Ketocyanine-Based Fluorescent Probe Revealing the Polarity Heterogeneity of Lipid Droplets and Enabling Accurate Diagnosis of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) has gradually become a pronoun for terrifying death owing to its high mortality rate. With the progression of HCC, lipid droplets (LDs) in HCC cells exhibit specific variations such as increased LDs number and decreased polarity, which can serve as the diagnostic target. However, developing an effective method to achieve HCC diagnosis and reveal LDs polarity heterogeneity is still a crucial challenge. Herein, the first high-performance LDs-targeting probe (1) is reported based on ketocyanine strategy with ultrasensitive polarity-responding ability and near-infrared emission. Probe 1 shows excellent sensitivity to polarity parameter Δf (0.027-0.290) with 808-fold fluorescence enhancement and the emission wavelength red-shifts 91 nm. In HCC cells, probe 1 shows a 2.5- to 5.9-fold fluorescence enhancement compared with normal and other cancer cells which exceeds clinical threshold of 2.0, indicating probe 1 can distinguish HCC cells. The LDs polarity heterogeneity is revealed and it displays a sequence, HCC cells < other cancer cells < normal cells, which may provide useful insight to engineer LDs-targeting probes for HCC cell discrimination. Finally, probe 1 realizes accurate HCC diagnosis on the cellular, organ, and in vivo levels, providing a satisfying tool for clinical HCC diagnosis and surgical navigation.

First Report of Bacterial Necrosis Caused by Pantoea vagans in Mango in China.

Mango (Mangifera indica L.) is one of the most important tropical fruits in China. Bacterial black spot is one of the primary factors limiting mango production and thus leads to huge economic losses (Bie et al. 2022). In June 2020, necrotic symptoms similar to bacterial black spot was observed with incidence 30% to 65% on mango cultivar Yuwen, Jinhuang, Tainong and Guifei in Baise, Guangxi, China. Typically, the lesions began as chlorotic spots that coalesced into an irregular shape, becoming black and slightly raised, with a yellow halo. Thirteen diseased samples collected from five orchards were cut into approximately 5-mm pieces, sterilized for 10 s with 75% ethanol, soaked with 2% NaClO for 1 min, and rinsed in sterilized water three times. The samples were then homogenized and a 10-fold serial dilution was made before plating onto Lysogeny broth (LB) agar. After incubation at 28°C for 3 days, one representative colony that was beige to yellow in color, round, convex and smooth with entire margins from each orchard was selected for further study. Genomic DNA was extracted to amplify the 16S rRNA gene (Lane et al. 1991). The resulting 16S rRNA sequences were compared in GenBank using BLASTn and shared at least 99% identity with Pantoea spp.. Furthermore, six housekeeping genes fusA, gyrB, leuS, pyrG, rplB and rpoB partial sequences of five isolates were amplified and sequenced (Delétoile et al. 2009). The sequences were deposited in GenBank (16S rRNA: OL413424 to OL413246, OP225727-OP225728; leuS: OL441796, OL441798 to OL441801; fusA, gyrB, leuS, pyrG, rplB and rpoB: OP272638-OP272662). The five bacterial isolates were classified as P. vagans based on the phylogenetic tree of the concatenated sequences and sequences derived from different Pantoea reference isolates inferred by maximum-likelihood using MegaX software (Kumar et al. 2018). Biochemical tests showed the isolates were Gram-negative, oxidase negative, and hydrogen oxidase positive, and could use D-glucose, D-fructose, L-rhamnose, D-galactose and D-mannitose as a carbon resource (Bradbury, 1986). Pathogenicity tests were performed on mango cv. Yuwen. The representative isolate was inoculated by infiltration with sterile needleless syringes on healthy leaves and spraying onto slightly scratched leaves with bacterial suspensions (OD600=0.1) respectively (Kutschera, et al. 2019). A Xanthomonas citri pv. mangiferaeindicae (Xcm) suspension and sterilized water were used as positive and negative controls, respectively. Inoculated plants were kept with 90 ± 5% relative humidity and 28 ± 1°C in the greenhouse for 1 week. Black to brown necrotic symptoms were observed on all leaves inoculated by infiltration except the negative control. These were observed in plants inoculated by spraying only after 2 weeks. Bacteria re-isolated from diseased tissues were consistent with the inoculated isolates and identified as P. vagans, fulfilling Koch's postulates. To date, P. vagans have been isolated from eucalyptus with bacterial blight and dieback, and maize with brown stalk rot (Brady et al. 2009). However, to our knowledge, this is the first report of P. vagans causing bacterial necrosis on mango in China. It was also found that some of the diseased samples were coinfected with P. vagans and Xcm in our investigation. Therefore, it is necessary to further study the infection mechanisms of this pathogen.

Design and Synthesis of Dipeptidomimetic Isocyanonaphthalene as Enhanced-Fluorescent Chemodosimeter for Sensing Mercury Ion and Living Cells.

A novel valine-based isocyanonaphthalene (NpI) was designed and synthesized by using an easy method and enabled the selective fluorescence detection of Hg2+. The chemodosimeter can display an immediate turn-on fluorescence response (500-fold) towards target metal ions upon the Hg2+-mediated conversion of isocyano to amino within NpI. Based on this specific reaction, the fluorescence-enhancement probe revealed a high sensitivity toward Hg2+ over other common metal ions and exhibited excellent aqueous solubility, good antijamming capability, high sensitivity (detection limit: 14.2 nM), and real-time detection. The response mechanism of NpI was supported by NMR spectroscopy, MS analysis and DFT theoretical calculation using various techniques. Moreover, a dipeptidomimetic NpI probe was successfully applied to visualize intracellular Hg2+ in living cells and monitor Hg2+ in real water samples with good recoveries and small relative standard deviations.

A benzothiazole-based ratiometric fluorescent probe for detection of formaldehyde and its applications for bioimaging.

We presented a new benzothiazole-based fluorescent probe for ratiometric sensing of formaldehyde. Upon treatment with formaldehyde, the alkylamine-functionalized probe can be converted to its aldehyde analogue via the target-mediated 2-aza-Cope rearrangement, which led to significant shifts in both absorption (from 392 to 452 nm) and emission (from 492 to 552 nm) bands. The sensing mechanism was confirmed by HPLC, UV/Vis and fluorescence spectroscopy. The probe is capable of sensing formaldehyde under physiological conditions with high selectivity over potentially competing biological analytes. The probe also displayed sensitive ratiometric fluorescence response (up to 35.7 fold) for formaldehyde with a low limit detection of 0.58 µM. Furthermore, the probe was successfully employed for ratiometric imaging of formaldehyde in living cells as well as in zebrafish.