The Effect of Pulsed Magnetic Fields on the Production of Reactive Oxygen Species by Neutrophils

Biophysics - It has been shown that pulsed magnetic fields (the first mode: pulse duration of 2 ms, frequency repetitions of 6.25 Hz, the pulse was bell-shaped; and the second mode: pulse duration...     

COPII collar defines the boundary between ER and ER exit site and does not coat cargo containers

COPII and COPI mediate the formation of membrane vesicles translocating in opposite directions within the secretory pathway. Live-cell and electron microscopy revealed a novel mode of function for COPII during cargo export from the ER. COPII is recruited to membranes defining the boundary between the ER and ER exit sites, facilitating selective cargo concentration. Using direct observation of living cells, we monitored cargo selection processes, accumulation, and fission of COPII-free ERES membranes. CRISPR/Cas12a tagging, the RUSH system, and pharmaceutical and genetic perturbations of ER-Golgi transport demonstrated that the COPII coat remains bound to the ER–ERES boundary during protein export. Manipulation of the cargo-binding domain in COPII Sec24B prohibits cargo accumulation in ERES. These findings suggest a role for COPII in selecting and concentrating exported cargo rather than coating Golgi-bound carriers. These findings transform our understanding of coat proteins’ role in ER-to-Golgi transport.     

The Crown Pearl: a draft genome assembly of the European freshwater pearl mussel Margaritifera margaritifera (Linnaeus, 1758)

Since historical times, the inherent human fascination with pearls turned the freshwater pearl mussel Margaritifera margaritifera (Linnaeus, 1758) into a highly valuable cultural and economic resource. Although pearl harvesting in M. margaritifera is nowadays residual, other human threats have aggravated the species conservation status, especially in Europe. This mussel presents a myriad of rare biological features, e.g. high longevity coupled with low senescence and Doubly Uniparental Inheritance of mitochondrial DNA, for which the underlying molecular mechanisms are poorly known. Here, the first draft genome assembly of M. margaritifera was produced using a combination of Illumina Paired-end and Mate-pair approaches. The genome assembly was 2.4 Gb long, possessing 105,185 scaffolds and a scaffold N50 length of 288,726 bp. The ab initio gene prediction allowed the identification of 35,119 protein-coding genes. This genome represents an essential resource for studying this species’ unique biological and evolutionary features and ultimately will help to develop new tools to promote its conservation.     

Disease modifying treatment of spinal cord injury with directly reprogrammed neural precursor cells in non-human primates

BACKGROUNDThe development of regenerative therapy for human spinal cord injury (SCI) is dramatically restricted by two main challenges: the need for a safe source of functionally active and reproducible neural stem cells and the need of adequate animal models for preclinical testing. Direct reprogramming of somatic cells into neuronal and glial precursors might be a promising solution to the first challenge. The use of non-human primates for preclinical studies exploring new treatment paradigms in SCI results in data with more translational relevance to human SCI.AIMTo investigate the safety and efficacy of intraspinal transplantation of directly reprogrammed neural precursor cells (drNPCs).METHODSSeven non-human primates with verified complete thoracic SCI were divided into two groups: drNPC group (n = 4) was subjected to intraspinal transplantation of 5 million drNPCs rostral and caudal to the lesion site 2 wk post injury, and lesion control (n = 3) was injected identically with the equivalent volume of vehicle.RESULTSFollow-up for 12 wk revealed that animals in the drNPC group demonstrated a significant recovery of the paralyzed hindlimb as well as recovery of somatosensory evoked potential and motor evoked potential of injured pathways. Magnetic resonance diffusion tensor imaging data confirmed the intraspinal transplantation of drNPCs did not adversely affect the morphology of the central nervous system or cerebrospinal fluid circulation. Subsequent immunohistochemical analysis showed that drNPCs maintained SOX2 expression characteristic of multipotency in the transplanted spinal cord for at least 12 wk, migrating to areas of axon growth cones.CONCLUSIONOur data demonstrated that drNPC transplantation was safe and contributed to improvement of spinal cord function after acute SCI, based on neurological status assessment and neurophysiological recovery within 12 wk after transplantation. The functional improvement described was not associated with neuronal differentiation of the allogeneic drNPCs. Instead, directed drNPCs migration to the areas of active growth cone formation may provide exosome and paracrine trophic support, thereby further supporting the regeneration processes.     

Buried Tracks: Ichnological Applications of High-Frequency Georadar

Recognition and sampling of traces in unconsolidated sands present a major challenge for ichnologists. This can be partially remedied through the application of high-resolution geophysical techniques, such as ground-penetrating radar (GPR or georadar), which uses electromagnetic impulse for continuous imaging of shallow subsurface. It addition to geological applications, GPR imaging has been used in several studies focused on animal traces as related to conservation of endangered fossorial species (Kinlaw et al., 2007 Kinlaw, A. E., Conyers, L. B. and Zajac, W. 2007. Use of ground penetrating radar to image burrows of the gopher tortoise (Gopherus polyphemus). Herpetological Review, 38: 50–56.  [Google Scholar]; Martin et al., 2011 Martin, A. J., Skaggs, S. A., Vance, R. K. and Greco, V. 2011. Ground-penetrating radar investigation of gopher-tortoise burrows: Refining the characterization of modern vertebrate burrows and associated commensal traces. Geological Society of America Abstracts with Programs, 43: 381 [Google Scholar]), slope and levee stability (Nichol et al., 2003 Nichol, D., Lenham, J. W. and Reynolds, J. M. 2003. Application of ground-penetrating radar to investigate the effects of badger setts on slope stability at St. Asaph Bypass, North Wales. Quarterly Journal of Engineering Geology and Hydrogeology, 36: 143–153.  [Google Scholar]; Di Prinzio et al., 2010 Di Prinzio, M, Bittelli, M., Castellarin, A. and Pisa, P. R. 2010. Application of GPR to the monitoring of river embankments. Journal of Applied Geophysics, 7: 53–61.  [Google Scholar]), and mapping of fossil tracks (Matthews et al., 2006 Matthews, N. A., Noble, T. A. and Breithaupt, B. H. 2006. “The application of photogrammetry, remote sensing and geographic information systems (GIS) to fossil resource management”. In Fossils from Federal Lands, Edited by: Lucas, S. G., Spielmann, J. A., Hester, P. M., Kenworthy, J. P. and Santucci, V. L. Vol. 34, 119–131. New Mexico Museum of Natural History and Science Bulletin.  [Google Scholar]; Aucoin and Hasbargen, 2010 Aucoin, C. D. and Hasbargen, L. 2010. Using GPR, GPS and close-range photography to map and characterize dinosaur tracks in the Connecticut River valley. Geological Society of America Abstracts with Programs, 42: 276 [Google Scholar]) and tracking surfaces (Webb, 2007 Webb, S. 2007. Further research of the Willandra Lakes fossil footprint site, southeastern Australia. Journal of Human Evolution, 52: 711–715. [Crossref], [PubMed], [Web of Science ®] , [Google Scholar]). Few efforts have been dedicated specifically to characterizing burrow and track characteristics (Stott, 1996 Stott, P. 1996. Ground-penetrating radar: a technique for investigating the burrow structure of fossorial vertebrates. Wildlife Research, 22: 519–530.  [Google Scholar]; Sensors & Software Inc., 2010 [compilation on geophysical projects related to animal burrows]; Buynevich and Hasiotis, 2011; Buynevich et al., 2011; Martin et al., 2011) and most of the above studies are published in journals not routinely accessed by ichnologists.     

CO2 Mediated Interaction in Yeast Stimulates Budding and Growth on Minimal Media

Here we show that carbon dioxide (CO₂) stimulates budding and shortens the lag-period of Saccharomyces cerevisiae cultures, grown on specific weak media. CO₂ can be both exogenous and secreted by another growing yeast culture. We also show that this effect can be observed only in the lag-period, and demonstrate minimal doses and duration of culture exposition to CO₂. Opposite to the effects of CO₂ sensitivity, previously shown for pathogens, where increased concentration of CO₂ suppressed mitosis and stimulated cell differentiation and invasion, here it stimulates budding and culture growth.     

Application of docking methodologies to modeled proteins

Protein docking is essential for structural characterization of protein interactions. Besides providing the structure of protein complexes, modeling of proteins and their complexes is important for understanding the fundamental principles and specific aspects of protein interactions. The accuracy of protein modeling, in general, is still less than that of the experimental approaches. Thus, it is important to investigate the applicability of docking techniques to modeled proteins. We present new comprehensive benchmark sets of protein models for the development and validation of protein docking, as well as a systematic assessment of free and template-based docking techniques on these sets. As opposed to previous studies, the benchmark sets reflect the real case modeling/docking scenario where the accuracy of the models is assessed by the modeling procedure, without reference to the native structure (which would be unknown in practical applications). We also expanded the analysis to include docking of protein pairs where proteins have different structural accuracy. The results show that, in general, the template-based docking is less sensitive to the structural inaccuracies of the models than the free docking. The near-native docking poses generated by the template-based approach, typically, also have higher ranks than those produces by the free docking (although the free docking is indispensable in modeling the multiplicity of protein interactions in a crowded cellular environment). The results show that docking techniques are applicable to protein models in a broad range of modeling accuracy. The study provides clear guidelines for practical applications of docking to protein models.     

DR3/LARD spliced mRNA variants’ frequency in colorectal cancer

Many variants of the DR3/LARD death receptor mRNA are derived during alternative splicing. Different DR3/LARD mRNAs encode the membrane and soluble forms of the receptor, which perform different functions. The frequency of the spliced mRNA variants of DR3/LARD was assessed by RT-PCR in patients with colorectal cancer and in cancer cell lines. Four forms of the DR3/LARD death receptor mRNA were detected with different frequencies in the studied samples. Two of them encoded the membrane molecules (LARD 1a mRNA and DR3β mRNA) and two other forms expressed the soluble forms of the receptor (LARD 3 mRNA and soluble DR3β mRNA). In the blood of healthy volunteers, 11 variants (spectra) of DR3/LARD mRNA forms were identified, and the full spectrum that included all four variants of DR3/LARD mRNA dominated. In blood and tumor center samples from patients with colon cancer, six spectra of DR3/LARD mRNA were found. The diversity of the DR3/LARD mRNA spectra was decreased in colon cancer patients due to the reduced frequency of soluble DR3β mRNA. In samples of tumor centers, the spectrum with the absence of only mRNA of the soluble DR3β form dominated. In the blood of patients, two spectra prevailed, i.e., the full spectrum and LARD 1a mRNA and LARD 3 mRNA. Only these two spectra of DR3/LARD mRNA were also found in cancer cell lines. Distinctions in the frequency of DR3/LARD mRNA spectra in healthy volunteers and patients with colorectal cancer can define the different susceptibility of immunocompetent and tumor cells to apoptosis signals.