete individual-based dynamics. This multi-dimensional diffusion process describes the dynamics of the population size as well as the proportion of each type in the population. We prove first that alleles extinctions occur successively and second that depending on population size dynamics near extinction, fixation can occur either before extinction almost surely, or not. The proofs of these different results rely on stochastic time changes, integrability of one-dimensional diffusion processes paths and multi-dimensional Girsanov’s tranform.

     

Modelling T-cell immunity against hepatitis C virus with liver organoids in a microfluidic coculture system

Hepatitis C virus (HCV) remains a global public health challenge with an estimated 71 million people chronically infected, with surges in new cases and no effective vaccine. New methods are needed to study the human immune response to HCV since in vivo animal models are limited and in vitro cancer cell models often show dysregulated immune and proliferative responses. Here, we developed a CD8⁺ T cell and adult stem cell liver organoid system using a microfluidic chip to coculture 3D human liver organoids embedded in extracellular matrix with HLA-matched primary human T cells in suspension. We then employed automated phase contrast and immunofluorescence imaging to monitor T cell invasion and morphological changes in the liver organoids. This microfluidic coculture system supports targeted killing of liver organoids when pulsed with a peptide specific for HCV non-structural protein 3 (NS3) (KLVALGINAV) in the presence of patient-derived CD8⁺ T cells specific for KLVALGINAV. This demonstrates the novel potential of the coculture system to molecularly study adaptive immune responses to HCV in an in vitro setting using primary human cells.     

Superinhibition of sarcoplasmic reticulum function by phospholamban induces cardiac contractile failure

To determine whether selective impairment of cardiac sarcoplasmic reticulum (SR) Ca(2+) transport may drive the progressive functional deterioration leading to heart failure, transgenic mice, overexpressing a phospholamban Val(49) --> Gly mutant (2-fold), which is a superinhibitor of SR Ca(2+)-ATPase affinity for Ca(2+), were generated, and their cardiac phenotype was examined longitudinally. At 3 months of age, the increased EC(50) level of SR Ca(2+) uptake for Ca(2+) (0.67 +/- 0.09 microm) resulted in significantly higher depression of cardiomyocyte rates of shortening (57%), relengthening (31%), and prolongation of the Ca(2+) signal decay time (165%) than overexpression (2-fold) of wild type phospholamban (68%, 64%, and 125%, respectively), compared with controls (100%). Echocardiography also revealed significantly depressed function and impaired beta-adrenergic responses in mutant hearts. The depressed contractile parameters were associated with left ventricular remodeling, recapitulation of fetal gene expression, and hypertrophy, which progressed to dilated cardiomyopathy with interstitial tissue fibrosis and death by 6 months in males. Females also had ventricular hypertrophy at 3 months but exhibited normal systolic function up to 12 months of age. These results suggest a causal relationship between defective SR Ca(2+) cycling and cardiac remodeling leading to heart failure, with a gender-dependent influence on the time course of these alterations.     

Feeding preferences and performance of an aquatic lepidopteran on macrophytes: plant hosts as food and habitat

Although host preferences in phytophagous insects may be generated by several factors, few studies have simultaneously examined several potential host choice determinants. In this study we tested the impact of the following potential host choice determinants on host preference of the semi-aquatic lepidopteran Munroessa gyralis (Pyralidae): growth on different host plants; protein content, polyphenolic content, toughness, and chemical extracts of different host plants; prior feeding experience; and predation pressure on the caterpillar by fishes. Two water lilies, Brasenia schreberi and Nymphaea odorata, were preferred in cafeteria-style feeding experiments over 14 other species of vascular plants. The most preferred water lily (Brasenia) also afforded the fastest growth relative to three other species on which growth was measured. Feeding preferences across species were unrelated to protein content, polyphenolic content, or toughness. Domiciles constructed by caterpillars from leaf fragments were protective from field assemblages of fishes, but domiciles made from preferred or unpreferred host species conferred no significant protection from fish in the laboratory. Caterpillars responded positively to chemical cues of water lilies, and prior feeding experience increased preference for an otherwise unpreferred water lily (Nuphar advena) within the life-span of individual caterpillars. M. gyralis is a generalist herbivore exhibiting modest preference induction and preferences for and among members of the family Nymphaeaceae. Our results suggest that relative growth rates, chemical cues, and previous feeding experience are important factors determining feeding preference. Protein content, polyphenolic content, and toughness appear less important, and the importance of fish predators remains in question. As pupation seems to occur exclusively on Nymphaea, we suggest that host use may be restricted due to life-stage-specific developmental constraints that are not apparent from the results of growth or preference assays. It is currently unknown how often specific life-stages may restrict host use, but our work suggests this as a potentially important area of inquiry.     

Spring colonisation of orchards by Anthonomus pomorum from adjacent forest borders

The early-season dispersal of the overwintered apple blossom weevil, Anthonomus pomorum (L.) (Coleoptera: Curculionidae), is a crucial stage in the colonisation of dwarf apple orchards adjacent to forests. We have conducted release-recapture studies with 1700 to 4000 marked weevils at two orchard sites in Switzerland over 2 years to characterise the spatial and temporal pattern of the dispersal process. The dispersal and colonisation of orchards in spring by overwintered weevils is dependent upon the prevailing temperature.An orientated dispersal from the forest border to the centre of the orchard was observed consistently, irrespective of the angle of the apple tree rows with respect to the forest border or of climatic conditions. The average dispersal distance of the weevils was 19 m. Approximately one third of the weevil population remained on the first tree encountered, the remainder of the population moved over short distances mainly along the tree rows. This dispersal pattern led to a strong edge effect with higher numbers of weevils occurring at the edges adjoining the forests as compared to the centre of orchards. The relevance of these findings to population dynamics and management of the pest is discussed.     

Plant sensitivity to low intensity 105 GHz electromagnetic radiation

Exposing seedlings of the flax, Linum usitatissimum L., to a variety of weak environmental stresses followed by a 2 day calcium deprivation, triggers the common response of production of epidermal meristems (actively dividing groups of cells) in the hypocotyl, which is the part of the stem between the root and the cotyledons (the pre-existing leaves in the embryo). This production reaches a plateau of 10-20 meristems after a month in the case of mechanical stimulation and cold shock. Recently, we have shown that radiation from a global system for mobile communication (GSM) telephone also triggers production of meristems with a plateau of around six meristems. Here, we show that a single 2 h exposure to radiation emitted at 105 GHz at non-thermal levels by a Gunn oscillator induces meristem production with kinetics similar to that induced by weak environmental stimuli and radiation from GSM telephone.     

The Arabidopsis F-box protein SLEEPY1 targets gibberellin signaling repressors for gibberellin-induced degradation

The nuclear DELLA proteins are highly conserved repressors of hormone gibberellin (GA) signaling in plants. In Arabidopsis thaliana, GA derepresses its signaling pathway by inducing proteolysis of the DELLA protein REPRESSOR OF ga1-3 (RGA). SLEEPY1 (SLY1) encodes an F-box-containing protein, and the loss-of-function sly1 mutant has a GA-insensitive dwarf phenotype and accumulates a high level of RGA. These findings suggested that SLY1 recruits RGA to the SCFSLY1 E3 ligase complex for ubiquitination and subsequent degradation by the 26S proteasome. In this report, we provide new insight into the molecular mechanism of how SLY1 interacts with the DELLA proteins for controlling GA response. By yeast two-hybrid and in vitro pull-down assays, we demonstrated that SLY1 interacts directly with RGA and GA INSENSITIVE (GAI, a closely related DELLA protein) via their C-terminal GRAS domain. The rga and gai null mutations additively suppressed the recessive sly1 mutant phenotype, further supporting the model that SCFSLY1 targets both RGA and GAI for degradation. The N-terminal DELLA domain of RGA previously was shown to be essential for GA-induced degradation. However, we found that this DELLA domain is not required for protein-protein interaction with SLY1 in yeast (Saccharomyces cerevisiae), suggesting that its role is in a GA-triggered conformational change of the DELLA proteins. We also identified a novel gain-of-function sly1-d mutation that increased GA signaling by reducing the levels of the DELLA protein in plants. This effect of sly1-d appears to be caused by an enhanced interaction between sly1-d and the DELLA proteins.     

Independent combinatorial effect of antisense oligonucleotides and RNAi-mediated specific inhibition of the recombinant rat P2X3 receptor

Synthetic 21-bp-long short interfering RNAs (siRNA) can stimulate sequence-specific mRNA degradation in mammalian cell cultures, a process referred to as RNA interference (RNAi). In the present study, the potential of RNAi was compared to the traditional antisense approach, acting mainly via RnaseH, for targeting the recombinant rat pain-related cation-channel P2X₃ expressed in CHO-K1 and a rat brain tumour-derived cell line, 33B. Downregulation of the P2X₃ receptor was evaluated at the mRNA, protein, and functional levels. In this study, four siRNA duplexes induced up to 95% sequence-specific inhibition of the P2X₃ mRNA, independent of the type of 2 nt 3′-overhang modification and the location of the targeted sequences. Furthermore, we detected and characterised an independent combinatorial effect of antisense oligonucleotides (ASOs) and RNAi-mediated specific inhibition of the P2X₃ receptor. Enhanced downregulation was observed only when siRNA was combined with nonhomologous ASO, targeting distant regions on the common P2X₃ mRNA. The two reagents resulted in more efficient downregulation of P2X₃ mRNA when administered in combination rather than separately. To our knowledge, this is the first investigation at the molecular level of the potential benefits of mixed antisense and RNAi-mediated treatment for inhibiting expression of a medically relevant pain-related gene.