Zaxinone Synthase overexpression modulates rice physiology and metabolism, enhancing nutrient uptake, growth and productivity.

The rice Zaxinone Synthase (ZAS) gene encodes a carotenoid cleavage dioxygenase (CCD) that forms the apocarotenoid growth regulator zaxinone in vitro. Here, we generated and characterized constitutive ZAS-overexpressing rice lines, to better understand ZAS role in determining zaxinone content and regulating growth and architecture. ZAS overexpression enhanced endogenous zaxinone level, promoted root growth and increased the number of productive tillers, leading to about 30% higher grain yield per plant. Hormone analysis revealed a decrease in strigolactone (SL) content, which we confirmed by rescuing the high-tillering phenotype through application of a SL analogue. Metabolomics analysis revealed that ZAS overexpressing plants accumulate higher amounts of monosaccharide sugars, in line with transcriptome analysis. Moreover, transgenic plants showed higher carbon (C) assimilation rate and elevated root phosphate, nitrate and sulphate level, enhancing the tolerance towards low phosphate (Pi). Our study confirms ZAS as an important determinant of rice growth and architecture and shows that ZAS regulates hormone homoeostasis and a combination of physiological processes to promote growth and grain yield, which makes this gene an excellent candidate for sustainable crop improvement.

Distinguishing the functions of canonical strigolactones as rhizospheric signals.

Strigolactones (SLs) act as regulators of plant architecture as well as signals in rhizospheric communications. Reduced availability of minerals, particularly phosphorus, leads to an increase in the formation and release of SLs that enable adaptation of root and shoot architecture to nutrient limitation and, simultaneously, attract arbuscular mycorrhizal fungi (AMF) for establishing beneficial symbiosis. Based on their chemical structure, SLs are designated as either canonical or non-canonical; however, the question of whether the two classes are also distinguished in their biological functions remained largely elusive until recently. In this review we summarize the latest advances in SL biosynthesis and highlight new findings pointing to rhizospheric signaling as the major function of canonical SLs.

Strigolactone biosynthesis in rice can occur via a 9-cis-3-OH-10'-apo-ß-carotenal intermediate.

Strigolactones (SLs) play a crucial role in regulating plant architecture and mediating rhizosphere interactions. They are synthesized from all-trans-ß-carotene converted into the intermediate carlactone (CL) via the intermediate 9-cis-ß-apo-10'-carotenal. Recent studies indicate that plants can also synthesize 3-OH-CL from all-trans-ß-zeaxanthin via the intermediate 9-cis-3-OH-ß-apo-10'-carotenal. However, the question of whether plants can form bioactive SLs from 9-cis-3-OH-ß-apo-10'-carotenal remains elusive. In this study, we supplied the 13 C-labeled 9-cis-3-OH-ß-apo-10'-carotenal to rice seedlings and monitored the synthesis of SLs using liquid chromatography-mass spectrometry (LC-MS) and Striga bioassay. We further validated the biological activity of 9-cis-3-OH-ß-apo-10'-carotenal-derived SLs using the ccd7/d17 SL-deficient mutant, which demonstrated increased Striga seed-germinating activity and partial rescue of tiller numbers and plant height. Our results establish 9-cis-3-OH-ß-apo-10'-carotenal as a significant SL biosynthetic intermediate with implications for understanding plant hormonal functions and potential applications in agriculture.

Abscisic acid inhibits germination of Striga seeds and is released by them likely as a rhizospheric signal supporting host infestation.

Seeds of the root parasitic plant Striga hermonthica undergo a conditioning process under humid and warm environments before germinating in response to host-released stimulants, particularly strigolactones (SLs). The plant hormone abscisic acid (ABA) regulates different growth and developmental processes, and stress response; however, its role during Striga seed germination and early interactions with host plants is under-investigated. Here, we show that ABA inhibited Striga seed germination and that hindering its biosynthesis induced conditioning and germination in unconditioned seeds, which was significantly enhanced by treatment with the SL analog rac-GR24. However, the inhibitory effect of ABA remarkably decreased during conditioning, confirming the loss of sensitivity towards ABA in later developmental stages. ABA measurement showed a substantial reduction of its content during the early conditioning stage and a significant increase upon rac-GR24-triggered germination. We observed this increase also in released seed exudates, which was further confirmed by using the Arabidopsis ABA-reporter GUS marker line. Seed exudates of germinated seeds, containing elevated levels of ABA, impaired the germination of surrounding Striga seeds in vitro and promoted root growth of a rice host towards germinated Striga seeds. Application of ABA as a positive control caused similar effects, indicating its function in Striga/Striga and Striga/host communications. In summary, we show that ABA is an essential player during seed dormancy and germination processes in Striga and acts as a rhizospheric signal likely to support host infestation.

Exploring Possible Diagnostic Precancerous Biomarkers for Oral Submucous Fibrosis: A Narrative Review.

Oral submucous fibrosis (OSF) stands as a progressive oral ailment, designated as a potentially malignant disorder. OSF has gained widespread recognition as a significant precursor to malignant transformation. In the pursuit of dependable, straightforward, and non-invasive diagnostic measures for the early detection of oral malignant progression, research has delved into potential diagnostic biomarkers of OSF. This comprehensive review delves into current investigations that explore the correlation between various biomarkers and OSF. The molecular biomarkers of OSF are categorized based on cytology and sampling methods. Moreover, this review encompasses pertinent studies detailing how these biomarkers are acquired and processed. Within this scope, we scrutinize four potential biomarkers that hold the promise of facilitating the development of diagnostic tools for detecting early-stage OSF.

Disruption of the rice 4-DEOXYOROBANCHOL HYDROXYLASE unravels specific functions of canonical strigolactones.

Strigolactones (SLs) regulate many developmental processes, including shoot-branching/tillering, and mediate rhizospheric interactions. SLs originate from carlactone (CL) and are structurally diverse, divided into a canonical and a noncanonical subfamily. Rice contains two canonical SLs, 4-deoxyorobanchol (4DO) and orobanchol (Oro), which are common in different plant species. The cytochrome P450 OsMAX1-900 forms 4DO from CL through repeated oxygenation and ring closure, while the homologous enzyme OsMAX1-1400 hydroxylates 4DO into Oro. To better understand the biological function of 4DO and Oro, we generated CRISPR/Cas9 mutants disrupted in OsMAX1-1400 or in both OsMAX1-900 and OsMAX1-1400. The loss of OsMAX1-1400 activity led to a complete lack of Oro and an accumulation of its precursor 4DO. Moreover, Os1400 mutants showed shorter plant height, panicle and panicle base length, but no tillering phenotype. Hormone quantification and transcriptome analysis of Os1400 mutants revealed elevated auxin levels and changes in the expression of auxin-related, as well as of SL biosynthetic genes. Interestingly, the Os900/1400 double mutant lacking both Oro and 4DO did not show the observed Os1400 architectural phenotypes, indicating their being a result of 4DO accumulation. Treatment of wild-type plants with 4DO confirmed this assumption. A comparison of the Striga seed germinating activity and the mycorrhization of Os900, Os900/1400, and Os1400 loss-of-function mutants demonstrated that the germination activity positively correlates with 4DO content while disrupting OsMAX1-1400 has a negative impact on mycorrhizal symbiosis. Taken together, our paper deciphers the biological function of canonical SLs in rice and reveals their particular contributions to establishing architecture and rhizospheric communications.

New Series of Zaxinone Mimics (MiZax) for Fundamental and Applied Research.

The apocarotenoid zaxinone is a recently discovered regulatory metabolite required for proper rice growth and development. In addition, zaxinone and its two mimics (MiZax3 and MiZax5) were shown to have a remarkable growth-promoting activity on crops and a capability to reduce infestation by the root parasitic plant Striga through decreasing strigolactone (SL) production, suggesting their potential for application in agriculture and horticulture. In the present study, we developed a new series of MiZax via structural modification of the two potent zaxinone mimics (MiZax3 and MiZax5) and evaluated their effect on plant growth and Striga infestation. In general, the structural modifications to MiZax3 and MiZax5 did not additionally improve their overall performance but caused an increase in certain activities. In conclusion, MiZax5 and especially MiZax3 remain the likely most efficient zaxinone mimics for controlling Striga infestation.

Disruption of the cytochrome CYP711A5 gene reveals MAX1 redundancy in rice strigolactone biosynthesis.

Strigolactones (SLs) inhibit shoot branching/tillering and are secreted by plant roots as a signal to attract symbiotic mycorrhizal fungi in the rhizosphere, particularly under phosphate starvation. However, SLs are also hijacked by root parasitic weeds as inducer for the germination of their seeds. There are around 35 natural SLs divided, based on their structures, into canonical and non-canonical SLs. Cytochrome P450 enzymes of the 711 clade, such as MORE AXILLARY GROWTH1 (MAX1) in Arabidopsis, are a major driver of SL structural diversity. Monocots, such as rice, contain several MAX1 homologs that participate in SL biosynthesis. To investigate the function of OsMAX1-1900 in planta, we generated CRISPR/Cas9 mutants disrupted in the corresponding gene. Characterizing of the generated mutants at metabolite and phenotype level suggests that OsMAX1-1900 loss-of-function does neither affect the SL pattern nor rice architecture, indicating functional redundancy among rice MAX1 homologs.

Perspectives on the metabolism of strigolactone rhizospheric signals.

Strigolactones (SLs) are a plant hormone regulating different processes in plant development and adjusting plant's architecture to nutrition availability. Moreover, SLs are released by plants to communicate with beneficial fungi in the rhizosphere where they are, however, abused as chemical cues inducing seed germination of root parasitic weeds, e.g. Striga spp., and guiding them towards host plants in their vicinity. Based on their structure, SLs are divided into canonical and non-canonical SLs. In this perspective, we describe the metabolism of root-released SLs and SL pattern in rice max1-900 mutants, which are affected in the biosynthesis of canonical SLs, and show the accumulation of two putative non-canonical SLs, CL+30 and CL+14. Using max1-900 and SL-deficient d17 rice mutants, we further investigated the metabolism of non-canonical SLs and their possible biological roles. Our results show that the presence and further metabolism of canonical and non-canonical SLs are particularly important for their role in rhizospheric interactions, such as that with root parasitic plants. Hence, we proposed that the root-released SLs are mainly responsible for rhizospheric communications and have low impact on plant architecture, which makes targeted manipulation of root-released SLs an option for rhizospheric engineering.

Canonical strigolactones are not the major determinant of tillering but important rhizospheric signals in rice.

Strigolactones (SLs) are a plant hormone inhibiting shoot branching/tillering and a rhizospheric, chemical signal that triggers seed germination of the noxious root parasitic plant Striga and mediates symbiosis with beneficial arbuscular mycorrhizal fungi. Identifying specific roles of canonical and noncanonical SLs, the two SL subfamilies, is important for developing Striga-resistant cereals and for engineering plant architecture. Here, we report that rice mutants lacking canonical SLs do not show the shoot phenotypes known for SL-deficient plants, exhibiting only a delay in establishing arbuscular mycorrhizal symbiosis, but release exudates with a significantly decreased Striga seed-germinating activity. Blocking the biosynthesis of canonical SLs by TIS108, a specific enzyme inhibitor, significantly lowered Striga infestation without affecting rice growth. These results indicate that canonical SLs are not the determinant of shoot architecture and pave the way for increasing crop resistance by gene editing or chemical treatment.

9-cis-ß-Apo-10'-carotenal is the precursor of strigolactones in planta.

MAIN CONCLUSION: 13C-isotope feeding experiments demonstrate that the apocarotenoid 9-cis-ß-apo-10'-carotenal is the precursor of several strigolactones in rice, providing a direct, in planta evidence for its role in strigolactone biosynthesis. Strigolactones (SLs) are plant hormone that regulates plant architecture and mediates rhizospheric communications. Previous in vitro studies using heterogously produced enzymes unraveled the conversion of all-trans-ß-carotene via the intermediate 9-cis-ß-apo-10'-carotenal into the SL precursor carlactone. However, a direct evidence for the formation of SLs from 9-cis-ß-apo-10'-carotenal is still missing. To provide this evidence, we supplied rice seedlings with 13C-labeled 9-cis-ß-apo-10'-carotenal and analyzed their SLs by LC-MS. Our results show that 9-cis-ß-apo-10'-carotenal is the SL precursor in planta and reveal, for the first time, the application of labeled long-chain apocarotenoids as a promising approach to investigate apocarotenoid metabolism and the genesis of carotenoid-derived growth regulators and signaling molecules.

Evaluation of the Biostimulant Activity of Zaxinone Mimics (MiZax) in Crop Plants.

Global food security is a critical concern that needs practical solutions to feed the expanding human population. A promising approach is the employment of biostimulants to increase crop production. Biostimulants include compounds that boost plant growth. Recently, mimics of zaxinone (MiZax) were shown to have a promising growth-promoting effect in rice (Oryza sativa). In this study, we investigated the effect of MiZax on the growth and yield of three dicot horticultural plants, namely, tomato (Solanum lycopersicum), capsicum (Capsicum annuum), and squash (Cucurbita pepo) in different growth environments, as well as on the growth and development of the monocot date palm (Phoenix dactylifera), an important crop in the Middle East. The application of MiZax significantly enhanced plant height, flower, and branch numbers, fruit size, and total fruit yield in independent field trials from 2020 to 2021. Importantly, the amount of applied MiZax was far less than that used with the commercial compound humic acid, a widely used biostimulant in horticulture. Our results indicate that MiZax have significant application potential to improve the performance and productivity of horticultural crops.

Towards precision sleep medicine: Self-attention GAN as an innovative data augmentation technique for developing personalized automatic sleep scoring classification.

It is very important to have good quality sleep, which can affect aspects such as memory consolidation, emotional regulation, learning, physical development, and quality of life. Diagnosing human sleep quality and problems quickly and accurately is an important issue for human well-being. Therefore, many automatic sleep scoring methods have been proposed. However, the methods have been developed using sleep data from different individuals or groups. The accuracies of these proposed methods might decrease, due to existing individual differences. In this study, the self-attention generative adversarial network (SAGAN) was applied as an advanced data augmentation technique to propose an improved personalized automatic sleep scoring classification. First, the spectrograms were converted from electroencephalography (EEG). Then, SAGAN was used to generate synthesized spectrograms for each subject. Finally, the real and synthesized spectrograms of each subject were utilized to train a personalized classifier. The averaged accuracy and standard deviation of the proposed method are 95.74% and 3.78%, respectively. Compared to the classifier trained with all subjects' training data, the average accuracy increased by 8.08%. The results proved that the generated spectrograms significantly improved the performance of the personalized automatic sleep scoring classification. The contributions of the proposed method were that made the medical staff and subjects save massive medical resources and time for manual recording and scoring.

Protocol for characterizing strigolactones released by plant roots.

The plant hormone strigolactones (SLs) are secreted by plant roots to act as rhizospheric signals. Here, we present a protocol for characterizing plant-released SLs. We first outline all necessary steps required for collection, processing, and analysis of plant root exudates using the C18 column for SL extraction, followed by liquid chromatography-mass spectrometry (LC-MS) for SL quantification. We then describe image processing by SeedQuant, an open-source artificial-intelligence-based software, for measuring the biological activity of SLs in inducing root parasitic plant seed germination. For complete details on the use and execution of this protocol, please refer to Wang et al. (2019) and Braguy et al. (2021).

A New Formulation for Strigolactone Suicidal Germination Agents, towards Successful Striga Management.

Striga hermonthica, a member of the Orobanchaceae family, is an obligate root parasite of staple cereal crops, which poses a tremendous threat to food security, contributing to malnutrition and poverty in many African countries. Depleting Striga seed reservoirs from infested soils is one of the crucial approaches to minimize subterranean damage to crops. The dependency of Striga germination on the host-released strigolactones (SLs) has prompted the development of the "Suicidal Germination" strategy to reduce the accumulated seed bank of Striga. The success of aforementioned strategy depends not only on the activity of the applied SL analogs, but also requires suitable application protocol with simple, efficient, and handy formulation for rain-fed African agriculture. Here, we developed a new formulation "Emulsifiable Concentration (EC)" for the two previously field-assessed SL analogs Methyl phenlactonoate 3 (MP3) and Nijmegen-1. The new EC formulation was evaluated for biological activities under lab, greenhouse, mini-field, and field conditions in comparison to the previously used Atlas G-1086 formulation. The EC formulation of SL analogs showed better activities on Striga germination with lower EC50 and high stability under Lab conditions. Moreover, EC formulated SL analogs at 1.0 µM concentrations reduced 89-99% Striga emergence in greenhouse. The two EC formulated SL analogs showed also a considerable reduction in Striga emergence in mini-field and field experiments. In conclusion, we have successfully developed a desired formulation for applying SL analogs as suicidal agents for large-scale field application. The encouraging results presented in this study pave the way for integrating the suicidal germination approach in sustainable Striga management strategies for African agriculture.

The growth mechanism and intriguing optical and electronic properties of few-layered HfS2.

Due to electronic properties superior to group VIB (Mo and W) transition metal dichalcogenides (TMDs), group IVB (Hf and Zr) TMDs have become intriguing materials in next-generation nanoelectronics. Therefore, the growth of few-layered hafnium disulfide (HfS2) on c-plane sapphire as well as on a SiO2/Si substrate has been demonstrated using chemical vapour deposition (CVD). The structural properties of HfS2 were investigated by recording X-ray diffraction patterns and Raman spectra. The XRD results reveal that the layers are well oriented along the (0001) direction and exhibit the high crystalline quality of HfS2. The Raman spectra confirm the in-plane and out-plane vibration of Hf and S atoms. Moreover, the HfS2 layers exhibit strong absorption in the UV to visible region. The HfS2 layer-based photodetector shows a photoresponsivity of ∼1.6, ∼0.38, and ∼0.21 µA W-1 corresponding to 9, 38, and 68 mW cm-2, respectively under green light illumination and is attributed to the generation of a large number of electron-hole pairs in the active region of the device. Besides, it also exhibits the highly crystalline structure of HfS2 at high deposition temperature. The PL spectrum shows a single peak at ∼1.8 eV and is consistent with the pristine indirect bandgap of HfS2 (∼2 eV). Furthermore, a few layered HfS2 back gate field-effect transistor (FET) is fabricated based on directly grown HfS2 on SiO2/Si, and the device exhibits p-type behaviour. Thus, the controllable and easy growth method opens the latest pathway to synthesize few layered HfS2 on different substrates for various electronic and optoelectronic devices.

Oxidized low-density lipoprotein induced hepatoma-derived growth factor upregulation mediates foam cell formation of cultured rat aortic vascular smooth muscle cells.

Vascular smooth muscle cells (SMCs) are important vascular components that are essential for the regulation of vascular functions during vascular atherosclerogenesis and vascular injury. Oxidized low-density lipoprotein (oxLDL) is known to induce SMC activation and foam cell transformation. This study characterized the role of hepatoma-derived growth factor (HDGF) in oxLDL-induced foam cell formation in cultured primary rat aortic SMCs. OxLDL exposure significantly increased HDGF expression and extracellular release. It also upregulated atherogenic regulators in SMCs, including TLR4, MyD88, LOX-1, and CD36. Exogenous HDGF stimulation not only increased the expression of cognate receptor nucleolin, but also the innate immunity regulators TLR4/MyD88 and lipid metabolism regulators, including LOX-1 and CD36. Oil red O staining showed that HDGF did not initiate, but enhanced oxLDL-driven foam cell formation in SMCs. Further signaling characterization demonstrated that oxLDL evoked activation of PI3K/Akt and p38 MAPK signaling pathways, both of which were involved in the upregulation of HDGF, LOX-1, and CD36 induced by oxLDL. Gene knockdown experiments using LOX-1 targeted siRNA demonstrated that LOX-1 expression was critical for oxLDL-induced HDGF upregulation, while HDGF gene depletion completely abolished oxLDL-triggered TLR4, LOX-1, and CD36 overexpression and foam cell formation in SMCs. These findings strongly suggest that oxLDL-induced HDGF upregulation participates in subsequent LOX-1 and CD36 expression in aortic SMCs and mechanistically contributes to the formation of SMC-derived foam cells. The oxLDL/LOX-1/HDGF axis may serve as a target for anti-atherogenesis therapy.

Hepatoma-derived growth factor enhances osteoblastic transformation of rat aortic vascular smooth muscle cells in vitro.

AIMS: Vascular smooth muscle cells (VSMCs) are important regulators of vascular functions and their conversion to osteoblasts is a key to development of vascular calcification. This study aimed to characterize in vitro effect of hepatoma-derived growth factor (HDGF) on phenotypic conversion of cultured aortic VSMCs into osteoblast-like cells. MATERIALS AND METHODS: Cell proliferation and migration assays were used to examine cell behaviors. Western blotting, alkaline phosphatase activity and calcium staining were used to evaluate osteoblastic marker expression and function, respectively. KEY FINDINGS: Recombinant HDGF treatment enhanced VSMC growth and motility. Treatment of osteogenic medium (OM) increased expression of not only HDGF but also osteoblastic markers, including Runx2 and osteopontin (OPN), while VSMC marker α-smooth muscle actin (α-SMA) declined. Coincidentally, HDGF and OM treatment alone stimulated signaling activities in both PI3K/Akt and MAPK pathways. Conversely, inhibition of Akt and p38 significantly blocked the OM-upregulated HDGF, Runx2, and OPN expression and NF-κB phosphorylation, but did not reversed the α-SMA downregulation, implicating the involvement of Akt and p38 activities in the osteoblastic transformation of VSMCs. Small interfering RNA-mediated HDGF gene silencing effectively prevented the Runx2 and OPN upregulation, alkaline phosphatase activation, and calcium deposition, but did not affect the α-SMA levels in the transformed cells, supporting the involvement of HDGF in regulation of Runx2 and OPN expression. SIGNIFICANCE: In conclusion, in synergism with other osteogenic factor, HDGF may promote the progression of osteobastic transformation of VSMCs via Akt and p38 signaling pathways and contribute to vascular calcification in arteriosclerosis. CHEMICAL COMPOUNDS STUDIED IN THIS STUDY: HDGF (PubChem CID:); LY294002 (PubChem CID: 3973); PD98059 (PubChem CID: 4713); SB203580 (PubChem CID: 176155); SB431542 (PubChem CID: 4521392); SP600125 (PubChem CID: 8515); Wortmannin (PubChem CID: 312145).

Development of a Blood Pressure Measurement Instrument with Active Cuff Pressure Control Schemes.

This paper presents an oscillometric blood pressure (BP) measurement approach based on the active control schemes of cuff pressure. Compared with conventional electronic BP instruments, the novelty of the proposed BP measurement approach is to utilize a variable volume chamber which actively and stably alters the cuff pressure during inflating or deflating cycles. The variable volume chamber is operated with a closed-loop pressure control scheme, and it is activated by controlling the piston position of a single-acting cylinder driven by a screw motor. Therefore, the variable volume chamber could significantly eliminate the air turbulence disturbance during the air injection stage when compared to an air pump mechanism. Furthermore, the proposed active BP measurement approach is capable of measuring BP characteristics, including systolic blood pressure (SBP) and diastolic blood pressure (DBP), during the inflating cycle. Two modes of air injection measurement (AIM) and accurate dual-way measurement (ADM) were proposed. According to the healthy subject experiment results, AIM reduced 34.21% and ADM reduced 15.78% of the measurement time when compared to a commercial BP monitor. Furthermore, the ADM performed much consistently (i.e., less standard deviation) in the measurements when compared to a commercial BP monitor.

Comparison of healthcare utilization and life-sustaining interventions between elderly patients with dementia and those with cancer near the end of life: A nationwide, population-based study in Taiwan.

AIM: Little is known about the pattern of healthcare services for end-of-life patients with dementia (PwD) in East Asia. We compared this pattern between PwD and cancer patients in their last year of life in Taiwan. METHODS: Taiwan's National Health Insurance Research Database was applied for this case-control analysis. The records of patients who had dementia and died between 2002 and 2011 were reviewed. The control group was decedents with cancer. The utilization of hospitalization, emergency department visits and life-sustaining interventions during the last year of life between the two groups were compared. RESULTS: Of the 2724 patients enrolled, 908 had dementia and 1816 had cancer. PwD were more likely to have a higher frequency of admission to hospital and intensive care unit, and longer stays compared with cancer patients. PwD had a higher risk of enteral tube insertion and feeding, endotracheal intubation and tracheostomy, mechanical ventilation, hemodialysis and cardiopulmonary resuscitation (OR 4.36, 95% CI 3.51-5.41), which was the highest among the selected procedures. CONCLUSIONS: PwD in their last year of life in Taiwan underwent aggressive interventions significantly more frequently than did their counterparts in Western countries. Providing comfort-centered care for better quality of life for end-of-life PwD is a priority of Taiwan's national health policy. Geriatr Gerontol Int 2017; 17: 2545-2551.