Stereocilia Membrane Deformation: Implications for the Gating Spring and Mechanotransduction Channel

AbstractIn hair cells, although mechanotransduction channels have been localized to tips of shorter stereocilia of the mechanically sensitive hair bundle, little is known about how force is transmitted to the channel. Here, we use a biophysical model of the membrane-channel complex to analyze the nature of the gating spring compliance and channel arrangement. We use a triangulated surface model and Monte Carlo simulation to compute the deformation of the membrane under the action of tip link force. We show that depending on the gating spring stiffness, the compliant component of the gating spring arises from either the membrane alone or a combination of the membrane and a tether that connects the channel to the actin cytoskeleton. If a bundle is characterized by relatively soft gating springs, such as those of the bullfrog sacculus, the need for membrane reinforcement by channel tethering then depends on membrane parameters. With stiffer gating springs, such as those from rat outer hair cells, the channel must be tethered for all biophysically realistic parameters of the membrane. We compute the membrane forces (resultants), which depend on membrane tension, bending modulus, and curvature, and show that they can determine the fate of the channel

Development of Standard Methods to Estimate Manure Production and Nutrient Characteristics From Dairy Cattle

Total collection data from many universities were pooled for statistical analyses to evaluate existing data from dairy animals and determine if one or more modifications were needed for Standard Table D384.1. Many of these studies were carried out to evaluate nutritional characteristics associated with different diets. The data collected were sufficient to evaluate total manure, total and volatile solids, and N excretion values. Some experiments had sufficient number of samples analyzed to evaluate P, K, Ca, Mg, Na, Cl, S, and micro-elements. Statistical analyses were conducted to determine if a single column value was appropriate and define the regression equations necessary to estimate excretions if the assumptions of the column inputs were not met. The results indicate that separate classifications are needed for replacement heifers and mature animals. Final classifications for replacement heifers included: milk fed calves, weaned calves weighing less than 274 kg, heifers weighing between 273 and 613 kg, and veal calves. Additionally, classifications for lactating and dry animals are needed. Previously, the estimates for manure production and nutrient excretion were based on body weight. These findings indicate that a better predictor for lactating cattle is daily milk production instead of body weight. Milk production drives feed intake in the lactating animal. It is most appropriate that estimated manure and nutrient excretion values reflect the relationship between feed intake, milk production (nutrient utilization) and nutrient excretion

Livestock grazing and vegetative filter strip buffer effects on runoff sediment, nitrate, and phosphorus losses

Livestock grazing in the Midwestern United States can result in significant levels of runoff sediment and nutrient losses to surface water resources. Some of these contaminants can increase stream eutrophication and are suspected of contributing to hypoxic conditions in the Gulf of Mexico. This research quantified effects of livestock grazing management practices and vegetative filter strip buffers on runoff depth and mass losses of total solids, nitrate-nitrogen (NO3-N), and ortho-phosphorus (PO4-P) under natural hydrologic conditions. Runoff data were collected from 12 rainfall events during 2001 to 2003 at an Iowa State University research farm in central Iowa, United States. Three vegetative buffers (paddock area:vegetative buffer area ratios of 1:0.2, 1:0.1, and 1:0 no buffer [control]) and three grazing management practices (continuous, rotational, and no grazing [control]) comprised nine treatment combinations (vegetative buffer ratio/grazing management practice) replicated in three 1.35 ha (3.34 ac) plot areas. The total 4.05 ha (10.02 ac) study area also included nine 0.4 ha (1.0 ac) paddocks and 27 vegetative buffer runoff collection units distributed in a randomized complete block design. The study site was established on uneven terrain with a maximum of 15% slopes and consisted of approximately 100% cool-season smooth bromegrass. Average paddock and vegetative buffer plant tiller densities estimated during the 2003 project season were approximately 62 million and 93 million tillers ha−1 (153 million and 230 million tillers ac−1), respectively. Runoff sample collection pipe leakage discovered and corrected during 2001 possibly reduced runoff depth and affected runoff contaminant mass losses data values. Consequently, 2001 runoff analysis results were limited to treatment comparisons within the 2001 season and were not compared with 2002 and 2003 data. Analysis results from 2001 showed no significant differences in average losses of runoff, total solids, NO3-N, and PO4-P among the nine vegetative buffer/grazing practice treatment combinations. Results from 2002 indicated significantly higher losses of runoff and total solids from 1:0 no buffer/rotational grazing and 1:0 no buffer/continuous grazing treatment combination plots, respectively, compared among other 2002 season treatment combinations. The 2003 results showed significantly higher runoff and total solids losses from 1:0 no buffer/no grazing treatment combination plots compared among all 2003 treatment combinations and from 1:0.1 vegetative buffer/no grazing treatment combination plots compared among all 2003 treatment combinations and with respective 2002 treatment combinations. However, the 2003 results indicated effective vegetative buffer performance with significantly lower runoff, total solids, and NO3-N losses from the larger 1:0.2 buffer area compared among the smaller 1:0.1 buffer area and 1:0 no buffer treatment combinations. The 2003 results also indicated a highly significant increase in losses of NO3-N from 1:0.1 buffer/no grazing treatment combination plots compared among other 2003 season treatment combinations and with respective 2002 treatment combinations. Overall results from this study suggest a shift from significantly higher 2002 season plot losses of continuous and rotational grazing treatment combinations to significantly higher 2003 season losses of no grazing treatment combinations. We speculate this shift to significantly higher runoff and contaminant losses from no grazing treatment combination plots during 2003 reflects the variability inherent to a complex and dynamic soil-water environment of livestock grazing areas. However, we also hypothesize the environmental conditions that largely consisted of a dense perennial cool-season grass type, high-relief landscape, and relatively high total rainfall depth may not necessarily include livestock grazing activities

Mouse genome-wide association studies and systems genetics uncover the genetic architecture associated with hepatic pharmacokinetic and pharmacodynamic properties of a constrained ethyl antisense oligonucleotide targeting Malat1

Antisense oligonucleotides (ASOs) have demonstrated variation of efficacy in patient populations. This has prompted our investigation into the contribution of genetic architecture to ASO pharmacokinetics (PK) and pharmacodynamics (PD). Genome wide association (GWA) and transcriptomic analysis in a hybrid mouse diversity panel (HMDP) were used to identify and validate novel genes involved in the uptake and efficacy of a single dose of a Malat1 constrained ethyl (cEt) modified ASO. The GWA of the HMDP identified two significant associations on chromosomes 4 and 10 with hepatic Malat1 ASO concentrations. Stabilin 2 (Stab2) and vesicle associated membrane protein 3 (Vamp3) were identified by ciseQTL analysis. HMDP strains with lower Stab2 expression and Stab2 KO mice displayed significantly lower PK than strains with higher Stab2 expression and the wild type (WT) animals respectively, confirming the role of Stab2 in regulating hepatic Malat1 ASO uptake. GWA examining ASO efficacy uncovered three loci associated with Malat1 potency: Small Subunit Processome Component (Utp11l) on chromosome 4, Rho associated coiled-coil containing protein kinase 2 (Rock2) and Aci-reductone dioxygenase (Adi1) on chromosome 12. Our results demonstrate the utility of mouse GWAS using the HMDP in detecting genes capable of impacting the uptake of ASOs, and identifies genes critical for the activity of ASOs in vivo

Hsp72 is targeted to the mitotic spindle by Nek6 to promote K-fiber assembly and mitotic progression

Hsp70 proteins represent a family of chaperones that regulate cellular homeostasis and are required for cancer cell survival. However, their function and regulation in mitosis remain unknown. In this paper, we show that the major inducible cytoplasmic Hsp70 isoform, Hsp72, is required for assembly of a robust bipolar spindle capable of efficient chromosome congression. Mechanistically, Hsp72 associates with the K-fiber-stabilizing proteins, ch-TOG and TACC3, and promotes their interaction with each other and recruitment to spindle microtubules (MTs). Targeting of Hsp72 to the mitotic spindle is dependent on phosphorylation at Thr-66 within its nucleotide-binding domain by the Nek6 kinase. Phosphorylated Hsp72 concentrates on spindle poles and sites of MT-kinetochore attachment. A phosphomimetic Hsp72 mutant rescued defects in K-fiber assembly, ch-TOG/TACC3 recruitment and mitotic progression that also resulted from Nek6 depletion. We therefore propose that Nek6 facilitates association of Hsp72 with the mitotic spindle, where it promotes stable K-fiber assembly through recruitment of the ch-TOG-TACC3 complex

Noninvasive Vascular Images for Face Transplant Surgical Planning

Objective: Face transplantation replaces substantial defects with anatomically identical donor tissues; preoperative vascular assessment relies on noninvasive imaging to separate and characterize the external carotid vessels and branches. The objective is to describe and illustrate vascular considerations for face transplantation candidates. Methods: Novel noninvasive imaging using computed tomography and magnetic resonance imaging over 3 spatial dimensions plus time was developed and tested in 4 face transplant candidates. Precontrast images assessed bones and underlying metal. Contrast media was used to delineate and separate arteries from veins. For computed tomography, acquisition over multiple time points enabled the computation of tissue perfusion metrics. Time-resolved magnetic resonance angiography was performed to separate arterial and venous phases. Results: The range of circulation times for the external carotid system was 6 to 14 seconds from arterial blush to loss of venous enhancement. Precontrast imaging provided a roadmap of bones and metal. Among the 4 patients, 3 had surgical clips, metal implants, or both within 1 cm of major vessels considered for surgery. Contrast-enhanced wide area detector computed tomographic data acquired in the axial mode separated these structures and provided arterial and venous images for planning the surgical anastomoses. Magnetic resonance imaging was able to distinguish between the large vessels from the external carotid systems. Conclusions: Vascular imaging maps are challenging in face transplantation because of the rapid circulation times and artifact from the initial injury, prior reconstructive attempts, or both. Nevertheless, face transplant candidates require high spatial and temporal resolution vascular imaging to determine those vessels appropriate for surgical anastomoses

Cross-Reactive Neuraminidase Antibodies Afford Partial Protection against H5N1 in Mice and Are Present in Unexposed Humans

BACKGROUND: A pandemic H5N1 influenza outbreak would be facilitated by an absence of immunity to the avian-derived virus in the human population. Although this condition is likely in regard to hemagglutinin-mediated immunity, the neuraminidase (NA) of H5N1 viruses (avN1) and of endemic human H1N1 viruses (huN1) are classified in the same serotype. We hypothesized that an immune response to huN1 could mediate cross-protection against H5N1 influenza virus infection. METHODS AND FINDINGS: Mice were immunized against the NA of a contemporary human H1N1 strain by DNA vaccination. They were challenged with recombinant A/Puerto Rico/8/34 (PR8) viruses bearing huN1 (PR8-huN1) or avN1 (PR8-avN1) or with H5N1 virus A/Vietnam/1203/04. Additional naïve mice were injected with sera from vaccinated mice prior to H5N1 challenge. Also, serum specimens from humans were analyzed for reactivity with avN1. Immunization elicited a serum IgG response to huN1 and robust protection against the homologous challenge virus. Immunized mice were partially protected from lethal challenge with H5N1 virus or recombinant PR8-avN1. Sera transferred from immunized mice to naïve animals conferred similar protection against H5N1 mortality. Analysis of human sera showed that antibodies able to inhibit the sialidase activity of avN1 exist in some individuals. CONCLUSIONS: These data reveal that humoral immunity elicited by huN1 can partially protect against H5N1 infection in a mammalian host. Our results suggest that a portion of the human population could have some degree of resistance to H5N1 influenza, with the possibility that this could be induced or enhanced through immunization with seasonal influenza vaccines

Evolutionary connectionism: algorithmic principles underlying the evolution of biological organisation in evo-devo, evo-eco and evolutionary transitions

The mechanisms of variation, selection and inheritance, on which evolution by natural selection depends, are not fixed over evolutionary time. Current evolutionary biology is increasingly focussed on understanding how the evolution of developmental organisations modifies the distribution of phenotypic variation, the evolution of ecological relationships modifies the selective environment, and the evolution of reproductive relationships modifies the heritability of the evolutionary unit. The major transitions in evolution, in particular, involve radical changes in developmental, ecological and reproductive organisations that instantiate variation, selection and inheritance at a higher level of biological organisation. However, current evolutionary theory is poorly equipped to describe how these organisations change over evolutionary time and especially how that results in adaptive complexes at successive scales of organisation (the key problem is that evolution is self-referential, i.e. the products of evolution change the parameters of the evolutionary process). Here we first reinterpret the central open questions in these domains from a perspective that emphasises the common underlying themes. We then synthesise the findings from a developing body of work that is building a new theoretical approach to these questions by converting well-understood theory and results from models of cognitive learning. Specifically, connectionist models of memory and learning demonstrate how simple incremental mechanisms, adjusting the relationships between individually-simple components, can produce organisations that exhibit complex system-level behaviours and improve the adaptive capabilities of the system. We use the term “evolutionary connectionism” to recognise that, by functionally equivalent processes, natural selection acting on the relationships within and between evolutionary entities can result in organisations that produce complex system-level behaviours in evolutionary systems and modify the adaptive capabilities of natural selection over time. We review the evidence supporting the functional equivalences between the domains of learning and of evolution, and discuss the potential for this to resolve conceptual problems in our understanding of the evolution of developmental, ecological and reproductive organisations and, in particular, the major evolutionary transitions