IFN-γ enhances protective efficacy against Nocardia seriolae infection in largemouth bass (Micropterus salmoides).

Introduction: Nocardia seriolae adversely impacts a diverse range of fish species, exhibiting significant pathogenic characteristics that substantially impede the progress of aquaculture. N. seriolae infects in fish has a long incubation period, and clinical symptoms are not obvious in the early stages. There is presently no viable and eco-friendly approach to combat the spread of the disease. According to reports, N. seriolae primarily targets macrophages in tissues after infecting fish and can proliferate massively, leading to the death of fish. Interferon-gamma (IFN-γ) is a crucial molecule that regulates macrophage activation, but little is known about its role in the N. seriolae prevention. Methods: IFN-γ was first defined as largemouth bass (Micropterus salmoides, MsIFN-γ), which has a highly conserved IFN-γ characteristic sequence through homology analysis. The recombinant proteins (rMsIFN-γ) were obtained in Escherichia coli (E. coli) strain BL21 (DE3). The inflammatory response-inducing ability of rMsIFN-γ was assessed in vitro using monocytes/macrophages. Meanwhile, the protective effect of MsIFN-γ in vivo was evaluated by N. seriolae infection largemouth bass model. Results: In the inflammatory response of the monocytes/macrophages activated by rMsIFN-γ, various cytokines were significantly increased. Interestingly, interleukin 1ß (IL-1ß) and tumor necrosis factor alpha (TNF-a) increased by 183- and 12-fold, respectively, after rMsIFN-γ stimulation. rMsIFN-γ improved survival by 42.1% compared with the control. The bacterial load in the liver, spleen and head kidney significantly decreased. rMsIFN-γ was also shown to better induce increased expression of IL-1ß, TNF-α, hepcidin-1(Hep-1), major histocompatibility complex I (MHCI), and MHC II in head kidney, spleen and liver. The histopathological examination demonstrated the transformation of granuloma status from an early necrotic foci to fibrosis in the infection period. Unexpectedly, the development of granulomas was successfully slowed in the rMsIFN-γ group. Discussion: This work paves the way for further research into IFN-γ of largemouth bass and identifies a potential therapeutic target for the prevention of N. seriolae.

Three-dimensional analysis of puncture needle path through safety triangle approach PLD and design of puncture positioning guide plate.

OBJECTIVE: In this study, the three-dimensional relationship between the optimal puncture needle path and the lumbar spinous process was discussed using digital technology. Additionally, the positioning guide plate was designed and 3D printed in order to simulate the surgical puncture of specimens. This plate served as an important reference for the preoperative simulation and clinical application of percutaneous laser decompression (PLD). METHOD: The CT data were imported into the Mimics program, the 3D model was rebuilt, the ideal puncture line N and the associated central axis M were developed, and the required data were measured. All of these steps were completed. A total of five adult specimens were chosen for CT scanning; the data were imported into the Mimics program; positioning guide plates were generated and 3D printed; a simulated surgical puncture of the specimens was carried out; an X-ray inspection was carried out; and an analysis of the puncture accuracy was carried out. RESULTS: (1) The angle between line N and line M was 42°~55°, and the angles between the line M and 3D plane were 1°~2°, 5°~12°, and 78°~84°, respectively; (2) As the level of the lumbar intervertebral disc decreases, the distance from point to line and point to surface changes regularly; (3) The positioning guide was designed with the end of the lumbar spinous process and the posterior superior iliac spine on both sides as supporting points. (4) Five specimens were punctured 40 times by using the guide to simulate surgical puncture, and the success rate was 97.5%. CONCLUSION: By analyzing the three-dimensional relationship between the optimal puncture needle path and the lumbar spinous process, the guide plate was designed to simulate surgical puncture, and the individualized safety positioning of percutaneous puncture was obtained.

Re-Rolling Treatment in the Fermentation Process Improves the Aroma Quality of Black Tea.

Aroma is a vital factor influencing tea quality and value. It is a challenge to produce a kind of black tea with a floral/fruity aroma, good taste, and without a green/grassy odor simultaneously using small- and medium-leaf tea species. In this study, the effect of re-rolling treatment on the aroma quality of small-leaf Congou black tea was investigated using the methods of the equivalent quantification of aroma and gas chromatography-mass spectrometry (GC-MS). Sensory evaluation showed that re-rolling treatment improved the aroma quality of Congou black tea by conferring upon it floral and fruity scents. In total, 179 volatile compounds were identified using GC-MS, of which 97 volatiles showed statistical differences (Tukey s-b(K), p < 0.05). Re-rolling treatment significantly reduced the levels of alcoholic fatty acid-derived volatiles (FADVs) and volatile terpenoid (VTs), but increased the levels of aldehydic and ester FADVs, most amino acid-derived volatiles (AADVs), carotenoid-derived volatiles (CDVs), alkene VTs, and some other important volatile compounds. Based on the odor characteristics and fold changes of differential volatile compounds, hexanoic acid, hexyl formate, cis-3-hexenyl hexanoate, (Z)-3-hexenyl benzoate, hexyl hexanoate, phenylacetaldehyde, benzyl alcohol, ß-ionone, α-ionone, dihydroactinidiolide, ipsenone, ß-farnesene, ß-octalactone, melonal, etc., were considered as the potential key odorants responsible for the floral and fruity scents of re-rolled black tea. In summary, this study provides a novel and simple processing technology to improve the aroma quality of small-leaf Congou black tea, and the results are beneficial to enriching tea aroma chemistry.

Characterization of the Sensory Properties and Quality Components of Huangjin Green Tea Based on Molecular Sensory-Omics.

Huangjin green tea (HJC) is one of the most famous regional green teas in China, and has gained attention for its unique flavor. Research on HJC has focused mainly on the synthesis of L-theanine, with fewer studies concentrating on sensory characteristics. In this study, molecular sensory science techniques, including color analysis, gas chromatography-ion mobility spectrometry, and E-tongue, were used to characterize the sensory properties of HJC, with Fuding Dabai and Anji Baicha teas used as conventional and high amino acid controls, respectively. The sensory characteristics and main quality components of HJC lie somewhere between these two other teas, and somewhat closer to the conventional control. They were difficult to distinguish by color, but significant differences exist in terms of volatile organic compounds (VOCs), E-tongue values on bitterness and astringency, and their contents of major taste components. VOCs such as (E)-2-octenal, linalool, ethyl acrylate, ethyl acetate, and 2-methyl-3-furanethiol were found to be the main differential components that contributed to aroma, significantly influencing the tender chestnut aroma of HJC. Free amino acids, tea polyphenols, and ester catechins were the main differential components responsible for taste, and its harmonious phenol-to-ammonia ratio was found to affect the fresh, mellow, heavy, and brisk taste of HJC.

Complete chloroplast genome sequence of Camellia sinensis: genome structure, adaptive evolution, and phylogenetic relationships.

The chloroplast (cp) genome holds immense potential for a variety of applications including species identification, phylogenetic analysis, and evolutionary studies. In this study, we utilized Illumina NovaSeq 6000 to sequence the DNA of Camellia sinensis L. cultivar 'Zhuyeqi', followed by the assembly of its chloroplast genome using SPAdes v3.10.1, with subsequent analysis of its features and phylogenetic placement. The results showed that the cp genome of 'Zhuyeqi' was 157,072 bp, with a large single-copy region (LSC, 86,628 bp), a small single-copy region (SSC,18,282 bp), and two inverted repeat regions (IR, 26,081 bp). The total AT and GC contents of the cp genome of 'Zhuyeqi' were observed to be 62.21% and 37.29%, respectively. The cp genome encoded 135 unique genes, including 90 protein-coding genes (CDS), 37 tRNA genes, and 8 rRNA genes. Moreover, 31 codons and 247 simple sequence repeats (SSRs) were identified. The cp genomes of 'Zhuyeqi' were found to be relatively conserved, with conservation observed in the IR region, which showed no evidence of inversions or rearrangements. The five regions with the largest variations were identified, with four regions (rps12, rps19, rps16, and rpl33) located in the LSC region and one divergent region (trnI-GAU) in the IR region. Phylogenetic analysis revealed that Camellia sinensis (KJ996106.1) was closely related to 'Zhuyeqi', indicating a close phylogenetic relationship between these two species. These findings could provide important genetic information for further research into breeding of tea tree, phylogeny, and evolution of Camellia sinensis.

Difference in Aroma Components of Black Teas Processed on Different Dates in the Spring Season.

Tea aroma greatly varies with the production date. This study investigated the aroma differences among black teas processed on different dates (March 23rd, April 8th, April 15th, April 27th, and May 7th) in the spring. A sensory evaluation showed that the black tea produced on April 15th had a strong and lasting sweet aroma and the highest score of 93.5. In total, 71 volatile compounds were identified, and alcohols were the predominant category, accounting for 60.98%. From March 23rd to May 7th, the total content of volatile compounds showed a parabolic change trend and reached its maximum on April 15th (715.27 µg/L); the flavor index first peaked on April 8th (23.25) and then gradually decreased. A multivariate statistical analysis showed that 39 volatile compounds were important, differential aroma components. An odor activity value (OAV) analysis showed that the predominant odorants were ß-ionone, ß-damascenone, linalool, (E)-ß-ocimene, and geraniol, all with values larger than 100. The total OAVs of undesirable odorants decreased and reached their minimum (70.4) on April 27th, while the total OAVs of pleasant odorants and the ratio of pleasant/undesirable odorants showed inverse changes and reached their maximum (2182.1 and 31.0, respectively) on April 27th. Based on the significance of differences and OAVs, linalool, (E)-ß-ocimene, geraniol, and (E,E)-2,4-nonadienal were considered as the key differential odorants. Combined with the sensory evaluation and the differences in aroma components, it was proposed that black teas produced around April 15th in the Hunan district are more likely to have a strong and lasting sweet aroma. This study will provide scientific guidance for the production of black tea in the Hunan district, China.

Distinct variation in taste quality of Congou black tea during a single spring season.

The quality of Congou black tea fluctuates greatly with the changing seasons. However, variations in the taste quality of Congou black tea manufactured during a single spring season are far from clear. Here, we analyzed the taste quality of HuangJinCha (HJC) Congou black tea using sensory evaluation and found the taste quality of black tea manufactured in the early spring was better than that manufactured in the late spring. Principal component analysis (PCA) and cluster analysis for the data from the electronic tongue confirmed the variation and revealed that April 4 may be the critical time point at which variations in taste quality become apparent. The contents of tea polyphenols (TP), total catechins (TC), total flavones, (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG), and gallic acid (GA) showed increasing trends, whereas total amino acids (TAA) declined over time. Moreover, the variations in total amino acids (r = 0.846) and total flavones (r = - 0.858) were highly significantly correlated with the average taste quality score (p < .01), suggesting these compounds were the primary factors responsible for the fluctuation in taste quality of Congou black tea processed during a single spring season.

A digital anatomic investigation of the safe triangle areas for L1-5 percutaneous minimally invasive discectomy.

PURPOSE: To reconstruct the three-dimensional safe triangle areas at L1-5 based on the computed tomography digital data, analyze the safe scopes for the puncture location and angles, and provide anatomic references for percutaneous lumbar discectomy. METHODS: Computed tomography data from patients and control group were imported from the database and anatomical reference parameters were measured in Mimics software. The rebuilt model was rotated clockwise along the M-axis to measure the inscribed circle radius of the safe triangle at different angles. Based on the outer diameter of the largest cannula, the safe angles were calculated. The distances between points on the projection of safe triangle-inscribed circle and the upper lumbar spinous process were measured. Similarly, while the safe triangle was on the left side, the model was contra-rotated to measure all the parameters. RESULTS: There was no significant difference between the patient and control group in both the least distance between the selected anatomical reference locations and the safe triangle-inscribed circle radius at L4-5. According to the series which had a largest cannula of 2.5 mm, the safe puncture angles increased with the descending disc levels. The optimal angles were 40°-45° for L1-2, 45°-50° for L2-3, 50° for L3-4, and 55° for L4-5 separately. The differences between genders in the distances of paired reference points were significant. CONCLUSIONS: Individual safe localization of the percutaneous puncture could be obtained by analyzing the three-dimensional relationship between the puncture localization and anatomical landmarks.

Black tea increased survival of Caenorhabditis elegans under stress.

The present study examined the effects of black tea (Camellia sinensis) extracts (BTE) in Caenorhabditis elegans under various abiotic stressors. Results showed BTE increased nematode resistance to osmosis, heat, and UV irradiation treatments. However, BTE could not increase nematodes' lifespan under normal culture conditions and MnCl2-induced toxicity at concentrations we used. Further studies showed that BTE decreased reactive oxygen species and up-regulated some antioxidant enzymes, including GSH-PX, and genes, such as gsh-px and sod-3. However, only a slight extension in mev-1 mutants mean lifespan was observed without significance. These results indicated that the antioxidant activity of BTE might be necessary but not sufficient to protect against aging to C. elegans. Moreover, BTE increased the mRNA level of stress-response genes such as sir-2.1 and sek-1. Our finding demonstrated BTE might increase heat and UV stress resistance in a sir.2.1-dependent manner. Taken together, BTE enhanced stress resistance with multiple mechanisms in C. elegans.