[Analysis of a case of Multiple pterygium syndrome due to a novel variant of CHRNG gene].

OBJECTIVE: To explore the clinical characteristics and genetic etiology of a child with multiple pterygium syndrome (MPS). METHODS: A child with MPS who was treated at the Orthopedics Department of Guangzhou Women and Children's Medical Center Affiliated to Guangzhou Medical University on August 19, 2020 was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and her parents were also collected. Whole exome sequencing (WES) was carried out for the child. Candidate variant was validated by Sanger sequencing of her parents and bioinformatic analysis. RESULTS: The child, an 11-year-old female, had a complain of "scoliosis found 8 years before and aggravated with unequal shoulder height for 1 year". WES results revealed that she has carried a homozygous c.55+1G>C splice variant of the CHRNG gene, for which both of her parents were heterozygous carriers. By bioinformatic analysis, the c.55+1G>C variant has not been recorded by the CNKI, Wanfang data knowledge service platform and HGMG databases. Analysis with Multain online software suggested that the amino acid encoded by this site is highly conserved among various species. As predicted with the CRYP-SKIP online software, the probability of activation and skipping of the potential splice site in exon 1 caused by this variant is 0.30 and 0.70, respectively. The child was diagnosed with MPS. CONCLUSION: The CHRNG gene c.55+1G>C variant probably underlay the MPS in this patient.

A novel mutation in ext2 caused hereditary multiple exostoses through reducing the synthesis of heparan sulfate.

Hereditary multiple exostoses (HME) is a rare skeletal disorder characterized by the formation of multiple benign cartilage-capped tumors, usually in the metaphyseal region of the long bones. Over 70% of HME cases arise from monoallelic mutations in either of the two genes encoding the heparan sulfate (HS) synthesis enzymes, ext1 and ext2. To identify more HME-associated mutations, genomic DNA from members of five independent consanguineous families with HME was sequenced with whole exome sequencing (WES). A novel heterozygous splice site mutation (c.1173+2T>A) in ext2 was detected in all three affected members of family V. Further study showed that the novel mutation caused exon 7 of ext2 mRNA to be skipped during splicing and caused a frameshift after the codon for Arg360, which results in the appearance of new 43 codons, followed by a termination codon. Although the resulting truncated protein was still localized to the Golgi, similar to the full-length EXT2, its HS synthesis activity decreased by 40%. In this study, a novel splice site mutation in ext2 was identified and suggested to be a pathogenic mutation of HME, which may expand the genetic etiology spectrum of HME and may be helpful for clinical genetic counseling and prenatal diagnosis.

Pomegranate Peel as a Source of Bioactive Compounds: A Mini Review on Their Physiological Functions.

The production and consumption of pomegranates have always been increasing owing to their taste and nutrition. However, during fruit processing, a large number of by-products are produced, such as peels and seeds, which can lead to environmental pollution problems if not handled properly. The pomegranate peel takes up about 26-30% of the total weight, while it contains abundant bioactive substances. This paper carries out a mini review of the characterization and physiological functions of key bioactive compounds in pomegranate peel, comprehensively assessing their effects on human health. The overview summarizes the main phenolic substances in pomegranate peel, including tannins, flavonoids, and phenolic acids. Dietary fiber and other bioactive substances such as alkaloids, minerals, and vitamins are also mentioned. These components act as antioxidants by improving oxidative biomarkers and scavenging or neutralizing reactive oxygen species, further contributing to their extensive functions like anti-inflammatory, anti-cancer, antibacterial, and cardiovascular protection. Overall, it is envisaged that through the deeper understanding of bioactive compounds in pomegranate peel, the waste sources can be better reused for physiological applications.

Molecular analysis of an actin gene, CarACT1, from chickpea (Cicer arietinum L.).

Actins are ubiquitous and highly conserved proteins that play key roles in cell formation and cellular activities. In this study, an actin gene was isolated from chickpea for the first time and designated as CarACT1 (for Cicer arietinum L. actin gene 1; Genbank accession no. EU529707). It encoded a putative protein with 377 amino acids and contained five exons and four introns within genomic DNA sequence. CarACT1 was localized in cytoplasm and showed high similarity to other well known actins from various species. Reverse transcription-polymerase chain reaction (RT-PCR) assay proved that CarACT1 transcripts were ubiquitously accumulated in all major organs, such as seedling roots, stems, leaves, flowers, young pods, and seeds, as well as in diverse developmental stages, such as leaf senescence, seed development and germination. Our results suggested that CarACT1 is an actin gene with physiological functions and may be served as a potential reference gene for transcription level of interesting genes in chickpea.

Characterization of a novel legumin alpha-amylase inhibitor from chickpea (Cicer arietinum L.) seeds.

A proteinaceous alpha-amylase inhibitor (CLAI) was purified from Cicer arietinum seeds. It had a molecular mass of 25.947 kDa and inhibited alpha-amylases from plants and mammals. Analysis of the amino acid sequence of a polypeptide from CLAI showed that it was different from other known alpha-amylase inhibitors, but had high identity to legumins from Cicer arietinum (100%) and Vicia faba var. minor (90%).

Comparative analysis of ESTs in response to drought stress in chickpea (C. arietinum L.).

Chickpea (Cicer arietinum L.) is an important pulse crop grown mainly in the arid and semi-arid regions. To identify the water-stress-induced genes, two non-normalized cDNA libraries were constructed from the seedling leaves of a drought-tolerant chickpea cultivar under PEG-treated and -nontreated conditions. About 2500 clones from each library were selected randomly for sequencing analysis. Based on IDEG6 online software analysis, 92 genes were differentially expressed, and these genes were involved in diverse biological progresses, such as metabolism, transcription, signal transduction, protein synthesis and others. Most of the up-regulated genes were related to drought tolerance, and the down-regulated genes were mainly involved in photosynthesis. The differential expression patterns of five functional unigenes were confirmed by quantitative real-time PCR (qPCR). The results will help in understanding the molecular basis of drought tolerance in chickpea.

Chemometric Analysis of the Volatile Compounds Generated by Aspergillus carbonarius Strains Isolated from Grapes and Dried Vine Fruits.

Ochratoxin A (OTA) contamination in grape production is an important problem worldwide. Microbial volatile organic compounds (MVOCs) have been demonstrated as useful tools to identify different toxigenic strains. In this study, Aspergillus carbonarius strains were classified into two groups, moderate toxigenic strains (MT) and high toxigenic strains (HT), according to OTA-forming ability. The MVOCs were analyzed by GC-MS and the data processing was based on untargeted profiling using XCMS Online software. Orthogonal projection to latent structures discriminant analysis (OPLS-DA) was performed using extract ion chromatogram GC-MS datasets. For contrast, quantitative analysis was also performed. Results demonstrated that the performance of the OPLS-DA model of untargeted profiling was better than the quantitative method. Potential markers were successfully discovered by variable importance on projection (VIP) and t-test. (E)-2-octen-1-ol, octanal, 1-octen-3-one, styrene, limonene, methyl-2-phenylacetate and 3 unknown compounds were selected as potential markers for the MT group. Cuparene, (Z)-thujopsene, methyl octanoate and 1 unknown compound were identified as potential markers for the HT groups. Finally, the selected markers were used to construct a supported vector machine classification (SVM-C) model to check classification ability. The models showed good performance with the accuracy of cross-validation and test prediction of 87.93% and 92.00%, respectively.

5000 h Multi-Factor Accelerated Aging Test of FRP Made Transmission Tower: Characterization, Thermal Decomposition and Reaction Kinetics Study.

Three kinds of fiber reinforced plastic (FRP) composites, including modified polyurethane resin (LGD), epoxy resin (E44) and modified unsaturated polyester resin (D33) glass-fiber reinforced plastics, were subjected to a 5000 h multi-factor accelerated aging test according to the power industry standard. To examine aging resistance and thermal stability of transmission towers made by these three composites, relevant bending properties, thermogravimetric analysis (TGA) and derivative thermogravimetry (DTG), activation energy, as well as microscopic morphology were revealed. The results showed that for these composites, bending modulus retention rates were higher than 94% under the aging test and that of the LGD was highest. Additionally, the onset degradation temperature, temperature at maximum rate of weight loss and T5% reduced at 5000 h, with D33 having highest value and lowest decline rate. The activation energy was calculated with the Bagchi, Coats-Redfern and Broido method, respectively. Although the activation energy of all composites decreased after test, the D33, LGD materials had the highest activation energy which enjoys slight decline. Analysis of the whole experimental results suggested that D33 and LGD composites have good aging resistance, whose basic performance could still perform well after 5000 h aging test, so they can be used to composite towers and applied to engineering practice.

Pyrosequencing reveals fungal communities in the rhizosphere of Xinjiang Jujube.

Fungi are important soil components as both decomposers and plant symbionts and play a major role in ecological and biogeochemical processes. However, little is known about the richness and structure of fungal communities. DNA sequencing technologies allow for the direct estimation of microbial community diversity, avoiding culture-based biases. We therefore used 454 pyrosequencing to investigate the fungal communities in the rhizosphere of Xinjiang jujube. We obtained no less than 40,488 internal transcribed spacer (ITS) rDNA reads, the number of each sample was 6943, 6647, 6584, 6550, 6860, and 6904, and we used bioinformatics and multivariate statistics to analyze the results. The index of diversity showed greater richness in the rhizosphere fungal community of a 3-year-old jujube than in that of an 8-year-old jujube. Most operational taxonomic units belonged to Ascomycota, and taxonomic analyses identified Hypocreales as the dominant fungal order. Our results demonstrated that the fungal orders are present in different proportions in different sampling areas. Redundancy analysis (RDA) revealed a significant correlation between soil properties and the abundance of fungal phyla. Our results indicated lower fungal diversity in the rhizosphere of Xinjiang jujube than that reported in other studies, and we hope our findings provide a reference for future research.

Cloning and characterization of a novel NAC family gene CarNAC1 from chickpea (Cicer arietinum L.).

The plant-specific NAC (for NAM, ATAF1,2 and CUC2) proteins have been found to play important roles in plant development and stress responses. In this study, a NAC gene CarNAC1 (for Cicer arietinum L. NAC gene 1) was isolated from a cDNA library constructed with chickpea seedling leaves treated by polyethylene glycol. CarNAC1 encoded a putative protein with 239 amino acids and contained 3 exons and 2 introns within genomic DNA sequence. CarNAC1 had a conserved NAC domain in the N-terminus and the CarNAC1:GFP (green fluorescent protein) fusion protein was localized in the nucleus of onion epidermal cells. Additionally, CarNAC1 exhibited the trans-activation activity which was mapped to the C-terminus. The CarNAC1 transcript was detected in many chickpea organs including seedling leaves, stems, roots, flowers, and young pods, but less accumulated in young seeds. CarNAC1 was induced by leaf age and showed changes in expression during seed development and germination. Furthermore, the expression of CarNAC1 was strongly induced by drought, salt, cold, wounding, H(2)O(2), ethephon, salicylic acid, indole-3-acetic acid, and gibberellin. Our results suggest that CarNAC1 encodes a novel NAC-domain protein and may be a transcriptional activator involved in plant development and various stress responses.

Characterization of a chickpea (Cicer arietinum L.) NAC family gene, CarNAC5, which is both developmentally- and stress-regulated.

It has been documented that the plant-specific NAC (for NAM, ATAF1,2 and CUC2) transcription factors play an important role in plant development and stress responses. In this study, a chickpea NAC gene CarNAC5 (for Cicer arietinum L. NAC gene 5) was isolated from a cDNA library from chickpea leaves treated by polyethylene glycol (PEG). CarNAC5, as a single/low copy gene, contained three exons and two introns within genomic DNA sequence and encoded a polypeptide with 291 amino acids. CarNAC5 protein had a conserved NAC domain in the N-terminus and showed high similarity to other NACs, especially ATAF subgroup members. The CarNAC5:GFP fusion protein was localized in the nucleus of onion epidermal cells. Furthermore, CarNAC5 protein activated the reporter genes LacZ and HIS3 in yeast. The transactivation activity was mapped to the C-terminal region. The transcripts of CarNAC5 appeared in many chickpea tissues including seedling leaves, stems, roots, flowers, seeds and pods, but mostly accumulated in flowers. Meanwhile, CarNAC5 was strongly expressed during seed maturation and in embryos of the early germinating seeds. It was also significantly induced by drought, heat, wounding, salicylic acid (SA), and indole-3-acetic acid (IAA) treatments. Our results suggest that CarNAC5 encodes a novel NAC-domain protein and acts as a transcriptional activator involved in plant developmental regulation and various stress responses.

A NAC transcription factor gene of Chickpea (Cicer arietinum), CarNAC3, is involved in drought stress response and various developmental processes.

NAC transcription factors have been found to play important roles in plant development and responses to environmental stresses. Based on two cDNA libraries constructed from the PEG-treated and -nontreated seedling leaves of chickpea, a NAC gene, CarNAC3, was isolated and characterized. The results indicated that CarNAC3 contained 285 amino acids and had a conserved NAC domain. It was localized in the nucleus and possessed trans-activation activity in the C-terminus. Phylogenetic analysis showed that CarNAC3 belonged to the NAP (NAC-like, activated by APETALA3/PISTILLATA) subgroup of the NAC protein family. CarNAC3 exhibited organ-specific expression and its induction was strongly dependent on leaf age. CarNAC3 showed differential expression patterns during seed development and germination, and could be significantly induced by drought stress, abscisic acid (ABA), ethephon (Et) and indole-3-acetic acid (IAA), but was inhibited by N-6-benzyl-adenine (6-BA). Our data suggest that CarNAC3 may be a transcriptional activator involved in drought stress response and various developmental processes.