Congressionally designated wilderness offers the highest level of land protection in the USA, comparable to category 1b wilderness as defined by the International Union for Conservation of Nature, yet wilderness areas face impacts of climate change. Researchers and managers increasingly recognize the immense challenge of stewarding wilderness in a changing climate. Although responding to climate change is considered a wicked problem, this framing has not been applied to wilderness contexts. We move theory into practice by using wicked problem framing and four illustrative cases to explore how the resist-accept-direct (RAD) framework can help untangle the wicked problem of addressing unprecedented ecological transformation in wilderness. The RAD framework clarifies a range of response options including resisting transformation, accepting it, or directing it toward preferred new conditions. As climate change continues to challenge wilderness managers, the RAD framework can help assess trade-offs and preserve wilderness character into the twenty-first century.
Leptospirosis is a widespread zoonotic disease caused by pathogenic Leptospira spp., affecting humans, domestic animals, and wildlife. Wild mammals are recognized as potential maintenance hosts for pathogenic Leptospira spp., and some species may act as asymptomatic reservoirs, representing a potential public health risk, especially in areas with increasing human-wildlife interaction. This cross-sectional study investigated the occurrence of pathogenic Leptospira spp. in wild mammals from Paraíba, Brazil, sampled between June 2022 and June 2023 using serological and molecular approaches. Blood samples from 60 animals were analyzed by the microscopic agglutination test (MAT), while tissue samples from 29 animals were evaluated by polymerase chain reaction (PCR) targeting the lipL32 gene, a marker specific for pathogenic Leptospira spp. Seroreactivity was detected in 9/60 animals (15.0%), with reactions against the serogroups Pomona, Hebdomadis, Grippotyphosa, Shermani, Patoc, and Pyrogenes. Pathogenic Leptospira DNA was detected in 6/29 animals (20.7%), with positive PCR results obtained from multiple tissues, including the kidneys, bladder, liver, brain, and reproductive organs. These findings demonstrate exposure to and infection by pathogenic Leptospira spp. in wild mammals from Paraíba and reinforce the importance of wildlife surveillance for a better understanding of the epidemiology of leptospirosis.
Hepatitis E virus (HEV) is responsible for emerging foodborne zoonotic infections in Europe, which are primarily associated with genotype 3 and more sporadically to genotype 4. While domestic pigs and wild boars are recognized as the principal reservoirs, the epidemiological role of wild ruminants remains less clearly defined. Knowledge of the health status of wildlife is essential, not only to minimize potential risks to human health associated with the consumption of game meat, but also to study the spread of zoonotic diseases in the territory. This study investigated the presence of HEV infections in wild ruminants from northwestern Italy between 2023 and 2025. A total of 883 liver samples were collected from red deer (Cervus elaphus, n = 256), roe deer (Capreolus capreolus, n = 378), fallow deer (Dama dama, n = 136), and alpine chamois (Rupicapra rupicapra, n = 113) in the regions of Liguria, Piedmont, and Aosta Valley. All samples were tested by one-step real-time RT-PCR. No positive samples were identified, indicating the absence of HEV infections in the investigated populations during the study period. However, the cross-sectional design, the analysis of liver samples only, and the lack of serological data did not allow assessment of previous exposure or temporal patterns of virus circulation. Overall, the results suggest that the investigated wild ruminant species in northwestern Italy are unlikely to play a significant role in HEV epidemiology in this geographic context. Continued integrated surveillance, including both molecular and serological approaches, is recommended to monitor potential changes in virus circulation in wildlife to further clarify their role in HEV epidemiology.
The selection of optimal conversion therapy for patients with initially unresectable colorectal liver metastases (CRLM) remains a significant clinical challenge. In patients with RAS wild-type disease, a key therapeutic decision involves whether to combine chemotherapy with an EGFR inhibitor (EGFRi) or bevacizumab. A systematic literature review and meta-analysis of randomised controlled trials (RCTs) published between 2015 and 2025 were conducted to compare chemotherapy plus either an EGFRi or bevacizumab as first-line treatment for patients with RAS wild-type metastatic colorectal cancer and liver-limited disease. Analyses encompassed both the overall population and subgroups defined by primary tumor location (PTL). Six RCTs, including data from 795 patients with CRLM, were identified. No statistically significant differences were found between EGFRi- and bevacizumab-based regimens for progression-free survival (PFS) and overall survival (OS). Four studies contributed to analyses of response rate (RR) and complete (R0) resection rate, while three studies contributed to the depth of response (DpR) analysis. EGFRi-based regimens demonstrated statistically significant improvements in both RR and DpR, with pooled odds ratios (ORs) of 1.98 and 1.70, respectively. No statistically significant difference in R0 resection rates was observed. Subgroup analyses by PTL revealed no statistically significant survival advantage for either agent in either left- or right-sided subgroups. This comprehensive meta-analysis identified no significant survival advantage for either anti-EGFRs or bevacizumab in patients with initially unresectable RAS wild-type CRLM. Further research is warranted to elucidate why the increased and deeper responses observed with anti-EGFRs do not result in higher R0 resection rates or improved survival.
First serological survey on rotavirus A (RVA) in domestic and wild pigeons.RVA widely distributed (∼80%) in domestic and feral pigeons but not in wild common wood pigeons.Almost all adult domestic pigeons (98.1%) possess anti-RVA antibodies.Antibody titres in vaccinated juvenile pigeons are similar to reconvalescents and adults.
Freshwater turtles are long-lived, abundant components of Australian aquatic ecosystems, yet little is known about the diversity and prevalence of zoonotic bacteria they may carry. In this study, we conducted one of the most comprehensive surveys to date of zoonotic microorganisms associated with wild Australian freshwater turtles. A total of 257 individuals representing 10 species across 6 genera were sampled from multiple riverine and wetland systems. Using next generation sequencing, we detected 18 confirmed zoonotic bacterial species spanning 10 genera, including Acinetobacter, Vibrio, Plesiomonas, Campylobacter, and Clostridium. Nearly half of all turtles (49.8%) were shedding at least one zoonotic taxon at the time of capture, with some individuals carrying multiple species simultaneously. Several additional isolates from genera with known zoonotic potential could not be resolved to species level, highlighting gaps in current reference databases and the need for improved genomic characterisation. No Salmonella spp. were detected, consistent with previous research suggesting a low risk of reptile-associated salmonellosis from Australian freshwater turtles. Shedding prevalence varied markedly among turtle species and locations, suggesting that host ecology and environmental conditions influence bacterial acquisition and persistence. Due to data sparsity for many taxa, analyses were limited to descriptive assessments. Overall, these findings demonstrate that Australian freshwater turtles harbour a diverse assemblage of bacteria with zoonotic relevance and may act as reservoirs within aquatic environments. Continued surveillance, integration of genomic tools, and consideration of human-wildlife interaction contexts, including cultural harvesting, will be important for assessing public health implications and informing conservation and biosecurity strategies.
Mutant C9orf72 has been extensively studied as a major genetic cause of amyotrophic lateral sclerosis and frontotemporal dementia, and is also known to generate Huntington's disease (HD)-like phenocopies. However, despite this strong disease association, the role of wild-type C9orf72 (wt-C9orf72) in neurodegeneration remains largely unexplored. HD is a neurodegenerative disease, and characterized by the accumulation of misfolded mutant Huntingtin (mHTT) and impaired proteostasis, yet the upstream mechanisms driving ubiquitin-proteasome system (UPS) dysfunction are not fully understood. Here, we identify a previously unrecognized modulatory role of wt-C9orf72 in regulating mHTT aggregation in experimental HD models. Analysis of public transcriptomic datasets reveal context-dependent C9ORF72 expression changes across HD-related human datasets, while C9orf72 levels are increased in R6/2 mouse brain lysates. Functional analyses reveal that overexpression of wt-C9orf72 increases mHTT aggregation and is accompanied by increased apoptotic signaling and reduced cell viability. Unbiased proteomic profiling identifies Stat1 as a key downstream effector. Mechanistically, wt-C9orf72 promotes Stat1 activation and nuclear translocation, leading to transcriptional upregulation of Isg15, a ubiquitin-like modifier. Elevated Isg15 disrupts UPS function, resulting in accumulation of polyubiquitinated proteins and impaired proteasomal degradation. Importantly, genetic suppression of Stat1 or Isg15 significantly attenuates C9orf72-associated mHTT aggregation, supporting a functional C9orf72-Stat1-Isg15 axis. Consistent with these cell-based findings, Stat1, phosphorylated Stat1 and Isg15 levels are elevated in the cortex and striatum of R6/2 HD mouse brains. Collectively, our findings identify a novel wt-C9orf72-Stat1-Isg15 axis that promotes proteasomal dysfunction and mHTT aggregation, providing new insights into wt-C9orf72-associated protein homeostasis.
The genus Pseudoconyza Cuatrec. (Inuleae, Asteraceae) is a monotypic genus widespread in Africa, Asia, North and Central America. Pseudoconyza viscosa (Mill.) D'Arcy is the only species of the genus, formerly known as Laggera aurita, although some authors report it as Blumea viscosa. The plant is largely utilised in the local popular medicine and although research has been carried out on its biological properties, its essential oil results are poorly investigated. This is the first report regarding the chemical composition of the essential oil (EO) from aerial parts of this plant, collected in Oman. The EO (Pv) was characterised by large amounts of sesquiterpene hydrocarbons (37.4%), with β-selinene (11.6%) as main metabolite of the oil, and oxygenated sesquiterpene (28.4%), although also good quantity of limonene (11.2%), belonging to monoterpene hydrocarbons (19.8%) was also observed. A comparison with previously published results regarding EOs of the Laggera species is discussed.
暂无摘要（点击查看详情）
HER2/ERBB2 kinase inhibition provides a useful case study for connecting curated bioactivity data, ligand-based modelling, chemical-space design, and structure-based triage. This study developed an applicability-domain-aware workflow for prioritizing scaffold-constrained HER2 kinase inhibitor analogs. Exact nanomolar IC50 records from ChEMBL target CHEMBL1824 were cleaned, converted to pIC50 values, aggregated at molecule level, and encoded as 2,048-bit Morgan fingerprints. Tree-ensemble classifiers gave the strongest ligand-based performance, and repeated scaffold-split validation was used to estimate scaffold-level generalization. Four HER2-associated Bemis-Murcko frameworks were selected for constrained R-group enumeration, yielding 500,000 valid analogs. Composite screening combined predicted HER2 activity, maximum-reference Tanimoto similarity as an applicability-domain measure, preferred physicochemical filters, and parent-scaffold priority, followed by Butina diversity selection of 100 docking-ready ligands. Synthetic-feasibility triage indicated that 94 of the 100 final ligands passed conservative RDKit-based medicinal-chemistry criteria. Wild-type docking, Prime MMGBSA rescoring, interaction-fingerprint comparison, and profiling against L755S, T798I, V777L, and V842I HER2 mutant receptors prioritized GEN_0015905 as the leading wild-type MMGBSA analog and highlighted GEN_0268162, GEN_0394004, and GEN_0015892 as follow-up candidates with smaller predicted wild-type-to-mutant score shifts. The workflow provides a reproducible computational route for selecting experimentally testable HER2 analogs while keeping model extrapolation, chemical diversity, and structure-based interpretation explicit. Repeated validation also showed the expected drop from random-split to scaffold-split performance, with random forest ROC-AUC decreasing from 0.973 ± 0.006 under repeated random splits to 0.956 ± 0.015 under repeated scaffold splits.
Colorectal cancer is a common gastrointestinal malignancy worldwide. KRAS gene mutation is an important molecular feature of colorectal cancer, but the clinical significance of different mutation sites remains unclear. This study aims to investigate the distribution characteristics of specific KRAS gene mutation sites and analyze their associations with clinicopathological parameters and survival outcomes in patients with colorectal cancer, thereby providing a basis for precise stratified diagnosis and treatment. A total of 550 patients with colorectal cancer who underwent surgical treatment at the Affiliated Cancer Hospital of Zhengzhou University from May 2018 to January 2020 were retrospectively included, and tumor tissue specimens were retained from all patients. Polymerase chain reaction (PCR) was first used to screen all specimens for KRAS gene mutations, and the overall mutation rate was calculated. High-throughput sequencing was then performed on 210 of these specimens using the Illumina Nova sequencing platform to determine the specific mutation types at codons 12 and 13 of exon 2 of the KRAS gene. Clinicopathological data, including age, sex, tumor stage, lymph node metastasis status, and primary lesion diameter, were collected. Survival data were obtained through outpatient follow-up and telephone follow-up, with the follow-up cutoff date of December 29, 2023. Univariate and multivariate Cox proportional hazards regression models were used to identify independent risk factors affecting prognosis. Variables were included based on a univariate analysis threshold of P<0.10 and clinically recognized key variables, such as tumor stage and lymph node metastasis. Among the 550 specimens, KRAS gene mutations were detected in 232 cases, with an overall mutation rate of 42.18%. Among the 210 specimens subjected to high-throughput sequencing, mutations were detected in 155 cases, with a mutation rate of 73.81%. Seven mutation types were identified. G12D was the most common mutation site (19.52%), followed by G12V (12.86%), G13D (12.86%), G12S (10.95%), G12A (8.57%), G12C (8.10%), and double mutations, including G12D+G12V and G12V+G13D, each accounting for 0.48%. Analysis of associations with clinical features showed that the proportion of patients with lymph node metastasis at initial diagnosis was 73.17% (30/41) among patients with the G12D mutation, which was significantly higher than that in patients with wild-type KRAS [38.18% (21/55), P<0.05]. However, the proportion of patients with a primary lesion diameter ≥4 cm was 36.59% in the G12D mutation group, which did not differ significantly from that in the wild-type group (25.45%, P>0.05). The proportions of patients with a primary lesion diameter ≥4 cm were 66.67%, 76.47%, 78.26%, and 66.67% among patients with G12A, G12C, G12S, and G13D mutations, respectively, all of which were significantly higher than that in the wild-type group (all P<0.05). However, the proportions of lymph node metastasis in these mutation groups did not differ significantly from that in the wild-type group (P>0.05). Cox proportional hazards regression analysis demonstrated that tumor stage Ⅲ-Ⅳ (HR=1.820, 95% CI 1.194 to 2.774, P=0.005), G12C mutation (HR=2.014, 95% CI 1.321 to 3.069, P=0.001), G12D mutation (HR=1.857, 95% CI 1.218 to 2.930, P=0.004), and G12V mutation (HR=1.751, 95% CI 1.149 to 2.669, P=0.009) were independent risk factors for death in patients with colorectal cancer. Different KRAS gene mutation sites are specifically associated with clinical features of colorectal cancer. The G12D mutation is associated with a high risk of lymph node metastasis, whereas G12A, G12C, G12S, and G13D mutations are associated with increased primary tumor size. G12C, G12D, and G12V mutations are independent prognostic risk factors and may serve as potential molecular markers for risk stratification and individualized treatment. 目的: 结直肠癌是全球高发消化道恶性肿瘤，KRAS基因突变是其重要的分子特征，但不同突变位点的临床意义差异尚未明确。本研究探讨KRAS基因特定突变位点的分布特征，分析其与结直肠癌患者临床病理参数、生存预后的关联，为精准分层诊疗提供依据。方法: 回顾性纳入2018年5月至2020年1月于郑州大学附属肿瘤医院接受手术治疗的550例结直肠癌患者，均留存肿瘤组织标本。采用聚合酶链反应(polymerase chain reaction，PCR)对所有标本进行KRAS基因突变筛查，计算总体突变率;采用Illumina Nova测序平台对其中的210例标本行高通量测序，明确KRAS基因第2号外显子第12、13位密码子的具体突变类型。收集患者年龄、性别、肿瘤分期、淋巴结转移状态、原发灶直径等临床病理资料，并通过门诊复查、电话随访获取生存数据，随访截止时间为2023年12月29日。采用单因素及多因素Cox比例风险回归模型，筛选影响患者预后的独立危险因素，变量纳入标准为单因素分析P<0.10及临床公认的关键变量(如肿瘤分期、淋巴结转移)。结果: 550例标本中，232例检出KRAS基因突变，总体突变率为42.18%。210例行高通量测序的标本中，155例检出突变，突变率为73.81%;共检出7种突变类型，其中G12D为最常见位点(19.52%)，其次为G12V(12.86%)、G13D(12.86%)、G12S(10.95%)、G12A(8.57%)、G12C(8.10%)、双突变(G12D+G12V、G12V+G13D，各占0.48%)。临床特征关联分析显示:G12D突变型患者初诊时淋巴结转移比例为73.17%(30/41)，显著高于野生型的38.18%(21/55;P<0.05)，而原发灶直径≥4 cm的比例(36.59%)与野生型(25.45%)比较，差异无统计学意义(P>0.05);G12A、G12C、G12S、G13D突变型患者原发灶直径≥4 cm的比例分别为66.67%、76.47%、78.26%、66.67%，均显著高于野生型(均P<0.05)，但淋巴结转移比例与野生型差异无统计学意义(P>0.05)。Cox比例风险回归分析结果显示:肿瘤分期Ⅲ~Ⅳ期(HR=1.820，95% CI 1.194~2.774，P=0.005)、G12C突变(HR=2.014，95% CI 1.321~3.069，P=0.001)、G12D突变(HR=1.857，95% CI 1.218~2.930，P=0.004)、G12V突变(HR=1.751，95% CI 1.149~2.669，P=0.009)是结直肠癌患者死亡的独立危险因素。结论: KRAS基因不同突变位点与结直肠癌临床特征存在特异性关联:G12D突变与淋巴结高转移风险相关，G12A、G12C、G12S、G13D突变与原发灶体积增大相关。G12C、G12D、G12V突变是患者预后的独立危险因素，可作为风险分层与个体化治疗的潜在分子标志物。.
Lactic acid bacteria (LAB), such as Limosilactobacillus fermentum are persistent contaminants in bioethanol fermentation facilities and inhibit Saccharomyces cerevisiae, reducing ethanol yields. Rising resistance to antibiotics such as virginiamycin makes contamination control increasingly difficult. Recombinant endolysins such as LysKB317 provide a promising alternative. To improve LysKB317 performance, five variants were engineered by duplicating and rearranging its enzymatically active (EAD) and cell binding (CBD) domains. These constructs were compared to the wild-type endolysin. Differential scanning calorimetry (DSC) and bacterial lytic assays demonstrated that domain shuffling altered bacteriolytic activity and thermostability. The wild-type LysKB317 exhibited the highest initial lytic activity (1.16 × 10-4 OD/s‧µM), whereas LysKB317EADx3-CBD displayed comparable bacteriolytic kinetics (7.99 × 10-5 OD/s‧µM) and LysKB317CBD-EAD-CBD (ΔCp = 24.66 kJ/mol‧K) exhibited the greatest thermostability. Despite reduced activity, both variants effectively control LAB contaminants and prevented stalled corn mash fermentation. Domain shuffling demonstrated effective bacterial control strategy for generating novel endolysins for biocontrol.
TDP-43 pathology is a hallmark of Amyotrophic Lateral Sclerosis (ALS), yet no therapeutic strategy effectively targets its upstream molecular consequences. Here, we investigated whether the anti-TDP-43 intrabody scFv B1 modulates neuroinflammatory and metabolic pathways in a preclinical ALS model, and whether these effects translate into functional benefit after symptom onset. Using phage display, we previously identified single-chain variable fragments (scFvs) binding TDP-43, including the candidate therapeutic scFv B1. In NSC-34 motor neuron-like cells overexpressing human wildtype TDP-43, B1 reduced NF-κB activation, consistent with disruption of TDP-43-driven inflammatory signaling. For in vivo assessment, B1 was delivered via AAV-CAP.B10 after symptom onset in the hTDP-43(WTxA315T) transgenic mouse model, enabling neuro-specific expression. Two cohorts were analyzed - longitudinal (nine months) and terminal (six months post-treatment) - through behavioral testing, PET imaging, metabolomics, transcriptomics, and plasma biomarker analyses. B1 achieved robust CNS expression and modulated several disease-relevant molecular pathways. RNA-sequencing revealed attenuation of NF-κB-related inflammatory signatures and partial normalization of metabolic and trophic gene expression. Metabolomic profiling identified shifts toward wild-type-like levels in oxidative stress, mitochondrial, and membrane phospholipid metabolites. Despite these molecular effects, symptomatic B1 administration did not improve motor behavior or reduce plasma neurofilament light chain (NfL) concentrations. Notably, plasma TDP-43 levels were stabilized, indicating systemic target engagement. Collectively, scFv B1 modulates upstream pathogenic processes associated with TDP-43 proteinopathy but is insufficient to reverse established neurodegeneration after symptom onset, underscoring the need for earlier and likely combinatorial intervention strategies in ALS.
THAP domain-containing protein 6 (THAP6) is a member of the THAP family and contains a conserved THAP zinc-finger domain, allowing it to regulate gene expression as a transcription factor. Previous studies have shown that THAP6 may be involved in macrophage activation and transcriptional regulation related to social stimuli; however, whether THAP6 affects emotional behavior and brain tissue structure has not yet been reported. This study aims to clarify the effects of Thap6 gene deletion on emotional behaviors in mice, preliminarily explore the effects of Thap6 gene knockout on mouse brain tissue, and provide experimental evidence for revealing the potential role of THAP6 in neuropsychiatric disorders. A whole-body Thap6 knockout mouse model was generated using CRISPR/Cas9 technology. Genotyping was performed to select heterozygous Thap6 knockout (Thap6+/-; n=8) and wild-type (WT; n=10) male C57BL/6J mice as study subjects. After the mice reached adulthood, motor and balance abilities were assessed using the rotarod test; general locomotor activity and exploratory ability were evaluated using the open-field test; depression-like behaviors were assessed using the sucrose preference test and tail suspension test; anxiety-like behavior was evaluated using the elevated plus-maze test; and social behavior was assessed using the three-chamber social test. After behavioral assessments, mouse brain tissues were collected, and hematoxylin and eosin (HE) staining and Nissl staining were used to observe changes in brain microstructure and neuronal number. Behavioral results showed that, compared with WT mice, Thap6+/- mice had a significantly lower percentage of entries into the open arms in the elevated plus maze test (t=2.516, P=0.024) and a significantly higher average movement speed in the open arms (t=3.045, P=0.013). In the three-chamber social test, the sociability index (t=4.350, P<0.001) and social preference index (t=2.732, P=0.014) of Thap6+/- mice were significantly lower than those of WT mice. No statistically significant differences were observed between the 2 groups in other behavioral indicators (all P>0.05). HE staining of brain tissues showed scattered abnormal cell morphology, loose cytoplasm, and blurred nuclei in the cortex of Thap6⁺/⁻ mice; loosely arranged neurons, enlarged intercellular spaces, and extensive vacuole formation in the amygdala; disordered arrangement of cerebellar Purkinje cells, unclear boundaries, swelling and dissolution of some nuclei, and cell loss; and loss of neuronal nucleoli and partial cellular pyknosis in the substantia nigra. In the hippocampus of Thap6⁺/⁻ mice, neurons were orderly arranged, with relatively good cell morphology, a larger number of cells, and compact arrangement. Nissl staining showed that the number of neurons in the hippocampal dentate gyrus region was significantly increased in Thap6⁺/⁻ mice (U=0, P=0.029), whereas morphological abnormalities and reduced neuronal numbers were observed in the cortex, amygdala, cerebellum, and substantia nigra (all U=0, P=0.029). Thap6 gene knockout leads to anxiety-like behavior and social behavioral deficits in mice. It also causes structural abnormalities and reductions in neuronal number across multiple brain regions. This study systematically reveals, for the first time, the important role of THAP6 in the regulation of emotional and social behaviors, provides new evidence for understanding the functional differentiation of THAP family proteins in the central nervous system, and suggests that Thap6 knockout mice may serve as a novel animal model for studying neuropsychiatric disorders related to anxiety and social deficits. 目的: THAP结构域蛋白6(THAP domain-containing protein 6，THAP6)是THAP家族成员之一，含保守的THAP锌指结构域，可作为转录因子调控基因表达。既往研究表明THAP6可能参与巨噬细胞激活及社会刺激相关的转录调控，但THAP6是否影响情绪和社交行为及脑组织结构，目前未见报道。本研究旨在明确Thap6基因缺失对小鼠情绪和社交行为的影响，初步探讨Thap6基因敲除对小鼠脑组织的影响，为揭示THAP6在神经精神疾病中的潜在作用提供实验依据。方法: 采用CRISPR/Cas9技术构建Thap6基因全身敲除小鼠模型，通过基因型鉴定筛选Thap6基因敲除杂合子(Thap6+/-，n=8)和野生型(wild-type，WT;n=10)C57BL/6J雄性小鼠作为研究对象。在小鼠成年后，采用转棒实验评估运动和平衡能力，采用旷场实验评估一般活动性和探索能力，采用糖水偏好实验和悬尾实验评估抑郁样行为，采用高架十字迷宫实验评估焦虑样行为，采用三箱社交实验评估社交行为。在行为评估结束后，收集小鼠脑组织，采用苏木精-伊红(hematoxylin and eosin，HE)染色和尼氏染色观察脑组织微结构和神经元数量的改变情况。结果: 行为学结果显示，与WT小鼠相比，Thap6+/-小鼠在高架十字迷宫中进入开放臂的次数百分比显著降低(t=2.516，P=0.024)、在开放臂中的平均运动速度显著增快(t=3.045，P=0.013);在三箱社交实验中，Thap6+/-小鼠的社交能力指数 (t=4.350，P<0.001)和社交偏好指数(t=2.732，P=0.014)均显著低于WT组;其他行为实验各指标2组间的差异均无统计学意义(均P>0.05)。脑组织HE染色结果显示，Thap6+/-小鼠皮质散在细胞形态异常、细胞质疏松、细胞核模糊;杏仁核神经元排列松散、细胞间隔增大、大量空泡形成;小脑浦肯野细胞排列不齐、边界不清、部分细胞核肿胀溶解、细胞缺失;黑质神经元核仁丢失、部分细胞固缩;Thap6+/-组小鼠海马神经元排列有序，细胞形态较好，数目较多，排列紧密。尼氏染色结果显示，Thap6+/-小鼠海马齿状回区域神经元数量显著增多(U=0，P=0.029)，皮质、杏仁核、小脑、黑质4个脑区均出现形态异常和神经元数量减少(均U=0，P=0.029)。结论: Thap6基因敲除导致小鼠焦虑样行为和社交行为缺陷，同时导致小鼠多个脑区脑结构异常和神经元数量减少。本研究首次系统揭示了THAP6在情绪与社交行为调控中的重要作用，为理解THAP家族蛋白在中枢神经系统中的功能分化提供了新证据，并提示Thap6基因敲除小鼠可能作为研究焦虑和社交障碍相关神经精神疾病的新型动物模型。.
Root system development is essential for water and nutrient acquisition but requires substantial carbon investment. Optimizing root architecture while minimizing carbon costs is therefore an important target for crop improvement. Here, we characterized a novel rice (Oryza sativa L.) mutant, truncated elongation growth 1 (teg1), which exhibits reduced seminal and crown root elongation but enhanced lateral root development. Histological analyses revealed that the shortened root phenotype was associated with reduced root apical meristem activity and decreased cell elongation. Rice possesses different lateral root types, with L-type lateral roots being thicker and capable of branching into smaller lateral roots, and S-type lateral roots being thinner and shorter than L-type lateral roots. Despite shorter seminal and crown roots, teg1 developed a higher density of L-type lateral roots and longer L-type lateral roots than the wild type. To investigate the interaction between root architecture and carbon allocation, teg1 was combined with the outstanding rooting 1 (our1) mutation, which promotes seminal, crown and lateral root elongation. The teg1 our1 double mutant exhibited increased L-type lateral root growth accompanied by reduced seminal and crown root growth while maintaining total root length. Carbon-14 tracer analysis revealed enhanced accumulation of newly assimilated carbon in elongating L-type lateral roots of the double mutant. Furthermore, the root-to-shoot dry weight ratio of the double mutant remained comparable to that of the wild type. These results suggest that teg1 promotes compensatory lateral root development and alters carbon allocation within the root system, providing a potential genetic resource for improving root system architecture in rice.
Protein adenylation (AMPylation) is a post-translational modification in which an adenosine monophosphate (AMP) group is covalently attached to target proteins by AMPylases using ATP as a donor. In metazoans, two conserved AMPylase families are known: FIC-domain proteins and SelO. The yeast Saccharomyces cerevisiae lacks a FIC-domain enzyme; its only known AMPylase is the mitochondrial SelO homologue, Fmp40, involved in redox signaling. We conducted the first comprehensive screen for AMPylated proteins in the mitochondrial proteome of S. cerevisiae analyzing both wild-type and fmp40Δ cells using quantitative mass spectrometry. We identified 124 AMPylated mitochondrial proteins in wild-type and 41 in fmp40Δ mitochondria, suggesting the existence of additional AMPylase(s) in yeast. Among the modified targets, seven ATP synthase subunits were AMPylated, many at sites also phosphorylated, underscoring complex PTM regulation of the enzyme. We demonstrated that substitutions of one such residue, serine 29 in the δ subunit (Atp16), to alanine or glutamic acid, altered ATP synthase activity and oxidative phosphorylation coupling under both fermentative and respiratory conditions. This regulation is crucial for maintaining mitochondrial membrane potential. Our study provides the first catalog of AMPylated mitochondrial proteins in yeast, establishing a foundation for future studies on mitochondrial AMPylation.
Heart failure (HF) resulting from chronic systolic overload is associated with increased burden on proteostasis and impaired proteasome function. Phosphorylation of proteasome subunit RPN6/PSMD11 at Ser14 (pS14-RPN6) mediates the activation of 26S proteasomes by PKA, but its significance in common forms of heart disease remains obscure. Hence, we investigated the impact of genetic blockade of pS14-RPN6 on cardiac remodeling and HF during systolic overload, a common condition occurring in hypertension and aortic stenosis. We detected marked increases in ubiquitin conjugates, along with elevated levels of RPN6 and pS14-RPN6 proteins in myocardial tissues from human patients with nonischemic HF. Similarly, myocardial pS14-Rpn6 was increased in wild-type mice 2 and 4 weeks after transverse aortic constriction (TAC). Compared with wild-type littermates, mice with genetic blockade of pS14-Rpn6 resulting from germline knock-in of Rpn6S14A (S14A mice) developed greater cardiac hypertrophy, fibrosis, left ventricular dysfunction, and lung congestion after TAC. Mechanistically, TAC induced comparable increases in 26S and 30S proteasomes at 2-week in both genotypes but, at 4-week post-TAC, the increases were remarkably attenuated in S14A mice. Meanwhile, TAC-induced increases in proteasome peptidase activities were significantly attenuated (2-week) or abolished (4-week) in S14A mice, accompanied by greater increases in total and K48-linked ubiquitin conjugates. Collectively, these findings demonstrate that pS14-RPN6 plays an essential role in upregulating myocardial proteasome activities and sustained upregulation of proteasome assembly, thereby assisting in maintaining proteostasis and protecting against maladaptive cardiac remodeling and HF during systolic overload.
Isocitrate dehydrogenase (IDH) mutation and 1p/19q codeletion are key molecular markers for glioma classification. Amide proton transfer weighted (APTw) and nuclear Overhauser effect-weighted (NOEw) markers showed promise for glioma characterization, by probing protein-related tissue properties. However, their interpretation is confounded by direct water saturation, macromolecules (semi-solid magnetization transfer-ssMT), and T1 relaxation. Here, we aimed to assess the performance of three APTw and NOEw metrics-uncorrected, spillover/ssMT-corrected (FMC), and fully spillover/ssMT- and T1-corrected (FMTC)-for glioma stratification. Fifty patients with suspected gliomas were prospectively enrolled (12 IDH-wild-type, 38 IDH-mutant, of which 21 with 1p/19q codeletion). Acquisitions were performed at 3 T using a 3D gradient echo readout with chemical exchange saturation transfer (B1 = 2 μT for APTw, 0.6 μT for NOEw; T1sat = 2 s), WASABI (WAter Shift And B1) for B0/B1 mapping, and saturation recovery for T1 mapping. Glioma subtypes were compared using metrics extracted from manually segmented masks, using two-tailed Mann-Whitney U tests, and the Benjamini-Hochberg false discovery rate (FDR) correction. Effect sizes were quantified using Cliff's δ with 95% bootstrap confidence intervals and classification performance was assessed by receiver operating characteristic analyses (area under the curve, AUC). IDH-mutant and wild-type gliomas differed significantly for the uncorrected APTw metric (p = 0.005, AUC = 0.79), with stronger discrimination following correction-APTw-FMC (p < 0.001, AUC = 0.94) and APTw-FMTC (p < 0.001, AUC = 0.96), both with large effect sizes. Only APTw-FMTC distinguished 1p/19q codeleted from non-codeleted gliomas before FDR correction (uncorrected p = 0.01, AUC = 0.74). The NOEw metrics did not differ between any molecular subgroups, likely due to limited sensitivity of this contrast at 3 T. These results suggest that correcting for fluid, ssMT, and T1 effects enhances the accuracy of APTw metrics, offering a more robust and biophysically grounded approach to noninvasive glioma diagnosis.
Nitrate consumption by ruminants may enrich nitrate-respiring Salmonella in the gut. To test whether tungstate, an inhibitor of nitrate reductase, may prevent nitrate-promoted enrichment of Salmonella, ruminal microbes inoculated with or without 104 colony-forming units (CFU)/mL of Salmonella Newport were incubated for 26 h under simulated rumen conditions with or without 10 mM nitrate, 100 mM tungstate, or their combination. Results indicated more nitrate was metabolized (p < 0.05) by ruminal populations supplemented with nitrate alone than with nitrate and tungstate combined, with means (± standard deviations) being 8.55 ± 0.74 and 3.97 ± 0.67 μmol nitrate/mL, respectively. Nitrite accumulations were affected (p < 0.05) by tungstate treatment, achieving 4.96 ± 0.26 and 0.18 ± 0.07 μmol/mL in populations supplemented with nitrate alone or combined with tungstate, respectively, when not inoculated with S. Newport and achieving 1.95 ± 0.18 and 0.06 ± 0.01 μmol/mL, respectively, when inoculated with S. Newport. Salmonella increased (p < 0.05) in populations treated with tungstate, alone or combined with nitrate (5.82 ± 0.10 and 5.57 ± 0.15 log10 CFU/mL, respectively), compared to controls or nitrate-only supplemented populations (1.63 ± 0.58 and 2.84 ± 0.21 log10 CFU/mL, respectively). In populations not inoculated with S. Newport, the addition of nitrate, tungstate, or their combination increased (p < 0.05) wild-type coliforms by 1.4 to 3.9 log10 units compared to untreated controls (2.67 ± 0.18 log10 CFU/mL). In S. Newport-inoculated populations, tungstate treatment, alone or combined with nitrate, increased (p < 0.05) coliforms by 2.8 to 3.2 log10 units compared to control and nitrate-only supplemented populations (3.06 ± 0.48 and 3.49 ± 0.06 log10 CFU/mL, respectively). Wildtype lactic acid bacteria were enriched by tungstate treatment and nitrate supplementation, alone or combined to 0.2 to 1.1 log10 units, compared to untreated controls (6.84 ± 0.03 and 6.68 ± 0.16 log10 CFU/mL, respectively). Methane production decreased by 70% (p < 0.05) in populations supplemented with nitrate, whether alone or combined with tungstate, compared to untreated or tungstate-only treated populations (29.85 ± 5.05 and 23.08 ± 5.49 μmol methane/mL of incubation fluid, respectively). These results indicate that tungstate treatment marginally decreased nitrate metabolism in the ruminal populations but surprisingly promoted Salmonella enrichment.
Selective breeding in aquaculture is necessary to establish food security and meet demand for sustainably produced protein. An informed selective breeding program requires understanding how population structure, environmental adaptation, and human activities shape natural genetic variation in wild conspecifics. Unfortunately, wild variation remains poorly characterised for many commercially important aquaculture species. Here, we conduct the first range-wide study of genomic population structure for the eastern oyster (Crassostrea virginica) across thousands of miles (Texas, USA to Eastern Canada) using a 200K SNP array. We integrate population structure analyses, genotype-environmental associations, and structural variant detection to identify adaptive loci and quantify human-mediated genetic impacts. Our data confirm two ancestral clusters with a phylogeographic break between the Gulf and Atlantic (FST = 0.16) and highlight patterns of substructure within each region. We find evidence of unexpected patterns of genomic variation in two locations: evidence of Gulf ancestry in a mid-Atlantic estuary (Chesapeake Bay), and evidence of Atlantic ancestry in a Gulf estuary (Apalachicola Bay). While we cannot definitively determine the causes of these unexpected patterns, we show that they are consistent with direct and indirect human impacts in these estuaries. Genotype-environment association analyses with in situ temperature and salinity measurements were used to identify putatively adaptive loci, including SNPs within large structural variants (> 1 Mb). Our results identified genomic targets for aquaculture breeding programs aimed at climate resilience, reveal complex patterns of human impacts in managed systems, and demonstrate how seascape genomics can be used to improve aquaculture outcomes.