Three consecutive cytosolic glycolysis enzymes modulate autophagic flux.

Autophagy serves as an important recycling route for the growth and survival of eukaryotic organisms in nutrient-deficient conditions. Since starvation induces massive changes in the metabolic flux that are coordinated by key metabolic enzymes, specific processing steps of autophagy may be linked with metabolic flux-monitoring enzymes. We attempted to identify carbon metabolic genes that modulate autophagy using VIGS screening of 45 glycolysis- and Calvin-Benson cycle-related genes in Arabidopsis (Arabidopsis thaliana). Here, we report that three consecutive triose-phosphate-processing enzymes involved in cytosolic glycolysis, triose-phosphate-isomerase (TPI), glyceraldehyde-3-phosphate dehydrogenase (GAPC), and phosphoglycerate kinase (PGK), designated TGP, negatively regulate autophagy. Depletion of TGP enzymes causes spontaneous autophagy induction and increases AUTOPHAGY-RELATED 1 (ATG1) kinase activity. TGP enzymes interact with ATG101, a regulatory component of the ATG1 kinase complex. Spontaneous autophagy induction and abnormal growth under insufficient sugar in TGP mutants are suppressed by crossing with the atg101 mutant. Considering that triose-phosphates are photosynthates transported to the cytosol from active chloroplasts, the TGP enzymes would be strategically positioned to monitor the flow of photosynthetic sugars and modulate autophagy accordingly. Collectively, these results suggest that TGP enzymes negatively control autophagy acting upstream of the ATG1 complex, which is critical for seedling development.

RGI-GOLVEN signaling promotes cell surface immune receptor abundance to regulate plant immunity.

Plant immune responses must be tightly controlled for proper allocation of resources for growth and development. In plants, endogenous signaling peptides regulate developmental and growth-related processes. Recent research indicates that some of these peptides also have regulatory functions in the control of plant immune responses. This classifies these peptides as phytocytokines as they show analogies with metazoan cytokines. However, the mechanistic basis for phytocytokine-mediated regulation of plant immunity remains largely elusive. Here, we identify GOLVEN2 (GLV2) peptides as phytocytokines in Arabidopsis thaliana. GLV2 signaling enhances sensitivity of plants to elicitation with immunogenic bacterial elicitors and contributes to resistance against virulent bacterial pathogens. GLV2 is perceived by ROOT MERISTEM GROWTH FACTOR 1 INSENSITIVE (RGI) receptors. RGI mutants show reduced elicitor sensitivity and enhanced susceptibility to bacterial infection. RGI3 forms ligand-induced complexes with the pattern recognition receptor (PRR) FLAGELLIN SENSITIVE 2 (FLS2), suggesting that RGIs are part of PRR signaling platforms. GLV2-RGI signaling promotes PRR abundance independent of transcriptional regulation and controls plant immunity via a previously undescribed mechanism of phytocytokine activity.

Synthesis of Isolated DNA Aptamer and Its Application of AC-Electrothermal Flow-Based Rapid Biosensor for the Detection of Dengue Virus in a Spiked Sample.

Dengue fever is an infectious disease caused by the dengue virus (DENV) and is transmitted by mosquitoes in tropical and subtropical regions. The early detection method at a low cost is essential. To address this, we synthesized the isolated DENV aptamer for fabricating a rapid electrochemical biosensor on a Au interdigitated microgap electrode (AuIMGE). The DENV aptamers were generated using the SELEX (systemic evolution of ligands by exponential enrichment) method for binding to DENV surface envelope proteins. To reduce the manufacturing cost, unnecessary nucleotide sequences were excluded from the isolation process of the DENV aptamer. To reduce the detection time, the alternating current electrothermal flow (ACEF) technique was applied to the fabricated biosensor, which can shorten the detection time to 10 min. The performance of the biosensor was evaluated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). In the diluted DENV protein solution, the linear range of the concentrations was from 1 pM to 1 µM and the LOD was 76.7 fM. Moreover, the proposed biosensor detected DENV in a diluted spiked sample at a linear range of 10-6 to 106 TCID50/mL, while the detection performance was proven with an LOD of 1.74 × 10-7 TCID50/mL along with high selectivity.

Construction of a rapid electrochemical biosensor consisting of a nanozyme/aptamer conjugate for waterborne microcystin detection.

Microcystin-LR (MC-LR) is a hepatotoxin generated by the excessive proliferation of cyanobacteria, which is a threat to humans and wildlife. Therefore, rapid detection of MC-LR is an important challenge. This study describes a rapid electrochemical biosensor comprising nanozymes and aptamers. Alternating current electrothermal flow (ACEF) significantly reduced the MC-LR detection period to 10 min. We also used MnO2/MC-LR aptamer conjugates to improve the sensitivity to MC-LR detection. Here, MnO2 amplified the electrochemical signal and the aptamer showed high selectivity for MC-LR. Under the optimal conditions, the limit of detection (LOD) and selectivity in freshwater were detected using cyclic voltammetry and differential pulse voltammetry. As a result, an LOD of 3.36 pg mL-1 was observed in the linear concentration range of 10 pg mL-1 to 1 µg mL-1. This study quickly and sensitively detected MC-LR in a situation where it causes serious damage worldwide. In addition, the ACEF technology introduction is the first example of MC-LR detection, suggesting a wide range of possibilities for MC-LR biosensors.

Effects of tympanic membrane perforation, middle ear cavity volume, and mastoid aeration on hearing impairment.

AIMS: To investigate the effects of the location and size of tympanic membrane (TM) perforation and middle ear cavity volume on conductive hearing loss in patients with TM perforation. METHODS: Data were collected via a retrospective medical chart review. RESULTS: We enrolled 128 patients with a mean age of 45.6 ± 10.1 years. The mean perforation size was 21.2 ± 8.6% of the TM area, and the mean air-bone gap (ABG) was 20.2 ± 8.6 dB HL on pure tone audiometry. Patients were divided into two groups based on mean ABG. Patients with a large ABG had a significantly larger TM perforation area and smaller mastoid volume. The TM perforation was most commonly located in the central section. However, regression analyses showed that the proportion of the perforated TM area was the only independent predictor of a large ABG (odds ratio, 1.053; 95% confidence interval, 1.022-1.085; p = 0.001). When we analyzed the frequencies in which hearing loss occurred due to TM perforation, we confirmed that hearing loss occurred mainly in the low-frequency range. CONCLUSION: In patients with TM perforation, conductive hearing loss occurs mainly at low frequencies and in proportion to the size of the TM perforation.

Effect of neoadjuvant chemotherapy on breast tissue composition: a longitudinal mammographic study with automated volumetric measurement.

OBJECTIVES: To investigate the effect of neoadjuvant chemotherapy (NAC) on breast tissue composition with mammographic automated volumetric measurement. METHODS: This retrospective study included 168 breast cancer patients who were treated with NAC and underwent serial mammography (pre-treatment, mid-treatment, and post-treatment) between January 2015 and October 2018. Automated volumetric measurements of the contralateral breast volume (BV), fibroglandular volume (FGV), and breast density (BD) were performed using Volpara software. BD grades were divided into 4 groups by Volpara density grade (VDG). The longitudinal changes in BV, FGV, BD, and their associated factors were evaluated. RESULTS: Repeated-measures analysis of variance demonstrated a significant reduction in BV, FGV, and BD over time (p < 0.001, p < 0.001, and p = 0.002, respectively). BV showed a greater reduction in the second half than in the first half (- 28.6 cm3 vs. - 15.2 cm3), BD showed a greater reduction in the first half than in the second half (- 0.8% vs. - 0.1%), and FGV steadily decreased (- 4.6 cm3 and - 3.9 cm3 in the first and second halves). On multivariable linear regression analysis, chemotherapy regimen was associated with BV change between pre- and post-treatment (p = 0.002); age (p = 0.024) and VDG (p = 0.027) were associated with FGV change; age (p = 0.037), VDG (p = 0.002), and chemotherapy regimen (p = 0.003) were associated with BD change. CONCLUSIONS: NAC affects breast tissue composition, reflected as reductions in BV, FGV, and BD. Mammography with automated volumetric measurement can capture quantitative changes in these breast tissue parameters during NAC. KEY POINTS: • Neoadjuvant chemotherapy (NAC) affects breast tissue composition with different patterns of reduction in breast volume, fibroglandular volume, and breast density. • Age, Volpara density grades, and NAC regimen were independent factors associated with breast density change between pre-treatment and post-treatment. • Mammography with automated volumetric measurement enables identification of longitudinal changes in breast tissue composition.

Functional characterization of chaperonin containing T-complex polypeptide-1 and its conserved and novel substrates in Arabidopsis.

Chaperonin containing T-complex polypeptide-1 (CCT) is an evolutionarily conserved chaperonin multi-subunit complex that mediates protein folding in eukaryotes. It is essential for cell growth and survival in yeast and mammals, with diverse substrate proteins. However, only a few studies on plant CCT have been reported to date, due to the essentiality of CCT subunit genes and the large size of the complex. Here, we have investigated the structure and function of the Arabidopsis CCT complex in detail. The plant CCT consisted of eight subunits that assemble to form a high-molecular-mass protein complex, shown by diverse methods. CCT-deficient cells exhibited depletion of cortical microtubules, accompanied by a reduction in cellular α- and ß-tubulin levels due to protein degradation. Cycloheximide-chase assays suggested that CCT is involved in the folding of tubulins in plants. Furthermore, CCT interacted with PPX1, the catalytic subunit of protein phosphatase 4, and may participate in the folding of PPX1 as its substrate. CCT also interacted with Tap46, a regulatory subunit of PP2A family phosphatases, but Tap46 appeared to function in PPX1 stabilization, rather than as a CCT substrate. Collectively, our findings reveal the essential functions of CCT chaperonin in plants and its conserved and novel substrates.

Does strength deficit correlate with shoulder function in patients with rotator cuff tears? Characteristics of massive tears.

BACKGROUND: The correlation between shoulder strength deficits and function in rotator cuff tears remains uncertain. This study aimed to determine the correlation between shoulder strength deficits and shoulder function evaluated by various clinical scoring systems. METHODS: A total of 262 patients (mean age, 59.67 years [standard deviation, 8.06 years]) who underwent full-thickness rotator cuff repair were included. Patients in group I (n = 188) had small to large rotator cuff tears, whereas those in group II (n = 74) had massive rotator cuff tears. Demographic factors, isokinetic test results, and shoulder function evaluated using various scoring systems were obtained. Correlation differences according to severity of the rotator cuff tear were evaluated. RESULTS: We found weak correlations between shoulder strength deficits (peak torque and total work) and clinical outcomes in patients with rotator cuff tears (r = -0.288). For patients in group I (nonmassive tears), we found a weaker correlation (r = -0.242) according to the tear pattern. However, shoulder strength deficits in group II patients (massive tears) were strongly correlated with American Shoulder and Elbow Surgeons (r = -0.598), Constant (r = -0.582), and Short Form 36 (r = -0.511) scores, especially regarding internal rotator strength deficits. CONCLUSIONS: Shoulder strength deficits measured via isokinetic testing and shoulder function were weakly correlated in patients with rotator cuff tears. However, shoulder strength deficits in patients with massive tears considerably worsened shoulder function and systemic disability, but not regional disability. In particular, internal rotator strength deficits were strongly correlated with poor shoulder function.

InsP6-sensitive variants of the Gle1 mRNA export factor rescue growth and fertility defects of the ipk1 low-phytic-acid mutation in Arabidopsis.

Myo-inositol-1,2,3,4,5,6-hexakisphosphate (InsP(6)), also known as phytic acid, accumulates in large quantities in plant seeds, serving as a phosphorus reservoir, but is an animal antinutrient and an important source of water pollution. Here, we report that Gle1 (GLFG lethal 1) in conjunction with InsP(6) functions as an activator of the ATPase/RNA helicase LOS4 (low expression of osmotically responsive genes 4), which is involved in mRNA export in plants, supporting the Gle1-InsP(6)-Dbp5 (LOS4 homolog) paradigm proposed in yeast. Interestingly, plant Gle1 proteins have modifications in several key residues of the InsP(6) binding pocket, which reduce the basicity of the surface charge. Arabidopsis thaliana Gle1 variants containing mutations that increase the basic charge of the InsP(6) binding surface show increased sensitivity to InsP(6) concentrations for the stimulation of LOS4 ATPase activity in vitro. Expression of the Gle1 variants with enhanced InsP(6) sensitivity rescues the mRNA export defect of the ipk1 (inositol 1,3,4,5,6-pentakisphosphate 2-kinase) InsP(6)-deficient mutant and, furthermore, significantly improves vegetative growth, seed yield, and seed performance of the mutant. These results suggest that Gle1 is an important factor responsible for mediating InsP(6) functions in plant growth and reproduction and that Gle1 variants with increased InsP(6) sensitivity may be useful for engineering high-yielding low-phytate crops.

C4/5 foraminal stenosis predicts C5 palsy after expansive open-door laminoplasty.

INTRODUCTION: Laminoplasty is frequently performed in cervical myelopathy patients, but can lead to unexpected postoperative C5 palsy. Although several studies have examined the pathogenesis and prevention of postoperative C5 palsy, many controversies remain and some radiological findings identified as risk factors were not correlated with our outcomes. PURPOSE: This study sought reliable radiological findings that predict C5 palsy after laminoplasty and examined why the predictors of C5 palsy did not apply in our series. METHODS: This study reviewed 116 patients who underwent open-door laminoplasty for cervical spondylotic myelopathy between January 2014 and April 2016, retrospectively. C5 palsy was defined as weakness of the deltoid muscle. We evaluated the incidence of C5 palsy, the preoperative C2-7 Cobb's angle, and Pavlov ratio at the C4/C5 level using simple cervical lateral radiographs. We also evaluated the existence of an ossified posterior longitudinal ligament, the transverse diameter of the C4/5 foramen, and increases in the anteroposterior diameter of the spinal canal at the C4/5 levels after surgery. Furthermore, a high signal intensity on T2-weighted images of the cervical spinal cord preoperatively was noted. RESULTS: Of the 116 patients, 16 cases were excluded and 100 cases were analyzed; postoperative C5 palsy occurred in 8 patients (8%). There were no significant differences between the two groups except the average diameter of the C4/5 foramen. A diameter of the C4/5 foramen of less than 2 mm was significantly related to C5 palsy in the binary logistic regression test. CONCLUSIONS: A smaller diameter of the C4/5 foramen was the only factor significantly correlated with C5 palsy after laminoplasty. The results suggest that C4/5 foraminal stenosis is one of the best predictors of C5 palsy, which might be related to ischemic/reperfusion injury of the C5 root nerve after laminoplasty.

Cell growth defect factor1/chaperone-like protein of POR1 plays a role in stabilization of light-dependent protochlorophyllide oxidoreductase in Nicotiana benthamiana and Arabidopsis.

Angiosperms require light for chlorophyll biosynthesis because one reaction in the pathway, the reduction of protochlorophyllide (Pchlide) to chlorophyllide, is catalyzed by the light-dependent protochlorophyllide oxidoreductase (POR). Here, we report that Cell growth defect factor1 (Cdf1), renamed here as chaperone-like protein of POR1 (CPP1), an essential protein for chloroplast development, plays a role in the regulation of POR stability and function. Cdf1/CPP1 contains a J-like domain and three transmembrane domains, is localized in the thylakoid and envelope membranes, and interacts with POR isoforms in chloroplasts. CPP1 can stabilize POR proteins with its holdase chaperone activity. CPP1 deficiency results in diminished POR protein accumulation and defective chlorophyll synthesis, leading to photobleaching and growth inhibition of plants under light conditions. CPP1 depletion also causes reduced POR accumulation in etioplasts of dark-grown plants and as a result impairs the formation of prolamellar bodies, which subsequently affects chloroplast biogenesis upon illumination. Furthermore, in cyanobacteria, the CPP1 homolog critically regulates POR accumulation and chlorophyll synthesis under high-light conditions, in which the dark-operative Pchlide oxidoreductase is repressed by its oxygen sensitivity. These findings and the ubiquitous presence of CPP1 in oxygenic photosynthetic organisms suggest the conserved nature of CPP1 function in the regulation of POR.

Pescadillo plays an essential role in plant cell growth and survival by modulating ribosome biogenesis.

Pescadillo (PES) is involved in diverse cellular processes such as embryonic development, ribosomal biogenesis, cell proliferation, and gene transcription in yeast and metazoans. In this study, we characterized cellular functions of plant PES in Nicotiana benthamiana, Arabidopsis, and tobacco BY-2 cells. A GFP fusion protein of PES is predominantly localized in the nucleolus, where its localization requires the N-terminal domain of PES. Silencing of plant PES led to growth arrest and acute cell death. PES interacts with plant homologs of BOP1 and WDR12 in the nucleolus, which are also nucleolar proteins involved in ribosome biogenesis of yeast and mammals. PES, BOP1, and WDR12 cofractionated with ribosome subunits. Depletion of any of these proteins led to defective biogenesis of the 60S ribosome large subunits and disruption of nucleolar morphology. PES-deficient plant cells also exhibited delayed maturation of 25S ribosomal RNA and suppressed global translation. During mitosis in tobacco BY-2 cells, PES is associated with the mitotic microtubules, including spindles and phragmoplasts, and PES deficiency disrupted spindle organization and chromosome arrangement. Collectively, these results suggest that plant PES has an essential role in cell growth and survival through its regulation of ribosome biogenesis and mitotic progression.

The PP2A regulatory subunit Tap46, a component of the TOR signaling pathway, modulates growth and metabolism in plants.

Tap42/α4, a regulatory subunit of protein phosphatase 2A, is a downstream effector of the target of rapamycin (TOR) protein kinase, which regulates cell growth in coordination with nutrient and environmental conditions in yeast and mammals. In this study, we characterized the functions and phosphatase regulation of plant Tap46. Depletion of Tap46 resulted in growth arrest and acute plant death with morphological markers of programmed cell death. Tap46 interacted with PP2A and PP2A-like phosphatases PP4 and PP6. Tap46 silencing modulated cellular PP2A activities in a time-dependent fashion similar to TOR silencing. Immunoprecipitated full-length and deletion forms of Arabidopsis thaliana TOR phosphorylated recombinant Tap46 protein in vitro, supporting a functional link between Tap46 and TOR. Tap46 depletion reproduced the signature phenotypes of TOR inactivation, such as dramatic repression of global translation and activation of autophagy and nitrogen mobilization, indicating that Tap46 may act as a positive effector of TOR signaling in controlling those processes. Additionally, Tap46 silencing in tobacco (Nicotiana tabacum) BY-2 cells caused chromatin bridge formation at anaphase, indicating its role in sister chromatid segregation. These findings suggest that Tap46, in conjunction with associated phosphatases, plays an essential role in plant growth and development as a component of the TOR signaling pathway.

Ultrasonographic evaluation of nail matrix topography for preservative nail surgery of nail unit melanoma.

Recently, functional or preservative surgery has been preferred for nail unit melanoma; however, complete resection of the nail unit, particularly the matrix, is challenging because of its complex structure. This study aims to measure the distance of important nail structures through ultrasonography. Herein, 14 patients without nail deformity were included. The length from the nail cuticle to the distal interphalangeal joint (distance X), to the attachment part of the extensor muscle (distance A), to the median proximal end of the nail matrix (distance B), and to the lateral proximal end of the nail matrix (distance C) were measured. In the axial plane, the length from the highest point of the nail plate to the bottom of the distal phalanx (distance Y) and to the lateral tip of the nail plate (distance D) were measured. On the first fingernail, third fingernail, first toenail, and third toenail, the mean ratio A:X, ratio B:X, ratio C:X, and ratio D:Y were 78.6%, 44.3%, 57.2%, 40.1%, and 84.6%; 55.9%, 64.9%, 40.2%, and 66.4%; 35.6%, 50.8%, 34.3%, and 81.9%; and 57.2%, 59.6%, and 31.7%, respectively. Nail units are often invisible to the naked eye; thus, this study will help identify the approximate scope of excision.

Complete genome sequence of Pseudomonas fluorescens IMGN2 isolated from undisturbed soil.

Here, we report the complete genome sequence of Pseudomonas fluorescens IMGN2, highlighting its biocontrol and plant growth-promoting capabilities. The genome analysis reveals genetic features that contribute to its potential in agricultural biotechnology, including genes related to secondary metabolite synthesis and plant-microbe interactions.

Draft genome sequence of Priestia megaterium strain IMGN3 derived from soil.

Priestia megaterium sp. strain IMGN3 was isolated from the soil in South Korea. Here, we report its draft genome sequence, comprising 12 contigs with a total sequence length of 5.64 Mbp. This genome will provide valuable resources for future genomic studies, particularly focusing on plant growth promotion and biocontrol.

Draft genome for Pseudomonas alkylphenolica IMGN1, isolated from soil.

Biocontrol using organisms like Pseudomonas alkylphenolica offers a viable alternative to chemical pesticides, enhancing plant growth and reducing environmental impact. This study details the genome of Pseudomonas alkylphenolica IMGN1, a strain known for promoting plant growth, advancing our understanding of biocontrol mechanisms.

Complete genome sequence of Bacillus paramobilis sp. strain IMGN7 from soil.

Here, we report the complete genome sequence of Bacillus paramobilis sp. strain IMGN7, which was isolated from soil in South Korea. Its complete genome size is 5.28 Mbp. This genome will provide various insights for further studies about their function as biocontrol agents, such as bioremediation and antibiosis.

Draft genome sequence of bacterium Bacillus proteolyticus strain IMGN4 from soil.

Here, we present the draft genome of Bacillus proteolyticus IMGN4, the gram-positive, soil-dwelling bacterium discovered in mountain Maemi, Republic of Korea in May 2019. The assembly resulted in 7 contigs, comprising a total of 6,063,502 base pairs and have 6,115 coding sequences.

Complete genome sequence of bacterium Peribacillus simplex strain IMGN11 from soil.

We report the complete genome sequence of Peribacillus simplex, a spore-forming bacterium originally classified within the Bacillus genus. Peribacillus simplex exhibits antibiotic, plant growth-promoting, and xenobiotic-degrading activities and resistance to environmental contamination. The genome sequence of Peribacillus simplex will provide insights into its capabilities and potential as a biocontrol agent.