פרסומים

Fungi transcriptionally upregulate expression of azole efflux pumps and ergosterol biosynthesis pathway genes when exposed to antifungal agents that target ergosterol biosynthesis. To date, these transcriptional responses have been shown to be dependent on the presence of the azoles and/or depletion of ergosterol. Using an inducible promoter to regulate , which encodes the major azole target, sterol 14α-demethylase, we were able to demonstrate that the CDR4 azole efflux pump can be transcriptionally activated by ergosterol biosynthesis inhibition even in the absence of azoles. By analyzing ergosterol deficient mutants, we demonstrate that the transcriptional responses by and, unexpectedly, genes encoding ergosterol biosynthesis enzymes ( genes) that are responsive to azoles, are not dependent on ergosterol depletion. Nonetheless, deletion of , which encodes C-8 sterol isomerase, also induced expression of . Deletion of also induced the expression of , the gene encoding C-14 sterol reductase, but not other tested genes which were responsive to inactivation. This indicates that inhibition of specific steps of ergosterol biosynthesis can result in different transcriptional responses, which is further supported by our results obtained using different ergosterol biosynthesis inhibitors. Together with the sterol profiles, these results suggest that the transcriptional responses by and genes are associated with accumulation of specific sterol intermediate(s). This was further supported by the fact that when the mutant was treated with ketoconazole, upstream inhibition overrode the effects by downstream inhibition on ergosterol biosynthesis pathway. Even though expression is associated with the accumulation of sterol intermediates, intra- and extracellular sterol analysis by HPLC-MS indicated that the transcriptional induction of did not result in efflux of the accumulated intermediate(s). This study demonstrates, by detailed genetic and chemical analysis, that transcriptional responses by a major efflux pump and genes of the ergosterol biosynthesis pathway to ergosterol biosynthesis inhibitors can be independent of the presence of the drugs and are linked with the accumulation of ergosterol intermediate(s).

Abstract The molecular mechanism regulating dormancy release in grapevine buds is as yet unclear. It was formerly proposed that dormancy is maintained by abscisic acid (ABA)-mediated repression of bud–meristem activity and that removal of this repression triggers dormancy release. It was also proposed that such removal of repression may be achieved via natural or artificial up-regulation of VvA8H-CYP707A4, which encodes ABA 8′-hydroxylase, and is the most highly expressed paralog in grapevine buds. The current study further examines these assumptions, and its experiments reveal that (a) hypoxia and ethylene, stimuli of bud dormancy release, enhance expression of VvA8H-CYP707A4 within grape buds, (b) the VvA8H-CYP707A4 protein accumulates during the natural transition to the dormancy release stage, and (c) transgenic vines overexpressing VvA8H-CYP707A4 exhibit increased ABA catabolism and significant enhancement of bud break in controlled and natural environments and longer basal summer laterals. The results suggest that VvA8H-CYP707A4 functions as an ABA degrading enzyme, and are consistent with a model in which the VvA8H-CYP707A4 level in the bud is up-regulated by natural and artificial bud break stimuli, which leads to increased ABA degradation capacity, removal of endogenous ABA-mediated repression, and enhanced regrowth. Interestingly, it also hints at sharing of regulatory steps between latent and lateral bud outgrowth.

The gold-standard treatment for childhood amblyopia remains patching or penalizing the fellow eye, resulting in an average of about a one line (0.1 logMAR) improvement in visual acuity following ≈120 h of patching in children 3–8 years old. However, compliance with patching and other treatment options is often poor. In contrast, fast-paced action video games can be highly engaging, and have been shown to yield broad-based improvements in vision and attention in adult amblyopia. Here, we pilot-tested a custom-made action video game to treat children with amblyopia. Twenty-one (n = 21) children (mean age 9.95 ± 3.14 [se]) with unilateral amblyopia (n = 12 anisometropic and n = 9 strabismic) completed 20 h of game play either monocularly, with the fellow eye patched (n = 11), or dichoptically, with reduced contrast to the fellow eye (n = 10). Participants were assessed for visual acuity (VA), stereo acuity and reading speed at baseline, and following 10 and 20 h of play. Additional exploratory analyses examined improvements after 6–10 weeks of completion of training (follow-up). Following 20 h of training, VA improved, on average, by 0.14 logMAR (≈38%) for the dichoptic group and by 0.06 logMAR (≈15%) for the monocular group. Similarly, stereoacuity improved by 0.07 log arcsec (≈17%) following dichoptic training, and by 0.06 log arcsec (≈15%) following monocular training. Across both treatment groups, 7 of the 12 individuals with anisometropic amblyopia showed improvement in stereoacuity, whereas only 1 of the 9 strabismic individuals improved. Most improvements were largely retained at follow-up. Our feasibility study therefore suggests that the action video game approach may be used as an effective adjunct treatment for amblyopia in children, achieving results similar to those of the gold-standard treatment in shorter duration.

Seminal roots constitute the initial wheat root system and provide the main route for water absorption during early stages of development. Seminal root number (SRN) varies among species. However, the mechanisms through which SRN is controlled and in turn contribute to environmental adaptation are poorly understood. Here, we show that SRN increased upon wheat domestication from 3 to 5 due to the activation of 2 root primordia that are suppressed in wild wheat, a trait controlled by loci expressed in the germinating embryo. Suppression of root primordia did not limit water uptake, indicating that 3 seminal roots is adequate to maintain growth during seedling development. The persistence of roots at their primordial state promoted seedling recovery from water stress through reactivation of suppressed primordia upon rehydration. Our findings suggest that under well-watered conditions, SRN is not a limiting factor, and excessive number of roots may be costly and maladaptive. Following water stress, lack of substantial root system suppresses growth and rapid recovery of the root system is essential for seedling recovery. This study underscores SRN as key adaptive trait that was reshaped upon domestication. The maintenance of roots at their primordial state during seedling development may be regarded as seedling protective mechanism against water stress.

Seminal roots constitute the initial wheat root system and provide the main route for water absorption during early stages of development. Seminal root number (SRN) varies among species. However, the mechanisms through which SRN is controlled and in turn contribute to environmental adaptation are poorly understood. Here, we show that SRN increased upon wheat domestication from 3 to 5 due to the activation of 2 root primordia that are suppressed in wild wheat, a trait controlled by loci expressed in the germinating embryo. Suppression of root primordia did not limit water uptake, indicating that 3 seminal roots is adequate to maintain growth during seedling development. The persistence of roots at their primordial state promoted seedling recovery from water stress through reactivation of suppressed primordia upon rehydration. Our findings suggest that under well-watered conditions, SRN is not a limiting factor, and excessive number of roots may be costly and maladaptive. Following water stress, lack of substantial root system suppresses growth and rapid recovery of the root system is essential for seedling recovery. This study underscores SRN as key adaptive trait that was reshaped upon domestication. The maintenance of roots at their primordial state during seedling development may be regarded as seedling protective mechanism against water stress.

A pulsed high energy and narrow bandwidth tunable Tm:YLF laser at the milli-Joule level is demonstrated. The spectral bandwidth was narrowed down to 0.15 nm FWHM, while 33 nm of tunability range between 1873 nm and 1906 nm was achieved using a pair of YAG Etalons. The laser was actively Q-switched using an acousto-optic modulator and mJ level pulse energy was measured along the whole tuning range at a repetition rate of 1 kHz. Up to 1.97 mJ of energy per pulse was achieved at a pulse duration of 37 ns at a wavelength of 1879 nm, corresponding to a peak-power of 53.2 kW and at a slope efficiency of 36 &\#x00025;. The combination of both high energy pulsed lasing and spectral tunability, while maintaining narrow bandwidth across the whole tunability range, enhances the laser abilities, which could enable new applications in the sensing, medical and material processing fields.

Treating M. abscessus infection is challenging due to the potent β-lactamase Bla Mab (Beta-lactamase of M. abscessus ) . Avibactam is a non-β-lactam, β-lactamase inhibitor shown to inhibit Bla Mab . We tested whether avibactem can render piperacillin effective against M. Abscessus. In-vitro, avibactam enhanced the activity of piperacillin by 16-32 fold, with no significant effect on meropenem. In an in-vivo Galleria mellonella model, meropenem and piperacillin/avibactam significantly decreased infection burden compared to untreated controls. Neither piperacillin nor avibactam alone had a significant effect.

A sol, aqueous solution-based ink is presented for fabrication of 3D transparent silica glass objects with complex geometries, by a simple 3D printing process conducted at room temperature. The ink combines a hybrid ceramic precursor that can undergo both the photopolymerization reaction and a sol-gel process, both in the solution form, without any particles. The printing is conducted by localized photopolymerization with the use of a low-cost 3D printer. Following printing, upon aging and densifying, the resulting objects convert from a gel to a xerogel and then to a fused silica. The printed objects, which are composed of fused silica, are transparent and have tunable density and refractive indices.

This paper characterizes the preferences over bounded infinite utility streams that satisfy the time-value of money principle and an additivity property, and preferences that in addition are impatient. Based on this characterization, the paper introduces a concept of optimization that is robust to a small imprecision in the specification of the preference, and proves that the set of feasible streams of payoffs of a finite Markov decision process admits such a robust optimization.

Summary Sensitization is a simple form of behavioral plasticity by which an initial stimulus, often signaling danger, leads to increased responsiveness to subsequent stimuli. Cross-modal sensitization is an important feature of arousal in many organisms, yet its molecular and neural mechanisms are incompletely understood. Here we show that in C. elegans, aversive mechanical stimuli lead to both enhanced locomotor activity and sensitization of aversive chemosensory pathways. Both locomotor arousal and cross-modal sensitization depend on the release of FLP-20 neuropeptides from primary mechanosensory neurons and on their receptor FRPR-3. Surprisingly, the critical site of action of FRPR-3 for both sensory and locomotor arousal is RID, a single neuroendocrine cell specialized for the release of neuropeptides that responds to mechanical stimuli in a FLP-20-dependent manner. Thus, FLP-20 peptides function as an afferent arousal signal that conveys mechanosensory information to central neurons that modulate arousal and other behavioral states.

Aflatoxins are poisonous byproducts of the soilborne fungus Aspergillus, involved in the decomposition of plant materials. Aflatoxins can be found in various food products, such as maize, sorghum, millet, rice and wheat. AFB1 is the most toxic of these, classified as a carcinogen and mutagen for both humans and animals. AFB1 has been detected in human cord blood and placenta; however, its toxic effect on sperm is less known. The current study examines sperm responses associated with AFB1 exposure. These included acrosome integrity and function, mitochondrial polarity, DNA fragmentation, fertilization competence and early embryonic development. Spermatozoa were obtained from bull ejaculate and epididymis and capacitated in vitro for 4h with 0, 0.1, 1, 10 and 100μM AFB1. Following capacitation, acrosome reaction (AR) was induced by Ca2+ ionophore. The integrity and functionality of sperm were examined simultaneously by florescent staining. A Halosperm DNA fragmentation kit was used to evaluate DNA integrity. An in-vitro culture system was used to evaluate fertilization competence and blastocyst formation rate, using bovine oocytes. Findings indicate dose-responsive variation among compartments to AFB1 exposure. Sperm viability, expressed by integrity of the plasma membrane, was lower in sperm isolated from ejaculate or epididymis after culturing with AFB1. Exposure to AFB1 reduced the proportion of sperm from the epididymis tail undergoing acrosome reaction induced by Ca2+ ionophore. AFB1 impaired mitochondrial membrane potential (ΔYm) in sperm isolated from ejaculate and the epididymis tail. Exposing ejaculated sperm to AFB1 increased the proportion of sperm with fragmented DNA and reduced the proportion of embryos that cleaved to the 2- to 4-cell stage, 42h postfertilization, however, the proportion of embryos that developed to blastocysts, 7days postfertilization, did not differ among groups. The findings explore the harmful effects of AFB1 on sperm viability, ΔΨm and DNA integrity associated with fertility competence. We postulate that AFB1-induced fragmentation in paternal DNA might have a carryover effect on the quality of developing embryos. Further evaluation for the quality of blastocysts derived from sperm exposed to AFB1 is warranted.