Dynamic relationships among pathways producing hydrocarbons and fatty acids of maize silk cuticular waxes.

The hydrophobic cuticle is the first line of defense between aerial portions of plants and the external environment. On maize (Zea mays L.) silks, the cuticular cutin matrix is infused with cuticular waxes, consisting of a homologous series of very long-chain fatty acids (VLCFAs), aldehydes, and hydrocarbons. Together with VLC fatty-acyl-CoAs (VLCFA-CoAs), these metabolites serve as precursors, intermediates, and end-products of the cuticular wax biosynthetic pathway. To deconvolute the potentially confounding impacts of the change in silk microenvironment and silk development on this pathway, we profiled cuticular waxes on the silks of the inbreds B73 and Mo17, and their reciprocal hybrids. Multivariate interrogation of these metabolite abundance data demonstrates that VLCFA-CoAs and total free VLCFAs are positively correlated with the cuticular wax metabolome, and this metabolome is primarily affected by changes in the silk microenvironment and plant genotype. Moreover, the genotype effect on the pathway explains the increased accumulation of cuticular hydrocarbons with a concomitant reduction in cuticular VLCFA accumulation on B73 silks, suggesting that the conversion of VLCFA-CoAs to hydrocarbons is more effective in B73 than Mo17. Statistical modeling of the ratios between cuticular hydrocarbons and cuticular VLCFAs reveals a significant role of precursor chain length in determining this ratio. This study establishes the complexity of the product-precursor relationships within the silk cuticular wax-producing network by dissecting both the impact of genotype and the allocation of VLCFA-CoA precursors to different biological processes and demonstrates that longer chain VLCFA-CoAs are preferentially utilized for hydrocarbon biosynthesis.

Excretion of triacylglycerol as a matrix lipid facilitating apoplastic accumulation of a lipophilic metabolite shikonin.

Plants produce a large variety of lipophilic metabolites, many of which are secreted by cells and accumulated in apoplasts. These compounds often play a role to protect plants from environmental stresses. However, little is known about how these lipophilic compounds are secreted into apoplastic spaces. In this study, we used shikonin-producing cultured cells of Lithospermum erythrorhizon as an experimental model system to analyze the secretion of lipophilic metabolites, taking advantage of its high production rate and the clear inducibility in culture. Shikonin derivatives are lipophilic red naphthoquinone compounds that accumulate exclusively in apoplastic spaces of these cells and also in the root epidermis of intact plants. Microscopic analysis showed that shikonin is accumulated in the form of numerous particles on the cell wall. Lipidomic analysis showed that L. erythrorhizon cultured cells secrete an appreciable portion of triacylglycerol (24-38% of total triacylglycerol), composed predominantly of saturated fatty acids. Moreover, in vitro reconstitution assay showed that triacylglycerol encapsulates shikonin derivatives with phospholipids to form lipid droplet-like structures. These findings suggest a novel role for triacylglycerol as a matrix lipid, a molecular component involved in the secretion of specialized lipophilic metabolites.

Development of toileting behaviour evaluation for Japanese older patients using wheelchairs in a hospital setting: a validation study.

BACKGROUND: Ageing limits the activities of daily living (ADLs). Among ADLs, a lack of toileting independence causes a decline in the quality of life, mental health, and social participation. Therefore, occupational therapists spend considerable time assessing toileting disability based on various assessment methods for toileting behaviour. However, these assessment methods have issues with the grading levels, number of items, and diseases covered, and they fail to evaluate toileting behaviour accurately and sensitively. Hence, this study developed a Toileting Behaviour Evaluation (TBE) on a 6-point ordinal scale for patients using wheelchairs, with 22 activity components for various diseases. METHODS: This study examined the reliability and validity of the TBE in acute and subacute hospitals in Japan. To this end, two occupational therapists assessed 50 patients for inter-rater reliability at different times and one assessed them twice within 7-10 days for intra-rater reliability using the TBE. Furthermore, occupational therapists assessed 100 patients for internal consistency using the TBE and for concurrent validity using the TBE and Functional Independence Measure (FIM). The patients had been diagnosed with various diseases. This study used the weighted kappa coefficient for statistical analysis of the inter-rater and intra-rater reliability, Cronbach's alpha coefficient for internal consistency, and Spearman's rank correlation coefficient for concurrent validity. We performed all statistical analyses using the IBM SPSS Statistics ver. 25 for Windows. All P-values < 0.05 were considered statistically significant. RESULTS: The minimum weighted kappa coefficients for the inter-rater and intra-rater reliability for each item were 0.67 and 0.79, respectively. Cronbach's alpha was 0.98 for the 22 items. The Spearman's rank correlation coefficient between the mean scores on the TBE and FIM for toilet-related items was 0.74 (P < .01). CONCLUSIONS: The TBE demonstrated good reliability and validity. This means that therapists can use it to identify impaired toileting behaviour. However, the relationship between impairments and each item of toileting behaviour should be explored in future studies. Additionally, studies should examine the creation of a specific index of functions of independence in each toileting behaviour.

[A Case of Complete Remission in a Patient with Synchronous Double Cancers of the Breast and Lung].

The patient is a 74-year-old woman. She had breast cancer(invasive ductal carcinoma, ER[+], PgR[+], HER2[-], Ki-67: 30-40%)and primary right lung cancer with lumbar metastasis, which led to the diagnosis synchronous double cancers of the breast and the lung. We decided to precede surgery for lung cancer because breast cancer was indicated hormonal receptor positive. Breast cancer is treated with anastrozole, thoracoscopic right upper lobectomy was performed for the lung cancer. Radiation therapy was performed for metastatic bone tumors. 13 months later, partial mastectomy sentinel lymph node biopsy performed. The histopathological diagnosis of breast cancer was pT2, pN0, cM0, pStage ⅡA, and histological response was Grade 2a. The remaining breast was treated radiation therapy. The breast cancer has not recurred and is doing well 6 months after surgery. As for primary lung cancer, 19 months have passed since surgery, and the patient is in complete remission without recurrence.

HEAT INDUCIBLE LIPASE1 Remodels Chloroplastic Monogalactosyldiacylglycerol by Liberating α-Linolenic Acid in Arabidopsis Leaves under Heat Stress.

Under heat stress, polyunsaturated acyl groups, such as α-linolenate (18:3) and hexadecatrienoate (16:3), are removed from chloroplastic glycerolipids in various plant species. Here, we showed that a lipase designated HEAT INDUCIBLE LIPASE1 (HIL1) induces the catabolism of monogalactosyldiacylglycerol (MGDG) under heat stress in Arabidopsis thaliana leaves. Using thermotolerance tests, a T-DNA insertion mutant with disrupted HIL1 was shown to have a heat stress-sensitive phenotype. Lipidomic analysis indicated that the decrease of 34:6-MGDG under heat stress was partially impaired in the hil1 mutant. Concomitantly, the heat-induced increment of 54:9-triacylglycerol in the hil1 mutant was 18% lower than that in the wild-type plants. Recombinant HIL1 protein digested MGDG to produce 18:3-free fatty acid (18:3-FFA), but not 18:0- and 16:0-FFAs. A transient assay using fluorescent fusion proteins confirmed chloroplastic localization of HIL1. Transcriptome coexpression network analysis using public databases demonstrated that the HIL1 homolog expression levels in various terrestrial plants are tightly associated with chloroplastic heat stress responses. Thus, HIL1 encodes a chloroplastic MGDG lipase that releases 18:3-FFA in the first committed step of 34:6 (18:3/16:3)-containing galactolipid turnover, suggesting that HIL1 has an important role in the lipid remodeling process induced by heat stress in plants.

Measuring material parameters using a tunable monochromatic terahertz wave source.

We measured complex refractive indices of materials that transmitted well in the terahertz (THz) range using a tunable, monochromatic pulsed THz wave source and a high-sensitivity semiconductor detector. This approach is based on simple monochromatic transmission power measurements. Contrast enhancement by refractive index cladding was employed to measure the complex refractive indices. The experimental results were in good agreement with previously reported values. These results suggested that our approach could be used to quickly analyze material properties, such as surface roughness, and to determine the complex refractive indices of liquids or gases. In addition, as an application of the findings obtained here, we applied an anti-reflection polymer film to the silicon prism that served as an output coupler of the THz wave source. The film reduced surface reflection by 25%.

Structural validity of the mealtime behaviour questionnaire for children with autism spectrum disorder in Japan.

[Purpose] Children with autism spectrum disorder (ASD) exhibit many problematic mealtime behaviours. Currently, there is no process for measuring the mealtime behaviours of children with ASD in Japan. Therefore, we developed the ASD-Mealtime Behaviour Questionnaire (ASD-MBQ) using the results of surveys measuring problematic mealtime behaviours in Japanese children with ASD aged 3-18 years. The objective of this study was to analyse the structural validity of the ASD-MBQ in Japan. [Participants and Methods] We recruited 378 children with ASD aged 3-18 years and performed a confirmatory factor analysis on the ASD-MBQ by using a five-factor structure. [Results] The confirmatory factor analysis demonstrated structural validity (χ2=796.5, degrees of freedom=265, comparative fit index=0.901, root mean square error of approximation [90% confidence interval]=0.073 [0.067-0.079]). [Conclusion] We have demonstrated the structural validity of the ASD-MBQ, which provided useful information for planning interventions and evaluations for children with ASD. Further studies need to consider cut-off score by age and inter-rater reliability.

Evaluation of psychometric properties of the activities of daily living scale of motor function used by caregivers using Rasch analysis.

[Purpose] We developed a prototype version of the Self-Assessment Burden Scale-Motor as a screening questionnaire for the need of care based on the evaluation of patients' activities of daily living. The questionnaire is comprised of seven items, each of which is scored on a 7-point Likert scale. This study aimed to examine the psychometric properties of the questionnaire using Rasch analysis. [Participants and Methods] A total of 200 individuals completed the questionnaire. Rasch analysis to investigate the rating scale structure and examine the structural validity and reliability of the scale. The unidimensionality of the items was examined using the mean square infit values and principal component analysis of residuals. The separation reliability of the scale was also examined. [Results] The rating scale structure can be improved by collapsing several categories (from seven categories to four categories). Unidimensionality was found for seven items. The separation reliability was acceptable for item calibrations and persons. [Conclusion] Inclusion of seven items with a 4-category rating scale was appropriate for the Self-Assessment Burden Scale-Motor questionnaire. Future studies should examine the intra-rater reliability and the criterion-related validity in more depth and develop a new scale to evaluate cognitive function.

[Stage â £ Breast Cancer with Difficulty in Initiating Chemotherapy-A Case Report].

We report a case of breast cancer(T4b[skin], N1, M1[lung], ER-, PR-, HER2 3+)in a 63-year-old woman with liver dysfunction of unknown cause(T-Bil 3.6mg/dL, ALP 3,483 U/L, AST 214 U/L, ALT 320 U/L, g / -GTP 1,943 U/L). Further- more, serum CA19-9(4,670 U/mL)and HbA1c(8.8%)levels were both elevated. First, she underwent chemotherapy with trastuzumab and capecitabine. Subsequently, liver dysfunction relieved gradually. CA19-9 and HbA1c levels were also decreased, but the tumor size was NC. Subsequently, trastuzumab, pertuzumab, and docetaxel were administered, as liver function became normal. The tumor shrank significantly after this treatment. Finally, she underwent mastectomy. Five years after the first visit, she has continued chemotherapy, with lung metastases almost scarred(CR).

Vector beam generation from vertical cavity surface emitting lasers.

Radially and azimuthally polarized beams in a single transverse mode are generated from a commercially available vertical cavity surface emitting laser (VCSEL) in an external cavity with a birefringent rutile lens, of which the c axis is parallel to the optical axis of the cavity, to select favorable polarization. Additionally, a vector Bessel-Gaussian beam is generated from a VCSEL, which is fabricated to oscillate with a linear polarization in a fixed direction in free running, in the same way. These results clearly show the potential ability of VCSELs to generate vector beams, which will be essential to space-division multiplexing in the future optical communication.

[Two Cases of Lung Metastasis from Breast Cancer Successfully Treated with Endocrine Therapy].

We report 2 cases of lung metastasis from breast cancer that were successfully treated with endocrine therapy.Case 1 is a 69-year-old woman with cirrhosis of the liver caused by hepatitis C.She underwent surgery for left breast cancer at the age of 58, and surgery for right breast cancer at the age of 65.Four years later, she was diagnosed with lung metastasis of breast cancer.She received letrozole and the treatment was effective.Because the severity of the pleural effusion increased 3 years later, fulvestrant was subsequently administered.As a result, the patient remained in good health for 1 year.She died 5 years later.Case 2 is a 72-year-old woman who underwent right breast cancer surgery 12 years previously.She complained of respiratory discomfort as a result of right pleural effusion from lung metastasis.She was hospitalized for cancer lymphangitis that had deteriorated.The patient was immediately treated with fulvestrant and her symptoms improved significantly; the pleural effusion also disappeared.Sixteen months later, no recurrence has been observed.

Production of mono- and sesquiterpenes in Camelina sativa oilseed.

MAIN CONCLUSION: Camelina was bioengineered to accumulate (4 S )-limonene and (+)-δ-cadinene in seed. Plastidic localization of the recombinant enzymes resulted in higher yields than cytosolic localization. Overexpressing 1-deoxy- d -xylulose-5-phosphate synthase ( DXS ) further increased terpene accumulation. Many plant-derived compounds of high value for industrial or pharmaceutical applications originate from plant species that are not amenable to cultivation. Biotechnological production in low-input organisms is an attractive alternative. Several microbes are well established as biotechnological production platforms; however, their growth requires fermentation units, energy input, and nutrients. Plant-based production systems potentially allow the generation of high-value compounds on arable land with minimal input. Here we explore whether Camelina sativa (camelina), an emerging low-input non-foodstuff Brassicaceae oilseed crop grown on marginal lands or as a rotation crop on fallow land, can successfully be refactored to produce and store novel compounds in seed. As proof-of-concept, we use the cyclic monoterpene hydrocarbon (4S)-limonene and the bicyclic sesquiterpene hydrocarbon (+)-δ-cadinene, which have potential biofuel and industrial solvent applications. Post-translational translocation of the recombinant enzymes to the plastid with concurrent overexpression of 1-deoxy-D-xylulose-5-phosphate synthase (DXS) resulted in the accumulation of (4S)-limonene and (+)-δ-cadinene up to 7 mg g(-1) seed and 5 mg g(-1) seed, respectively. This study presents the framework for rapid engineering of camelina oilseed production platforms for terpene-based high-value compounds.

[A case of locally advanced breast cancer that responded to preoperative chemotherapy].

A 51-year-old woman presented with a huge tumor in the left breast. The tumor occupied the whole breast, accompanied with a 10-cm wide red discoloration of the skin, subsequently followed by a massive axillary lymph node metastasis. The patient was diagnosed with locally advanced breast cancer, graded as T4cN2aM0, StageIIIB. After preoperative chemotherapy, even though the tumor could not be observed on a CT scan, it was assumed to persist in the skin as discoloration was observed. Bt+Ax+Mj+Mi(level 1+2) and a full-thickness skin graft transplantation were performed. Pathological findings revealed a benign remnant of the active cancer in the skin. Four months after the operation, local recurrence was observed in the skin of the chest wall. Therefore, radiotherapy was performed.

Examination of reliability and validity of the Self-Assessment Burden Scale-Motor for community-dwelling older adults in Japan: a validation study.

Background: The aging society in Japan is progressing rapidly compared with that in the United States and European countries. Aging limits activities of daily living (ADL) in older adults, declining their lives and functions at home. Therefore, improving their ADL to effectively support their functioning at home for as long as possible is vital. Consequently, supporters need to have a common understanding, be promptly aware of the decline in ADL, and quickly introduce rehabilitation. The Functional Independence Measure (FIM) and Barthel Index (BI) are the main scales used to quantitatively assess ADL. However, previous studies have reported that FIM requires specialized knowledge for evaluation, and BI does not appropriately capture changes in ADL. The Self-Assessment Burden Scale-Motor (SAB-M) was developed as a scale for family caregivers to appropriately assess changes in ADL in older adults. Previous studies using the SAB-M have confirmed its reliability and validity in hospitalized patients as assessed by their family caregivers. Therefore, this study aimed to investigate the reliability and validity of the SAB-M among community-dwelling older adults as assessed by their family caregivers. Methods: This study included community-dwelling older adults who received home-visit rehabilitation at the first author's facility between October 2020 and December 2020 in Japan. Following previous studies, the SAB-M was used by family caregivers to assess 20 older adults twice for intra-rater reliability. Furthermore, 168 older adults were evaluated by family caregivers for internal consistency using the SAB-M. For criterion validity, the SAB-M was used for the assessment by family caregivers, and therapists used the FIM-Motor (FIM-M). This study used the weighted kappa, Cronbach's alpha, and Spearman's rank correlation coefficients for the statistical analysis of intra-rater reliability, internal consistency, and criterion validity, respectively. Results: The weighted kappa coefficient for the total score was 0.98 (p < 0.01) and individual item, it was 0.93 for feeding (p < 0.01), 0.91 for bathing (p < 0.01), 0.98 for dressing (p < 0.01), 0.94 for transfer (p < 0.01), 0.94 for walking/wheelchair (p < 0.01), 0.95 for stairs (p < 0.01), and 0.96 for bladder management (p < 0.01). The Cronbach's alpha was 0.93 for the seven items. The Spearman's rank correlation coefficient between the SAB-M and FIM-M scores was 0.91 (p < 0.01). Conclusion: The SAB-M has sufficient reliability and validity among community-dwelling older adults. Family caregivers can routinely assess changes in the ADL of community-dwelling older adults using the SAB-M, enabling them to promptly consider introducing rehabilitation when older adults' ADL declines. Therefore, implementing SAB-M helps older adults live and function at home for as long as possible.

Network analysis for gene discovery in plant-specialized metabolism.

Recent omics technologies provide information on multiple components of biological networks. Web-based data mining tools are continuously being developed. Because genes involved in specialized (secondary) metabolism are often co-ordinately regulated at the transcriptional level, a number of gene discovery studies have been successfully conducted using network analysis, especially by integrating gene co-expression network analysis and metabolomic investigation. In addition, next-generation sequencing technologies are currently utilized in functional genomics investigations of Arabidopsis and non-model plant species including medicinal plants. Systems-based approaches are expected to gain importance in medicinal plant research. This review discussed network analysis in Arabidopsis and gene discovery in plant-specialized metabolism in non-model plants.

Atomic structure of salutaridine reductase from the opium poppy (Papaver somniferum).

The opium poppy (Papaver somniferum L.) is one of the oldest known medicinal plants. In the biosynthetic pathway for morphine and codeine, salutaridine is reduced to salutaridinol by salutaridine reductase (SalR; EC 1.1.1.248) using NADPH as coenzyme. Here, we report the atomic structure of SalR to a resolution of ∼1.9 Šin the presence of NADPH. The core structure is highly homologous to other members of the short chain dehydrogenase/reductase family. The major difference is that the nicotinamide moiety and the substrate-binding pocket are covered by a loop (residues 265-279), on top of which lies a large "flap"-like domain (residues 105-140). This configuration appears to be a combination of the two common structural themes found in other members of the short chain dehydrogenase/reductase family. Previous modeling studies suggested that substrate inhibition is due to mutually exclusive productive and nonproductive modes of substrate binding in the active site. This model was tested via site-directed mutagenesis, and a number of these mutations abrogated substrate inhibition. However, the atomic structure of SalR shows that these mutated residues are instead distributed over a wide area of the enzyme, and many are not in the active site. To explain how residues distal to the active site might affect catalysis, a model is presented whereby SalR may undergo significant conformational changes during catalytic turnover.

Skotomorphogenesis exploits threonine to promote hypocotyl elongation.

Mobilisation of seed storage reserves is important for seedling establishment in Arabidopsis. In this process, sucrose is synthesised from triacylglycerol via core metabolic processes. Mutants with defects in triacylglycerol-to-sucrose conversion display short etiolated seedlings. We found that whereas sucrose content in the indole-3-butyric acid response 10 (ibr10) mutant was significantly reduced, hypocotyl elongation in the dark was unaffected, questioning the role of IBR10 in this process. To dissect the metabolic complexity behind cell elongation, a quantitative-based phenotypic analysis combined with a multi-platform metabolomics approach was applied. We revealed that triacylglycerol and diacylglycerol breakdown were disrupted in ibr10, resulting in low sugar content and poor photosynthetic ability. Importantly, batch-learning self-organised map clustering revealed that threonine level was correlated with hypocotyl length. Consistently, exogenous threonine supply stimulated hypocotyl elongation, indicating that sucrose levels are not always correlated with etiolated seedling length, suggesting the contribution of amino acids in this process.

Transcriptomic, Hormonomic and Metabolomic Analyses Highlighted the Common Modules Related to Photosynthesis, Sugar Metabolism and Cell Division in Parthenocarpic Tomato Fruits during Early Fruit Set.

Parthenocarpy, the pollination-independent fruit set, can raise the productivity of the fruit set even under adverse factors during the reproductive phase. The application of plant hormones stimulates parthenocarpy, but artificial hormones incur extra financial and labour costs to farmers and can induce the formation of deformed fruit. This study examines the performance of parthenocarpic mutants having no transcription factors of SlIAA9 and SlTAP3 and sldella that do not have the protein-coding gene, SlDELLA, in tomato (cv. Micro-Tom). At 0 day after the flowering (DAF) stage and DAFs after pollination, the sliaa9 mutant demonstrated increased pistil development compared to the other two mutants and wild type (WT). In contrast to WT and the other mutants, the sliaa9 mutant with pollination efficiently stimulated the build-up of auxin and GAs after flowering. Alterations in both transcript and metabolite profiles existed for WT with and without pollination, while the three mutants without pollination demonstrated the comparable metabolomic status of pollinated WT. Network analysis showed key modules linked to photosynthesis, sugar metabolism and cell proliferation. Equivalent modules were noticed in the famous parthenocarpic cultivars 'Severianin', particularly for emasculated samples. Our discovery indicates that controlling the genes and metabolites proffers future breeding policies for tomatoes.

Food Lipidomics for 155 Agricultural Plant Products.

Lipids exhibit functional bioactivities based on their polar and acyl chain properties; humans obtain lipids from dietary plant product intake. Therefore, the identification of different molecular species facilitates the evaluation of biological functions and nutrition levels and new phenotype-modulating lipid structures. As a rapid screening strategy, we performed untargeted lipidomics for 155 agricultural products in 58 species from 23 plant families, wherein product-specific lipid diversities were shown using computational mass spectrometry. We characterized 716 lipid species, for which the profiles revealed the National Center for Biotechnology Information-established organismal classification and unique plant tissue metabotypes. Moreover, we annotated unreported subclasses in plant lipidology; e.g., triacylglycerol estolide (TG-EST) was detected in rice seeds (Oryza sativa) and several plant species. TG-EST is known as the precursor molecule producing the fatty acid ester of hydroxy fatty acid, which lowers ambient glycemia and improves glucose tolerance. Hence, our method can identify agricultural plant products containing valuable lipid ingredients.

Crystallization and preliminary X-ray diffraction analysis of salutaridine reductase from the opium poppy Papaver somniferum.

The opium poppy Papaver somniferum is the source of the narcotic analgesics morphine and codeine. Salutaridine reductase (SalR; EC 1.1.1.248) reduces the C-7 keto group of salutaridine to the C-7 (S)-hydroxyl group of salutaridinol in the biosynthetic pathway that leads to morphine in the opium poppy plant. P. somniferum SalR was overproduced in Escherichia coli and purified using cobalt-affinity and size-exclusion chromatography. Hexagonal crystals belonging to space group P6(4)22 or P6(2)22 were obtained using ammonium sulfate as precipitant and diffracted to a resolution of 1.9 A.