Cercospora polygonatum, a new species causing gray leaf spot disease in Polygonatum cyrtonema.

This study identified a new species (Cercospora Polygonatum) that causes gray leaf spot (GLS) disease in cultivated Polygonatum cyrtonema. This fungal species was isolated from the affected region of GLS on P. cyrtonema leaves. Pathogenicity bioassays were conducted based on Koch's postulates. Morphology was examined based on the features of conidiomata, conidiogenous loci, conidia/conidiophores, and conidiogenous cells. The rDNA internal transcribed spacer region, calmodulin, translation elongation factor 1-alpha, and histone genes were subjected to phylogenetic analysis using MrBayes tool via in Phylosuite. Bootstrap support analysis for phylogenetic placement confirmed the new species, which was significantly different from the closely related species C. senecionis-walkeri and C. zeae-maydis. The morphological characteristics also supported this finding, with the conidiogenous of C. polygonatum being considerably shorter than those of C. senecionis-walkeri or C. zeae-maydis. In addition, C. polygonatum was distinguished by its cultural characteristics. As this fungus was isolated from P. cyrtonema, it was named C. polygonatum F.Q. Yin, M. Liu&W. L. Ma, sp. nov. The type specimen (H8-2) was preserved at the China General Microbiological Culture Collection Center. This is the first report of GLS caused by C. polygonatum on P. cyrtonema leaves in China. The current study enriches the knowledge regarding Cercospora sp., contributes to the identification of a species causing GLS in P. cyrtonema, and provides useful information for the effective management of this disease.

First report of Coniella quercicola causing leaf spot on Elaeocarpus decipiens in China.

Elaeocarpus decipiens Hemsl., is a member of the Elaeocarpaceae family. It is a broad-leaved evergreen tree (Zhang et al. 2021) and has been widely used in landscape and gardens virescence. In March 2022, leaf spots were observed on E. decipiens leaves with 30-40% disease incidence and about 25 of disease index in Wanzhou District (30°32'N; 108°22'E) of Chongqing. Lesions showed light yellowish brown in color, black fruiting body in the center (Sporodochium), and surrounded by a purplish red halo at the interface between healthy and diseased tissues. The tissue interface of the lesions were cut into small pieces (5×5 mm), sterilized with 75% (vol. -/vol.) ethanol solution for 30 s, and 3% (vol. -/vol.) sodium hypochlorite solution for 3 min, and rinsed three times with sterile water. The sterile leaf tissues were placed on potato dextrose agar (PDA) medium in petri dishes and incubated for 5 days at 28°C in the dark, and produced thirty three uniform fungal colonies with in shape and color. The colonies had petal-shaped edges, with whitish or light pink hyphae, and black sporophores were observed at 14 days after inoculation. Sporodochium were ellipsoidal to globose with a size of 121.7 ~ 232.6 × 97.2 ~ 179.6 µm (n = 40). Conidiogenous cells were simple, tapering, hyaline, and smooth, 8 ~ 16 × 5.3 ~ 13.5 µm in size Its apex was surrounded by a gelatinous coating. Conidia were hyaline, slightly curved to naviculate, rounded to acute apex, smooth-walled, aseptate, and were 9 ~ 14.2 × 1.7 ~ 2.6 µm in size (n = 40). These morphological of the cultures are consistent with those of Coniella sp. reported by Alvarez et al. (2016). The genomic DNA of representative isolates DY4, DY24, and DY28 were extracted. The internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (TEF1), and large subunit ribosomal RNA (LSU) were amplified with primers ITS1/ITS4 (White et al., 1990), EF728/EF986 (Rehner et al., 2005), and LR0R/LR5 (Vilgalys et al., 1990). The sequences were submitted to NCBI GenBank (https://www.ncbi.nlm.nih.gov/). BLASTn searches showed that the ITS (OQ926882-84), TEF1 (OR282454-56), and LSU (OQ926945-47) sequences had the highest similarity to Coniella quercicola with 99% (596/613, 597/613, and 593/613) identity for ITS (KX833595); 94% (315/536, 323/536, and 322/536) identity for TEF1 (KX833698); and 99% (933/898, 871/898, and 932/898) identity for LSU (KX833414), respectively. Phylogenetic analysis was performed using maximum likelihood method in MEGA 11.0 (Tamura et al., 2021), and the phylogenetic tree revealed a 100 % sequence similarity to the C. quercicola CBS 283.76 (ITS, KX833594; TEF1, KX833697; LSU, KX833413) and C. quercicola CBS 904.69. In the pathogenicity test, nine healthy plants of E. decipiens (five-year-old) were selected to use, 10 µL of spore suspension (106 conidia ml-1) were sprayed on the surface of four leaves per plant (six plants in total), and the other three plants were sprayed with sterile distilled water as controls. All plants were placed in a greenhouse with 95±1% relative humidity at 28°C for penetration of the cultures in an alternating dark (12 h) and light (12 h). At 5 days after inoculation, circular lesions symptoms were observed, whereas control plants remained asymptomatic. The fungus was reisolated from diseased leaf tissue and identified as Coniella quercicola according to the methods described as above. Previously, C. quercicola has been reported as a pathogen on Eucalyptus cloeziana in China (Zou et al., 2023), and Quercus robur in Netherlands (Alvarez et al., 2016). To our knowledge, this is the first report of C. quercicola causing leaf spot on E. decipiens in China. This study provides a basis for further elucidating the pathogenic mechanism, and the development of effective management for this disease.

Acceptor Engineering Produces Ultrafast Nonradiative Decay in NIR-II Aza-BODIPY Nanoparticles for Efficient Osteosarcoma Photothermal Therapy via Concurrent Apoptosis and Pyroptosis.

Small-molecule photothermal agents (PTAs) with intense second near-infrared (NIR-II, 1,000 to 1,700 nm) absorption and high photothermal conversion efficiencies (PCEs) are promising candidates for treating deep-seated tumors such as osteosarcoma. To date, the development of small-molecule NIR-II PTAs has largely relied on fabricating donor-acceptor-donor (D-A-D/D') structures and limited success has been achieved. Herein, through acceptor engineering, a donor-acceptor-acceptor (D-A-A')-structured NIR-II aza-boron-dipyrromethene (aza-BODIPY) PTA (SW8) was readily developed for the 1,064-nm laser-mediated phototheranostic treatment of osteosarcoma. Changing the donor groups to acceptor groups produced remarkable red-shifts of absorption maximums from first near-infrared (NIR-I) regions (~808 nm) to NIR-II ones (~1,064 nm) for aza-BODIPYs (SW1 to SW8). Furthermore, SW8 self-assembled into nanoparticles (SW8@NPs) with intense NIR-II absorption and an ultrahigh PCE (75%, 1,064 nm). This ultrahigh PCE primarily originated from an additional nonradiative decay pathway, which showed a 100-fold enhanced decay rate compared to that shown by conventional pathways such as internal conversion and vibrational relaxation. Eventually, SW8@NPs performed highly efficient 1,064-nm laser-mediated NIR-II photothermal therapy of osteosarcoma via concurrent apoptosis and pyroptosis. This work not only illustrates a remote approach for treating deep-seated tumors with high spatiotemporal control but also provides a new strategy for building high-performance small-molecule NIR-II PTAs.

Biodegradable silica nanocapsules enable efficient nuclear-targeted delivery of native proteins for cancer therapy.

Cell nucleus is the desired subcellular organelle of many therapeutic drugs. Although numerous nanomaterial-based methods have been developed which could facilitate nuclear-targeted delivery of small-molecule drugs, few are known to be capable of delivering exogenous native proteins. Herein, we report a convenient and highly robust approach for effective nuclear-targeted delivery of native proteins/antibodies by using biodegradable silica nanocapsules (BSNPs) that were surface-modified with different nuclear localization signals (NLS) peptides. We found that, upon gaining entry to mammalian cells via endocytosis, such nanocapsules (protein@BSNP-NLS) could effectively escape from endolysosomal vesicles with the assistance of an endosomolytic peptide (i.e., L17E), accumulate in cell nuclei and release the encapsulated protein cargo with biological activities. Cloaked with HeLa cell membrane, DNase@BSNP-NLS/L17E-M (with L17E encapsulated) homologously delivered functional proteins to cancer cell nuclei in tumor-xenografted mice. In vitro and in vivo anti-tumor properties, such as long blood circulation time and effective tumor growth inhibition, indicate that the nuclear-targeted cell-membrane-cloaked BSNPs (DNase@BSNP-NLS/L17E-M) platform is a promising therapeutic approach to nuclear related diseases.

Porous Silicon Nanocarriers with Stimulus-Cleavable Linkers for Effective Cancer Therapy.

Porous silicon nanoparticles (pSiNPs) are widely utilized as drug carriers due to their excellent biocompatibility, large surface area, and versatile surface chemistry. However, the dispersion in pore size and biodegradability of pSiNPs arguably have hindered the application of pSiNPs for controlled drug release. Here, a step-changing solution to this problem is described involving the design, synthesis, and application of three different linker-drug conjugates comprising anticancer drug doxorubicin (DOX) and different stimulus-cleavable linkers (SCLs) including the photocleavable linker (ortho-nitrobenzyl), pH-cleavable linker (hydrazone), and enzyme-cleavable linker (ß-glucuronide). These SCL-DOX conjugates are covalently attached to the surface of pSiNP via copper (I)-catalyzed alkyne-azide cycloaddition (CuAAC, i.e., click reaction) to afford pSiNP-SCL-DOXs. The mass loading of the covalent conjugation approach for pSiNP-SCL-DOX reaches over 250 µg of DOX per mg of pSiNPs, which is notably twice the mass loading achieved by noncovalent loading. Moreover, the covalent conjugation between SCL-DOX and pSiNPs endows the pSiNPs with excellent stability and highly controlled release behavior. When tested in both in vitro and in vivo tumor models, the pSiNP-SCL-DOXs induces excellent tumor growth inhibition.

Nonmetallic SERS-based immunosensor byintegrating MoS2 nanoflower and nanosheet towards the direct serum detection of carbohydrate antigen 19-9.

Recently, nonmetallic substrates have stimulated great interest in surface-enhanced Raman scattering (SERS)-based immunoassay owing to their good uniformity, stability, and biocompatibility. In this context, a novel nonmetallic SERS-based immunoassay mediated by two-dimensional molybdenum disulfide (MoS2) was delivered for the sensitive and specific monitoring of carbohydrate antigen 19-9 (CA19-9). The effective enrichment of molecules on the large active surfaces of MoS2 as well as potential 532-nm laser-induced charge transfer resonances between them engendered desirable enhancement factor values at the level of 105. Intriguingly, a sandwich immunocomplex combined MoS2 nanoflower and nanosheet exhibited not only a wide linear range from 5 × 10-4 to 1 × 102 IU·mL-1 but also a limit of detection as low as 3.43 × 10-4 IU·mL-1 towards CA19-9. More meaningful, the analytical result for clinical patient serum sample was basically compared with the conventional chemiluminescent immunoassay. Such a novel nonmetallic SERS-based immunosensor with desirable biocompatibility and sensitivity is promising for clinical diagnosis.

A computational and experimental investigation of donor-acceptor BODIPY based near-infrared fluorophore for in vivo imaging.

TD-DFT quantum calculation was performed to predict and/or illustrate the electronic transition, the related absorption and emission maxima of some pyrrole-difluoroboron derivatives with different electron donor-acceptor unit or π-conjugated degree. Upon the calculated results, a new near infrared (NIR) fluorophore (abbreviated as TPBD-BP) was designed and fabricated through linking triphenylamine and pyrrole-difluoroboron units to benzothiadiazole (BTD) backbone. The fluorescence of TPBD-BP in solid state centered at 932 nm, which was 985 nm for TPBD-BP nanoparticles (TPBD-BP dots) encapsulated in PEG-6000. The fluorescence of TPBD-BP in both solid state and dots exhibited off-peak tail emission to NIR-II region (extended to 1300 nm). The TPBD-BP dots showed excellent water solubility, biocompatibility and aggregation induced emission (AIE), which was suitable to be applied in vivo imaging. NIR-II emission signal of TPBD-BP dots can be observed in the reproductive organ of normal nude mice after tail vein injection. This attractive combination of computational and experimental investigation would help to develop new-typed small-molecular NIR fluorophores.

Activation of WNT7b autocrine eases metastasis of colorectal cancer via epithelial to mesenchymal transition and predicts poor prognosis.

BACKGROUND: Aberrant activation of the Wnt/ß-catenin signaling pathway is one of the most frequent abnormalities in human cancer, including colorectal cancer (CRC). Previous studies revealed pivotal functions of WNT family members in colorectal cancer, as well as their prognostic values. Nevertheless, the prognostic role and mechanisms underlying WNT7b in colorectal cancer development remains unclear. METHODS: In this study, WNT7b expression was measured by immunohistochemical staining of 100 cases of surgically resected human colorectal cancerous tissues as well as matched adjacent normal tissues constructed as tissue microarrays. In vitro studies, we attempted to substantiate the WNT7b expressional pattern previously found in immunohistochemistry staining. We used the colorectal cancer cell-line HCT116 and normal colorectal cell-line FHC for immunofluorescence staining and nuclear/cytoplasmic separated western blotting. We measured epithelial-mesenchymal transition (EMT) markers and migration capacity of HCT116 in the context of WNT7b knocked-down using short interfering RNA. Finally, clinical and prognostic values of WNT7b activation levels were examined. RESULTS: WNT7b was expressed in the nucleus in adjacent normal tissues. In CRC tissues, nuclear expression of WNT7b was similar; however, membrane and cytoplasmic expression was strikingly enhanced. Consistently, in vitro analysis confirmed the same expression pattern of WNT7b. Compared with FHC cells, HCT116 cells displayed higher levels of WNT7b membrane and cytoplasmic enrichment, as well as higher migration capacity with a sensitized EMT process. Either partial knockdown of WNT7b or blockade of the Wnt/ß-catenin signaling pathway reversed EMT process and inhibited the migration of HCT116 cells. Finally, elevated secretion levels of WNT7b were significantly associated with lymphatic and remote metastasis and predicted worse prognosis in the CRC cohort. CONCLUSION: In summary, we demonstrated that the activation of WNT7b autocrine probably contributes to CRC metastasis by triggering EMT process through the Wnt/ß-catenin signaling pathway. High levels of WNT7b autocrine secretion predicts poor outcome in patients with CRC. This molecule is a promising candidate for clinical CRC treatments.

Immunoassay of Tumor Markers Based on Graphene Surface-Enhanced Raman Spectroscopy.

With a unique chemical enhancement capability, graphene oxide is exceptionally suitable to serve as an alternative surface-enhanced Raman scattering (SERS) substrate, which is usually defined as GERS. However, such a GERS matrix with both promising uniformity and molecule enrichment ability has not been applied in the quantitative detection of tumor markers. Herein, an ultrasensitive and specific immunoassay of carcinoembryonic antigens mediated by the GERS matrix was demonstrated. With the assistance of Au NRs as immunoprobes, a reliable limit of detection as low as 3.01 pg/mL was finally achieved because of the collective effect of chemical and electromagnetic enhancements. Meanwhile, a calibration curve with a high R2 value of 0.996 was obtained in the range from 1 µg/mL to 10 pg/mL. These results exhibit that the GERS matrix has a great promise for participating in immunoassay, which may pave a potential avenue for the real utilization of the SERS technique in clinical biomedicine.

Theoretical Design of Near-Infrared Al3+ Fluorescent Probes Based on Salicylaldehyde Acylhydrazone Schiff Base Derivatives.

The aim of this paper is to design near-infrared (NIR) Al3+ fluorescent probes based on a Schiff base to extend their applications in biological systems. By combining benzo[h]quinoline unit and salicylaldehyde acylhydrazone, we designed two new Schiff base derivatives. According to theoretical simulations on previous experimental Al3+ probes, we obtained the appropriate theoretical approaches to describe the properties of these fluorescent probes. By employing such approaches on our newly designed molecules, it is found that the new molecules have high selectivity toward Al3+ and that their corresponding Al3+ complexes can emit NIR fluorescence. As a result, they are expected to be potential NIR Al3+ fluorescent probes.