Filamentous Growth Research Articles

Target site selection in CAST systems

CRISPR Biology Exciting genomic engineering possibilities exist for natural integration systems called transposons, which have coopted CRISPR/Cas systems. An unexplained feature of these systems involves how they direct insertions in a single orientation at a precise distance from the programmed target sequence. Park et al. show that orientation information is communicated to the transposase TnsB using the unidirectional growth of a helical filament made up of an AAA+ protein, TnsC. ATP hydrolysis trims the filament to a minimal unit that is marked by TniQ and defined by the Cas12k protein to provide spacing information. This finding may help future engineering of these systems for therapeutic applications. Science , abi8976, this issue p. [768][1] [1]: /lookup/doi/10.1126/science.abi8976

The NDR kinase-MOB complex FgCot1-Mob2 regulates polarity and lipid metabolism in Fusarium graminearum.

Members of the NDR (nuclear Dbf2-related) protein-kinase family are essential for cell differentiation and polarized morphogenesis. However, their functions in plant pathogenic fungi are not well understood. Here, we characterized the NDR kinase FgCot1 and its activator FgMob2 in Fusarium graminearum, a major pathogen causing Fusarium head blight (FHB) in wheat. FgCot1 and FgMob2 formed a NDR kinase-MOB protein complex. Localization assays using FgCot1-GFP or FgMob2-RFP constructs showed diverse subcellular localizations, including cytoplasm, septum, nucleus and hyphal tip. ΔFgcot1 and ΔFgmob2 exhibited serious defects in hyphal growth, polarity, fungal development and cell wall integrity as well as reduced virulence in planta. In contrast, lipid droplet accumulation was significantly increased in these two mutants. Phosphorylation of FgCot1 at two highly conserved residues (S462 and T630) as well as five new sites synergistically contributed its role in various cellular processes. In addition, non-synonymous mutations in two MAPK (mitogen-activated protein kinase) proteins, FgSte11 and FgGpmk1, partially rescued the growth defect of ΔFgmob2, indicating a functional link between the FgCot1-Mob2 complex and the FgGpmk1 signalling pathway in regulating filamentous fungal growth. These results indicated that the FgCot1-Mob2 complex is critical for polarity, fungal development, cell wall organization, lipid metabolism and virulence in F. graminearum.

The SAGA and NuA4 component Tra1 regulates Candida albicans drug resistance and pathogenesis.

Candida albicans is the most common cause of death from fungal infections. The emergence of resistant strains reducing the efficacy of first-line therapy with echinocandins, such as caspofungin calls for the identification of alternative therapeutic strategies. Tra1 is an essential component of the SAGA and NuA4 transcriptional co-activator complexes. As a PIKK family member, Tra1 is characterized by a C-terminal phosphoinositide 3-kinase domain. In Saccharomyces cerevisiae, the assembly and function of SAGA and NuA4 are compromised by a Tra1 variant (Tra1Q3) with three arginine residues in the putative ATP-binding cleft changed to glutamine. Whole transcriptome analysis of the S. cerevisiae tra1Q3 strain highlights Tra1's role in global transcription, stress response, and cell wall integrity. As a result, tra1Q3 increases susceptibility to multiple stressors, including caspofungin. Moreover, the same tra1Q3 allele in the pathogenic yeast C. albicans causes similar phenotypes, suggesting that Tra1 broadly mediates the antifungal response across yeast species. Transcriptional profiling in C. albicans identified 68 genes that were differentially expressed when the tra1Q3 strain was treated with caspofungin, as compared to gene expression changes induced by either tra1Q3 or caspofungin alone. Included in this set were genes involved in cell wall maintenance, adhesion, and filamentous growth. Indeed, the tra1Q3 allele reduces filamentation and other pathogenesis traits in C. albicans. Thus, Tra1 emerges as a promising therapeutic target for fungal infections.

Linearity improvement of HfOx-based memristor with multilayer structure

The limitation of traditional Von Neumann architecture could be resolved by machine learning training in neuromorphic computing. However, the nonlinearity characteristic during conductance modulation in memristor severely restricts its application in neuromorphic computing. To improve the analog switching including linearity of the hafnium oxide memristors, Ti metal layer has been inserted on hafnium oxide by using the redox reaction on the interface to achieve more gradual switching. Furthermore, a new multilayer structure device is fabricated utilizing the Ti/HfOx interface characteristic, which enhanced the electrical characteristic and the conductance modulation linearity to 98.4%, amplitude and symmetry have also been improved. The conductive mechanism of segmented growth of conductive filaments by adjusting the oxygen vacancy concentration gradient was analyzed and further characterized by XPS and TEM. The results of this paper will exalt conductance modulation linearity of hafnium oxide memristors for application to neuromorphic computing systems.

Spatiotemporal control of pathway sensors and cross-pathway feedback regulate a differentiation MAPK pathway in yeast.

Mitogen-activated protein kinase (MAPK) pathways control cell differentiation and the response to stress. In Saccharomyces cerevisiae, the MAPK pathway that controls filamentous growth (fMAPK) shares components with the pathway that regulates the response to osmotic stress (HOG). Here, we show that the two pathways exhibit different patterns of activity throughout the cell cycle. The different patterns resulted from different expression profiles of genes encoding mucin sensors that regulate the pathways. Cross-pathway regulation from the fMAPK pathway stimulated the HOG pathway, presumably to modulate fMAPK pathway activity. We also show that the shared tetraspan protein Sho1p, which has a dynamic localization pattern throughout the cell cycle, induced the fMAPK pathway at the mother-bud neck. A Sho1p-interacting protein, Hof1p, which also localizes to the mother-bud neck and regulates cytokinesis, also regulated the fMAPK pathway. Therefore, spatial and temporal regulation of pathway sensors, and cross-pathway regulation, control a MAPK pathway that regulates cell differentiation in yeast.

The Complex Genetic Basis and Multilayered Regulatory Control of Yeast Pseudohyphal Growth.

Eukaryotic cells are exquisitely responsive to external and internal cues, achieving precise control of seemingly diverse growth processes through a complex interplay of regulatory mechanisms. The budding yeast Saccharomyces cerevisiae provides a fascinating model of cell growth in its stress-responsive transition from planktonic single cells to a filamentous pseudohyphal growth form. During pseudohyphal growth, yeast cells undergo changes in morphology, polarity, and adhesion to form extended and invasive multicellular filaments. This pseudohyphal transition has been studied extensively as a model of conserved signaling pathways regulating cell growth and for its relevance in understanding the pathogenicity of the related opportunistic fungus Candida albicans, wherein filamentous growth is required for virulence. This review highlights the broad gene set enabling yeast pseudohyphal growth, signaling pathways that regulate this process, the role and regulation of proteins conferring cell adhesion, and interesting regulatory mechanisms enabling the pseudohyphal transition.

Structural basis for target site selection in RNA-guided DNA transposition systems.

CRISPR-associated transposition systems allow guide RNA-directed integration of a single DNA cargo in one orientation at a fixed distance from a programmable target sequence. We used cryo-electron microscopy (cryo-EM) to define the mechanism that underlies this process by characterizing the transposition regulator, TnsC, from a type V-K CRISPR-transposase system. In this scenario, polymerization of adenosine triphosphate-bound TnsC helical filaments could explain how polarity information is passed to the transposase. TniQ caps the TnsC filament, representing a universal mechanism for target information transfer in Tn7/Tn7-like elements. Transposase-driven disassembly establishes delivery of the element only to unused protospacers. Finally, TnsC transitions to define the fixed point of insertion, as revealed by structures with the transition state mimic ADP•AlF3 These mechanistic findings provide the underpinnings for engineering CRISPR-associated transposition systems for research and therapeutic applications.

Blocking lithium dendrite growth in solid-state batteries with an ultrathin amorphous Li-La-Zr-O solid electrolyte

Lithium dendrites have become a roadblock in the realization of solid-state batteries with lithium metal as high-capacity anode. The presence of surface and bulk defects in crystalline electrolytes such as the garnet Li7La3Zr2O12 (LLZO) facilitates the growth of these hazardous lithium filaments. Here we explore the amorphous phase of LLZO as a lithium dendrite shield owing to its grain-boundary-free microstructure, stability against lithium metal, and high electronic insulation. By tuning the lithium stoichiometry, the ionic conductivity can be increased by 4 orders of magnitude while retaining a negligible electronic conductivity. In symmetric cells, plating-stripping shows no signs of lithium penetration up to 3.2 mA cm−2. The dense conformal nature of the films enables microbatteries with an electrolyte thickness of only 70 nm, which can be cycled at 10C for over 500 cycles. The application of this material as a coating on crystalline LLZO lowers the interface resistance and increases the critical current density. The effectiveness of the amorphous Li-La-Zr-O as dendrite blocking layer can accelerate the development of better solid-state batteries.

Phytohormone biosynthesis and signaling pathways of mosses.

Most known phytohormones regulate moss development. We present a comprehensive view of the synthesis and signaling pathways for the most investigated of these compounds in mosses, focusing on the model Physcomitrium patens. The last 50years of research have shown that most of the known phytohormones are synthesized by the model moss Physcomitrium patens (formerly Physcomitrella patens) and regulate its development, in interaction with responses to biotic and abiotic stresses. Biosynthesis and signaling pathways are best described in P. patens for the three classical hormones auxins, cytokinins and abscisic acid. Furthermore, their roles in almost all steps of development, from early filament growth to gametophore development and sexual reproduction, have been the focus of much research effort over the years. Evidence of hormonal roles exist for ethylene and for CLE signaling peptides, as well as for salicylic acid, although their possible effects on development remain unclear. Production of brassinosteroids by P. patens is still debated, and modes of action for these compounds are even less known. Gibberellin biosynthesis and signaling may have been lost in P. patens, while gibberellin precursors such as ent-kaurene derivatives could be used as signals in a yet to discover pathway. As for jasmonic acid, it is not used per se as a hormone in P. patens, but its precursor OPDA appears to play a corresponding role in defense against abiotic stress. We have tried to gather a comprehensive view of the biosynthesis and signaling pathways for all these compounds in mosses, without forgetting strigolactones, the last class of plant hormones to be reported. Study of the strigolactone response in P. patens points to a novel signaling compound, the KAI2-ligand, which was likely employed as a hormone prior to land plant emergence.

Growth control agent for filamentous fungi: FDM based 3D printed cubes for suspended Aspergillus sojae fermentation

The main problem for submerged fermentation of filamentous fungi is the nutrition limitation with high cell density or cell leakage by the uncontrollable hyphae, clusters, or pellets. There are several techniques such as microparticle, immobilization, pH shifting, substrate limitation etc. for controlling filamentous fungi growth on submerged fermentation. In this research, FDM (Fused Deposition Modelling) based 3D printed cubes is used for growth control agent of recombinant Aspergillus sojae for the first time. Lattice structure sizes, number of cubes and pH were chosen to be main factors of fermentation in order to study the combine effect of the factors on A. sojae fermentation. The results revealed that specific activity values are improved from 2045.96 U/mg (the highest control activity) to 3291.67 U/mg with lower pellet sizes and controllable growth. FDM based 3D printed cubes was successfully controlled the recombinant Aspergillus sojae fermentation and enhanced β-mannanase production. In addition, this research was also showed that FDM based 3D printed cubes also have the potential to be used as immobilization materials like SLS based 3D printed products in further research.

Effect of Operating Conditions on Membrane Fouling in Pilot-Scale MBRs; Filaments Growth, Diminishing Dissolved Oxygen and Recirculation Rate of the Activated Sludge.

This is the first study that examines the effect of operating conditions on fouling of Membrane Bio-Reactors (MBRs), which treat municipal wastewater in field conditions, with specific regard to the controlled development of filamentous microorganisms (or filaments). The novelty of the present work is extended to minimize the dissolved oxygen (DO) in recirculated activated sludge for improving the process of denitrification. For this purpose, two pilot-scale MBRs were constructed and operated in parallel: (i) Filament-MBR, where an attempt was made to regulate the growth of filaments by adjustment of DO, the Food-to-Microorganisms (F/M) ratio and temperature, and (ii) Control-MBR, where a gentle stirring tank was employed for the purpose of zeroing the DO in the recycled sludge. Results showed that low temperature (<15 °C) slightly increased the number of filaments in the Filament-MBR which, in turn, decreased the Trans-Membrane Pressure (TMP). As the Soluble Microbial Products (SMP) and the colloids are considered to be the basic foulants of membranes in MBR systems, specific attention was directed to keep their concentration at low values in the mixed liquor. The low F/M ratio in the aeration tanks which preceded the membrane tank was achieved to keep the SMP proteins and carbohydrates at very low values in the mixed liquor, i.e., less than 6 mg/L. Moreover, as a result of the low recirculation rate (2.6∙Qin), good aggregation of the produced excess sludge was achieved, and low concentration of colloids with a size ≤50 nm (nearly the membranes’ pore size used for filtration/separation) was measured, accounted for maximum 15% of the total colloids. Additionally, the increase in filamentous population at the Filament-MBR contributed to the further reduction of colloids in the mixed liquor at 7.9%, contributing beneficially to the reduction of TMP and of membrane fouling. The diminishing of DO in the recirculated sludge improved denitrification, and resulted in lower concentrations of Ν-NO3− and TN in the effluent of the Control-MBR. Furthermore, the recirculation rate of Qr = 2.6∙Qin, in comparison with Qr = 4.3∙Qin, resulted in improved performance regarding the removal of N-NH4+. Finally, high organics removal and ammonium nitrification was observed in the effluent of both pilots, since COD and Ν-ΝH4+ concentrations were generally in the range of 10–25 mg/L and <0.1 mg/L, respectively.

#45: Phenotypic heterogeneity among isolates of Candida albicans from specific anatomical niches in VLBW premature infants

Abstract Background Disseminated candidiasis is associated with high morbidity and mortality among very low birth weight (VLBW) infants. Research about C. albicans traits which lead to colonization versus invasive disease is lacking. Ability to grow in a filamentous form is a known virulence trait of C. albicans. Nevertheless, significant heterogeneity in filamentous growth in C. albicans clinical isolates has been documented (Hirakawa et. al). However, the clinical isolates investigated were obtained from varied tissue niches from hosts with differing risk factors, which could account for the variation in filamentation. A collection of C. albicans isolates prospectively collected from VLBW infants and their mothers demonstrated vertical transmission of C. albicans from mother to infant (Bliss et. al). We used the infants’ isolates from this collection to investigate filamentation amongst isolates obtained from the same tissue niche and host type. Methods The standard C. albicans reference strain in the field, SC5314 was chosen for a positive control as it demonstrates strong filamentation and high virulence. A strain lacking Efg1, a regulatory transcription factor that promotes filamentation, produces hypofilamentous colonies and was used as the negative control. Reference strains and the clinical isolates were grown on two types of stimulation media at 37 C for 7 days after which filamentation was scored. Results Filamentation development varied widely among isolates. Of the strains which were acquired through vertical transmission, 5 of 11 strains showed strong filamentation, 5 of 11 had an intermediate phenotype, and 1 did not display any filamentation. Three strains were acquired through horizontal transmission (RO10, RO54, and RO56). Of these, one showed strong filamentation whereas the other two failed to form filaments. None of the neonatal isolates showed colony wrinkling, a prominent phenotype in the reference strain SC5314. Strain RO61, an isolate associated with candidemia showed strong filamentation. In contrast, strain RO39, which was associated with dermatitis, was non-filamentous. Conclusions We observed phenotypic heterogeneity with respect to filament development among clinical isolates obtained from the same niche (rectal) in similar hosts (VLBW premature infants). This is consistent with the variation seen in Hirakawa et al and indicates that phenotypic variation is not a function of host risk factors nor clinical niche. Filamentation was not required for vertical transmission. No conclusions can be drawn based on this observation with only 2 strains, but it is interesting to note that the isolate which caused candidemia and death of the patient was highly filamentous whereas the isolate that caused dermatitis was not. Further investigation of these strains, including genomic sequencing and additional phenotypic evaluations, will explore the underlying factors leading to transmission and virulence of C. albicans in VLBW premature infants.

Single-molecule imaging of LexA degradation in Escherichia coli elucidates regulatory mechanisms and heterogeneity of the SOS response.

The bacterial SOS response stands as a paradigm of gene networks controlled by a master transcriptional regulator. Self-cleavage of the SOS repressor, LexA, induces a wide range of cell functions that are critical for survival and adaptation when bacteria experience stress conditions1, including DNA repair2, mutagenesis3,4, horizontal gene transfer5–7, filamentous growth, and the induction of bacterial toxins8–12, toxin-antitoxin systems13, virulence factors6,14, and prophages15–17. SOS induction is also implicated in biofilm formation and antibiotic persistence11,18–20. Considering the fitness burden of these functions, it is surprising that the expression of LexA-regulated genes is highly variable across cells10,21–23 and that cell subpopulations induce the SOS response spontaneously even in the absence of stress exposure9,11,12,16,24,25. Whether this reflects a population survival strategy or a regulatory inaccuracy is unclear, as are the mechanisms underlying SOS heterogeneity. Here, we developed a single-molecule imaging approach based on a HaloTag fusion to directly monitor LexA inside live Escherichia coli cells, demonstrating the existence of 3 main states of LexA: DNA-bound stationary molecules, free LexA and degraded LexA species. These analyses elucidate the mechanisms by which DNA-binding and degradation of LexA regulate the SOS response in vivo. We show that self-cleavage of LexA occurs frequently throughout the population during unperturbed growth, rather than being restricted to a subpopulation of cells, which causes substantial cell-to-cell variation in LexA abundances. LexA variability underlies SOS gene expression heterogeneity and triggers spontaneous SOS pulses, which enhance bacterial survival in anticipation of stress.

Cdc42 regulates reactive oxygen species production in the pathogenic yeast Candida albicans

Across eukaryotes, Rho GTPases such as Rac and Cdc42 play important roles in establishing cell polarity, which is a key feature of cell growth. In mammals and filamentous fungi, Rac targets large protein complexes containing NADPH oxidases (NOX) that produce reactive oxygen species (ROS). In comparison, Rho GTPases of unicellular eukaryotes were believed to signal cell polarity without ROS, and it was unclear whether Rho GTPases were required for ROS production in these organisms. We document here the first example of Rho GTPase–mediated post-transcriptional control of ROS in a unicellular microbe. Specifically, Cdc42 is required for ROS production by the NOX Fre8 of the opportunistic fungal pathogen Candida albicans. During morphogenesis to a hyphal form, a filamentous growth state, C. albicans FRE8 mRNA is induced, which leads to a burst in ROS. Fre8-ROS is also induced during morphogenesis when FRE8 is driven by an ectopic promoter; hence, Fre8 ROS production is in addition controlled at the post-transcriptional level. Using fluorescently tagged Fre8, we observe that the majority of the protein is associated with the vacuolar system. Interestingly, much of Fre8 in the vacuolar system appears inactive, and Fre8-induced ROS is only produced at sites near the hyphal tip, where Cdc42 is also localized during morphogenesis. We observe that Cdc42 is necessary to activate Fre8-mediated ROS production during morphogenesis. Cdc42 regulation of Fre8 occurs without the large NOX protein complexes typical of higher eukaryotes and therefore represents a novel form of ROS control by Rho GTPases.

Mitochondrial perturbation reduces susceptibility to xenobiotics through altered efflux in Candida albicans.

Candida albicans is a leading human fungal pathogen, which can cause superficial infections or life-threatening systemic disease in immunocompromised individuals. The ability to transition between yeast and filamentous forms is a major virulence trait of C. albicans, and a key regulator of this morphogenetic transition is the molecular chaperone Hsp90. To explore the mechanisms governing C. albicans morphogenesis in response to Hsp90 inhibition, we performed a functional genomic screen using the gene replacement and conditional expression collection to identify mutants that are defective in filamentation in response to the Hsp90 inhibitor, geldanamycin. We found that transcriptional repression of genes involved in mitochondrial function blocked filamentous growth in response to the concentration of the Hsp90 inhibitor used in the screen, and this was attributable to increased resistance to the compound. Further exploration revealed that perturbation of mitochondrial function reduced susceptibility to two structurally distinct Hsp90 inhibitors, geldanamycin and radicicol, such that filamentous growth was restored in the mitochondrial mutants by increasing the compound concentration. Deletion of two representative mitochondrial genes, MSU1 and SHY1, enhanced cellular efflux and reduced susceptibility to diverse intracellularly acting compounds. Additionally, screening a C. albicans efflux pump gene deletion library implicated Yor1 in the efflux of geldanamycin and Cdr1, in the efflux of radicicol. Deletion of these transporter genes restored sensitivity to Hsp90 inhibitors in MSU1 and SHY1 homozygous deletion mutants, thereby enabling filamentation. Taken together, our findings suggest that mitochondrial dysregulation elevates cellular efflux and consequently reduces susceptibility to xenobiotics in C. albicans.

LncRNA DINOR is a virulence factor and global regulator of stress responses in Candida auris.

The emergent fungal pathogen Candida auris exhibits high resistance to antifungal drugs and environmental stresses, impeding treatment and decontamination1-3. The fungal factors mediating this stress tolerance are largely unknown. In the present study, we performed piggyBac, transposon-mediated, genome-wide mutagenesis and genetic screening in C. auris, and identified a mutant that grew constitutively in the filamentous form. Mapping the transposon insertion site revealed the disruption of a long non-coding RNA, named DINOR for DNA damage-inducible non-coding RNA. Deletion of DINOR caused DNA damage and an upregulation of genes involved in morphogenesis, DNA damage and DNA replication. The DNA checkpoint kinase Rad53 was hyperphosphorylated in dinorΔ mutants, and deletion of RAD53 abolished DNA damage-induced filamentation. DNA-alkylating agents, which cause similar filamentous growth, induced DINOR expression, suggesting a role for DINOR in maintaining genome integrity. Upregulation of DINOR also occurred during exposure to the antifungal drugs caspofungin and amphotericin B, macrophages, H2O2 and sodium dodecylsulfate, indicating that DINOR orchestrates multiple stress responses. Consistently, dinorΔ mutants displayed increased sensitivity to these stresses and were attenuated for virulence in mice. Moreover, genome-wide genetic interaction studies revealed links between the function of DINOR and TOR signalling, an evolutionarily conserved pathway that regulates the stress response. Identification of the mechanism(s) by which DINOR regulates stress responses in C. auris may provide future opportunities for the development of therapeutics.

Improved uniformity in resistive switching behaviors based on PMMA films with embedded carbon quantum dots

The growth and rupture of conductive filaments act a crucial part in the reliability of resistive switching behaviors. The random growth and rupture of conductive filaments are the primary reason for the instability of set/reset reproducibility. Hence, we propose a method that embedded carbon quantum dots (CQDs) in polymethylmethacrylate (PMMA) to fabricate the Ag/PMMA&amp;CQDs/FTO resistive switching device. Five different concentrations of CQDs are embedded in PMMA to regulate the resistive switching properties, and the resistive memory characteristics of the optimal group are systematically studied. The optimal group exhibits excellent switching repeatability, low set/reset voltages, and stable forming voltage, which is much better than PMMA without CQDs. Furthermore, we employ the COMSOL software to build a simulation model for exploring the influence of CQDs on the internal electric field of PMMA, which proved that the introduction of CQDs might have a favorable effect on the orderly growth of conductive filaments.

Insights into Mechanism of Aβ42 Fibril Growth on Surface of Graphene Oxides: Oxidative Degree Matters

The filamentous β-amyloid deposition has been regarded as the hallmark pathology of Alzheimer's disease (AD). Nanomaterials such as graphene oxides (GOs) have achieved significant progress in the therapy of AD, but the molecular pathway of the growth propagation remains challenging to investigate, especially on the surfaces of materials. The thermodynamics and kinetics of fibril elongation on GO surfaces with different oxidative degrees have been investigated by a combination of in vitro experiments and simulations. ThT kinetics, calorimetric measurements, and TEM observations suggest that low oxidative GO-10 promotes the fibril elongation, while both high oxidative GO-20 and GO-40 inhibit the fibril elongation. Computational results reveal that the apparent regulation behaviors of GOs on filament growth depend on the balance between the promoting effect by templating the incoming of monomers and the retarding effect by capturing the monomer during docking and locking phases through hydrogen bonding. This work will promote the understanding of the interplay between biomolecules and materials, thus providing new thoughts for the rational design of novel materials for amyloidosis therapy.

Accelerostat study in conventional and microfluidic bioreactors to assess the key role of residual glucose in the dimorphic transition of Yarrowia lipolytica in response to environmental stimuli

Yarrowia lipolytica, with a diverse array of biotechnological applications, is able to grow as ovoid yeasts or filamentous hyphae depending on environmental conditions. This study has explored the relationship between residual glucose levels and dimorphism in Y. lipolytica. Under pH stress conditions, the morphological and physiological characteristics of the yeast were examined during well-controlled accelerostat cultures using both a 1 L-laboratory scale and a 1 mL-microfluidic bioreactor. The accelerostat mode, via a smooth increase of dilution rate (D), enabled the cell growth rate to increase gradually up to the cell wash-out (D ≥μmax of the strain), which was accompanied by a progressive increase in residual glucose concentration. The results showed that Y. lipolytica maintained an ovoid morphology when residual glucose concentration was below a threshold value of around 0.35−0.37 mg L−1. Transitions towards more elongated forms were triggered at this threshold and progressively intensified with the increase in residual glucose levels. The effect of cAMP on the dimorphic transition was assessed by the exogenous addition of cAMP and the quantification of its intracellular levels during the accelerostat. cAMP has been reported to be an important mediator of environmental stimuli that inhibit filamentous growth in Y. lipolytica by activating the cAMP-PKA regulatory pathway. It was confirmed that the exogenous addition of cAMP inhibited the mycelial morphology of Y. lipolytica, even with glucose concentrations exceeding the threshold level. The results suggest that dimorphic responses in Y. lipolytica are regulated by sugar signaling pathways, most likely via the cAMP-PKA dependent pathway.

Modulation of the Conductive Filament Growth During the Electroforming Process in Metal Oxide-Based RRAM

Metal oxide-based Resistive Random-Access Memories (RRAM) exhibit multiple intermediate resistive states, due to the formation/rupture of a conductive filament (CF) inside a switching layer. While there have been extensive studies on the resistive switching behavior caused by a pre-existing CF, the initial stage of the formation and growth of a CF, i.e., the electroforming process has been less understood. In this work, we developed a physical model to fundamentally understand the formation and growth behavior of a CF in a metal oxide layer constrained by the top/bottom electrodes. It is revealed that the CF formation is assisted by the supply of oxygen vacancies at the anode and the oxygen vacancies transport in the oxide layer. The effects of the external conditions (the oxygen exchange rate j 0 and the applied voltage) and the physical properties of the oxides (the electrical σ and thermal conductivity k th ) on the CF morphology evolution, current density evolution, and local temperature and electrical potential distribution have been systematically explored. It is found that larger oxygen vacancy exchange rate and relatively small voltage sweep rate result in a more uniform CF. Furthermore, by choosing oxides with lower Lorenz number (ratio of k th and σ ), CF with more homogeneous morphology can be realized. This work provides a fundamental understanding of the kinetic behaviors of the CF formation and growth during the electroforming process, and could potentially guide the oxide and electrode materials selection, synthesis and working conditions for switching to achieve improved RRAM stability and functionality.