CRISPR/Cas9-Mediated Generation of Mutant Lines in Medicago truncatula Indicates a Symbiotic Role of MtLYK10 during Nodule Formation.

CRISPR/Cas9 systems are commonly used for plant genome editing; however, the generation of homozygous mutant lines in Medicago truncatula remains challenging. Here, we present a CRISPR/Cas9-based protocol that allows the efficient generation of M. truncatula mutants. Gene editing was performed for the LysM receptor kinase gene MtLYK10 and two major facilitator superfamily transporter genes. The functionality of CRISPR/Cas9 vectors was tested in Nicotiana benthamiana leaves by editing a co-transformed GUSPlus gene. Transformed M. truncatula leaf explants were regenerated to whole plants at high efficiency (80%). An editing efficiency (frequency of mutations at a given target site) of up to 70% was reached in the regenerated plants. Plants with MtLYK10 knockout mutations were propagated, and three independent homozygous mutant lines were further characterized. No off-target mutations were identified in these lyk10 mutants. Finally, the lyk10 mutants and wild-type plants were compared with respect to the formation of root nodules induced by nitrogen-fixing Sinorhizobium meliloti bacteria. Nodule formation was considerably delayed in the three lyk10 mutant lines. Surprisingly, the size of the rare nodules in mutant plants was higher than in wild-type plants. In conclusion, the symbiotic characterization of lyk10 mutants generated with the developed CRISPR/Cas9 protocol indicated a role of MtLYK10 in nodule formation.

Effect of Different Compatibilizers on the Mechanical, Flame Retardant, and Rheological Properties of Highly Filled Linear Low-Density Polyethylene/Magnesium Hydroxide Composites.

Maleic anhydride-modified homopolymerized polypropylene (PP-g-MAH) and maleic anhydride-modified polyolefin elastomer (POE-g-MAH) were used as bulking agents to improve the poor processing and mechanical properties of highly filled composites due to high filler content. In this study, a series of linear low-density polyethylene (LLDPE)/magnesium hydroxide (MH) composites were prepared by the melt blending method, and the effects of the compatibilizer on the mechanical properties, flame retardancy, and rheological behavior of the composites were investigated. The addition of the compatibilizer decreased the limiting oxygen index (LOI) values of the composites, but they were all greater than 30.00%, which belonged to the flame retardant grade. Mechanical property tests showed that the addition of the compatibilizer significantly increased the tensile and impact strengths of the LLDPE/60MH (MH addition of 60 wt%) composites. Specifically, the addition of 5 wt% POE-g-MAH increased 154.07% and 415.47% compared to the LLDPE/60MH composites, respectively. The rotational rheology test showed that the addition of the compatibilizer could effectively improve the processing flow properties of the composites. However, due to the hydrocarbon structure of the compatibilizer, its flame retardant properties were adversely affected. This study provides a strategy that can improve the processing and mechanical properties of highly filled composites.

An extracellular ß-N-acetylhexosaminidase of Medicago truncatula hydrolyzes chitooligosaccharides and is involved in arbuscular mycorrhizal symbiosis but not required for nodulation.

Establishment of symbiosis between plants and arbuscular mycorrhizal (AM) fungi depends on fungal chitooligosaccharides (COs) and lipo-chitooligosaccharides (LCOs). The latter are also produced by nitrogen-fixing rhizobia to induce nodules on leguminous roots. However, host enzymes regulating structure and levels of these signals remain largely unknown. Here, we analyzed the expression of a ß-N-acetylhexosaminidase gene of Medicago truncatula (MtHEXO2) and biochemically characterized the enzyme. Mutant analysis was performed to study the role of MtHEXO2 during symbiosis. We found that expression of MtHEXO2 is associated with AM symbiosis and nodulation. MtHEXO2 expression in the rhizodermis was upregulated in response to applied chitotetraose, chitoheptaose, and LCOs. M. truncatula mutants deficient in symbiotic signaling did not show induction of MtHEXO2. Subcellular localization analysis indicated that MtHEXO2 is an extracellular protein. Biochemical analysis showed that recombinant MtHEXO2 does not cleave LCOs but can degrade COs into N-acetylglucosamine (GlcNAc). Hexo2 mutants exhibited reduced colonization by AM fungi; however, nodulation was not affected in hexo2 mutants. In conclusion, we identified an enzyme, which inactivates COs and promotes the AM symbiosis. We hypothesize that GlcNAc produced by MtHEXO2 may function as a secondary symbiotic signal.

Effect of Functionalized Polyethylene Wax on the Melt Processing and Properties of Highly Filled Magnesium Hydroxide/Linear Low-Density Polyethylene Composites.

The poor processing and rheological properties of highly filled composites caused by the high loading of fillers can be improved with the use of maleic anhydride grafted polyethylene wax (PEWM) as compatibilizer and lubricant. In this study, two PEWMs with different molecular weights were synthesized by melt grafting, and their compositions and grafting degrees were characterized by Fourier transform infrared (FTIR) spectroscopy and acid-base titration. Subsequently, magnesium hydroxide (MH)/linear low-density polyethylene (LLDPE) composites with 60 wt% of MH were prepared using polyethylene wax (PEW) and PEWM, respectively. The equilibrium torque and melt flow index tests indicate that the processability and fluidity of MH/MAPP/LLDPE composites are significantly improved with the addition of PEWM. The addition of PEWM with a lower molecular weight leads to a substantial reduction in viscosity. The mechanical properties are also increased. The limiting oxygen index (LOI) test and cone calorimeter test (CCT) show that both PEW and PEWM have adverse effects on flame retardancy. This study provides a strategy to simultaneously improve the processability and mechanical properties of highly filled composites.

Nanocatalysts as fast and powerful medical intervention: Bridging cochlear implant therapies and advanced modelling using Hidden Markov Models (HMMs) for effective treatment of inner ear infections.

As human population growth and waste from technologically advanced industries threaten to destabilise our delicate ecological equilibrium, the global spotlight intensifies on environmental contamination and climate-related changes. These challenges extend beyond our external environment and have significant effects on our internal ecosystems. The inner ear, which is responsible for balance and auditory perception, is a prime example. When these sensory mechanisms are impaired, disorders such as deafness can develop. Traditional treatment methods, including systemic antibiotics, are frequently ineffective due to inadequate inner ear penetration. Conventional techniques for administering substances to the inner ear fail to obtain adequate concentrations as well. In this context, cochlear implants laden with nanocatalysts emerge as a promising strategy for the targeted treatment of inner ear infections. Coated with biocompatible nanoparticles containing specific nanocatalysts, these implants can degrade or neutralise contaminants linked to inner ear infections. This method enables the controlled release of nanocatalysts directly at the infection site, thereby maximising therapeutic efficacy and minimising adverse effects. In vivo and in vitro studies have demonstrated that these implants are effective at eliminating infections, reducing inflammation, and fostering tissue regeneration in the ear. This study investigates the application of hidden Markov models (HMMs) to nanocatalyst-loaded cochlear implants. The HMM is trained on surgical phases in order to accurately identify the various phases associated with implant utilisation. This facilitates the precision placement of surgical instruments within the ear, with a location accuracy between 91% and 95% and a standard deviation between 1% and 5% for both sites. In conclusion, nanocatalysts serve as potent medicinal instruments, bridging cochlear implant therapies and advanced modelling utilising hidden Markov models for the effective treatment of inner ear infections. Cochlear implants loaded with nanocatalysts offer a promising method to combat inner ear infections and enhance patient outcomes by addressing the limitations of conventional treatments.

Refining feasibility assessment of endoscopic ear surgery: a radiomics model utilizing machine learning on external auditory canal CT scans.

BACKGROUND: Feasibility assessment of endoscopic ear surgery (EES) relies solely on subjective evaluation by surgeons. OBJECTIVE: Extracting radiomic features from preoperative CT images of the external auditory canal, we aim to classify EES patients into easy and difficult groups and improve accuracy in determining surgery feasibility. METHODS: 85 patients' external auditory canal CT scans were collected and 139 radiomic features were extracted using PyRadiomics. The most relevant features were selected and three machine learning algorithms (logistic regression, support vector machine, and random forest) were compared using K-fold cross-validation (k = 5) to predict surgical feasibility. RESULTS: The best-performing machine learning model, the support vector machine (SVM), was selected to predict the difficulty of EES. The proposed model achieved a high accuracy of 86.5%, and F1 score of 84.6%. The area under the ROC curve was 0.93, indicating good discriminatory power. CONCLUSIONS AND SIGNIFICANCE: The proposed machine learning model provides a reliable and accurate method for classifying patients undergoing otologic surgery based on preoperative imaging data. The model can help clinicians to better prepare for challenging surgical cases and optimize treatment plans for individual patients.

Transcanal endoscopic management of isolated congenital middle ear malformations.

BACKGROUND: A few studies have reported transcanal endoscopic management of isolated congenital middle ear malformations (CMEMs). OBJECTIVE: The purpose of this study is to describe our surgical experience in endoscopic ear surgery for isolated CMEMs and evaluate the surgical effect of hearing reconstruction. METHODS: From January 2017 to January 2022, a retrospective study was performed on 36 patients (37 ears) with isolated CMEMs who all underwent endoscopic surgery. Demographic data, high-resolution computed tomography (HRCT) findings, intraoperative findings, surgical management and audiometric data were recorded. RESULTS: Anomalies were categorized according to the Teunissen and Cremers classification system: 8 ears were categorized as class I, 8 ears as class II, 19 ears as class III and 2 ears as class IV. The air conduction pure tone average (AC-PTA) of 37 cases was 61.5 ± 8.6 dB preoperatively and 29.6 ± 6.9 dB postoperatively (p < 0.001). The mean preoperative air-bone gap (ABG) significantly decreased from 43.1 ± 8.7 dB to 12.8 ± 5.5 dB postoperatively. 36 of 37 cases (97%) met the criteria for successful operation. CONCLUSION: Isolated CMEMs are mainly manifested as aplasia of the stapes' superstructure and dysplasia of the long process of the incus. Transcanal endoscopic surgery seems a safe technique for the management of isolated CMEMs.

The Medicago truncatula hydrolase MtCHIT5b degrades Nod factors of Sinorhizobium meliloti and cooperates with MtNFH1 to regulate the nodule symbiosis.

Nod factors secreted by nitrogen-fixing rhizobia are lipo-chitooligosaccharidic signals required for establishment of the nodule symbiosis with legumes. In Medicago truncatula, the Nod factor hydrolase 1 (MtNFH1) was found to cleave Nod factors of Sinorhizobium meliloti. Here, we report that the class V chitinase MtCHIT5b of M. truncatula expressed in Escherichia coli can release lipodisaccharides from Nod factors. Analysis of M. truncatula mutant plants indicated that MtCHIT5b, together with MtNFH1, degrades S. meliloti Nod factors in the rhizosphere. MtCHIT5b expression was induced by treatment of roots with purified Nod factors or inoculation with rhizobia. MtCHIT5b with a fluorescent tag was detected in the infection pocket of root hairs. Nodulation of a MtCHIT5b knockout mutant was not significantly altered whereas overexpression of MtCHIT5b resulted in fewer nodules. Reduced nodulation was observed when MtCHIT5b and MtNFH1 were simultaneously silenced in RNA interference experiments. Overall, this study shows that nodule formation of M. truncatula is regulated by a second Nod factor cleaving hydrolase in addition to MtNFH1.

Selective Removal of Boron from Aqueous Solutions Using ECH@NGM Aerogels with Excellent Hydrophilic and Mechanical Properties: Performance and Response Surface Methodology Analysis.

The remediation of environmental boron contamination has received extensive research attention. The adsorbent ECH@NGM aerogel with high hydrophilic and mechanical properties was synthesized to remove boron. The ECH@NGM aerogel had a high adsorption capacity of 81.11 mg/g, which was 14.50% higher than that of commercial boron-selective resin Amberlite IRA743. The Freundlich model and pseudo-second-order model described the adsorption behavior well. In addition, the response surface methodology (RSM) could predict the experimental outcomes and optimize the reaction conditions, and X-ray photoelectron spectroscopy (XPS) and control tests were utilized to investigate probable adsorption mechanisms. These data showed that the B ← N coordination bond was the primary adsorption force. The adsorbent had good resistance to interference from coexisting salts, high reusability, good adsorption performance even after five reuse cycles, and a high desorption rate in a relatively short time. The adsorption performance in real brines could be maintained at 80%. Therefore, this work not only provided ECH@NGM aerogels for the removal of boron from brine but also elucidated the main adsorption processes between N-containing adsorbents and boron, facilitating the design of future adsorbents for boron removal.

Transparent dual-band ultraviolet photodetector based on graphene/p-GaN/AlGaN heterojunction.

Versatile applications have driven a desire for dual-band detection that enables seeing objects in multiple wavebands through a single photodetector. In this paper, a concept of using graphene/p-GaN Schottky heterojunction on top of a regular AlGaN-based p-i-n mesa photodiode is reported for achieving solar-/visible-blind dual-band (275 nm and 365 nm) ultraviolet photodetector with high performance. The highly transparent graphene in the front side and the polished sapphire substrate at the back side allows both top illumination and back illumination for the dual band detection. A system limit dark current of 1×10-9 A/cm2 at a negative bias voltage up to -10 V has been achieved, while the maximum detectivity obtained from the detection wavebands of interests at 275 nm and 365 nm are ∼ 9.0 ×1012 cm·Hz1/2/W at -7.5 V and ∼8.0 × 1011 cm·Hz1/2/W at +10 V, respectively. Interestingly, this new type of photodetector is dual-functional, capable of working as either photodiode or photoconductor, when switched by simply adjusting the regimes of bias voltage applied on the devices. By selecting proper bias, the device operation mode would switch between a high-speed photodiode and a high-gain photoconductor. The device exhibits a minimum rise time of ∼210 µs when working as a photodiode and a maximum responsivity of 300 A/W at 6 µW/cm2 when working as a photoconductor. This dual band and multi-functional design would greatly extend the utility of detectors based on nitrides.

Repumping atomic media for an enhanced sensitivity atomic magnetometer.

Atomic vapour magnetometers sense the local magnetic field strength by measuring the resulting precession rate of a well-defined quantum state. An essential prerequisite for this approach is a requirement to drive the media into this quantum state, which is frequently achieved via optical pumping. In real-world alkali-metal atoms, with their multiplicity of ground states, the optical pumping process is necessarily lossy, with a large fraction of the atoms being lost to quantum states that do not contribute to the useful magnetically sensitive signal. This consequently reduces the sensitivity of all optically-pumped atomic sensors. Here we theoretically and experimentally study the population changes of the quantum ground states of 87Rb during optical pumping. We use this understanding to develop a repumping scheme that allows us to increase the number of atoms that are contributing to the useful magnetic sensing output. Unlike prior schemes, our approach delivers this improved sensitivity with significantly suppressed fictitious magnetic fields associated with the repumping, which would otherwise reduce the accuracy of the sensor. When operated at Earth's field strength (∼50µT), the repumped sensor shows a magnetic sensitivity of 200 fT/Hz, that is nearly three times higher than the non-repumped version.

In2S3 Quantum Dots: Preparation, Properties and Optoelectronic Application.

Low-dimensional semiconductors exhibit remarkable performances in many device applications because of their unique physical, electrical, and optical properties. In this paper, we report a novel and facile method to synthesize In2S3 quantum dots (QDs) at atmospheric pressure and room temperature conditions. This involves the reaction of sodium sulfide with indium chloride and using sodium dodecyl sulfate (SDS) as a surfactant to produce In2S3 QDs with excellent crystal quality. The properties of the as-prepared In2S3 QDs were investigated and photodetectors based on the QDs were also fabricated to study the use of the material in optoelectronic applications. The results show that the detectivity of the device stabilizes at ~ 1013 Jones at room temperature under 365 nm ultraviolet light irradiation at reverse bias voltage.

Role of the Nod Factor Hydrolase MtNFH1 in Regulating Nod Factor Levels during Rhizobial Infection and in Mature Nodules of Medicago truncatula.

Establishment of symbiosis between legumes and nitrogen-fixing rhizobia depends on bacterial Nod factors (NFs) that trigger symbiosis-related NF signaling in host plants. NFs are modified oligosaccharides of chitin with a fatty acid moiety. NFs can be cleaved and inactivated by host enzymes, such as MtNFH1 (MEDICAGO TRUNCATULA NOD FACTOR HYDROLASE1). In contrast to related chitinases, MtNFH1 hydrolyzes neither chitin nor chitin fragments, indicating a high cleavage preference for NFs. Here, we provide evidence for a role of MtNFH1 in the symbiosis with Sinorhizobium meliloti Upon rhizobial inoculation, MtNFH1 accumulated at the curled tip of root hairs, in the so-called infection chamber. Mutant analysis revealed that lack of MtNFH1 delayed rhizobial root hair infection, suggesting that excess amounts of NFs negatively affect the initiation of infection threads. MtNFH1 deficiency resulted in nodule hypertrophy and abnormal nodule branching of young nodules. Nodule branching was also stimulated in plants expressing MtNFH1 driven by a tandem CaMV 35S promoter and plants inoculated by a NF-overproducing S. meliloti strain. We suggest that fine-tuning of NF levels by MtNFH1 is necessary for optimal root hair infection as well as for NF-regulated growth of mature nodules.

A single amino acid substitution in a chitinase of the legume Medicago truncatula is sufficient to gain Nod-factor hydrolase activity.

The symbiotic interaction between nitrogen-fixing rhizobia and legumes depends on lipo-chitooligosaccharidic Nod-factors (NFs). The NF hydrolase MtNFH1 of Medicago truncatula is a symbiotic enzyme that hydrolytically inactivates NFs with a C16 : 2 acyl chain produced by the microsymbiont Sinorhizobium meliloti 1021. MtNFH1 is related to class V chitinases (glycoside hydrolase family 18) but lacks chitinase activity. Here, we investigated the substrate specificity of MtNFH1-related proteins. MtCHIT5a and MtCHIT5b of M. truncatula as well as LjCHIT5 of Lotus japonicus showed chitinase activity, suggesting a role in plant defence. The enzymes failed to hydrolyse NFs from S. meliloti. NFs from Rhizobium leguminosarum with a C18 : 4 acyl moiety were neither hydrolysed by these chitinases nor by MtNFH1. Construction of chimeric proteins and further amino acid replacements in MtCHIT5b were performed to identify chitinase variants that gained the ability to hydrolyse NFs. A single serine-to-proline substitution was sufficient to convert MtCHIT5b into an NF-cleaving enzyme. MtNFH1 with the corresponding proline-to-serine substitution failed to hydrolyse NFs. These results are in agreement with a substrate-enzyme model that predicts NF cleavage when the C16 : 2 moiety is placed into a distinct fatty acid-binding cleft. Our findings support the view that MtNFH1 evolved from the ancestral MtCHIT5b by gene duplication and subsequent symbiosis-related neofunctionalization.

Far off-resonance laser frequency stabilization using multipass cells in Faraday rotation spectroscopy.

We propose a far off-resonance laser frequency stabilization method by using multipass cells in Rb Faraday rotation spectroscopy. Based on the detuning equation, if multipass cells with several meters optical path length are used in the conventional Faraday spectroscopy, the detuning of the lock point can be extended much further from the alkali metal resonance. A plate beam splitter was used to generate two different Faraday signals at the same time. The transmitted optical path length was L=50 mm and the reflected optical path length was 2L=100 mm. When the optical path length doubled, the detuning of the lock points moved further away from the atomic resonance. The temperature dependence of the detuning of the lock point was also analyzed. A temperature-insensitive lock point was found near resonance when the cell temperature was between 110°C and 130°C. We achieved an rms fluctuation of 0.9 MHz/23 h at a detuning of 0.5 GHz. A frequency drift of 16 MHz/h at a detuning of -5.6 GHz and 4 MHz/h at a detuning of -5.2 GHz were also obtained for the transmitted and reflected light Faraday signal.

Light intensity stabilization based on the second harmonic of the photoelastic modulator detection in the atomic magnetometer.

The fluctuations of the probe light intensity seriously affect the performance of the sensitive atomic magnetometer. Here we propose a novel method for the intensity stabilization based on the second harmonic component of the photoelastic modulator (PEM) detection in the atomic magnetometer. The method not only could be used to eliminate the intensity fluctuations of the laser source, but also remove the fluctuations from the optical components caused by the environment. A relative fluctuation of the light intensity of 0.035% was achieved and the corresponding fluctuation of the output signal of the atomic magnetometer has decreased about two orders of magnitude from 4.06% to 0.041%. As the scheme proposed here only contains optical devices and does not require additional feedback controlled equipments, it is especially suitable for the integration of the atomic magnetometer.

Study of the operation temperature in the spin-exchange relaxation free magnetometer.

We study the influence of the cell temperature on the sensitivity of the spin-exchange relaxation free (SERF) magnetometer and analyze the possibility of operating at a low temperature. Utilizing a 25 × 25 × 25 mm(3) Cs vapor cell with a heating temperature of 85 °C, which is almost half of the value of potassium, we obtain a linewidth of 1.37 Hz and achieve a magnetic field sensitivity of 55 fT/Hz(1/2) in a single channel. Theoretical analysis shows that fundamental sensitivity limits of this device with an active volume of 1 cm(3) could approach 1 fT/Hz(1/2). Taking advantage of the higher saturated vapor pressure, SERF magnetometer based on Cs opens up the possibility for low cost and portable sensors and is particularly appropriate for lower temperature applications.

Light-shift measurement and suppression in atomic spin gyroscope.

We present a method to determine and suppress the light shift in an atomic spin gyroscope. This method doesn't require additional drive source or frequency modulation, and it is based on the dynamics of an atomic spin gyroscope to determine a clean curve as a function of the frequency of the pump beam that predicts the zero light shift. We experimentally validate the method in a Cs-(129)Xe atomic spin gyroscope and verify the results through numerical simulations. This method can also be applied to an atomic spin magnetometer based on the spin-exchange relaxation-free exchange that experiences light shift. The method is useful for atomic spin devices because it can improve long-term performance and reduce the influence of the laser.