Phenyldivinylsulfonamides for the construction of antibody-drug conjugates with controlled four payloads.

Phenyldivinylsulfonamides emerged from a series of divinylsulfonamides, demonstrating their ability to effectively re-bridge disulfide bonds. This kind of linkers was attached to monomethyl auristatin E (MMAE) and further conjugated with a model antibody, trastuzumab. After optimization, the linker 20 can deliver stable and highly homogenous DAR (Drug-to-Antibody Ratio) four antibody-drug conjugates (ADCs). The method was also applicable for other IgG1 antibodies to obtain ADCs with controlled four payloads. Moreover, the MMAE-bearing ADC is potent, selective and efficacious against target cell lines.

Structural and functional basis of the selectivity filter as a gate in human TRPM2 channel.

Transient receptor potential melastatin 2 (TRPM2), a Ca2+-permeable cation channel, is gated by intracellular adenosine diphosphate ribose (ADPR), Ca2+, warm temperature, and oxidative stress. It is critically involved in physiological and pathological processes ranging from inflammation to stroke to neurodegeneration. At present, the channel's gating and ion permeation mechanisms, such as the location and identity of the selectivity filter, remain ambiguous. Here, we report the cryo-electron microscopy (cryo-EM) structure of human TRPM2 in nanodisc in the ligand-free state. Cryo-EM map-guided computational modeling and patch-clamp recording further identify a quadruple-residue motif as the ion selectivity filter, which adopts a restrictive conformation in the closed state and acts as a gate, profoundly contrasting with its widely open conformation in the Nematostella vectensis TRPM2. Our study reveals the gating of human TRPM2 by the filter and demonstrates the feasibility of using cryo-EM in conjunction with computational modeling and functional studies to garner structural information for intrinsically dynamic but functionally important domains.

Decision Tree Searching Strategy to Boost the Identification of Cross-Linked Peptides.

We describe an efficient decision tree searching strategy (DTSS) to boost the identification of cross-linked peptides. The DTSS approach allows the identification of a wealth of complementary information to facilitate the construction of more protein-protein interaction networks for human cell lysate, which was tested by the use of a recently reported cross-linking data set (ACS Cent. Sci. 2019, 5, 1514-1522). A variant of the PhoX-linker, named pDSPE, was synthesized and applied to cross-link Escherichia coli cell lysate to demonstrate that the acquisition of doubly charged ions can significantly improve identification results. The method can be seamlessly integrated to other search engines to maximize the number of identified cross-links.

Cuprous oxide nanoparticles inhibit prostate cancer by attenuating the stemness of cancer cells via inhibition of the Wnt signaling pathway.

Disordered copper metabolism plays a critical role in the development of various cancers. As a nanomedicine containing copper, cuprous oxide nanoparticles (CONPs) exert ideal antitumor pharmacological effects in vitro and in vivo. Prostate cancer is a frequently diagnosed male malignancy prone to relapse, and castration resistance is the main reason for endocrine therapy failure. However, whether CONPs have the potential to treat castration-resistant prostate cancer is still unknown. Here, using the castration-resistant PC-3 human prostate cancer cell line as a model, we report that CONPs can selectively induce apoptosis and inhibit the proliferation of cancer cells in vitro and in vivo without affecting normal prostate epithelial cells. CONPs can also attenuate the stemness of cancer cells and inhibit the Wnt signaling pathway, both of which highlight the great potential of CONPs as a new clinical castration-resistant prostate cancer therapy.

Large-scale identification of encystment-related proteins and genes in Pseudourostyla cristata.

The transformation of a ciliate into cyst is an advance strategy against an adverse situation. However, the molecular mechanism for the encystation of free-living ciliates is poorly understood. A large-scale identification of the encystment-related proteins and genes in ciliate would provide us with deeper insights into the molecular mechanisms for the encystations of ciliate. We identified the encystment-related proteins and genes in Pseudourostyla cristata with shotgun LC-MS/MS and scale qRT-PCR, respectively, in this report. A total of 668 proteins were detected in the resting cysts, 102 of these proteins were high credible proteins, whereas 88 high credible proteins of the 724 total proteins were found in the vegetative cells. Compared with the vegetative cell, 6 specific proteins were found in the resting cyst. However, the majority of high credible proteins in the resting cyst and the vegetative cell were co-expressed. We compared 47 genes of the co-expressed proteins with known functions in both the cyst and the vegetative cell using scale qRT-PCR. Twenty-seven of 47 genes were differentially expressed in the cyst compared with the vegetative cell. In our identifications, many uncharacterized proteins were also found. These results will help reveal the molecular mechanism for the formation of cyst in ciliates.

Proteomic approach to reveal the proteins associated with encystment of the ciliate Euplotes encysticus.

In order to identify and reveal the proteins related to encystment of the ciliate Euplotes encysticus, we analyzed variation in the abundance of the proteins isolated from the resting cyst comparing with proteins in the vegetative cell. 2-D electrophoresis, MALDI-TOF MS techniques and Bioinformatics were used for proteome separation, quantification and identification. The comparative proteomics studies revealed 26 proteins with changes on the expression in the resting cysts, including 12 specific proteins and 14 differential proteins. 12 specific proteins and 10 out of the 14 differential proteins were selected and identified by MALDI-TOF MS. The identified specific proteins with known functions included type II cytoskeletal 1, keratin, Nop16 domain containing protein, protein arginine n-methyltransferase, epsilon-trimethyllysine hydroxylase and calpain-like protein. The identified differential proteins with known functions included Lysozyme C, keratinocyte growth factor, lysozyme homolog AT-2, formate acetyltransferase, alpha S1 casein and cold-shock protein. We discussed the functions of these proteins as well as their contribution in the process of encystment. These identified proteins covered a wide range of molecular functions, including gene regulation, RNA regulation, proteins degradation and oxidation resistance, stress response, material transport and cytoskeleton organization. Therefore, differential expression of these proteins was essential for cell morphological and physiological changes during encystment. This suggested that the peculiar proteins and differential proteins might play important roles in the process of the vegetative cells transforming into the resting cysts. These observations may be novel findings that bring new insights into the detailed mechanisms of dormancy.

Spatial and temporal effects on seed dispersal and seed predation of Musa acuminata in southern Yunnan, China.

Wild bananas are abundant in tropical areas and many ecologists have observed that the succession process is quicker following increased disturbance. This study was conducted to analyze animal-seed interactions and their effects on the seed fate of a wild banana species (Musa acuminata) in tropical southern Yunnan (China) through experiments considering spatial (site and habitat) and temporal (seasons) variation. The largest proportion of fruits (81%) was removed by frugivorous seed dispersers, especially by bats at nighttime. Only 13% of the fruits were removed by climbing seed predators (different species of rats). In the exclosure treatment, rodents accounted for a significantly higher total artificially exposed seed removal number than ants, but with spatial and temporal differences. The highest seed predation rate by rodents (70%) was found in forest with wild banana stands, corresponding with the highest rodent diversity (species numbers and abundance) among the habitat types. In contrast, the seed removal number by ants (57%) was highest in the open land habitats, but there was no close correlation with ant diversity. Seed removal numbers by ants were significantly higher in the dry compared to the rainy season, but rodent activity showed no differences between seasons. The overall results suggest that the largest proportion of seeds produced by wild banana are primarily dispersed by bats. Primary seed dispersal by bats at nighttime is essential for wild banana seeds to escape seed predation.

[Effect of Ligustrazine on type I, III collagen synthesis in lung tissue of silica-treated rat].

OBJECTIVE: To study the effect of Ligustrazine on type I, III collagen synthesis in lung tissue of silica-treated rat. METHODS: The 128 experimental rats were randomly divided into control, silica and Ligustrazine group. 1ml silica (50g/L) was injected intratracheally in silica group and Ligustrazine group, while Ligustrazine group were injected intraperitoneally 50mg/(kg x d) Ligustrazine. Samples were collected on the 1st, 3rd, 7th, 14th, 21st, 28th day after injected silica. Type I , III collagen on paraffin-embedded lung sections were stained with sirius red, detected by polarized light microscopy and quantified by Image-Pro Plus. RESULTS: On the 3rd day after silica instillation, type III collagen began to appear, while type I collagen formed on the 7th day. Thereafter Type I, III collagen increased progressively. Compared with control group, type I, III collagen expression of silica group increased. At different time points type I, III collagen expression of Ligustrazine group decreased than silica group. There was significant difference of type I collagen area percentage on the 7th, 14th, 28th day, type III collagen area percentage on the 28th day between Ligustrazine group and silica group. CONCLUSION: Silica dioxide could induce the increase of type I, III collagen production in lung tissue, and their stages are different. Ligustrazine could suppress type I, III collagen production in lung tissue of silica-treated rat, and it's mechanism would be studied.