Involvement of NADH Oxidase in Biofilm Formation in Streptococcus sanguinis

Biofilms play important roles in microbial communities and are related to infectious diseases. Here, we report direct evidence that a bacterial nox gene encoding NADH oxidase is involved in biofilm formation. A dramatic reduction in biofilm formation was observed in a Streptococcus sanguinis nox mutant under anaerobic conditions without any decrease in growth. The membrane fluidity of the mutant bacterial cells was found to be decreased and the fatty acid composition altered, with increased palmitic acid and decreased stearic acid and vaccenic acid. Extracellular DNA of the mutant was reduced in abundance and bacterial competence was suppressed. Gene expression analysis in the mutant identified two genes with altered expression, gtfP and Idh, which were found to be related to biofilm formation through examination of their deletion mutants. NADH oxidase-related metabolic pathways were analyzed, further clarifying the function of this enzyme in biofilm formation

Spatiotemporal patterns and determinants of renewable energy innovation: Evidence from a province-level analysis in China

China’s renewable energy innovation is essential for realizing its carbon neutrality targets and the low-carbon transition, but few studies have spatially examined its characteristics and spillover effects. To fill the research gap, this study investigates its distribution and trends from a spatiotemporal dimension and focuses on the spatial effects of the influencing factors to identify those that have a significant impact on renewable energy innovation by using China’s provincial panel data from 2006 to 2019. The results show the following findings. (1) Renewable energy innovation shows distinct spatial differences across China’s provinces such that it is high in the east and south and low in the west and north, which exhibits spatial locking and path-dependence. (2) There is a positive spatial correlation with renewable energy innovation. (3) R&D investment and GDP per capita significantly promote renewable energy innovation, but the former effect is mainly observed in the local area, whereas the latter shows spatial effects. More market-oriented policies should be taken for the improvement of renewable energy innovation and the establishment of regional coordination mechanisms are proposed

Large-Scale Green Supplier Selection Approach under a Q-Rung Interval-Valued Orthopair Fuzzy Environment

As enterprises pay more and more attention to environmental issues, the green supply chain management (GSCM) mode has been extensively utilized to guarantee profit and sustainable development. Greensupplierselection(GSS),whichisakeysegmentofGSCM,hasbeeninvestigated to put forward plenty of GSS approaches

Construction and validation of an aging-related gene signature predicting the prognosis of pancreatic cancer

Background: Pancreatic cancer is a malignancy with a high mortality rate and worse prognosis. Recently, public databases and bioinformatics tools make it easy to develop the prognostic risk model of pancreatic cancer, but the aging-related risk signature has not been reported. The present study aimed to identify an aging-related risk signature with potential prognostic value for pancreatic cancer patients.Method: Gene expression profiling and human clinical information of pancreatic cancer were derived from The Cancer Genome Atlas database (TCGA). Aging-related gene sets were downloaded from The Molecular Signatures Database and aging-related genes were obtained from the Human Ageing Genomic Resources database. Firstly, Gene set enrichment analysis was carried out to investigate the role of aging process in pancreatic cancer. Secondly, differentially expressed genes and aging-related prognostic genes were screened on the basis of the overall survival information. Then, univariate COX and LASSO analysis were performed to establish an aging-related risk signature of pancreatic cancer patients. To facilitate clinical application, a nomogram was established to predict the survival rates of PCa patients. The correlations of risk score with clinical features and immune status were evaluated. Finally, potential therapeutic drugs were screened based on the connectivity map (Cmap) database and verified by molecular docking. For further validation, the protein levels of aging-related genes in normal and tumor tissues were detected in the Human Protein Atlas (HPA) database.Result: The genes of pancreatic cancer were markedly enriched in several aging-associated signaling pathways. We identified 14 key aging-related genes related to prognosis from 9,020 differentially expressed genes and establish an aging-related risk signature. This risk model indicated a strong prognostic capability both in the training set of TCGA cohort and the validation set of PACA-CA cohort and GSE62452 cohort. A nomogram combining risk score and clinical variables was built, and calibration curve and Decision curve analysis (DCA) have proved that it has a good predictive value. Additionally, the risk score was tightly linked with tumor immune microenvironment, immune checkpoints and proinflammatory factors. Moreover, a candidate drug, BRD-A47144777, was screened and verified by molecular docking, indicating this drug has the potential to treat PCa. The protein expression levels of GSK3B, SERPINE1, TOP2A, FEN1 and HIC1 were consistent with our predicted results.Conclusion: In conclusion, we identified an aging-related signature and nomogram with high prediction performance of survival and immune cell infiltration for pancreatic cancer. This signature might potentially help in providing personalized immunotherapy for patients with pancreatic cancer

Antitumor activity of celastrol nanoparticles in a xenograft retinoblastoma tumor model

Zhanrong Li,1,* Xianghua Wu,1,* Jingguo Li,2 Lin Yao,1 Limei Sun,1 Yingying Shi,1 Wenxin Zhang,1 Jianxian Lin,1 Dan Liang,1 Yongping Li1 1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, 2School of Chemistry and Chemical Engineering, Sun Yat-Sen University, Guangzhou, People's Republic of China*These authors contributed equally to this workBackground: Celastrol, a Chinese herbal medicine, has shown antitumor activity against various tumor cell lines. However, the effect of celastrol on retinoblastoma has not yet been analyzed. Additionally, the poor water solubility of celastrol restricts further therapeutic applications. The goal of this study was to evaluate the effect of celastrol nanoparticles (CNPs) on retinoblastoma and to investigate the potential mechanisms involved.Methods: Celastrol-loaded poly(ethylene glycol)-block-poly(ε-caprolactone) nanopolymeric micelles were developed to improve the hydrophilicity of celastrol. The 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulf-ophenyl)-2H tetrazolium monosodium salt (WST-8) assay was used to determine the inhibitory effect of CNPs on SO-Rb 50 cell proliferation in vitro. Immunofluorescence was used to evaluate the apoptotic effect of CNPs on nuclear morphology, and flow cytometry was used to quantify cellular apoptosis. The expression of Bcl-2, Bax, NF-κB p65, and phospo-NF-κB p65 proteins was assessed by Western blotting. A human retinoblastoma xenograft model was used to evaluate the inhibitory effects of CNPs on retinoblastoma in NOD-SCID mice. Hematoxylin and eosin staining was used to assess the apoptotic effects of CNPs on retinoblastoma.Results: CNPs inhibit the proliferation of SO-Rb 50 cells in a dose- and time-dependent manner with an IC50 of 17.733 µg/mL (celastrol-loading content: 7.36%) after exposure to CNPs for 48 hours. CNPs induce apoptosis in SO-Rb 50 cells in a dose-dependent manner. The expression of Bcl-2, NF-κB p65, and phospo-NF-κB p65 proteins decreased after exposure to CNPs 54.4 µg/mL for 48 hours. Additionally, the Bax/Bcl-2 ratio increased, whereas the expression of Bax itself was not significantly altered. CNPs inhibit the growth of retinoblastoma and induce apoptosis in retinoblastoma cells in mice.Conclusion: CNPs inhibit the growth of retinoblastoma in mouse xenograft model by inducing apoptosis in SO-Rb 50 cells, which may be related to the increased Bax/Bcl-2 ratio and the inhibition of NF-κB. CNPs may represent a potential alternative treatment for retinoblastoma.Keywords: apoptosis, SO-Rb 50 cells, poly(ethylene glycol)-block-poly(ε-caprolactone), nanopolymeric micelles, celastrol nanoparticles&nbsp

A Dopa Decarboxylase Modulating the Immune Response of Scallop Chlamys farreri

BACKGROUND: Dopa decarboxylase (DDC) is a pyridoxal 5-phosphate (PLP)-dependent enzyme that catalyzes the decarboxylation of L-Dopa to dopamine, and involved in complex neuroendocrine-immune regulatory network. The function for DDC in the immunomodulation remains unclear in invertebrate. METHODOLOGY: The full-length cDNA encoding DDC (designated CfDDC) was cloned from mollusc scallop Chlamys farreri. It contained an open reading frame encoding a polypeptide of 560 amino acids. The CfDDC mRNA transcripts could be detected in all the tested tissues, including the immune tissues haemocytes and hepatopancreas. After scallops were treated with LPS stimulation, the mRNA expression level of CfDDC in haemocytes increased significantly (5.5-fold, P<0.05) at 3 h and reached the peak at 12 h (9.8-fold, P<0.05), and then recovered to the baseline level. The recombinant protein of CfDDC (rCfDDC) was expressed in Escherichia coli BL21 (DE3)-Transetta, and 1 mg rCfDDC could catalyze the production of 1.651±0.22 ng dopamine within 1 h in vitro. When the haemocytes were incubated with rCfDDC-coated agarose beads, the haemocyte encapsulation to the beads was increased significantly from 70% at 6 h to 93% at 24 h in vitro in comparison with that in the control (23% at 6 h to 25% at 24 h), and the increased haemocyte encapsulation was repressed by the addition of rCfDDC antibody (which is acquired via immunization 6-week old rats with rCfDDC). After the injection of DDC inhibitor methyldopa, the ROS level in haemocytes of scallops was decreased significantly to 0.41-fold (P<0.05) of blank group at 12 h and 0.47-fold (P<0.05) at 24 h, respectively. CONCLUSIONS: These results collectively suggested that CfDDC, as a homologue of DDC in scallop, modulated the immune responses such as haemocytes encapsulation as well as the ROS level through its catalytic activity, functioning as an indispensable immunomodulating enzyme in the neuroendocrine-immune regulatory network of mollusc

Novi VP2/VP3 rekombinantni senekavirus A izoliran u sjevernoj Kini

Senecavirus A (SVA), previously called the Seneca Valley virus, is the only member of the genus Senecavirus within the family Picornaviridae. This virus was discovered as a serendipitous finding in 2002 and named Seneca Valley virus 001 (SVV-001). SVA is an emerging pathogen that can cause vesicular lesions and epidemic transient neonatal a sharp decline in swine. In this study, an SVA strain was isolated from a pig herd in Shandong Province in China and identified as SVA-CH-SDFX-2022. The full-length genome was 7282 nucleotides (nt) in length and contained a single open reading frame (ORF), excluding the poly (A) tails of the SVA isolates. Phylogenetic analysis showed that the isolate shares its genomic organization, resembling and sharing high nucleotide identities of 90.5% to 99.6%, with other previously reported SVA isolates. The strain was proved by in vitro characterization and the results demonstrate that the virus has robust growth ability in vitro. The recombination event of the SVA-CH-SDFX-2022 isolate was found and occurred between nts 1836 and 2710, which included the region of the VP2 (partial), and VP3 (partial) genes. It shows the importance of faster vaccine development and a better understanding of virus infection and spread because of increased infection rates and huge economic losses. This novel incursion has substantial implications for the regional control of vesicular transboundary diseases, and will be available for further study of the epidemiology of porcine SVA. Our findings provide useful data for studying SVA in pigs.Senekavirus A (SVA), prije nazivan virusom doline Seneca Valley, jedini je pripadnik roda senekavirusa u porodici Picornaviridae. Virus je slučajno otkriven 2002. i nazvan virusom doline Seneca 001 (SVV-001). SVA je novi patogen koji može uzrokovati vezikularne lezije i prolaznu epidemiju novorođene prasadi s naglim gubicima u proizvodnji. U ovom je istraživanju soj SVA izoliran u populaciji svinja iz provincije Shandong u Kini i identificiran kao SVA-CHSDFX-2022. Kompletni genom izolata SVA imao je 7282 nukleotida (nt) u dužini i sadržavao je jedan otvoreni okvir za očitavanje (ORF), bez poli-A repova. Filogenetska je analiza pokazala da izolat u velikoj mjeri sadržava genomsku organizaciju i nukleotidne identitete, od 90,5 % do 99,6 %, s drugim poznatim SVA izolatima. Karakterizacija virusa je pokazala da ima veliku sposobnost rasta in vitro. Pronađena je rekombinacija izolata SVA-CH-SDFX-između nukleotida 1836 i 2710 što je uključilo regiju gena VP2 (parcijalno) i gena VP3 (parcijalno). Zbog visoke stope infektivnosti i golemih ekonomskih gubitaka važan je brži razvoj cjepiva i bolje razumijevanje zaraze. Rezultati ovog istraživanja pružaju korisne podatke za proučavanje SVA virusa, posebno s obzirom na njegovu epidemiologiju u svinja i regionalnu prekograničnu kontrolu vezikularnih bolesti

Analiza genskih varijacija rekombinantnog soja dobivenog iz triju linija virusa-2 reproduktivnog i respiratornog sindroma svinja

Since the rise of the porcine reproductive and respiratory syndrome virus (PRRSV) in China, gene mutations have frequently occurred. To understand the current prevalence and evolution of PRRSV in Shandong Province, 1,528 samples suspected of PRRSV were collected from local pig farms of different sizes. The complete genome sequence of the PRRSV strain SDLY-27 was determined by next-generation sequencing (NGS) technology. The genomic sequence of SDLY-27 was 15,363 nucleotides (nt) in length, comparative analysis of the whole genome sequence suggested that the homology between SDLY 27 and 81 PRRSV strains from China and other countries in genbank was 61.9 ~ 96.4%. This study is the first to detect recombinants from multiple recombination events among the Lineage 8 (JXA1-like strains), Lineage 5 (RespPRRSV-MLV and VR2332 strains) and Sublineage 1.5 (NADC34-like strains) in Shandong, China, and provides new data for the epidemiological study of PRRSV in China. This study enriches the epidemiological data on PRRSV in Shandong Province, China. It provides an important reference for the development of new vaccines and for the prevention and control of PRRSV in China.Usporedno sa širenjem virusa reproduktivnog i respiratornog sindroma svinja (PRRSV) u Kini, sve su češće bile i njegove genske mutacije. Kako bi se ustanovila trenutačna prevalencija i evolucija PRRSV-a u pokrajini Shandong, s lokalnih farmi prikupljeno je 1528 uzoraka svinja različitih kategorija za koje je postojala sumnja na zarazu PRRSVom. Kompletan genomski slijed soja SDLY-27 PRRSV-a određen je tehnologijom sekvenciranja sljedeće generacije (NGS). Slijed je imao dužinu od 15 363 nukleotida (nt), a komparativna analiza cijeloga genomskog slijeda uputila je na to da je homolognost između sojeva SDLY 27 i 81 PRRSV-a iz Kine i uzoraka u banci gena iz drugih zemalja 61,9~96,4%. Ovo je prvo istraživanje koje je otkrilo rekombinantne sojeve iz višestrukih rekombinacija među linijama 8 (sojevi nalik na JXA1), 5 (sojevi RespPRRSV-MLV i VR2332) i podlinije 1,5 (sojevi nalik na NADC34) u Shandongu, Kina.Kao takvo, istraživanje pruža nove podatke o epidemiologiji PRRSV-a u Kini, posebno u pokrajini Shandong, a ujedno predstavlja i važnu referenciju za razvoj novih cjepiva te prevenciju i kontrolu bolesti uzrokovane navedenim virusom

PathNarratives: Data annotation for pathological human-AI collaborative diagnosis

Pathology is the gold standard of clinical diagnosis. Artificial intelligence (AI) in pathology becomes a new trend, but it is still not widely used due to the lack of necessary explanations for pathologists to understand the rationale. Clinic-compliant explanations besides the diagnostic decision of pathological images are essential for AI model training to provide diagnostic suggestions assisting pathologists practice. In this study, we propose a new annotation form, PathNarratives, that includes a hierarchical decision-to-reason data structure, a narrative annotation process, and a multimodal interactive annotation tool. Following PathNarratives, we recruited 8 pathologist annotators to build a colorectal pathological dataset, CR-PathNarratives, containing 174 whole-slide images (WSIs). We further experiment on the dataset with classification and captioning tasks to explore the clinical scenarios of human-AI-collaborative pathological diagnosis. The classification tasks show that fine-grain prediction enhances the overall classification accuracy from 79.56 to 85.26%. In Human-AI collaboration experience, the trust and confidence scores from 8 pathologists raised from 3.88 to 4.63 with providing more details. Results show that the classification and captioning tasks achieve better results with reason labels, provide explainable clues for doctors to understand and make the final decision and thus can support a better experience of human-AI collaboration in pathological diagnosis. In the future, we plan to optimize the tools for the annotation process, and expand the datasets with more WSIs and covering more pathological domains

Cathepsin K activity controls cardiotoxin‐induced skeletal muscle repair in mice

Abstract Background: Cathepsin K (CatK) is a widely expressed cysteine protease that has gained attention because of its enzymatic and non‐enzymatic functions in signalling. Here, we examined whether CatK‐deficiency (CatK−/−) would mitigate injury‐related skeletal muscle remodelling and fibrosis in mice, with a special focus on inflammation and muscle cell apoptosis. Methods: Cardiotoxin (CTX, 20 μM/200 μL) was injected into the left gastrocnemius muscle of male wild‐type (CatK+/+) and CatK−/− mice, and the mice were processed for morphological and biochemical studies. Results: On post‐injection Day 14, CatK deletion ameliorated muscle interstitial fibrosis and remodelling and performance. At an early time point (Day 3), CatK−/− reduced the lesion macrophage and leucocyte contents and cell apoptosis, the mRNA levels of monocyte chemoattractant protein‐1, toll‐like receptor‐2 and toll‐like receptor‐4, and the gelatinolytic activity related to matrix metalloproteinase‐2/‐9. CatK deletion also restored the protein levels of caspase‐3 and cleaved caspase‐8 and the ratio of the BAX to the Bcl‐2. Moreover, CatK deficiency protected muscle fibre laminin and desmin disorder in response to CTX injury. These beneficial muscle effects were mimicked by CatK‐specific inhibitor treatment. In vitro experiments demonstrated that pharmacological CatK inhibition reduced the apoptosis of C2C12 mouse myoblasts and the levels of BAX and caspase‐3 proteins induced by CTX. Conclusions: These results demonstrate that CatK plays an essential role in skeletal muscle loss and fibrosis in response to CTX injury, possibly via a reduction of inflammation and cell apoptosis, suggesting a novel therapeutic strategy for the control of skeletal muscle diseases by regulating CatK activity