Filamentous Morphology Research Articles

Super-resolution microscopy reveals significant impact of M2e-specific monoclonal antibodies on influenza A virus filament formation at the host cell surface

Influenza A virions are highly pleomorphic, exhibiting either spherical or filamentous morphology. The influenza A virus strain A/Udorn/72 (H3N2) produces copious amounts of long filaments on the surface of infected cells where matrix protein 1 (M1) and 2 (M2) play a key role in virus filament formation. Previously, it was shown that an anti-M2 ectodomain (M2e) antibody could inhibit A/Udorn/72 virus filament formation. However, the study of these structures is limited by their small size and complex structure. Here, we show that M2e-specific IgG1 and IgG2a mouse monoclonal antibodies can reduce influenza A/Udorn/72 virus plaque growth and infectivity in vitro. Using Immuno-staining combined with super-resolution microscopy that allows us to study structures beyond the diffraction limit, we report that M2 is localized at the base of viral filaments that emerge from the membrane of infected cells. Filament formation was inhibited by treatment of A/Udorn/72 infected cells with M2e-specific IgG2a and IgG1 monoclonal antibodies and resulted in fragmentation of pre-existing filaments. We conclude that M2e-specific IgGs can reduce filamentous influenza A virus replication in vitro and suggest that in vitro inhibition of A/Udorn/72 virus replication by M2e-specific antibodies correlates with the inhibition of filament formation on the surface of infected cells.

Spinning of Photocatalytic Fiber as Splittable Segmented-Pie Bi-Component Fibers for Antibacterial Textiles.

The increase of public concern regarding hygienic living environments has accelerated research on anti-microbial modifications for many commodity and technical textile products. This study attempted to produce a durable self-cleaning and antibacterial textile material with a relatively low production cost using inorganic photocatalytic nanoparticles embedded in the textile fibers. The spinning of segmented-pie bi-component fibers was conducted using compositions of polyethylene and nylon 6 with 1.0% of TiO₂ as photocatalytic particulate filler. A lab-scale double-extrusion fiber spinning system was used to spin the filament fibers with differing winding speeds of 300 m/min, 500 m/m, and 700 m/m. The extruded round fibers were split into minuscule pie-shaped filaments, which significantly increased their photocatalytic surface areas. The compositions and morphologies of the split filaments and the TiO₂ particles in the filaments were characterized by optical microscopic images. Scanning electron microscopic images were used to investigate particles secured on the filament surfaces. The antibacterial effects of the fibers were assessed using a modified method based on the standard method, AATCC100: Assessment of Antibacterial Finishes on Textile Materials, by adding a visible light source above the test samples. The filament samples were tested with the two test organisms of Staphylococcus aureus and Klebsiella pneumoniae. The numbers of bacteria present were determined, and the percent reduction for the specimens with fibers embedded with TiO₂ and that with fibers without TiO₂ were calculated. The segmented-pie bi-component fiber with 1.0% TiO₂ showed enhanced bioactivity against both species of tested bacteria.

Regulation of Candida albicans Hyphal Morphogenesis by Endogenous Signals.

Candida albicans is a human commensal fungus that is able to assume several morphologies, including yeast, hyphal, and pseudohyphal. Under a range of conditions, C. albicans performs a regulated switch to the filamentous morphology, characterized by the emergence of a germ tube from the yeast cell, followed by a mold-like growth of branching hyphae. This transition from yeast to hyphal growth has attracted particular attention, as it has been linked to the virulence of C. albicans as an opportunistic human pathogen. Signal transduction pathways that mediate the induction of the hyphal transcription program upon the imposition of external stimuli have been extensively investigated. However, the hyphal morphogenesis transcription program can also be induced by internal cellular signals, such as inhibition of cell cycle progression, and conversely, the inhibition of hyphal extension can repress hyphal-specific gene expression, suggesting that endogenous cellular signals are able to modulate hyphal gene expression as well. Here we review recent developments in the regulation of the hyphal morphogenesis of C. albicans, with emphasis on endogenous morphogenetic signals.

Whole-Ecosystem Experiments Reveal Varying Responses of Phytoplankton Functional Groups to Epilimnetic Mixing in a Eutrophic Reservoir

Water column mixing can influence community composition of pelagic phytoplankton in lakes and reservoirs. Previous studies suggest that low mixing favors cyanobacteria, while increased mixing favors green algae and diatoms. However, this shift in community dominance is not consistently achieved when epilimnetic mixers are activated at the whole-ecosystem scale, possibly because phytoplankton community responses are mediated by mixing effects on other ecosystem processes. We conducted two epilimnetic mixing experiments in a small drinking water reservoir using a bubble-plume diffuser system. We measured physical, chemical, and biological variables before, during, and after mixing and compared the results to an unmixed reference reservoir. We observed significant increases in the biomass of cyanobacteria (from 0.8 ± 0.2 to 2.4 ± 1.1 μg L−1, p = 0.008), cryptophytes (from 0.7 ± 0.1 to 1.9 ± 0.6 μg L−1, p = 0.003), and green algae (from 3.8 to 4.4 μg L−1, p = 0.15) after our first mixing event, likely due to increased total phosphorus from entrainment of upstream sediments. After the second mixing event, phytoplankton biomass did not change but phytoplankton community composition shifted from taxa with filamentous morphology to smaller, rounder taxa. Our results suggest that whole-ecosystem dynamics and phytoplankton morphological traits should be considered when predicting phytoplankton community responses to epilimnetic mixing.

Enzyme-Identified Phosphorus Limitation Linked to More Rapid Headloss Accumulation in Drinking Water Biofilters.

Drinking water biofilters can improve water quality by transforming contaminants or their precursors, but they also can develop headloss more rapidly than do abiotic filters. Phosphorus supplementation has been proposed as one strategy to lengthen biofilter run times, but the impact of this strategy in field tests has been mixed. The current bench-scale study found that severe phosphorus limitation, as indicated by a high phosphatase to total glycosidase activity ratio (PHO:GLY), led to 230% higher headloss accumulation rate when particles were loaded onto the biofilters as compared to the same experiment performed under a mild phosphorus limitation. Phosphorus limitation was associated with higher concentrations of extracellular polymeric substances, lower biomass concentrations, a more filamentous biofilm morphology, and increased relative abundance of Hyphomicrobiaceae (a family of stalked bacteria) on the biofilter media. These differences in the biofilm likely contributed to higher headloss. This work suggests that phosphorus supplementation could improve biofilter hydraulics in the field if the biofilter is severely phosphorus limited, which was indicated by a PHO:GLY greater than 154 under the conditions tested in this study.

Vimentin disruption by lipoxidation and electrophiles: Role of the cysteine residue and filament dynamics

The intermediate filament protein vimentin constitutes a critical sensor for electrophilic and oxidative stress, which induce extensive reorganization of the vimentin cytoskeletal network. Here, we have investigated the mechanisms underlying these effects. In vitro, electrophilic lipids, including 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) and 4-hydroxynonenal (HNE), directly bind to vimentin, whereas the oxidant diamide induces disulfide bond formation. Mutation of the single vimentin cysteine residue (Cys328) blunts disulfide formation and reduces lipoxidation by 15d-PGJ2, but not HNE. Preincubation with these agents differentially hinders NaCl-induced filament formation by wild-type vimentin, with effects ranging from delayed elongation and increased filament diameter to severe impairment of assembly or aggregation. Conversely, the morphology of vimentin Cys328Ser filaments is mildly or not affected. Interestingly, preformed vimentin filaments are more resistant to electrophile-induced disruption, although chemical modification is not diminished, showing that vimentin (lip)oxidation prior to assembly is more deleterious. In cells, electrophiles, particularly diamide, induce a fast and drastic disruption of existing filaments, which requires the presence of Cys328. As the cellular vimentin network is under continuous remodeling, we hypothesized that vimentin exchange on filaments would be necessary for diamide-induced disruption. We confirmed that strategies reducing vimentin dynamics, as monitored by FRAP, including cysteine crosslinking and ATP synthesis inhibition, prevent diamide effect. In turn, phosphorylation may promote vimentin disassembly. Indeed, treatment with the phosphatase inhibitor calyculin A to prevent dephosphorylation intensifies electrophile-induced wild-type vimentin filament disruption. However, whereas a phosphorylation-deficient vimentin mutant is only partially protected from disorganization, Cys328Ser vimentin is virtually resistant, even in the presence of calyculin A. Together, these results indicate that modification of Cys328 and vimentin exchange are critical for electrophile-induced network disruption.

Small but mighty: microorganisms offer inspiration for mine remediation and waste stabilisation

Understanding the natural microbiological mechanisms that promote iron cycling in iron ore mine environments may provide novel tools for the remediation of the fragile, iron-rich duricrust ecosystems associated with these environments as well as provide a solution for the stabilisation of hillslopes and tailings (waste) dams. A diverse array of microfossils is frequently identified throughout metrescale duricrusts (canga; >50 wt.% Fe) that cap iron ore deposits in Brazil, shedding light on the intimate role of microorganisms in the evolution of these crusts. Nanoscale secondary ion mass spectrometry revealed that carbon and nitrogen biosignatures are occasionally preserved, and typically associated with the cell envelope structures of microfossils. The microfossils are 1-5 mm in length, with filamentous and rod-shaped morphologies commonly preserved1,2. When examined using backscatter electron scanning electron microscopy, canga shows a complex microstructure from repeated dissolution and re-precipitation of iron oxide minerals. Geochronology3, geochemistry4 and microbiology5 provide insights into the past and present-day role of microorganisms in the evolution of canga. These dynamic biogeochemical processes in canga contribute to the continuous formation of new iron cements, preserving some of world's longest-lived continuously exposed surfaces. Harnessing and accelerating the biogeochemical cycling of iron may contribute to the development of novel technologies for mine remediation and waste stabilisation.

ONS-donor ligand based Pt(II) complexes display extremely high anticancer potency through autophagic cell death pathway

The current study unveils ONS-donor ligand based Pt(II) complexes with unusual anticancer potency showing higher anticancer effect as compared to cisplatin. This series of Pt(II)(R-salicylaldimine)Cl (C1a-C4a) (R = 5-H, 5-CH3, F, 3-CH3O) complexes were prepared in single step in good isolated yields from commercially available materials. The chloride ancillary ligand of “a” series (C1a-C4a) was replaced with 4-picoline and “b” series of four complexes Pt(II)(R-salicylaldimine)(4-picoline)BF4 (C1b-C4b) (R = 5-H, 5-CH3, F, 3-CH3O) was obtained. All these complexes were characterized by different structure elucidation techniques. Among these, the structures of C1a, C2a, C2b and C3b were determined in solid state by single crystal X-ray analysis. We found quick aquation of “a” series of complexes in DMSO/water mixture that was well investigated by 1H NMNR, LCMS and ESI-MS, while “b” series of these complexes was quite stable over a month as described by the 1H NMNR in DMSO/D2O mixture. This ONS-donor ligand based class of Pt(II) complexes showed unusual anticancer potency in non-small cell lung cancer A549, colorectal cancer HT-29 and triple negative breast cancer MDA-MB-231 cells. These Pt(II) complexes induced PARP cleavage and significantly inhibited colony formation ability of cancer cells. Mechanistically, we found reduced aggressive growth of cancer cells by the induction of autophagic cell death via LC3-I/LC3-II expression and recruitment of LC3B to autophagosomal membrane. These complexes induced p21 expression, that suggested their potentials to suppress cell cycle progression. Significant activation of Caspase3/7-dependent apoptotic signaling was observed in cancer cells treated with these Pt(II) complexes. Morphological changes of cancer cells suggested their potentials to modulate epithelial-mesenchymal-transition (EMT) like features of cancer cells. Gel electrophoresis study revealed their interaction with plasmid DNA. Similarly, strong growth retardation effect and filamentous morphology was observed in Escherichia coli (E. coli). These ONS-donor Pt(II) complexes possessed strong anticancer effect in multiple human cancer cells via activation of multiple pathways for apoptotic and autophagic cell death.

The toxin from a ParDE toxin-antitoxin system found in Pseudomonas aeruginosa offers protection to cells challenged with anti-gyrase antibiotics.

SummaryToxin‐antitoxin systems are mediators of diverse activities in bacterial physiology. For the ParE‐type toxins, their reported role of gyrase inhibition utilized during plasmid‐segregation killing indicates they are toxic. However, their location throughout chromosomes leads to questions about function, including potential non‐toxic outcomes. The current study has characterized a ParDE system from the opportunistic human pathogen Pseudomonas aeruginosa (Pa). We identified a protective function for this ParE toxin, PaParE, against effects of quinolone and other antibiotics. However, higher concentrations of PaParE are themselves toxic to cells, indicating the phenotypic outcome can vary based on its concentration. Our assays confirmed PaParE inhibition of gyrase‐mediated supercoiling of DNA with an IC50 value in the low micromolar range, a species‐specificity that resulted in more efficacious inhibition of Escherichia coli derived gyrase versus Pa gyrase, and overexpression in the absence of antitoxin yielded an expected filamentous morphology with multi‐foci nucleic acid material. Additional data revealed that the PaParE toxin is monomeric and interacts with dimeric PaParD antitoxin with a KD in the lower picomolar range, yielding a heterotetramer. This work provides novel insights into chromosome‐encoded ParE function, whereby its expression can impart partial protection to cultures from selected antibiotics.

Preliminary analysis of Chloroflexi populations in full-scale UASB methanogenic reactors.

The phylum Chloroflexi is frequently found in high abundance in methanogenic reactors, but their role is still unclear as most of them remain uncultured and understudied. Hence, a detailed analysis was performed in samples from five up-flow anaerobic sludge blanket (UASB) full-scale reactors fed different industrial wastewaters. Quantitative PCR show that the phylum Chloroflexi was abundant in all UASB methanogenic reactors, with higher abundance in the reactors operated for a long period of time, which presented granular biomass. Both terminal restriction fragment length polymorphism and 16S rRNA gene amplicon sequencing revealed diverse Chloroflexi populations apparently determined by the different inocula. According to the phylogenetic analysis, the sequences from the dominant Chloroflexi were positioned in branches where no sequences of the cultured representative strains were placed. Fluorescent insitu hybridization analysis performed in two of the reactors showed filamentous morphology of the hybridizing cells. While members of the Anaerolineae class within phylum Chloroflexi were predominant, their diversity is still poorly described in anaerobic reactors. Due to their filamentous morphology, Chloroflexi may have a key role in the granulation in methanogenic UASB reactors. Our results bring new insights about the diversity, stability, dynamics and abundance of this phylum in full-scale UASB reactors which aid in understanding their function within the reactor biomass. However, new methodological approaches and analysis of bulking biomass are needed to completely unravel their role in these reactors. Combining all this knowledge with reactor operational parameters will allow to understand their participation in granulation and bulking episodes and design strategies to prevent Chloroflexi overgrowth.

Propeptide genesis by Kex2-dependent cleavage of yeast wall protein 1 (Ywp1) of Candida albicans.

Candida albicans is a prevalent fungal resident and opportunistic pathogen of humans, exhibiting a variety of ovoid and filamentous morphologies. Anchored within the cell wall of the ovoid yeast form of C. albicans is an abundant glycoprotein termed yeast wall protein 1 (Ywp1). Ywp1 has an antiadhesive effect that may facilitate yeast cell dispersal; it also contributes to the masking of the glucan matrix of the yeast cell wall, potentially providing shielding from recognition by the human immune system. Mature Ywp1 consists of an O-glycosylated core of 378 amino acids associated with an N-glycosylated propeptide that originates from an N-terminal segment of Ywp1. A tribasic (-RRR-) sequence in the immature Ywp1 polypeptide is separated by 8 amino acids from a dibasic (-KR-) sequence that is a canonical site for cleavage by the intracellular endopeptidase Kex2, and cleavage occurs at both of these sites to generate an 11 kilodalton (kDa) propeptide that remains strongly associated with the mature core of Ywp1. Previous studies demonstrated an absence of the 11 kDa propeptide in strains lacking Kex2, but the presence of lesser amounts of a 12 kDa propeptide ostensibly (and paradoxically) arising from cleavage at the dibasic site. Subsequent studies of wild type strains, however, suggested that post-secretion cleavages were carried out in vitro by acid proteases in unbuffered cultures to generate the 12 kDa propeptide. Here, intact and Gfp-tagged Ywp1 are utilized to show that neither of the two multibasic sites is normally cleaved in the absence of Kex2, but that uncleaved Ywp1 is still N-glycosylated and subsequently anchored to the cell wall. This furthers our understanding of the multistep cleavage of this highly conserved sequence, as well as the possible contributions of the cleaved propeptide to the maturation and functioning of Ywp1.

Identification of an anti-Gram-negative bacteria agent disrupting the interaction between lipopolysaccharide transporters LptA and LptC

IntroductionThe emergence of drug-resistant Gram-negative bacteria is a serious clinical problem that causes increased morbidity and mortality. However, the slow discovery of new antibiotics is unable to meet the need for treating bacterial infections caused by drug-resistant strains. Lipopolysaccharide (LPS) is synthesized in the cytoplasm and transported to the cell envelope by the LPS transport (Lpt) system. LptA and LptC form a complex that transports LPS from the inner membrane to the outer membrane. MethodsThis study performed a screen for agents that disrupt the transport of LPS in Gram-negative bacteria Escherichia coli. It established a yeast two-hybrid system to detect LptA-LptC interaction and used this system to identify a compound, IMB-881, that blocks this interaction and shows antibacterial activity. ResultsThis study demonstrated that the IMB-881 compound specifically binds to LptA to disrupt LptA-LptC interaction using surface plasmon resonance assay. Overproduction of LptA protein but not that of LptC lowered the antibacterial activity of IMB-881. Strikingly, Escherichia coli cells accumulated ‘extra’ membrane material in the periplasm and exhibited filament morphology after treatment with IMB-881. ConclusionThis study successfully identified, by using a yeast two-hybrid system, an antibacterial agent that likely blocks LPS transport in Gram-negative bacteria.

Analysis of tail bits generation of multilevel storage in resistive switching memory**Project supported by the Ministry of Science and Technology of China (Grant Nos. 2016YFA0203800, 2016YFA0201803, and 2018YFB0407502), the National Natural Science Foundation of China (Grant Nos. 61522408, 61334007, and 61521064), Beijing Municipal Science & Technology Commission Program, China (Grant No. Z161100000216153), and Huawei Data Center Technology Laboratory.

The tail bits of intermediate resistance states (IRSs) achieved in the SET process (IRSS) and the RESET process (IRSR) of conductive-bridge random-access memory were investigated. Two types of tail bits were observed, depending on the filament morphology after the SET/RESET operation. (i) Tail bits resulting from lateral diffusion of Cu ions introduced an abrupt increase of device resistance from IRS to ultrahigh-resistance state, which mainly happened in IRSS. (ii) Tail bits induced by the vertical diffusion of Cu ions showed a gradual shift of resistance toward lower value. Statistical results show that more than 95% of tail bits are generated in IRSS. To achieve a reliable IRS for multilevel cell (MLC) operation, it is desirable to program the IRS in RESET operation. The mechanism of tail bit generation that is disclosed here provides a clear guideline for the data retention optimization of MLC resistive random-access memory cells.

Interfacial Adhesion and Mechanical Properties of PET Fabric/PVC Composites Enhanced by SiO₂/Tributyl Citrate Hybrid Sizing.

Poly(ethylene terephthalate) (PET) fabric-reinforced polyvinyl chloride (PVC) composites have a wide range of applications, but the interface bonding of PET fabric/PVC composites has remained a challenge. In this work, a new in-situ SiO2/tributyl citrate sizing agent was synthesized according to the principle of “similar compatibility.” The developed sizing agent was used as a PET surface modifier to enhance the interfacial performance of PET fabric/PVC composites. The morphology and structure of the PET filaments, the wettability and tensile properties of the PET fabric, the interfacial adhesion, and the tensile and tearing properties of the PET fabric/PVC composites were investigated. Experimental results showed that many SiO2 nanoparticles were scattered on the surface of the modified PET filaments. Moreover, the surface roughness of the modified PET filaments remarkably increased in comparison with that of the untreated PET filaments. The contact angle of the modified PET filaments was also smaller than that of the untreated ones. The peeling strength of the modified PET fabrics/PVC composites was 0.663 N/mm, which increased by 62.50% in comparison with the peeling strength of the untreated ones (0.408 N/mm). This work provides a new approach to the surface modification of PET and improves the properties of PET fabric/PVC composites.

In situ visualisation of the abundant Chloroflexi populations in full-scale anaerobic digesters and the fate of immigrating species.

Anaerobic digestion is a key process for the conversion of waste organics to biogas for energy and is reliant on the synergistic activities of complex microbial communities. Members of the phylum Chloroflexi are often found to be abundant in these systems, yet little is known of their role, with most members yet to be cultured or identified. The aim of this study was to characterize the Chloroflexi communities present in full-scale anaerobic digesters receiving excess sludge from wastewater treatment plants. The core genus-level-phylotypes were identified from extensive 16S rRNA gene amplicon sequencing surveys of 19 full-scale systems over a 6 year period. The T78 and Leptolinea, and the RB349 and SJA-170, were found to be the most abundant genera of mesophilic and thermophilic digesters, respectively. With the exception of Leptolinea, these phylotypes are known only by their 16S rRNA gene sequence, and their morphology and metabolic potentials are not known. Fluorescence in situ hybridisation (FISH) probes were designed for these phylotypes, with their application revealing a similar thin filamentous morphology, indicating a possible role for these organisms in maintaining floc structure. The new FISH probes provide a useful tool for future efforts to characterize these organisms in situ. FISH also suggests that immigrating Chloroflexi species die off in the anaerobic digester environment and their high abundance in anaerobic digesters, observed with DNA based sequencing surveys, was quite possibly due to the persistence of their DNA after their death. This observation is important for the interpretation of popular DNA-based sequencing methods applied for the characterisation of communities with substantial immigration rates, such as anaerobic digesters.

The vacuolated morphology of chordoma cells is dependent on cytokeratin intermediate filaments.

Notochordal cells (NCs), characterized by their vacuolated morphology and coexpression of cytokeratin and vimentin intermediate filaments (IFs), form the immature nucleus pulposus (NP) of the intervertebral disc. As humans age, NCs give way to mature NP cells, which do not possess a vacuolated morphology and typically only express vimentin IFs. In light of their concomitant loss, we investigated the relationship between cytosolic vacuoles and cytokeratin IFs, specifically those containing cytokeratin-8 proteins, using a human chordoma cell line as a model for NCs. We demonstrate that the chemical disruption of IFs with acrylamide, F-actin with cytochalasin-D, and microtubules with nocodazole all result in a significant (p < 0.001) decrease in vacuolation. However, vacuole loss was the greatest in acrylamide-treated cells. Examination of the individual roles of vimentin and cytokeratin-8 IFs in the existence of vacuoles was accomplished using small interfering RNA-mediated RNA interference to knock down either vimentin or cytokeratin-8 expression. Reduction of cytokeratin-8 expression was associated with a less-vacuolated cell morphology. These data demonstrate that cytokeratin-8 IFs are involved in stabilizing vacuoles and that their diminished expression could play a role in the loss of vacuolation in NCs during aging. A better understanding of the NCs may assist in preservation of this cell type for NP maintenance and regeneration.

Theoretical investigation of the Ag filament morphology in conductive bridge random access memories

Conductive bridge random access memories (CBRAMs) usually involve active Ag or Cu metals, where the formation of metal filaments accounts for the low resistance state. For the application of neuromorphic computation, it is highly desirable to develop artificial neurons and synapses, which utilize the complicated volatile or nonvolatile resistive switching phenomena, respectively. This can be achieved by controlling the morphology and stability of the filaments, which requires a deep understanding of the filament formation and disruption mechanisms. Using ab initio calculations, we explored the physical mechanism behind various Ag filament morphologies and growth modes, using GeSe, ZrO2, SiO2, and a-Si as the examples. The roles of Ag and Ag+ stability inside the dielectric, the migration barrier of Ag+, and the Ag+ solvation effect have been investigated in detail. A comprehensive model has been proposed, which in particular could explain the diverse Ag filament morphology experimentally observed in sputtered SiO2 and PECVD SiO2. Our theoretical approach can serve as a pre-screening method in designing new solid-state electrolyte materials of CBRAM, aiming at new functionalities in neuromorphic computation or in-memory logic computing.

Effect of wettability and surface free energy of collection substrates on the structure and morphology of dry-spun cellulose nanofibril filaments

Utilization of cellulose nanofibrils (CNF) for filament production is a comparatively new approach, which can broaden nanocellulose applications by providing continuous long filaments suitable for composite and textile applications. Methods are proposed for the spinning of filaments from cellulose nanofibril suspensions, which are mainly categorized into two groups, i.e. dry-spinning and wet-spinning. For the dry-spinning method, the substrate on which the filament is spun will have a significant influence on the properties of the filament because the as-spun CNF will initially contact it. In this work, the influence of different collection substrates on the properties of CNF filament was studied. Filaments with an average diameter of 0.1 mm were spun on different collection substrates including Teflon tape, Teflon film and glass. The influence of adding oil, as a friction modifier, on the surface properties of the substrate and the structure of the resultant filaments was evaluated. Wettability of each substrate in the absence of oil, and the presence of a thin layer and a thick layer of oil was studied. Circularity of the filaments was measured using image analysis of the cross sections and was correlated with the surface properties of the substrates. It was found that while the surface properties of the collection substrate were related to the uniformity and cross section circularity of the filaments, those spun on a thick layer of oil had the best circular shape irrespective of the substrate material. The results of this study can be used to optimize dry-spinning of CNF filaments.

Morphological transitions of elastic filaments in shear flow

The morphological dynamics, instabilities, and transitions of elastic filaments in viscous flows underlie a wealth of biophysical processes from flagellar propulsion to intracellular streaming and are also key to deciphering the rheological behavior of many complex fluids and soft materials. Here, we combine experiments and computational modeling to elucidate the dynamical regimes and morphological transitions of elastic Brownian filaments in a simple shear flow. Actin filaments are used as an experimental model system and their conformations are investigated through fluorescence microscopy in microfluidic channels. Simulations matching the experimental conditions are also performed using inextensible Euler-Bernoulli beam theory and nonlocal slender-body hydrodynamics in the presence of thermal fluctuations and agree quantitatively with observations. We demonstrate that filament dynamics in this system are primarily governed by a dimensionless elasto-viscous number comparing viscous drag forces to elastic bending forces, with thermal fluctuations playing only a secondary role. While short and rigid filaments perform quasi-periodic tumbling motions, a buckling instability arises above a critical flow strength. A second transition to strongly deformed shapes occurs at a yet larger value of the elasto-viscous number and is characterized by the appearance of localized high-curvature bends that propagate along the filaments in apparent "snaking" motions. A theoretical model for the as yet unexplored onset of snaking accurately predicts the transition and explains the observed dynamics. We present a complete characterization of filament morphologies and transitions as a function of elasto-viscous number and scaled persistence length and demonstrate excellent agreement between theory, experiments, and simulations.

LA POTENCIA VOLUMÉTRICA COMO UN PARÁMETRO CONFIABLE PARA EL ESCALAMIENTO DE MATRACES AGITADOS A BIORREACTOR DE TANQUE AGITADO: PRODUCCIÓN DE UNA GLICOPROTEINA RECOMBINANTE POR Streptomyces lividans

In Streptomyces lividans, filamentous morphology varies according to hydrodynamics/aeration culture conditions, playing important roles in the production, secretion, and post-translational modification of recombinant glycoproteins. Volumetric power input (P/V) is normally used to scale-up bioprocesses. Here, we analyzed the effect of P/V on S. lividans growth and morphology as well as the production and O-mannosylation of the recombinant glycoprotein APA from Mycobacterium tuberculosis. Previously, we measured the P/V in conventional normal (NF, 0.20 kW/m3) and coiled (CF, 0.44 kW/m3) shake flasks. Here, these P/V were matched in 1.0 L bioreactors by adjusting the stirring speed. Similar specific growth rates were observed between shake flask and bioreactor cultures. In addition, similar rAPA yields (~25%) were observed in both the bioreactor and NF cultures, but these were lower than the yield in the CF culture (~36%). Furthermore, up to seven mannose residues were attached to the C-terminal of rAPA in both bioreactor cultures, which was one more than in the NF and CF cultures. However, at similar P/V, the morphology and biomass kinetics of growth were not similar. Our data indicate that scaling-up the production of recombinant glycoproteins using S. lividans by matching the P/V can be successful in terms of some, but not all stequeometric parameters, suggesting that the cellular responses could be differently affected by hydrodynamics/aeration differences between shake flasks and bioreactors.