Preparation, characterization, and biological activity of Cinnamomum cassia essential oil nano-emulsion.

To solve the problems of low bioavailability and unstable properties of Cinnamomum cassia Essential oil (CCEO), encapsulation technology was introduced as an effective means to improve its shortcomings. In this study, Cinnamomum cassia Essential oil nano-emulsion (CCEO-NE) was successfully synthesized by the oil-in-water method and characterized by standard analytical methods, including dynamic light scattering (DLS), Scanning electron microscopy (SEM), and Transmission electron microscopy (TEM). The results show that the synthesized CCEO is spherical, smooth in surface, and uniform in shape, with an average particle size of 221.8 ± 1.95 nm, which is amorphous. In this experiment, by simulating the digestion of CCEO-NE in the gastrointestinal tract, it was found that CCEO-NE was undigested in the oral cavity, mainly in the stomach, followed by the small intestine. By understanding the digestion of CCEO-NE, we can improve the potential of CCEO bioavailability in food and drug applications. In addition, through the study of ABTS and DPPH free radicals by CCEO and CCEO-NE, it was found that the antioxidant activity of CCEO-NE was more potent than that of CCEO. When the concentration of CCEO-NE and CCEO is 400 µg/mL, the DPPH free radical scavenging rate is 92.03 ± 0.548% and 80.46 ± 5.811%, respectively. In comparison, ABTS free radical scavenging rate is 90.35 ± 0.480% and 98.44 ± 0.170% when the concentration of CCEO- NE, and CCEO is 75 µg/mL, respectively. The antibacterial test shows that CCEO-NE can inhibit both Gram-positive and Gram-negative bacteria. Among them, CCEO-NE has a stronger antibacterial ability than CCEO, and the maximum inhibition zone diameter of CCEO can reach 15 mm, while that of CCEO-NE can reach 18 mm. Meanwhile, SEM and TEM showed that CCEO-NE treatment destroyed the ultrastructure of bacteria. Generally speaking, we know the situation of CCEO in the gastrointestinal tract. CCEO-NE has more potent antioxidant and antibacterial ability than CCEO. Our research results show that whey protein is an effective packaging strategy that can improve the effectiveness, stability, and even bioavailability of CCEO in various applications, including food and health care industries.

SlBES1 promotes tomato fruit softening through transcriptional inhibition of PMEU1.

Fruit softening indicated by firmness determines the texture, transportability, and shelf life of tomato products. However, the regulatory mechanism underlying firmness formation in tomato fruit is poorly understood. Here, we report the regulatory role of SlBES1, an essential component of brassinosteroid hormone signaling, in tomato fruit softening. We found that SlBES1 promotes fruit softening during tomato fruit ripening and postharvest storage. RNA-seq analysis suggested that PMEU1, which encodes a pectin methylesterase, might participate in SlBES1-mediated softening. Biochemical and immunofluorescence assays indicated that SlBES1 inhibited PMEU1-related pectin de-methylesterification. Further molecular and genetic evidence verified that SlBES1 directly binds to the E-box of PMEU1 to repress its expression, leading to fruits softening. Loss-of-function SlBES1 mutant generated by CRISPR-Cas9 showed firmer fruits and longer shelf life during postharvest storage without other quality alteration. Collectively, our results indicated the potential of manipulating SlBES1 to regulate firmness without negative consequence on visual and nutrition quality.

Regulation of fruit ripening by the brassinosteroid biosynthetic gene SlCYP90B3 via an ethylene-dependent pathway in tomato.

The essential role of ethylene in fruit ripening has been thoroughly studied. However, the involvement of brassinosteroids (BRs) in the regulation of fruit ripening and their relationship with the ethylene pathway are poorly understood. In the current study, we found that BRs were actively synthesized during tomato fruit ripening. We then generated transgenic lines overexpressing or silencing SlCYP90B3, which encodes a cytochrome P450 monooxygenase that catalyzes the rate-limiting step of BR synthesis. The expression level of SlCYP90B3 was positively related to the contents of bioactive BRs as well as the ripening process in tomato fruit, including enhanced softening and increased soluble sugar and flavor volatile contents. Both carotenoid accumulation and ethylene production were strongly correlated with the expression level of SlCYP90B3, corroborated by the altered expression of carotenoid biosynthetic genes as well as ethylene pathway genes in transgenic tomato fruits. However, the application of the ethylene perception inhibitor 1-methycyclopropene (1-MCP) abolished the promotion effect of SlCYP90B3 overexpression on carotenoid accumulation. Taken together, these results increase our understanding of the involvement of SlCYP90B3 in bioactive BR biosynthesis as well as fruit ripening in tomato, thus making SlCYP90B3 a target gene for improvement of visual, nutritional and flavor qualities of tomato fruits with no yield penalty.

Chorea and the effectiveness of steroids in a patient with the syndrome of transient headache with neurologic deficits and cerebrospinal fluid lymphocytosis: a case report.

BACKGROUND: The clinical manifestations of the transient headache and neurologic deficits with cerebrospinal fluid lymphocytosis (HaNDL) syndrome are variable, and involuntary movements have not been reported. The etiology and treatment of the syndrome are not entirely clarified. CASE: A 25-year-old female presented with recurrent transient headache, involuntary movements (left upper extremity chorea) and paralysis. Lumbar punctures showed intracranial hypertension and cerebrospinal fluid pleocytosis. Symptoms and intracranial hypertension were relieved after administration of steroids. DISCUSSION/CONCLUSION: Chorea may also present in the HaNDL syndrome. Steroids might relieve the symptoms of the HaNDL syndrome, which indicates the possible treatment strategy for HaNDL and supports the infectious or postinfectious autoimmune etiology hypothesis.

Identification of a novel mutation in PLA2G6 gene and phenotypic heterogeneity analysis of PLA2G6-related neurodegeneration.

INTRODUCTION: This study reports a novel mutation site of the phospholipase A2 group VI (PLA2G6) gene, and analyzes the information of 67 previously published cases to elucidate PLA2G6 phenotype-genotype variations. METHODS: We collected clinical data and examined gene mutation sites from one Chinese patient with adult-onset ataxia and her family. Next-generation sequencing (NGS) and Sanger sequencing were used to verify possible mutations. PolyPhen-2, SIFT, and MutationTaster were used to predict their pathogenicity. For analyzing the distribution frequency of the mutation, 597 healthy controls were recruited. We also analyzed the clinical and genetic information of 67 cases from 23 studies in Pubmed database. RESULTS: A novel compound heterozygous mutation of the PLA2G6 gene, c.1648delC and c.991G > T, was found in the Chinese patient, and classified as pathogenic. The c.1648delC variation was absent in ExAC, 1000G, dbSNP databases and the 597 healthy controls. Of the 67 cases, 29 presented ataxia. The signs of cerebellar atrophy appeared in the MRIs of most patients, while signs of iron accumulation were absent in older-aged patients with a compound heterozygous mutation. Thirty-eight patients showed no ataxia. A negative or mild extrapyramidal symptom accompanied by a low age, a homogenous mutation, while moderate or severe extrapyramidal symptoms were associated with an old age and a compound heterozygous mutation. CONCLUSION: A novel compound heterozygous mutation of the PLA2G6 gene, c.1648delC and c.991G > T, is associated with adult onset ataxia. Phenotype-genotype variations of PLA2G6 are predicted to be caused by the loss of protein or enzyme activity of phospholipase-2.

Comparative transcriptomics uncovers alternative splicing and molecular marker development in radish (Raphanus sativus L.).

BACKGROUND: Alternative splicing (AS) plays important roles in gene expression and proteome diversity. Single nucleotide polymorphism (SNP) and insertion/deletion (InDel) are abundant polymorphisms and co-dominant inheritance markers, which have been widely used in germplasm identification, genetic mapping and marker-assisted selection in plants. So far, however, little information is available on utilization of AS events and development of SNP and InDel markers from transcriptome in radish. RESULTS: In this study, three radish transcriptome datasets were collected and aligned to the reference radish genome. A total of 56,530 AS events were identified from three radish genotypes with intron retention (IR) being the most frequent AS type, which accounted for 59.4% of the total expressed genes in radish. In all, 22,412 SNPs and 9436 InDels were identified with an average frequency of 1 SNP/17.9 kb and 1 InDel/42.5 kb, respectively. A total of 43,680 potential SSRs were identified in 31,604 assembled unigenes with a density of 1 SSR/2.5 kb. The ratio of SNPs with nonsynonymous/synonymous mutations was 1.05:1. Moreover, 35 SNPs and 200 InDels were randomly selected and validated by Sanger sequencing, 83.9% of the SNPs and 70% of the InDels exhibited polymorphism among these three genotypes. In addition, the 15 SNPs and 125 InDels were found to be unevenly distributed on 9 linkage groups. Furthermore, 40 informative InDel markers were successfully used for the genetic diversity analysis on 32 radish accessions. CONCLUSIONS: These results would not only provide new insights into transcriptome complexity and AS regulation, but also furnish large amount of molecular marker resources for germplasm identification, genetic mapping and further genetic improvement of radish in breeding programs.

Genome-Wide Characterization of the MADS-Box Gene Family in Radish (Raphanus sativus L.) and Assessment of Its Roles in Flowering and Floral Organogenesis.

The MADS-box gene family is an important transcription factor (TF) family that is involved in various aspects of plant growth and development, especially flowering time and floral organogenesis. Although it has been reported in many plant species, the systematic identification and characterization of MADS-box TF family is still limited in radish (Raphanus sativus L.). In the present study, a comprehensive analysis of MADS-box genes was performed, and a total of 144 MADS-box family members were identified from the whole radish genome. Meanwhile, a detailed list of MADS-box genes from other 28 plant species was also investigated. Through the phylogenetic analysis between radish and Arabidopsis thaliana, all the RsMADS genes were classified into two groups including 68 type I (31 Mα, 12 Mß and 25Mγ) and 76 type II (70 MIKCC and 6 MIKC∗). Among them, 41 (28.47%) RsMADS genes were located in nine linkage groups of radish from R1 to R9. Moreover, the homologous MADS-box gene pairs were identified among radish, A. thaliana, Chinese cabbage and rice. Additionally, the expression profiles of RsMADS genes were systematically investigated in different tissues and growth stages. Furthermore, quantitative real-time PCR analysis was employed to validate expression patterns of some crucial RsMADS genes. These results could provide a valuable resource to explore the potential functions of RsMADS genes in radish, and facilitate dissecting MADS-box gene-mediated molecular mechanisms underlying flowering and floral organogenesis in root vegetable crops.

Carbon dioxide (CO2) ablation as a remedy for hepatocellular carcinoma.

BACKGROUND/AIMS: The purpose of this research was to investigate the application of carbon dioxide (CO2) ablation and determine if carbon dioxide plays a role in treating hepatocellular carcinoma. METHODOLOGY: Twelve mice with subcutaneous xenografts of a hepatocellular cancer cell line were randomly separated into 2 groups. One group underwent CO2 ablation with their liver tumor in vivo while the other group in vitro, respectively. All mice were killed on day 28. Hematoxylin-eosin (HE) staining was performed on the mice to estimate the pathological change. RESULTS: Just after 4 weeks, no significant difference in tumor size was detected between the two groups. The dimensions of the principal tumor varied from 2-3 cm with an average size of 2.2 cm in the greatest dimension. HE staining demonstrated an array of construct damage and necrosis in both groups. CONCLUSIONS: CO2 ablation could be an important treatment in the management of cancer.