OsTKPR2 is part of a sporopollenin-producing metabolon required for exine formation in rice.

The sporopollenin polymer is a major component of the pollen exine. Fatty acid derivatives synthesized in the tapetum are among the precursors of sporopollenin. Progress has been made to understand sporopollenin metabolism in rice; however, the underlying molecular mechanisms remain elusive. We found that OsTKPR2 and OsTKPR1 share a similar expression pattern, and their coding proteins have a similar subcellular localization and enzyme activities towards reduced tetraketide α-pyrone and hydroxylated tetraketide α-pyrone. Unexpectedly, OsTKPR1pro:OsTKPR2-eGFP could not rescue the phenotype of ostkpr1-4. Three independent ostkpr2 mutant lines generated by CRISPR/Cas9 displayed reduced male fertility to various extents which were correlated with the severity of gene disruptions. Notably, the anther cuticle, Ubisch bodies, and pollen development were affected in the ostkpr2-1 mutant, where a thinner pollen exine was noticed. OsTKPR1 and OsTKPR2 were integrated into a metabolon including OsACOS12 and OsPKS2, which resulted in a significant increased enzymatic efficiency when both OsTKPR1 and OsTKPR2 were present, indicating the mutual dependence of OsTKPR2 and OsTKPR1 for their full biochemical activities. Thus, our results demonstrated that OsTKPR2 is required for anther and pollen development where an OsTKPR2-containing metabolon is functional during rice sporopollenin synthesis. Furthermore, the cooperation and possible functional divergence between OsTKPR2 and OsTKPR1 is also discussed.

Spillover-feedback effects of social, economic, and environmental footprints based on the "Belt and Road Initiative".

The Belt and Road Initiative (BRI) links China to markets and sources of raw materials around the world while stimulating economic growth in developing countries and leading to widespread and occasionally unexpected social and environmental impacts. Considering that few studies have focused on the trade-embodied social and environmental impacts and two-way impact mechanisms throughout the BRI, this study, for the first time, uses social-economic-environmental footprints and the corresponding indicators of ecological efficiency and environmental stress to methodically measure the spatial distribution and regional transfer among three major regions comprising China (CHN), countries along the Belt and Road countries except China (CBR), and the rest of world (REW) from the standpoint of spillover and feedback effects, to better coordinate sustainable development between environment, economy, and society of the BRI. The results indicated the importance of spillover-feedback effects. Its significance lies not in comparison with the multiplier effect or the rates to TF, but in the indication of the two-way influence between regions through the numerical value of spillover-feedback effects of different regions and different industries. We identified that CBR and REW displayed the highest regional correlation in environment-economic aspects, followed by CHN and REW. The biggest contribution to the total footprint stemmed from the multiplier effects, suggesting the possibility that initiatives (e.g., BRI) could reduce the multiplier effects through internal drive optimization to offset the negative impact of spillover-feedback effects between regions (e.g., pollution transfer). Regarding employment, the feedback effects in CBR were the highest, suggesting that the indirect response due to the world's final demand pulled the largest employment in CBR. Moreover, REW was the net exporter of WF, CF, and EF, while the net importer of VAF and TOF. Furthermore, if ecological efficiency and environmental stress indicators were not considered, the stress of regional water resource use and carbon emission will be underestimated to a certain extent. For example, I-AGR of CHN and CBR in water resources stress, and III-HIS and V-EGW of CHN in carbon emissions stress. Finally, the proportions of different industries should be considered to evade misestimating the level of cleaner production in various industries during policymaking. Elucidating these mechanisms is essential to support decision-making that leads to sustainable development.

Novel Toll/IL-1 Receptor Homologous Region Adaptors Act as Negative Regulators in Amphioxus TLR Signaling.

Studies have shown that the basal chordate amphioxus possesses an extraordinarily complex TLR system, including 39 TLRs and at least 40 Toll/IL-1R homologous region (TIR) adaptors. Besides homologs to MyD88 and TIR domain-containing adaptor molecule (TICAM), most amphioxus TIR adaptors exhibit domain architectures that are not observed in other species. To reveal how these novel TIR adaptors function in amphioxus Branchiostoma belcheri tsingtauense (bbt), four representatives, bbtTIRA, bbtTIRB, bbtTIRC, and bbtTIRD, were selected for functional analyses. We found bbtTIRA to show a unique inhibitory role in amphioxus TICAM-mediated pathway by interacting with bbtTICAM and bbt receptor interacting protein 1b, whereas bbtTIRC specifically inhibits the amphioxus MyD88-dependent pathway by interacting with bbtMyD88 and depressing the polyubiquitination of bbt TNFR-associated factor 6. Although both bbtTIRB and bbtTIRD are located on endosomes, the TIR domain of bbtTIRB can interact with bbtMyD88 in the cytosol, whereas the TIR domain of bbtTIRD is enclosed in endosome, suggesting that bbtTIRD may be a redundant gene in amphioxus. This study indicated that most expanded TIR adaptors play nonredundant regulatory roles in amphioxus TLR signaling, adding a new layer to understanding the diversity and complexity of innate immunity at basal chordate.

Polydatin Inhibits Hepatocellular Carcinoma Cell Proliferation and Sensitizes Doxorubicin and Cisplatin through Targeting Cell Mitotic Machinery.

Polydatin (PD) is a natural compound with anticancer activities, but the underlying mechanisms remain largely unclear. To understand how PD inhibited hepatocellular carcinoma (HCC), we studied PD treatments in HCC HepG2 and SK-HEP1 cells, and normal liver HL-7702 cells. PD selectively blocked the proliferation of HCC cells but showed low toxicity in normal cells, while the effects of doxorubicin (DOX) and cisplatin (DDP) on HCC and normal liver cells were opposite. In the cotreatment studies, PD synergistically improved the inhibitory activities of DOX and DDP in HCC cells but alleviated their toxicity in HL-7702 cells. Furthermore, RNA-seq studies of PD-treated HepG2 cells revealed multiple altered signaling pathways. We identified 1679 Differentially Expressed Genes (DEGs) with over a 2.0-fold change in response to PD treatment. Integrative analyses using the DEGs in PD-treated HepG2 cells and DEGs in a TCGA dataset of HCC patients revealed five PD-repressed DEGs regulating mitotic spindle midzone formation. The expression of these genes showed significantly positive correlation with poor clinical outcomes of HCC patients, suggesting that mitotic machinery was likely a primary target of PD. Our findings improve the understanding of PD's anticancer mechanisms and provide insights into developing effective clinical approaches in HCC therapies.

Mechanisms regulating DMTF1ß/γ expression and their functional interplay with DMTF1α.

The cyclin D binding myb­like transcription factor 1 (DMTF1), a haplo­insufficient tumor suppressor gene, has 3 alternatively spliced mRNA isoforms encoding DMTF1α, ß and γ proteins. Previous studies have indicated a tumor suppressive role of DMTF1α and the oncogenic activity of DMTF1ß, while the function of DMTF1γ remains largely undetermined. In the present study, the mechanisms regulating DMTF1 isoform expression were investigated and the functional interplay of DMTF1ß and γ with DMTF1α was characterized. It was found that specific regions of DMTF1ß and γ transcripts can impair their mRNA integrity or stability, and thus reduce protein expression levels. Additionally, DMTF1ß and γ proteins exhibited a reduced stability compared to DMTF1α and all 3 DMTF1 isoforms were localized in the nuclei. Two basic residues, K52 and R53, in the DMTF1 isoforms determined their nuclear localization. Importantly, both DMTF1ß and γ could associate with DMTF1α and antagonize its transactivation of the ARF promoter. Consistently, the ratios of both DMTF1ß/α and γ/α were significantly associated with a poor prognoses of breast cancer patients, suggesting oncogenic roles of DMTF1ß and γ isoforms in breast cancer development.

Rethinking environmental stress from the perspective of an integrated environmental footprint: Application in the Beijing industry sector.

Individual footprint indicators are limited in that they usually only address one specific environmental aspect. For this reason, assessments involving multiple footprint indicators are preferred. However, the interpretation of a profile of footprint indicators can be difficult as the relative importance of the different footprint results is not readily discerned by decision-makers. In this study, a time series (1997-2012) of carbon, water and land footprints was calculated for industry sectors in the Beijing region using input-output analysis. An integrated environmental footprint (IEF) was subsequently developed using normalization and entropy weighting. The results show that steep increases in environmental footprint have accompanied Beijing's rapid economic development. In 2012, the Primary Industry had the largest IEF (8.32); however, the Secondary Industry had the greatest increase over the study period, from 0.19 to 6.37. For the Primary Industry, the greatest contribution to the IEF came from the land footprint. For the Secondary and Tertiary Industries, the water footprint was most important. Using the IEF, industry sectors with low resource utilization efficiency and high greenhouse gas emissions intensity can be identified. As such, the IEF can help to inform about industry sectors which should be given priority for modernization as well as the particular footprints that require priority attention in each sector. The IEF can also be helpful in identifying industry sectors that could be encouraged to expand within the Beijing region as they are especially efficient in terms of value adding relative to IEF. Other industries, over time, may be better located in other regions that do not face the same environmental pressures as Beijing.