A non-invasive artificial intelligence model for identifying axillary pathological complete response to neoadjuvant chemotherapy in breast cancer: a secondary analysis to multicenter clinical trial.

BACKGROUND: This study aims to develop a stacking model for accurately predicting axillary lymph node (ALN) response to neoadjuvant chemotherapy (NAC) using longitudinal MRI in breast cancer. METHODS: We included patients with node-positive breast cancer who received NAC following surgery from January 2012 to June 2022. We collected MRIs before and after NAC, and extracted radiomics features from the tumour, peritumour, and ALN regions. The Mann-Whitney U test, least absolute shrinkage and selection operator, and Boruta algorithm were used to select features. We utilised machine learning techniques to develop three single-modality models and a stacking model for predicting ALN response to NAC. RESULTS: This study consisted of a training cohort (n = 277), three external validation cohorts (n = 313, 164, and 318), and a prospective cohort (n = 81). Among the 1153 patients, 60.62% achieved ypN0. The stacking model achieved excellent AUCs of 0.926, 0.874, and 0.862 in the training, external validation, and prospective cohort, respectively. It also showed lower false-negative rates (FNRs) compared to radiologists, with rates of 14.40%, 20.85%, and 18.18% (radiologists: 40.80%, 50.49%, and 63.64%) in three cohorts. Additionally, there was a significant difference in disease-free survival between high-risk and low-risk groups (p < 0.05). CONCLUSIONS: The stacking model can accurately predict ALN status after NAC in breast cancer, showing a lower false-negative rate than radiologists. TRIAL REGISTRATION NUMBER: The clinical trial numbers were NCT03154749 and NCT04858529.

Noninvasive Artificial Intelligence System for Early Predicting Residual Cancer Burden during Neoadjuvant Chemotherapy in Breast Cancer.

OBJECTIVE: To develop an artificial intelligence (AI) system for the early prediction of residual cancer burden (RCB) scores during neoadjuvant chemotherapy (NAC) in breast cancer. SUMMARY BACKGROUND DATA: RCB III indicates drug resistance in breast cancer, and early detection methods are lacking. METHODS: This study enrolled 1048 patients with breast cancer from four institutions, who were all receiving NAC. Magnetic resonance images were collected at the pre- and mid-NAC stages, and radiomics and deep learning features were extracted. A multitask AI system was developed to classify patients into three groups (RCB 0-I, II, and III ) in the primary cohort (PC, n=335). Feature selection was conducted using the Mann-Whitney U- test, Spearman analysis, least absolute shrinkage and selection operator regression, and the Boruta algorithm. Single-modality models were developed followed by model integration. The AI system was validated in three external validation cohorts. (EVCs, n=713). RESULTS: Among the patients, 442 (42.18%) were RCB 0-I, 462 (44.08%) were RCB II and 144 (13.74%) were RCB III. Model-I achieved an area under the curve (AUC) of 0.975 in the PC and 0.923 in the EVCs for differentiating RCB III from RCB 0-II. Model-II distinguished RCB 0-I from RCB II-III, with an AUC of 0.976 in the PC and 0.910 in the EVCs. Subgroup analysis confirmed that the AI system was consistent across different clinical T stages and molecular subtypes. CONCLUSIONS: The multitask AI system offers a noninvasive tool for the early prediction of RCB scores in breast cancer, supporting clinical decision-making during NAC.

Atomic-Limit Mott Insulator in [4]Triangulene Frameworks.

Triangulene, one unique class of zigzag-edged triangular graphene molecules, has attracted tremendous research interest. In this work, as an ultimate phase of the Mott insulator, we present the realization of the atomic-limit Mott insulator in experimentally synthesized [4]triangulene frameworks ([4]-TGFs) from first-principles calculations. The frontier molecular orbitals of the nonmagnetic [4]triangulene consist of three coupled corner modes. After the isolated [4]triangulene is assembled into [4]-TGF, one special enantiomorphic flat band is created through the coupling of these corner modes, which is identified to be a second-order topological insulator with half-filled topological corner states at the Fermi level. Moreover, [4]-TGF prefers an antiferromagnetic ground state under Hubbard interactions, which further splits these metallic zero-energy states into an atomic-limit Mott insulator with spin-polarized corners. Since the fractional filling of topological corner states is a smoking-gun signature of higher-order topology, our results demonstrate a universal approach to explore the atomic-limit Mott insulators in higher-order topological materials.

Identifying topological corner states in two-dimensional metal-organic frameworks.

Due to the diversity of molecular building blocks, the two-dimensional (2D) metal-organic frameworks (MOFs) are ideal platforms to realize exotic lattice models in condensed matter theory. In this work, we demonstrate the universal existence of topological corner states in 2D MOFs with a star lattice configuration, and confirm the intriguing higher-order nontrivial topology in the energy window between two Kagome-bands, or between Dirac-band and four-band. Furthermore, combining first-principles calculations and scanning tunneling microscopy measurements, the unique topological corner state is directly identified in monolayer Ni3(HITP)2 (HITP = 2,3,6,7,10,11-hexaiminotriphenylene) grown on the Au(111) substrate. Our results not only illustrate the first organic topological state in the experiments, but also offer an exciting opportunity to study higher-order topology in 2D MOFs with the large insulating band gap.

Pseudospin Polarized Dual-Higher-Order Topology in Hydrogen-Substituted Graphdiyne.

Although the topological band theory is applicable to both Fermionic and bosonic systems, the same electronic and phononic topological phases are seldom reported in one natural material. In this work, we show the presence of a dual-higher-order topology in hydrogen-substituted graphdiyne (H-GDY) by first-principles calculations. The intriguing enantiomorphic flat-bands are realized in both electronic and phononic bands of H-GDY, which is confirmed to be an organic 2D second-order topological insulator (SOTI). Most importantly, we found that the topological corner states are pseudospin polarized in H-GDY, exhibiting a clockwise or counterclockwise texture perpendicular to the radial direction. Our results not only identify the existence of the dual-higher-order topology in covalent organic frameworks but also uncover a unique pseudospin polarization-coordinate locking relation, further extending the well-known spin-momentum locking relation in conventional topological insulators.

3D-imaging and quantitative assessment for size-related penetration of HfO2 nanoparticles in breast cancer tumor by synchrotron radiation microcomputed tomography.

The development of quantitative analytical methods to assess the heterogeneous distribution and penetration of nanodrugs in solid tumors is of great importance for anticancer nanomedicine. Herein, Expectation-Maximization (EM) iterate algorithm and threshold segmentation methods were used to visualize and quantify the spatial distribution patterns, penetration depth and diffusion features of two-sized hafnium oxide nanoparticles (s-HfO2 NPs in 2 nm and l-HfO2 NPs in 50 nm sizes) in mouse models of breast cancer using synchrotron radiation micro-computed tomography (SR-µCT) imaging technique. The three-dimensional (3D) SR-µCT images were reconstructed based on the EM iterate algorithm thus clearly displayed the size-related penetration and distribution within the tumors after intra-tumoral injection of HfO2 NPs and X-ray irradiation treatment. The obtained 3D animations clearly show that a considerable amount of s-HfO2 and l-HfO2 NPs diffused into tumor tissues at 2 h post-injection and displayed the obvious increase in the tumor penetration and distribution area within the tumors at day 7 after combination with low-dose X-ray irradiation treatment. A thresholding segmentation for 3D SR-µCT image was developed to assess the penetration depth and quantity of HfO2 NPs along the injection sites in tumors. The developed 3D-imaging techniques revealed that the s-HfO2 NPs presented more homogeneous distribution pattern, diffused more quickly and penetrated more deeply within tumor tissues than the l-HfO2 NPs did. Whereas, the low-dose X-ray irradiation treatment greatly enhanced the wide distribution and deep penetration of both s-HfO2 and l-HfO2 NPs. This developed method may provide quantitative distribution and penetration information for the X-ray sensitive high-Z metal nanodrugs in the cancer imaging and therapy.

Bi-Functionalized Transferrin@MoS2-PEG Nanosheets for Improving Cellular Uptake in HepG2 Cells.

Pre-coating with a protein corona on the surface of nanomaterials (NMs) is an important strategy for reducing non-specific serum protein absorption while maintaining targeting specificity. Here, we present lipoic acid-terminated polyethylene glycol and transferrin bi-functionalized MoS2 nanosheets (Tf@MoS2-PEG NSs) as a feasible approach to enhance cellular uptake. Tf@MoS2-PEG NSs can maintain good dispersion stability in cell culture medium and effectively protect MoS2 NSs from oxidation in ambient aqueous conditions. Competitive adsorption experiments indicate that transferrin was more prone to bind MoS2 NSs than bovine serum albumin (BSA). It is noteworthy that single HepG2 cell uptake of Tf@MoS2-PEG presented a heterogeneous distribution pattern, and the cellular uptake amount spanned a broader range (from 0.4 fg to 2.4 fg). Comparatively, the intracellular Mo masses in HepG2 cells treated with BSA@MoS2-PEG and MoS2-PEG showed narrower distribution, indicating homogeneous uptake in the single HepG2 cells. Over 5% of HepG2 cells presented uptake of the Tf@MoS2-PEG over 1.2 fg of Mo, about three-fold that of BSA@MoS2-PEG (0.4 fg of Mo). Overall, this work suggests that Tf coating enhances the cellular uptake of MoS2 NSs and is a promising strategy for improving the intracellular uptake efficiency of cancer cells.

Synthesizing Cr-Based Two-Dimensional Conjugated Metal-Organic Framework Through On-Surface Substitution Reaction.

A single-layer Cr3 (HITP)2 (HITP = 2,3,6,7,10,11-hexaiminotriphenylene) conjugated metal-organic framework (c-MOF) is synthesized under ultrahigh vacuum conditions by substituting Cr for Ni in Ni3 (HITP)2 template. As revealed by low-temperature scanning tunneling microscopy and scanning tunneling spectroscopy, while codeposition of Cr atoms and 2,3,6,7,10,11-hexaaminotriphenylene precursors produces irregular branches, crystalline Cr3 (HITP)2 frameworks are obtained by depositing Cr atoms to the Ni3 (HITP)2 templates. The density functional theory calculations reveal that the binding energy between Cr and HITP ligands is much higher than that for Ni, which hampers the growth of crystalline Cr3 (HITP)2 frameworks through direct coordination assembly but makes the substitution reaction energetically favorable. This work demonstrates a new strategy to prepare high-quality early-transition-metal-based c-MOFs under ultrahigh vacuum conditions.

Investigation on demands for license skill training among grassroots public health personnel in Hohhot City / 预防医学

Objective@#To investigate the needs for license skill training among grassroots public health personnel in Hohhot City, so as to provide the advice for improvements in training among grassroots public health personnel in Hohhot City. @*Methods@#A total of 1 802 staff were sampled from 108 grassroots healthcare institutions in four districts, four counties and one banner of Hohhot City using a multi-stage stratified random sampling method. Participants' demographics and five dimensions of training needs, including professional knowledge, professional skills, basic public health service guidelines, epidemiological survey and development of interventions, were collected through questionnaire surveys, and factors affecting the training demands were identified using a generalized linear mixed-effect model.@*Results@#The respondents included 789 men (43.78%) and 958 individuals at ages of 31 to 50 years (53.16%). There were 1 379 respondents (76.53%) that wished to receive training on professional knowledge, 1 312 respondents (72.81%) that wished to receive training on professional skills, 1 012 respondents (56.16%) that wished to receive training on basic public health service guidelines, 333 respondents (18.48%) that wished to receive training on epidemiological survey and 206 respondents (11.43%) that wished to receive training on development of interventions. Participants at ages of 41 to 50 years and 61 years and older, participants with a specialty in nursing, and participants that worked on construction of resident archives and health management of patients with tuberculosis had higher demands for training on professional knowledge; participants that worked on construction of resident archives, children healthcare management and health management of patients with chronic diseases had higher demands for training on professional skills; participants with a specialty in general practice, preventive medicine and public health, participants that worked on construction of resident archives, health management of pregnant and lying-in women, health management of elderly people and health management of patients with severe mental disorders, report and response of infectious diseases and emergency public health events had higher demands for training on basic public health service guidelines; participants that worked on vaccination, report and response of infectious diseases and emergency public health events and assisted management of health and family planning supervision had higher demands for training on epidemiological surveys; participants that worked on health management of patients with chronic diseases, assisted management of health and family planning supervision, and participants with experiences of clinical practices had higher demands for training on development of interventions (all P<0.05). @*Conclusions@#The grassroots public health personnel have a high demand for license skill training in Hohhot City, and age-, specialty- and job-specific training is required.

Intrinsic Second-Order Topological Insulator in Two-Dimensional Covalent Organic Frameworks.

As an intriguing topological phase, higher-order topological insulators have attracted tremendous attention, but the candidate materials are limited in artificial and inorganic systems. In this work, we propose a universal approach to search for two-dimensional (2D) second-order topological insulators (SOTIs) in covalent organic frameworks (COFs) with C3 symmetric cores. The underlying mechanism is illustrated through tight-binding calculations in a star lattice, showing the 2D SOTI in an overlooked energy window between two Kagome-bands with four types of nontrivial band structures. The emergence of the unique topological edge and corner states can be understood from the Su-Schrieffer-Heeger model. Furthermore, using the frontier orbital of the monomer building block as an indicator, the 2D SOTI is directly confirmed in three realistic COFs by first-principles calculations. Our results not only extend the concept of organic topological insulators from first-order to second-order but also demonstrate the universal existence of intrinsic higher-order topology in 2D COFs.

Prevention of taxane-associated acute pain syndrome with etoricoxib for patients with breast cancer: A phase II randomised trial.

BACKGROUND: For patients with breast cancer who receive docetaxel chemotherapy, taxane-associated acute pain syndrome (T-APS), considered a form of neural pathology, is a significant clinical problem. We evaluated the effect of prophylactic etoricoxib on T-APS in patients with breast cancer. MATERIALS AND METHODS: We conducted a phase II randomised trial including 144 patients with breast cancer receiving four cycles of docetaxel-based chemotherapy. Patients were randomised in the ratio 1:1 to receive prophylactic etoricoxib (60 mg, Day 1 to Day 8) or no prophylactic treatment. The primary end-point was the overall incidence of T-APS across all cycles. Secondary end-points included the incidence of severe pain (greater than 5 on a scale 0-10); severity and duration of T-APS; Functional Assessment of Cancer Therapy-Breast subscale; chronic sensory and motor neurotoxicity and adverse events. RESULTS: The overall incidence of T-APS across all cycles of chemotherapy in the etoricoxib group was 57.1%, while that in the control group was 91.5% (P < 0.001). The incidences of severe T-APS were 11.4% and 54.9% for the etoricoxib and control groups, respectively (P < 0.001). The mean Functional Assessment of Cancer Therapy-Breast subscale score of the etoricoxib group (103.79-107.24) was significantly higher than that of the control group (93.88-96.71) (P = 0.001 at cycle 1 and P < 0.001 at cycles 2-4). After four cycles of docetaxel chemotherapy, the etoricoxib group demonstrated a significantly higher mean Functional Assessment of Cancer Treatment Neurotoxicity subscale score than the control group (38.46, 95% CI: 37.63-39.29; 34.59, 95% CI: 33.73-35.45, respectively; P < 0.001). Electromyography showed that most peripheral sensory nerves in the etoricoxib group had significantly improved action potential amplitudes and conduction velocities compared with those in the control group. CONCLUSION: Prophylactic use of etoricoxib could significantly reduce the incidence and severity of docetaxel-induced acute pain syndrome and potentially decrease docetaxel-induced peripheral neuropathy. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04565600.

Performance of tidal and non-tidal mangrove constructed wetlands in treating maricultural wastewater.

This study aims to evaluate the removal efficiency of nitrogen and phosphorus in the tidal and non-tidal constructed wetlands with typical mangrove (Aegiceras corniculatum) as a wetland plant model to treat simulated marine wastewater. The results showed that the average removals of NO2--N, NO3--N, NH4+-N, TN and TP were 88.4, 80.5, 81.4, 79.7 and 40.8%, respectively, in the non-tidal subsurface flow (HF) mangrove wetland, and 65.3, 61.3, 90.6, 60.1 and 19.2% in the tidal (TF) mangrove wetland, and 11.4, 64.6, 68.7, 56.6 and 16.3% in the non-tidal free water surface (FWS) mangrove wetland, respectively. Moreover, it was observed that the composition of microbial communities in the HF mangrove wetland was beneficial to the nitrogen cycle and has more quantitative associations of N-metabolism genes. The results indicated that non-tidal HF mangrove wetland has a stable and an effective capacity for potential treatment of marine wastewater compared with the non-tidal FWS mangrove wetland and tidal TF mangrove wetland.

A Model Incorporating Axillary Tail Position on Mammography for Preoperative Prediction of Non-sentinel Lymph Node Metastasis in Patients with Initial cN+ Breast Cancer after Neoadjuvant Chemotherapy.

RATIONALE AND OBJECTIVES: This study aimed to develop a model incorporating axillary tail position on mammography (AT) for the prediction of non-sentinel Lymph Node (NSLN) metastasis in patients with initial clinical node positivity (cN+). METHODS AND MATERIALS: The study reviewed a total of 257 patients with cN+ breast cancer who underwent both sentinel lymph node biopsy (SLNB) and axillary lymph node dissection (ALND) following neoadjuvant chemotherapy (NAC). A logistic regression model was developed based on these factors and the results of post-NAC AT and axillary ultrasound (AUS). RESULTS: Four clinical factors with p<0.1 in the univariate analysis, including ycT0(odds ratio [OR]: 4.84, 95% confidence interval [CI]: 2.13-11.91, p<0.001), clinical stage before NAC (OR: 2.68, 95%CI: 1.15-6.58, p=0.025), estrogen receptor (ER) expression (OR: 3.29, 95%CI: 1.39-8.39, p=0.009), and HER2 status (OR: 0.21, 95%CI: 0.08-0.50, p=0.001), were independent predictors of NSLN metastases. The clinical model based on the above four factors resulted in the area under the curve (AUC) of 0.82(95%CI: 0.76-0.88) in the training set and 0.83(95% CI: 0.74-0.92) in the validation set. The results of post-NAC AUS and AT were added to the clinical model to construct a clinical imaging model for the prediction of NSLN metastasis with AUC of 0.87(95%CI: 0.81-0.93) in the training set and 0.89(95%CI: 0.82-0.96) in the validation set. CONCLUSIONS: The study incorporated the results of post-NAC AT and AUS with other clinal factors to develop a model to predict NSLN metastasis in patients with initial cN+ before surgery. This model performed excellently, allowing physicians to select patients for whom unnecessary ALND could be avoided after NAC.

Effects of Surgery on Prognosis of Young Women With Operable Breast Cancer in Different Marital Statuses: A Population-Based Cohort Study.

BACKGROUND: The influence of surgical approaches [including mastectomy, breast-conserving therapy (BCT) and post-mastectomy breast reconstruction (PMBR) on prognosis of young women (<40 years old) with operable breast cancer has not been determined yet, and this might vary in patients with different marital statuses. Therefore, we aimed to investigate the effect of surgery on survival outcomes for young women with operable breast cancer in different marital statuses. METHODS: We used the Surveillance, Epidemiology, and End Results (SEER) database to identify young women with operable breast cancer between 2004 and 2016, who underwent mastectomy, BCT or PMBR. We assessed overall survival (OS) and breast cancer-specific survival (BCSS) using the Kaplan-Meier method and hazard ratios using multivariate Cox proportional hazard regression. RESULTS: Compared to mastectomy, both of BCT and PMBR conferred better OS (BCT: HR = 0.79, 95%CI: 0.69-0.90, p <0.001; PMBR: HR = 0.70, 95%CI: 0.63-0.78, p <0.001) and BCSS (BCT: HR = 0.79, 95%CI: 0.69-0.91, p = 0.001; PMBR: HR = 0.73, 95%CI: 0.65-0.81, p <0.001), but there was no significant difference of survival between BCT and PMBR group. The survival benefit of BCT compared to mastectomy remained significant in unmarried young women (OS: HR = 0.68, 95%CI: 0.55-0.83, p <0.001; BCSS: HR = 0.69, 95%CI: 0.56-0.86, p = 0.001) but not in the married (OS: HR = 0.89, 95%CI: 0.75-1.05, p = 0.177; BCSS: HR = 0.89, 95%CI: 0.75-1.05, p = 0.161), while no matter married or not, PMBR group had better OS and BCSS than mastectomy group but not BCT group. CONCLUSION: Both of BCT and PMBR had improved survival compared to mastectomy for young women with operable breast cancer. The survival benefit of BCT compared to mastectomy remained significant in unmarried patients but not in married patients.

Electronegativity Assisted Synthesis of Magnetically Recyclable Ni/NiO/g-C3N4 for Significant Boosting H2 Evolution.

A magnetically recyclable Ni/NiO/g-C3N4 photocatalyst with significantly enhanced H2 evolution efficiency was successfully synthesized by a simple ethanol-solvothermal treatment. The presence of electronegative g-C3N4 is found to be the key factor for Ni0 formation in ternary Ni/NiO/g-C3N4, which provides anchoring sites for Ni2+ absorption and assembling sites for Ni0 nanoparticle formation. The metallic Ni0, on one side, could act as an electron acceptor enhancing carrier separation and transfer efficiency, and on the other side, it could act as active sites for H2 evolution. The NiO forms a p-n heterojunction with g-C3N4, which also promotes carrier separation and transfer efficiency. The strong magnetic property of Ni/NiO/g-C3N4 allows a good recyclability of catalyst from aqueous solution. The optimal Ni/NiO/g-C3N4 showed a full-spectrum efficiency of 2310 µmol·h-1·g-1 for hydrogen evolution, which is 210 times higher than that of pure g-C3N4. This ethanol solvothermal strategy provides a facile and low-cost synthesis of metal/metal oxide/g-C3N4 for large-scale application.

Niche separation and weak interactions in the high tidal zone of saltmarsh-mangrove mixing communities.

Saltmarsh-mangrove ecotones occur at the boundary of the natural geographic distribution of mangroves and salt marshes. Climate warming and species invasion can also drive the formation of saltmarsh-mangrove mixing communities. How these coastal species live together in a "new" mixed community is important in predicting the dynamic of saltmarsh-mangrove ecosystems as affected by ongoing climate change or human activities. To date, the understanding of species interactions has been rare on adult species in these ecotones.Two typical coastal wetlands were selected as cases to understand how mangrove and saltmarsh species living together in the ecotones. The leaves of seven species were sampled from these coastal wetlands based on their distribution patterns (living alone or coexisting) in the high tidal zone, and seven commonly used functional traits of these species were analyzed.We found niche separation between saltmarsh and mangrove species, which is probably due to the different adaptive strategies they adopted to deal with intertidal environments.Weak interactions between coexisting species were dominated in the high tidal zone of the two saltmarsh-mangrove communities, which could be driven by both niche differentiation and neutral theory.Synthesis. Our field study implies a potential opportunity to establish a multispecies community in the high tidal zone of saltmarsh-mangrove ecotones, where the sediment was characterized by low salinity and high nitrogen.

Higher Temperatures Do Not Always Achieve Better Antibiotic Resistance Gene Removal in Anaerobic Digestion of Swine Manure.

This study employed high-throughput quantitative PCR and 16S rRNA sequencing to evaluate the effect of temperature and residual antibiotics on the dynamics of antibiotic resistance genes (ARGs) and microbial communities during anaerobic digestion of swine manure. The abundances of total ARGs and 16S rRNA genes significantly decreased in all of four treatments (25°C, 37°C, and 37°C with 50 mg of wet weight antibiotics of body weight, and 55°C). The abundances of most ARG types were significantly correlated with those of the 16S rRNA gene and transposase gene (P < 0.01). However, the abundances of total ARGs at 55°C were much higher than those of other treatments. Meanwhile, the microbial communities at 55°C, where the Streptococcus pathogen remained at a relatively high abundance and cellulose degraders and hydrogen producers, such as Ethanoligenens and Coprococcus bacteria, increased, were markedly different from those of other treatments. Redundancy analysis indicates that temperature, pH, and the genus Streptococcus had the highest explanation for ARG variation among experimental factors, chemical properties, and representative genera, respectively. Network analysis further showed that the genus Streptococcus contributed greatly to the higher ARG abundance at 55°C. The moderate antibiotic residue only caused a slight and transitory inhibition for microbially diverse populations and promotion for ARG abundance, probably due to the degradation of antibiotics and microbial adaptability. Our results clarify the cooperativity of gene transfer-related items on ARG variation and intensively prove that higher temperature cannot always achieve better ARG removal in anaerobic digestion unless pathogens and gene transfer elements are more efficiently inhibited.IMPORTANCE Antibiotic resistance genes (ARGs) are frequently detected with high abundance in manure-applied soils. Anaerobic digestion is one of widely used processes for animal waste treatment. Thus, it is critical to understand the potential of anaerobic digestion to attenuate ARGs. Although some previous studies recommended thermophilic digestion for ARG removal, they did not get sufficient evidence to support this view. The antibiotics applied to animals are mostly excreted through feces and urine because of incomplete metabolism. It is indispensable to know whether residual antibiotics in manure will hinder ARG attenuation in anaerobic digesters. The significance of our research is in comprehensively understanding the evolution and mechanism of ARGs in anaerobic digestion of swine manure affected by temperature and residual antibiotics, which will allow the development of an ARG elimination strategy before their release into the environment.

Reproductive isolation is mediated by pollen incompatibility in sympatric populations of two Arnebia species.

To explore uncertain aspects of the processes that maintain species boundaries, we evaluated contributions of pre- and postpollination reproductive isolation mechanisms in sympatric populations of Arnebia guttata and A. szechenyi. For this, we investigated their phylogenetic relationships, traits, microenvironments, pollinator visits, action of natural selection on floral traits, and the outcome of hand pollination between the two species. Phylogenetic analysis indicates that A. szechenyi is a derived species that could be closely related to A. guttata, and both could be diploid species. Arnebia guttata flowers have larger parts than A. szechenyi flowers, but smaller nectar guides. Soil supporting A. szechenyi had higher water contents than soil supporting neighboring populations of A. guttata (in accordance with their geographical distributions). The pollinators shared by the two species preferred A. szechenyi flowers, but interspecific visitations were frequent. We found evidence of conflicting selection pressures on floral tube length, flower diameter and nectar guide size mediated via male fitness, and on flower diameter and floral tube diameter via female fitness. Hand-pollination experiments indicate complete pollen incompatibility between the two species. Our results suggest that postpollination prezygotic mechanisms are largely responsible for reproductive isolation of sympatric populations of the two Arnebia species.