Molecular-scale topographic cues induce the orientation and directional movement of fibroblasts on two-dimensional collagen surfaces

Collagen fibres within the extracellular matrix lend tensile strength to tissues and form a functional scaffold for cells. Cells can move directionally along the axis of fibrous structures, in a process important in wound healing and cell migration. The precise nature of the structural cues within the collagen fibrils that can direct cell movement are not known. We have investigated the structural features of collagen that are required for directional motility of mouse dermal fibroblasts, by analysing cell movement on two-dimensional collagen surfaces. The surfaces were prepared with aligned fibrils of collagen type I, oriented in a predefined direction. These collagen-coated surfaces were generated with or without the characteristic 67 nm D-periodic banding. Quantitative analysis of cell morphodynamics showed a strong correlation of cell elongation and motional directionality with the orientation of D-periodic collagen microfibrils. Neither directed motility, nor cell body alignment, was observed on aligned collagen lacking D-periodicity, or on D-periodic collagen in the presence of peptide containing an RGD motif. The directional motility of fibroblast cells on aligned collagen type I fibrils cannot be attributed to contact guidance, but requires additional structural information. This allows us to postulate a physiological function for the 67 nm periodicity.     

Impairment of nuclear pores in bovine herpesvirus 1-infected MDBK cells

Herpesvirus capsids originating in the nucleus overcome the nucleocytoplasmic barrier by budding at the inner nuclear membrane. The fate of the resulting virions is still under debate. The fact that capsids approach Golgi membranes from the cytoplasmic side led to the theory of fusion between the viral envelope and the outer nuclear membrane, resulting in the release of capsids into the cytoplasm. We recently discovered a continuum from the perinuclear space to the Golgi complex implying (i) intracisternal viral transportation from the perinuclear space directly into Golgi cisternae and (ii) the existence of an alternative pathway of capsids from the nucleus to the cytoplasm. Here, we analyzed the nuclear surface by high-resolution microscopy. Confocal microscopy of MDBK cells infected with recombinant bovine herpesvirus 1 expressing green fluorescent protein fused to VP26 (a minor capsid protein) revealed distortions of the nuclear surface in the course of viral multiplication. High-resolution scanning and transmission electron microscopy proved the distortions to be related to enlargement of nuclear pores through which nuclear content including capsids protrudes into the cytoplasm, suggesting that capsids use impaired nuclear pores as gateways to gain access to the cytoplasmic matrix. Close examination of Golgi membranes, rough endoplasmic reticulum, and outer nuclear membrane yielded capsid-membrane interaction of high identity to the budding process at the inner nuclear membrane. These observations signify the ability of capsids to induce budding at any cell membrane, provided the fusion machinery is present and/or budding is not suppressed by viral proteins.     

Hindlimb unloading alters nitric oxide and autonomic control of resting arterial pressure in conscious rats

After periods of microgravity or bed rest, individuals often exhibit reduced Vo(2 max), hypovolemia, cardiac and vascular effects, and autonomic dysfunction. Recently, alterations in expression of vascular and central nervous system NO synthase (NOS) have been observed in hindlimb-unloaded (HU) rats, a model used to simulate physiological effects of microgravity or bed rest. We examined the effects of 14 days of hindlimb unloading on hemodynamic responses to systemic NOS inhibition in conscious control and HU rats. Because differences in NO and autonomic regulation might occur after hindlimb unloading, we also evaluated potential differences in resting autonomic tone and effects of NOS inhibition after autonomic blockade. Administration of nitro-L-arginine methyl ester (L-NAME; 20 mg/kg iv) increased mean arterial pressure (MAP) to similar levels in control and HU rats. However, the change in MAP in response to L-NAME was less in HU rats, that had an elevated baseline MAP. In separate experiments, atropine (1 mg/kg iv) increased heart rate (HR) in control but not HU rats. Subsequent administration of the ganglionic blocker hexamethonium (30 mg/kg iv) decreased MAP and HR to a greater extent in HU rats. Administration of L-NAME after autonomic blockade increased MAP in both groups to a greater extent compared with intact conditions. However, the pressor response to L-NAME was still reduced in HU rats. These data suggest that hindlimb unloading in rats reduces peripheral NO as well as cardiac parasympathetic tone. Along with elevations in sympathetic tone, these effects likely contribute to alterations in vascular control and changes in autonomic reflex function following spaceflight or bed rest.     

Protein biomarkers for subtyping breast cancer and implications for future research

Introduction: Breast cancer subtypes are currently defined by a combination of morphologic, genomic, and proteomic characteristics. These subtypes provide a molecular portrait of the tumor that aids diagnosis, prognosis, and treatment escalation/de-escalation options. Gene expression signatures describing intrinsic breast cancer subtypes for predicting risk of recurrence have been rapidly adopted in the clinic. Despite the use of subtype classifications, many patients develop drug resistance, breast cancer recurrence, or therapy failure.

Areas covered: This review provides a summary of immunohistochemistry, reverse phase protein array, mass spectrometry, and integrative studies that are revealing differences in biological functions within and between breast cancer subtypes. We conclude with a discussion of rigor and reproducibility for proteomic-based biomarker discovery.

Expert commentary: Innovations in proteomics, including implementation of assay guidelines and standards, are facilitating refinement of breast cancer subtypes. Proteomic and phosphoproteomic information distinguish biologically functional subtypes, are predictive of recurrence, and indicate likelihood of drug resistance. Actionable, activated signal transduction pathways can now be quantified and characterized. Proteomic biomarker validation in large, well-designed studies should become a public health priority to capitalize on the wealth of information gleaned from the proteome.     

Bone in the breast? Long term toxicity 21 years after interstitial brachytherapy as a boost

An 81-year-old patient developed an exulcerous tumor in her left breast 21 years after breast cancer treatment with lumpectomy and adjuvant radiotherapy. At the time of the initial treatment 21 years ago, whole breast irradiation was performed with a prescribed dose of 48?Gy and a maximal dose of 69?Gy. In addition, the patient received a 14.7?Gy boost with multicatheter brachytherapy as partial breast irradiation.In general, fat necrosis after radiotherapy, surgery or trauma is a minor problem for patients, but can lead to diagnostic difficulties. The incidence varies: the literature indicates that it occurs in up to 34% of cases. The direct pathogenesis is not clear; it can be due to high radiation dose to the breast, dosimetric inhomogeneities or surgical complications (seromas and inflammation).The tumor in the case described here, occurring more than two decades after the primary treatment, is a rarity in this extent and is an unusual clinical, radiological, and histological finding. It provides a good example of the need for an individualized approach to treatment.     

Elevated CO2 and water addition enhance nitrogen turnover in grassland plants with implications for temporal stability

Temporal variation in soil nitrogen (N) availability affects growth of grassland communities that differ in their use and reuse of N. In a 7‐year‐long climate change experiment in a semi‐arid grassland, the temporal stability of plant biomass production varied with plant N turnover (reliance on externally acquired N relative to internally recycled N). Species with high N turnover were less stable in time compared to species with low N turnover. In contrast, N turnover at the community level was positively associated with asynchrony in biomass production, which in turn increased community temporal stability. Elevated CO₂ and summer irrigation, but not warming, enhanced community N turnover and stability, possibly because treatments promoted greater abundance of species with high N turnover. Our study highlights the importance of plant N turnover for determining the temporal stability of individual species and plant communities affected by climate change.     

Land management change greatly impacts biofuels’ greenhouse gas emissions

Harvesting corn stover for biofuel production may decrease soil organic carbon (SOC) and increase greenhouse gas (GHG) emissions. Adding additional organic matter into soil or reducing tillage intensity, however, could potentially offset this SOC loss. Here, using SOC and life cycle analysis (LCA) models, we evaluated the impacts of land management change (LMC), that is, stover removal, organic matter addition, and tillage on spatially explicit SOC level and biofuels’ overall life cycle GHG emissions in US corn–soybean production systems. Results indicate that under conventional tillage (CT), 30% stover removal (dry weight) may reduce baseline SOC by 0.04 t C ha^?1 yr^?1 over a 30‐year simulation period. Growing a cover crop during the fallow season or applying manure, on the other hand, could add to SOC and further reduce biofuels’ life cycle GHG emissions. With 30% stover removal in a CT system, cover crop and manure application can increase SOC at the national level by about 0.06 and 0.02 t C ha^?1 yr^?1, respectively, compared to baseline cases without such measures. With contributions from this SOC increase, the life cycle GHG emissions for stover ethanol are more than 80% lower than those of gasoline, exceeding the US Renewable Fuel Standard mandate of 60% emissions reduction in cellulosic biofuels. Reducing tillage intensity while removing stover could also limit SOC loss or lead to SOC gain, which would lower stover ethanol life cycle GHG emissions to near or under the mandated 60% reduction. Without these organic matter inputs or reduced tillage intensity, however, the emissions will not meet this mandate. More efforts are still required to further identify key practical LMCs, improve SOC modeling, and accounting for LMCs in biofuel LCAs that incorporate stover removal.     

Measuring progress in status of land under forest landscape restoration using abiotic and biotic indicators

The paper suggests a minimum set of abiotic and biotic threshold indicators and progress indicators for forest landscape restoration (FLR), then also briefly discusses progress indicators of pressures and project outputs. FLR aims to restore multiple functions of forests at a landscape scale. It is predicated on the hypothesis that restoration produces enabling conditions for ecosystem services, including regulating services such as carbon sequestration and pollination, and provisioning services such as food and energy. As FLR gains greater uptake, it is increasingly important to monitor progress. The types of indicators required are influenced by the degree of forest loss and degradation. To measure the status of land under restoration, one or more abiotic and biotic threshold indicators are required, measuring the return of enabling conditions for restoration (soil quality, water, etc.), along with progress indicators measuring the reemergence of the ecosystem services. Although all elements of the proposed monitoring framework are well known, compiling them into a coherent system, suitable for application in a wide range of conditions, will take much further development.     

Rhamnosidase activity of selected probiotics and their ability to hydrolyse flavonoid rhamnoglucosides

Bioprocess and Biosystems Engineering - Bioavailability of flavonoids is low, especially when occurring as rhamnoglucosides. Thus, the hydrolysis of rutin, hesperidin, naringin and a mixture of...