Mitochondrial gene expression analysis reveals aberrant transcription of cox3 in Gossypium barbadense CMS line H276A.

Cotton cytoplasmic male sterility (CMS) has been extensively studied; however, information regarding its molecular mechanisms has not yet been disclosed. Therefore, to explore the molecular mechanism of pollen abortion of cotton CMS line H276A, transcript profiles of 30 mitochondrial protein-encoding genes at tetrad stage were conducted with northern blot and a differential expression gene cox3 was identified. Quantitative reverse-transcribed PCR (qRT-PCR) analysis indicated that the expression level of cox3 in the CMS line H276A was 0.39-fold compared to its maintainer line H276B. In addition, the immunoblot analysis revealed that the amount of COX3 protein was decreased to 59.38% in CMS line H276A. The 5` and 3` terminals of the transcript of cox3 in two materials were determined simultaneously with circularized RNA reverse-transcribed PCR (CR-RT-PCR). The data indicated that seven 5` end of transcript of cox3 in H276A (-451/-464/-465/-467/-471/-472/-508 respect to ATG) were identified which were different from that of H276B (-411/-412). A total of 15 single nucleotide polymorphisms (SNPs) was detected by clone sequencing analysis of upstream of cox3. To our knowledge, we are the first to comprehensively analyze the transcripts of the mitochondrial genome in the cotton CMS line and to identify the 5` and 3` terminals of the transcript of cox3 in cotton. Our data will provide a framework for a better understanding of molecular mechanisms of CMS and mitochondrial gene expression in cotton.

Transfer behavior of pesticides from honeysuckle into tea infusions: Establishment of an empirical model for transfer rate prediction.

Affected by some external conditions and internal factors, pesticides can be transferred from tea into its infusion, causing subsequent damage to humans as tea infusion is generally consumed. This study aimed to explore the inherent regularity in transfer behavior of 23 pesticides belonging to different classes from honeysuckle to its tea infusion, and to understand the effects of external brewing conditions and internal physicochemical parameters of the pesticides on their transfer rates. Results indicated that the transfer rates (Rt) of pesticides from honeysuckle into tea solutions increased with prolonged brewing time, or adding a cover on a container, but decreased with increasing the times of infusion. In addition, the transfer potential of these pesticides greatly depended on their physicochemical properties but not their type. The pesticides with high water solubility and low water partition coefficient (LogKow, e.g., omethoate) were more easily transferred into tea infusions than those with low water solubility and high LogKow (e.g., chlorpyrifos). Compared the tea brewing in a covered container, the empirical models obtained in an uncovered cup predicted the transfer behavior and drinking risk of pesticides potentially introduced into honeysuckle and its tea infusion. The linear equation was as follow: Rt = 10.756 LogWS + 7.517, R = 0.8771. In practice, honeysuckle should be brewed in an uncovered cup within a short brewing time, and the first tea infusion should be abandoned to reduce the transfer percentage of pesticides. This study provided beneficial references for pesticide application in honeysuckle plantation to establish realistic maximum residue limits of multi-pesticides in honeysuckle tea and related products.

First report of root rot caused by Fusarium solani on roselle in Nanning, Guangxi, China.

Roselle (Hibiscus sabdariffa L.) is an annual herbaceous plant in the Malvaceae family with high anthocyanin and is widely cultivated in Nanning, Guangxi of China due to its economic and nutritional importance. In August 2021, a severe root rot disease with incidence of 42.4% (860 plants in the field) was observed in roselle plants in an open-field crop in Nanning (108°33"E, 22°84"N), Guangxi, China. The roots of the diseased plants were discolored and rotten, and the xylem became black, extending along the main root to the junction of the rhizomes. The above-ground symptoms were leaf yellowing, vascular tissue browning, wilting and death. Three diseased samples were rinsed thoroughly with sterile distilled water, cut with a sterile scalpel into approximately 0.5-cm pieces, surface disinfested with 75% ethanol for one minute, rinsed 3-4 times with sterile water, and finally incubated on potato dextrose agar (PDA) at 28 °C in the dark for 3 days. Emerging colonies were transferred to new PDA two-three times until a single colony was obtained. The aerial mycelium was initially white, turning pale yellow after 5 days of growth on PDA. Microscopic observations revealed that microconidia were hyaline and ovoid with sizes of 5.13 to 15.12 and 2.50 to 4.20 µm (average 9.02, 3.32 µm, n=30). Macroconidia were falciform with 3- to 4-septate, with sizes of 19.08 to 24.35 and 4.5 to 8.00 µm (average 24.35 and 5.23 µm, n=30). The morphological characteristics of the microscopic images were identical to those described for Fusarium solani (Leslie and Summerell 2006). A representative isolate (GXRST29) was selected for DNA extraction for further characterization. The internal transcribed spacer rRNA regions (ITSs), beta tubulin gene sequence and a fragment of the translation elongation factor 1-alpha (EF 1-α) gene sequence were amplified using the primer pairs ITS1/ITS4 (Chehri 2014), Bt-1/Bt-2 (Wang et al. 2014) and EF1-F/EF2-R (O'Donnell et al. 2010), respectively. PCR products were sequenced and deposited in GenBank (accession Nos. OL314654, ON157430 and ON157431, respectively). BLASTn analysis showed that the ITS sequence had 96.99% homology with sequence of F. solani (NR 163531), and 99.26% for Fusarium cf. solani (MG775565) obtained from Homo sapiens. The beta tubulin sequence had 97.96% similarity with BLAST sequence of F. solani (MN295052.1) and EF 1-α gene had 100% identity to published F. solani (MN977912.1). The fungus was identified as F. solani. Five roselle plants at the 5-leaf stage were artificially inoculated by root dipping into a 106-107-mL-1 spore suspension of the isolated GXRST29 for pathogenicity testing. The experiment was conducted three times, and the negative controls were replaced with sterile water. Compared to the control, the growth of plants was significantly inhibited, leaves turned yellow, plants dwarfed and wilted, and roots decayed three days post-inoculation. One week post-inoculation, all plants exhibited symptoms similar to those observed in the field, and F. solani was steadily reisolated from those diseased plants, while no positive isolations were obtained in the controls. F. solani has been reported to cause root rot on roselle in Upper Egypt (Hassan et al. 2014) and lisianthus in China (Xiao et al. 2018). To our knowledge, however, this is the first report of Fusarium wilt caused by F. solani in roselle plants in Nanning, Guangxi, China, and could result in severe crop losses.

Aptasensors for mycotoxins in foods: Recent advances and future trends.

Mycotoxin contamination in foods has posed serious threat to public health and raised worldwide concern. The development of simple, rapid, facile, and cost-effective methods for mycotoxin detection is of urgent need. Aptamer-based sensors, abbreviated as aptasensors, with excellent recognition capacity to a wide variety of mycotoxins have attracted ever-increasing interest of researchers because of their simple fabrication, rapid response, high sensitivity, low cost, and easy adaptability for in situ measurement. The past few decades have witnessed the rapid advances of aptasensors for mycotoxin detection in foods. Therefore, this review first summarizes the reported aptamer sequences specific for mycotoxins. Then, the recent 5-year advancements in various newly developed aptasensors, which, according to the signal output mode, are divided into electrochemical, optical and photoelectrochemical categories, for mycotoxin detection are comprehensively discussed. A special attention is taken on their strengths and limitations in real-world application. Finally, the current challenges and future perspectives for developing novel highly reliable aptasensors for mycotoxin detection are highlighted, which is expected to provide powerful references for their thorough research and extended applications. Owing to their unique advantages, aptasensors display a fascinating prospect in food field for safety inspection and risk assessment.

Nucleo-cytoplasmic interactions affect the 5' terminal transcription of mitochondrial genes between the isonuclear CMS line UG93A and its maintainer line UG93B of kenaf (Hibiscus cannabinus).

Gene expression and translation in plant mitochondria remain poorly understood due to the complicated transcription of its mRNA. In this study, we report the 5' and 3' RNA extremities and promoters of five mitochondrial genes, atp1, atp4, atp6, atp9, and cox3. The results reveal that four genes (atp1, atp4, atp6, and cox3) are transcribed from multiple initiation sites but with a uniform transcript at the 3' end, indicating that heterogeneity of the 5' end is a common feature in the transcription of kenaf mitochondrial genes. Furthermore, we found that the transcription initiation sites of these four genes are significantly different in UG93A, UG93B, and the F1 hybrid. These data indicate that nuclear loci and unknown transcription factors within the mitochondria of different cytoplasmic types may be involved in mitochondrial transcription. Promoter architecture analysis showed that the promoter core sequences are conserved in the kenaf mitochondrial genome but are highly divergent, suggesting that these elements are essential for the promoter activity of mitochondrial genes in kenaf. Our results reveal that the heterogeneity of the 5' end and uniformity at the 3' end are common transcriptional features of mitochondrial genes. These data provide essential information for understanding the transcription of mitochondrial genes in kenaf and can be used as a reference for other plants.

Identification of differentially expressed genes and pathways in isonuclear kenaf genotypes under salt stress.

Salinity is a potential abiotic stress and globally threatens crop productivity. However, the molecular mechanisms underlying salt stress tolerance with respect to cytoplasmic effect, gene expression, and metabolism pathway under salt stress have not yet been reported in isonuclear kenaf genotypes. To fill this knowledge gap, growth, physiological, biochemical, transcriptome, and cytoplasm changes in kenaf cytoplasmic male sterile (CMS) line (P3A) and its iso-nuclear maintainer line (P3B) under 200 mM sodium chloride (NaCl) stress and control conditions were analyzed. Salt stress significantly reduced leaf soluble protein, soluble sugars, proline, chlorophyll content, antioxidant enzymatic activity, and induced oxidative damage in terms of higher MDA contents in both genotypes. The reduction of these parameters resulted in a reduced plant growth compared with control. However, P3A was relatively more tolerant to salt stress than P3B. This tolerance of P3A was further confirmed by improved physio-biochemical traits under salt stress conditions. Transcriptome analysis showed that 4256 differentially expressed genes (DEGs) between the two genotypes under salt stress were identified. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that photosynthesis, photosynthesis antenna-protein, and plant hormone signal transduction pathways might be associated with the improved NaCl stress tolerance in P3A. Conclusively, P3A cytoplasmic male sterile could be a potential salt-tolerant material for future breeding program of kenaf and can be used for phytoremediation of salt-affected soils. These data provide a valuable resource on the cytoplasmic effect of salt-responsive genes in kenaf and salt stress tolerance in kenaf.

Development of a ZnCdS@ZnS quantum dots-based label-free electrochemiluminescence immunosensor for sensitive determination of aflatoxin B1 in lotus seed.

In this study, we designed a ZnCdS@ZnS quantum dots (QDs)-based label-free electrochemiluminescence (ECL) immunosensor for sensitive determination of aflatoxin B1 (AFB1). A Nafion solution assembled abundant QDs on the surface of a Au electrode as ECL signal probes, with specially coupled anti-AFB1 antibodies as the capturing element. As the reduction reaction between S2O82- in the electrolyte and QDs on the electrode led to ECL emission, the decreased ECL signals resulting from target AFB1 in the samples were recorded for quantification. We evaluated electrochemical impedance spectroscopy and ECL measurements along each step in the construction of the proposed immunosensor. After systematic optimization of crucial parameters, the ECL immunosensor exhibited a good sensitivity, with a low detection limit of 0.01 ng/mL for AFB1 in a wide concentration range of 0.05-100 ng/mL. Testing with lotus seed samples confirmed the satisfactory selectivity, stability, and reproducibility of the developed ECL immunosensor for rapid, efficient, and sensitive detection of AFB1 at trace levels in complex matrices. This study provides a powerful and universal analytical platform for a variety of analytes that can be used in broad applications for real-time analysis, such as food and traditional Chinese medicine safety testing, environmental pollution monitoring, and disease diagnostics. Graphical abstract Development of a ZnCdS@ZnS quantum dots based label-free electrochemiluminescence immunosensor for sensitive detection of aflatoxin B1 in lotus seed.

Cytometric Microbead Magnetic Suspension Array for High-Throughput Ultrasensitive Detection of Aflatoxin B1.

High-throughput and low-cost detection of mycotoxins in complex matrices is becoming increasingly urgent but it is still challenging to perform ultrasensitive analyses. Here we report a green and practical cytometric microbead magnetic suspension array (CBMSA) strategy for rapid and economical detection of aflatoxin B1 (AFB1) in multiple batches of lotus seed samples. The protocol included (1) fabrication of suspension array chips by immobilizing biotin-modified bovine serum albumin-AFB1 (antigen) onto the surface of streptavidin-coated magnetic microbeads in a multiwell array, (2) indirect immunocompetition of antigen and target of AFB1 in lotus seed samples with the specific antibodies, (3) rapid magnetic separation regardless of complex pretreatment steps, and (4) ultrasensitive fluorescence detection of fluorescein isothiocyanate-labeled goat anti-mouse immunoglobulin G (FITC-IgG) probes. After systematic optimization of some crucial parameters, the developed CBMSA assay allowed for ultrasensitive detection of AFB1 with limit of detection as low as 7.8125 pg·kg-1. For high-throughput analysis, the CBMSA technique was capable of on-site co-instantaneous detection of 50-100 samples in one operation within 30 s, only needing a small amount (50 µL) of solution, which is much cheaper, greener, and more user-friendly than conventional techniques. Moreover, CBMSA with magnetic separation is free of multiple centrifugation and cleanup steps to avoid unpredictable loss of targets. Since various capture and fluorescent probes can be randomly constructed and bound onto the surface of magnetic microbeads to establish an ultrasensitive detection system, the CBMSA technique is very promising for more trace analytes in complex matrices and for broad point-of-need applications, such as drug screening and real-time high-throughput analysis.

Comparative Transcriptome Analysis Reveals the Cause for Accumulation of Reactive Oxygen Species During Pollen Abortion in Cytoplasmic Male-Sterile Kenaf Line 722HA.

Cytoplasmic male sterility (CMS) is a maternally inherited trait used for hybrid production in plants, a novel kenaf CMS line 722HA was derived from the thermo-sensitive male-sterile mutant 'HMS' by recurrent backcrossing with 722HB. The line 722HA has great potential for hybrid breeding in kenaf. However, the underlying molecular mechanism that controls pollen abortion in 722HA remains unclear, thus limiting the full utilization of this line. To understand the possible mechanism governing pollen abortion in 722HA, cytological, transcriptomic, and biochemical analyses were carried out to compare the CMS line 722HA and its maintainer line 722HB. Cytological observations of the microspore development revealed premature degradation of the tapetum at the mononuclear stage, which resulted in pollen dysfunction. The k-means clustering analysis of differentially expressed genes (DEGs) revealed that these genes are related to processes associated with the accumulation of reactive oxygen species (ROS), including electron transport chain, F1F0-ATPase proton transport, positive regulation of superoxide dismutase (SOD), hydrogen peroxide catabolic, and oxidation-reduction. Biochemical analysis indicated that ROS-scavenging capability was lower in 722HA than in 722HB, resulting in an accumulation of excess ROS, which is consistent with the transcriptome results. Taken together, these results demonstrate that excessive ROS accumulation may affect the normal development of microspores. Our study provides new insight into the molecular mechanism of pollen abortion in 722HA and will promote further studies of kenaf hybrids.

[Research progress of pretreatment technology for mycotoxin detection in Chinese materia medica and complex matrices].

In the processes of planting, harvest, transport and storage, improper treatment of Chinese materia medica (CMM) and foodstuffs and agricultural products will result in fungal growth and mycotoxins contamination, which will not only directly affect the quality, safety and efficacy of these complex matrices, but also seriously threaten the consumers' health and lives. Therefore, the establishment of high-throughout analytical methods with high sensitivity for the determination of mycotoxins in CMM and foodstuffs and agricultural products at trace levels will provide reliable references for reducing the risk of mycotoxin exposure in humans. Due to the matrix complexity of CMM and foodstuffs and agricultural products, highly-effective pretreatment technologies are necessary for the establishment of such analytical techniques. In this review, the current extraction and purification methods commonly used for the detection of mycotoxins were summarized, the importance of pretreatment techniques for the precise quantification of mycotoxins in complex matrices such as Chinese herbal medicines was highlighted, as well as the development tendency about the pretreatment techniques for mycotoxins in complex matrices in the future was proposed.

Comparative Analysis of the Cytology and Transcriptomes of the Cytoplasmic Male Sterility Line H276A and Its Maintainer Line H276B of Cotton (Gossypium barbadense L.).

In this study, the tetrad stage of microspore development in a new cotton (Gossypium barbadense L.) cytoplasmic male sterility (CMS) line, H276A, was identified using paraffin sections at the abortion stage. To explore the molecular mechanism underlying CMS in cotton, a comparative transcriptome analysis between the CMS line H276A and its maintainer line H276B at the tetrad stage was conducted using an Illumina HiSeq 4000 platform. The comparison of H276A with H276B revealed a total of 64,675 genes, which consisted of 59,255 known and 5420 novel genes. An analysis of the two libraries with a given threshold yielded a total of 3603 differentially expressed genes (DEGs), which included 1363 up- and 2240 down-regulated genes. Gene Ontology (GO) annotation showed that 2171 DEGs were distributed into 38 categories, and a Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that 2683 DEGs were classified into 127 groups. Thirteen DEGs were randomly selected and detected by quantitative reverse-transcribed PCR (qRT-PCR), and the results indicated that the transcriptome sequencing results were reliable. The bioinformatic analysis results in conjunction with previously reported data revealed key DEGs that might be associated with the male sterility features of H276A. Our results provide a comprehensive foundation for understanding anther development and will accelerate the study of the molecular mechanisms of CMS in cotton.

Effects of radiotherapy on nasopharyngeal carcinoma cell invasiveness.

Radiotherapy is widely used in the treatment of nasopharyngeal carcinoma (NPC), whereas its effects on the NPC growth, survival, and metastases have not been completely evaluated. Here, we compared the detected metastatic NPC tissues after radiotherapy (m-NPC) to the resected primary NPC tissues prior to radiotherapy (p-NPC). We detected higher levels of Snail2 protein, but not mRNA in m-NPC, compared to p-NPC. In vitro, a modest irradiation on NPC cells resulted in significant cell death, but increased Snail2 protein, but mRNA levels in the surviving NPC cells. Bioinformatics analyses showed that miR-613, which was significantly decreased in NPC cells after irradiation, targeted the 3'-UTR of Snail2 mRNA to inhibit its translation. Moreover, miR-613 overexpression inhibited Snail2-mediated cell invasiveness, while miR-613 depletion increased Snail2-mediated cell invasiveness in NPC cells. Finally, we detected significantly lower levels of miR-613 in m-NPC, compared to p-NPC. Together our data suggest that although radiotherapy induced NPC cell death, it may increase Snail2-mediated NPC cell invasiveness through downregulating miR-613.

Gel-Based Triboelectric Nanogenerators for Flexible Sensing: Principles, Properties, and Applications.

The rapid development of the Internet of Things and artificial intelligence technologies has increased the need for wearable, portable, and self-powered flexible sensing devices. Triboelectric nanogenerators (TENGs) based on gel materials (with excellent conductivity, mechanical tunability, environmental adaptability, and biocompatibility) are considered an advanced approach for developing a new generation of flexible sensors. This review comprehensively summarizes the recent advances in gel-based TENGs for flexible sensors, covering their principles, properties, and applications. Based on the development requirements for flexible sensors, the working mechanism of gel-based TENGs and the characteristic advantages of gels are introduced. Design strategies for the performance optimization of hydrogel-, organogel-, and aerogel-based TENGs are systematically summarized. In addition, the applications of gel-based TENGs in human motion sensing, tactile sensing, health monitoring, environmental monitoring, human-machine interaction, and other related fields are summarized. Finally, the challenges of gel-based TENGs for flexible sensing are discussed, and feasible strategies are proposed to guide future research.

Sex difference in alcohol consumption associated with colorectal cancer risk in Quzhou, China: A nested case-control study.

Objective: Colorectal cancer (CRC) incidence has been increasing worldwide over time. This study investigated whether drinking was associated with CRC risk. Methods: We designed a case-control study nested in a mass CRC screening program in Quzhou, China. Cases were newly diagnosed CRC in 2020-2022. Controls were randomly sampled using frequency match. Drinking variables included drinking status, frequency, duration, and others. Logistic regressions were used to estimate odds ratio (OR) and 95 % confidence interval (CI). Results: The crude OR (cOR) (95 % CI) of drinking between 153 cases and 650 controls was 1.46 (0.99, 2.16) in current drinkers, 3.31 (1.44, 7.60) in former drinkers, 1.82 (1.21, 2.74) in drinking 6-7 days/week, and 3.48 (1.29, 9.37) in drinking 1-19 years. Stratifying by sex, all drinking variables in women but not all in men were consistently associated with CRC risk. The adjusted OR (aOR) (95 % CI) was 1.01 (0.59, 1.74) in current drinking men, 2.27 (0.78, 6.64) in former drinking men, and 4.24 (1.61, 11.13) in current drinking women. The aOR (95 % CI) of drinking whisky was 0.19 (0.04, 0.83), 1.89 (0.86, 4.17), 2.25 (1.05, 4.83), and 1.82 (0.85, 3.92) in men drinking ≤0.5, >0.5-≤1.0, >1.0-≤1.5, and >1.5 Liter/week (P trend = 0.011), and 3.80 (1.03, 14.00) and 9.92 (2.01, 49.00) in women drinking ≤0.5 and >0.5 Liter/week (P trend = 0.001), respectively. Conclusions: There was sex difference in drinking associated with increased risk of CRC which association was stronger in women than that in men. Men's association between drinking whisky and CRC risk was J-shaped.

A simple and rapid method for isolating high-quality RNA from kenaf with high polysaccharide and polyphenol contents.

The extraction of high-quality RNA from kenaf is essential for genetic and molecular biology research. However, the presence of high levels of polysaccharide and polyphenol compounds in kenaf poses challenges for RNA isolation. We proposed a simplified, time-saving and cost-effective method for isolating high quantities of RNA from various kenaf tissues. This method exhibited superior efficiency in RNA isolation compared with the conventional cetyltrimethylammonium bromide method and demonstrated greater adaptability to different samples than commercial kits. Furthermore, the high-quality RNA obtained from this method was successfully utilized for RT-PCR, real-time RT-PCR and northern blot analysis. Moreover, this proposed protocol also enables the acquisition of both high-quality and -quantity gDNA through RNase A treatment. In addition, the effectiveness of this approach in isolating high-quality RNA from other plant species has been experimentally confirmed.

Multi-mycotoxin detection and human exposure risk assessment in medicinal foods.

Mycotoxin contamination in medicinal foods has attracted increasing global attention. In this study, a simple and sensitive ultrasonication assisted one-step extraction based ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS) method was developed for simultaneous detection of multi-mycotoxins in five kinds of medicinal foods rich in starch. Under optimal conditions, the developed technique displayed excellent analytical performances. Limits of detection and quantitation for the six mycotoxins were 0.04-0.25 ng/mL and 0.10-0.67 ng/mL, respectively. Average recoveries at three fortified levels ranged from 75.33 % to 118.0 %. Real-world application in 103 batches of medicinal foods displayed that 58 samples were positive with one or more mycotoxins at an occurrence rate of 56.31 % (58/103). Coix seed gave the highest positive rate of 96.15 %, followed by Lily (90 %), Chinese yam (50 %), Lotus seed (34.04 %) and Malt (30 %). Zearalenone had the highest positive rate of 28.16 % with contents in 5 Coix seeds exceeding the maximum residue limit (MRL), followed by aflatoxin B1 of 27.18 % (28/103) with contents in 7 Coix seed and 10 Lotus seeds over its MRL, and ochratoxin A (OTA) of 11.65 % with contents in 1 Lotus seed and 5 Lily samples greater than its MRL. Exposure risk assessment indicated that Coix seed and Lotus seeds that were susceptible to aflatoxins posed great threats to human health. Long-term consumption of Lily that was easily contaminated with OTA were also harmful. This work provides a robust platform for multi-mycotoxin monitoring in medicinal foods to protect the consumers from potential health risks.

Antibody responses to an inactivated SARS-CoV-2 vaccine in individuals aged from 50 to 102 years.

Objectives: To assess antibody responses to an inactivated SARS-CoV-2 vaccine in individuals aged 50 and older. Methods: We conducted a post-market cross-sectional seroepidemiology study. We recruited 4,632 vaccinated individuals aged 50 and older, measured their total serum SARS-CoV-2-specific antibody (TA), and collected correlates. The primary outcome was the geometric mean titer (GMT) of TA, and the secondary outcome was the decline of TA with age. Univariate, bivariate, and multivariate analyses were used to examine the associations of the TA GMT with age, and trend analyses were used to test whether their associations were significant. Results: All participants had a detectable TA, which was generally at a low level across all age groups. The TA GMT (95% CI) in AU/mL was 3.05 (2.93, 3.18); the corresponding arithmetic mean (95% CI) was 17.77 (16.13, 19.42) in all participants and 4.33 (3.88, 4.84), 3.86 (3.49, 4.28), 3.24 (2.92, 3.59), 2.77 (2.60, 2.96), and 2.65 (2.48, 2.83) in the age groups of 50-54, 55-59, 60-64, 65-74, and 75 years or older, respectively. The TA GMT decreased with age with a P trend < 0.001. The TA GMT was significantly lower in those with hypertension or diabetes compared to those with neither. Conclusion: The inactivated SARS-CoV-2 vaccine is effective in individuals aged 50 and older. This is the first study that has found an inverse dose-response relationship between ages and the low-level TAs. Older people, especially those with chronic diseases, should get the SARS-CoV-2 vaccine, and their vaccination frequency, dose, and method may need to be different from those of younger people.

Impact of Gender on STEAM Education in Elementary School: From Individuals to Group Compositions.

Gender differences are essential factors influencing collaborative learning at both individual and group levels. However, few studies have systematically investigated their impact on student performance in the innovative context of STEAM education, particularly in the elementary school setting. To address this research need, this study examined the learning behaviors of 91 sixth graders in a STEAM program, who were classified into three gender groupings, namely, boy-only, girl-only, and mixed-gender groups, and further compared their performance in terms of cognition, interaction, and emotion by both gender and gender group type. The results show that, compared to individual gender differences, the gender group type had a greater impact on students' behavioral performance during STEAM education. While all gender groupings had specific advantages, mixed-gender groups proved to be the most preferable, with benefits such as enhanced higher-order thinking, interaction, and emotional expression. Moreover, the study revealed that both boys and girls acted differently when working with the opposite gender in mixed-gender groups. These research findings have several implications for facilitating STEAM learning in co-ed elementary schools.

Single-Molecule Real-Time Sequencing to Explore the Mycobiome Diversity in Malt.

This study determined the composition of fungal communities and characterized the enriched fungal species in raw and roasted malts via the third-generation PacBio-based full-length single-molecule real-time (SMRT) sequencing of the full-length amplicon of the internal transcribed spacer (ITS) region. In total, one kingdom, six phyla, 23 classes, 56 orders, 120 families, 188 genera, 333 species, and 780 operational taxonomic units (OTUs) were detected with satisfactory sequencing depth and sample size. Wickerhamomyces (56%), Cyberlindnera (15%), Dipodascus (12%), and Candida (6.1%) were characterized as the dominant genera in the raw malts, and Aspergillus (35%), Dipodascus (21%), Wickerhamomyces (11%), and Candida (3.5%) in the roasted malts. Aspergillus proliferans, Aspergillus penicillioides, and Wickerhamomyces anomalus represented the crucial biomarkers causing intergroup differences. Correlation analysis regarding environmental factors indicated that the water activity (aw) of the samples affected the composition of the fungal communities in the malts. In practice, special attention should be paid to the mycotoxin-producing fungi, as well as other fungal genera that are inversely correlated with their growth, to ensure the safe use of malt and its end products. IMPORTANCE Fungal contamination and secondary metabolite accumulation in agricultural products represent a global food safety challenge. Although high-throughput sequencing (HTS) is beneficial for explaining fungal communities, it presents disadvantages, such as short reads, species-level resolution, and uncertain identification. This work represents the first attempt to characterize the fungal community diversity, with a particular focus on mycotoxin-producing fungi, in malt via the third-generation PacBio-based full-length SMRT sequencing of the ITS region, aiming to explore and compare the differences between the fungal communities of raw and roasted malts. The research is beneficial for developing effective biological control and conservation measures, including improving the roasting conditions, monitoring the environmental humidity and aw, and effectively eliminating and degrading fungi in the industry chain according to the diverse fungal communities determined, for the safe use of malts and their end products, such as beers. In addition, the third-generation SMRT sequencing technology allows highly efficient analysis of fungal community diversity in complex matrices, yielding fast, high-resolution long reads at the species level. It can be extended to different research fields, updating modern molecular methodology and bioinformatics databases.