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Abstract The chromatin modifier GRAIN WEIGHT 6a (GW6a) enhances rice grain size and yield. However, little is known about its gene network determining grain size. Here, we report that MITOGEN-ACTIVED PROTEIN KINASE 6 (OsMAPK6) and E3 ligase CHANG LI GENG 1 (CLG1) interact with and target GW6a for phosphorylation and ubiquitylation, respectively. Unexpectedly, however, in vitro and in vivo assays reveal that both of the two post-translational modifications stabilize GW6a. Furthermore, we uncover two major GW6a phosphorylation sites (serine142 and threonine186) targeted by OsMAPK6 serving an important role in modulating grain size. In addition, our genetic and molecular results suggest that the OsMAPK6-GW6a and CLG1-GW6a axes are crucial and operate in a non-additive manner to control grain size. Overall, our findings identify a previously unknown mechanism by which phosphorylation and ubiquitylation non-additively stabilize GW6a to enhance grain size, and reveal correlations and interactions of these posttranslational modifications during rice grain development.

Summary: Integrins play critical roles in connecting the extracellular matrix and actin. While the upregulation of integrins is thought to promote cancer stemness and metastasis, the mechanisms underlying their upregulation in cancer stem cells (CSCs) remain poorly understood. Herein, we show that USP22 is essential in maintaining breast cancer cell stemness by promoting the transcription of integrin β1 (ITGB1). Both genetic and pharmacological inhibition of USP22 largely impaired breast CSCs self-renewal and prevented their metastasis. Reconstitution of integrin β1 partially rescued USP22-null breast cancer metastasis. USP22 functions as a bona fide deubiquitinase to protect the proteasomal degradation of the forkhead box M1 (FoxM1), a transcription factor for tumoral ITGB1 gene transcription. Immunohistochemistry staining detected a positive correlation among USP22, FoxM1, and integrin β1 in human breast cancers. Collectively, our study identifies the USP22-FoxM1-integrin β1 signaling axis as critical for cancer stemness and offers a potential target for antitumor therapy.

Estimation of key indicators in rivers was usually conducted with use of monitoring data of Internet of Things. Currently, it has been a more practical demand to extract key indexes in rivers from the perspective of visual remote sensing, rather than data-driven perspective. As consequence, this study aims to explore the deep learning-based key indicators extraction method from remote sensing images of rivers. First of all, a large amount of river-related remote sensing images, including high-resolution satellite images and aerial photographs, are collected. Then, U-Net structure is utilized as the backbone network to realize semantic segmentation via multimodal feature fusion. On this basis, fine-grained vision features are extracted to estimate values of key indicators. Finally, the width and flow velocity of the river are identified and verified. Using deep convolutional neural networks and recurrent neural networks for feature extraction and modeling, the model can infer and estimate relevant information of rivers by learning key features from images. Empirically, we have also carried out some experiments on real-world remote sensing images to evaluate the proposal. The results indicate that our method performs well in extracting key indicators of rivers, and has higher accuracy compared to traditional methods. In addition, we also conduct sensitivity analysis on the model and find that it has certain stability to factors that affect river characteristics, such as geographical environment and climate conditions.

In this study, the extraction, purification and metabolic enzyme inhibition potential of Selaginella doederleinii were investigated. In order to extract the total biflavonoids from S. doederleinii (SDTBs), the optimum extraction process was obtained by optimizing the ultrasonic extraction parameters using response-surface methodology. This resulted in a total biflavonoid content of 22.26 ± 0.35 mg/g. Purification of the S. doederleinii extract was carried out using octadecylsilane (ODS), and the transfer rate of the SDTBs was 82.12 ± 3.48% under the optimum purification conditions. We determined the effect of the SDTBs on α-glucosidase (AG), α-amylase and xanthine oxidase (XOD) and found that the SDTBs had an extremely potent inhibitory effect on AG, with an IC50 value of 57.46 μg/mL, which was much lower than that of the positive control. Meanwhile, they also showed significant inhibition of XOD and α-amylase, with IC50 values of 289.67 μg/mL and 50.85 μg/mL, respectively. In addition, molecular docking studies were carried out to understand the nature of the action of the biflavonoids on AG and XOD. The results showed that robustaflavone had the lowest binding energy to AG (−11.33 kcal/mol) and XOD (−10.21 kcal/mol), while, on the other hand, amentoflavone showed a good binding affinity to AG (−10.40 kcal/mol) and XOD (−9.962 kcal/mol). Moreover, molecular dynamics simulations verified the above results.

This paper aims to present a comprehensive assessment of the evolution of environmental law in Puerto Rico from 1897 to 2021. The study identifies significant legislative developments, with a surge in federal and local laws post-1960, correlating with heightened environmental awareness. Political dynamics, notably party affiliations, significantly influence the level of legal protection afforded to the environment. A new classification system is introduced to analyze laws based on ecosystem and resource protection. Distinct periods of legislative activity are identified, with federal laws predominant from 1960 to 1990 and state laws from 1990 to 2021, revealing a unique pattern of legislative ''pulses.'' Federal laws primarily address coastal ecosystems and pollution mitigation, while state laws focus on land ecosystems, reserves, and resource management. The study establishes a statistically significant correlation between the passage of time and the number of enacted laws, indicating an upward trend in environmental legislation. Additionally, it highlights the impact of political parties on lawmaking, with variations in legislative output observed under different party control scenarios. The study found that the legislative output in both the US and Puerto Rico was affected by party control. Specifically, when the Republican Party or the local Popular Democratic Party controls the House of Representatives or the Senate, the output of environmental legislation decreases. On the other hand, the study suggests that the US President's party identity has a lesser effect on enacting environmental laws. The study underscores the intricate relationship between politics and environmental policy, emphasizing the significance of political dynamics in shaping environmental law. The findings offer valuable insights for policymakers, environmentalists, and stakeholders, aiding in developing more effective strategies for sustainable development and conservation in Puerto Rico. Recommendations include utilizing the comprehensive table of environmental regulations to detect agency overlaps and investigate the correlation between legal protection and ecosystem health.

Abstract. Integrins are a family of transmembrane receptors that connect the extracellular matrix and actin skeleton, which mediate cell adhesion, migration, signal transduction, and gene transcription. As a bi-directional signaling molecule, integrins can modulate many aspects of tumorigenesis, including tumor growth, invasion, angiogenesis, metastasis, and therapeutic resistance. Therefore, integrins have a great potential as antitumor therapeutic targets. In this review, we summarize the recent reports of integrins in human hepatocellular carcinoma (HCC), focusing on the abnormal expression, activation, and signaling of integrins in cancer cells as well as their roles in other cells in the tumor microenvironment. We also discuss the regulation and functions of integrins in hepatitis B virus-related HCC. Finally, we update the clinical and preclinical studies of integrin-related drugs in the treatment of HCC.

Persistent human papillomavirus (HPV) infections is necessary for the development of cervical cancers. An increasing number of retrospective studies have found the depletion of Lactobacillus microbiota in the cervico-vagina facilitate HPV infection and might be involved in viral persistence and cancer development. However, there have been no reports confirming the immunomodulatory effects of Lactobacillus microbiota isolated from cervico-vaginal samples of HPV clearance in women. Using cervico-vaginal samples from HPV persistent infection and clearance in women, this study investigated the local immune properties in cervical mucosa. As expected, type I interferons, such as IFN-α and IFN-β, and TLR3 globally downregulated in HPV+ persistence group. Luminex cytokine/chemokine panel analysis revealed that L. jannaschii LJV03, L. vaginalis LVV03, L. reuteri LRV03, and L. gasseri LGV03 isolated from cervicovaginal samples of HPV clearance in women altered the host's epithelial immune response, particularly L. gasseri LGV03. Furthermore, L. gasseri LGV03 enhanced the poly (I:C)-induced production of IFN by modulating the IRF3 pathway and attenuating poly (I:C)-induced production of proinflammatory mediators by regulating the NF-κB pathway in Ect1/E6E7 cells, indicating that L. gasseri LGV03 keeps the innate system alert to potential pathogens and reduces the inflammatory effects during persistent pathogen infection. L. gasseri LGV03 also markedly inhibited the proliferation of Ect1/E6E7 cells in a zebrafish xenograft model, which may be attributed to an increased immune response mediated by L. gasseri LGV03.

ABSTRACT Widespread bacterial resistance among Gram-negative bacteria is rapidly depleting our antimicrobial arsenal. Adjuvants that enhance the bactericidal activity of existing antibiotics provide a way to alleviate the resistance crisis, as new antimicrobials are becoming increasingly difficult to develop. The present work with Escherichia coli revealed that neutralized lysine (lysine hydrochloride) enhances the bactericidal activity of β-lactams in addition to increasing bacteriostatic activity. When combined, lysine hydrochloride and β-lactam increased expression of genes involved in the tricarboxylic acid (TCA) cycle and raised reactive oxygen species (ROS) levels; as expected, agents known to mitigate bactericidal effects of ROS reduced lethality from the combination treatment. Lysine hydrochloride had no enhancing effect on the lethal action of fluoroquinolones or aminoglycosides. Characterization of a tolerant mutant indicated involvement of the FtsH/HflkC membrane-embedded protease complex in lethality enhancement. The tolerant mutant, which carried a V86F substitution in FtsH, exhibited decreased lipopolysaccharide levels, reduced expression of TCA cycle genes, and reduced levels of ROS. Lethality enhancement by lysine hydrochloride was abolished by treating cultures with Ca2+ or Mg2+, cations known to stabilize the outer membrane. These data, plus damage observed by scanning electron microscopy, indicate that lysine stimulates β-lactam lethality by disrupting the outer membrane. Lethality enhancement of β-lactams by lysine hydrochloride was also observed with Acinetobacter baumannii and Pseudomonas aeruginosa, thereby suggesting that the phenomenon is common among Gram-negative bacteria. Arginine hydrochloride behaved in a similar way. Overall, the combination of lysine or arginine hydrochloride and β-lactam offers a new way to increase β-lactam lethality with Gram-negative pathogens. IMPORTANCE Antibiotic resistance among Gram-negative pathogens is a serious medical problem. The present work describes a new study in which a nontoxic nutrient increases the lethal action of clinically important β-lactams. Elevated lethality is expected to reduce the emergence of resistant mutants. The effects were observed with significant pathogens (Escherichia coli, Acinetobacter baumannii, and Pseudomonas aeruginosa), indicating widespread applicability. Examination of tolerant mutants and biochemical measurements revealed involvement of endogenous reactive oxygen species in response to outer membrane perturbation. These lysine hydrochloride–β-lactam data support the hypothesis that lethal stressors can stimulate the accumulation of ROS. Genetic and biochemical work also revealed how an alteration in a membrane protease, FtsH, abolishes lysine stimulation of β-lactam lethality. Overall, the work presents a method for antimicrobial enhancement that should be safe, easy to administer, and likely to apply to other nutrients, such as arginine.

In this study, the deep eutectic solvent based ultrasound-assisted extraction (DES-UAE) was investigated for the efficient and environmentally friendly extraction of Selaginella chaetoloma total biflavonoids (SCTB). As an extractant for optimization, tetrapropylaminium bromide-1,4-butanediol (Tpr-But) was employed for the first time. 36 DESs were created, with Tpr-But producing the most effective results. Based on response surface methodology (RSM), the greatest extraction rate of SCTB was determined to be 21.68 ± 0.78 mg/g, the molar ratio of HBD to HBA was 3.70:1, the extraction temperature was 57 °C, and the water content of DES was 22 %. In accordance with Fick's second rule, a kinetic model for the extraction of SCTB by DES-UAE has been derived. With correlation coefficients 0.91, the kinetic model of the extraction process was significantly correlated with the general and exponential equations of kinetics, and some important kinetic parameters such as rate constants, energy of activation and raffinate rate were determined. In addition, molecular dynamics simulations were used to study the extraction mechanisms generated by different solvents. Comparing the effect of several extraction methods on S.chaetoloma using ultrasound-assisted extraction and conventional methods, together with SEM examination, revealed that DES-UAE not only saved time but also enhanced SCTB extraction rate by 1.5–3 folds. SCTB demonstrated superior antioxidant activity in three studies in vitro. Furthermore, the extract could suppress the growth of A549, HCT-116, HepG2, and HT-29 cancer cells. Alpha-Glucosidase (AG) inhibition experiment and molecular docking studies suggested that SCTB exhibited strong inhibitory activity against AG and potential hypoglycemic effects. The results of this study indicated that a Tpr-But-based UAE method was suitable for the efficient and environmentally friendly extraction of SCTB, and also shed light on the mechanisms responsible for the increased extraction efficiency, which could aid in the application of S.chaetoloma and provide insight into the extraction mechanism of DES.

IntroductionSince its discovery in 1999, a substantial body of research has shown that iASPP is highly expressed in various kinds of tumors, interacts with p53, and promotes cancer cell survival by antagonizing the apoptotic activity of p53. However, its role in neurodevelopment is still unknown.MethodsWe studied the role of iASPP in neuronal differentiation through different neuronal differentiation cellular models, combined with immunohistochemistry, RNA interference and gene overexpression, and studied the molecular mechanism involved in the regulation of neuronal development by iASPP through coimmunoprecipitation coupled with mass spectrometry (CoIP-MS) and coimmunoprecipitation (CoIP).ResultsIn this study, we found that the expression of iASPP gradually decreased during neuronal development. iASPP silencing promotes neuronal differentiation, while its overexpression inhibited neurite differentiation in a variety of neuronal differentiation cellular models. iASPP associated with the cytoskeleton-related protein Sptan1 and dephosphorylated the serine residues in the last spectrin repeat domain of Sptan1 by recruiting PP1. The non-phosphorylated and phosphomimetic mutant form of Sptbn1 inhibited and promoted neuronal cell development respectively.ConclusionOverall, we demonstrate that iASPP suppressed neurite development by inhibiting phosphorylation of Sptbn1.

Extreme climate events such as storms and severe droughts are becoming more frequent under the warming climate. In the tropics, excess rainfall carried by hurricanes causes massive flooding and threatens ecosystems and human society. We assessed recent major floodings on the tropical island of Puerto Rico after Hurricane Maria in 2017 and Hurricane Fiona in 2022, both of which cost billions of dollars damages to the island. We analyzed the Sentinel-1 synthetic aperture radar (SAR) images right after the hurricanes and detected surface inundation extent by applying a random forest classifier. We further explored hurricane rainfall patterns, flow accumulation, and other possible drivers of surface inundation at watershed scale and discussed the limitations. An independent validation dataset on flooding derived from high-resolution aerial images indicated a high classification accuracy with a Kappa statistic of 0.83. The total detected surface inundation amounted to 10,307 ha after Hurricane Maria and 7949 ha after Hurricane Fiona for areas with SAR images available. The inundation patterns are differentiated by the hurricane paths and associated rainfall patterns. We found that flow accumulation estimated from the interpolated Fiona rainfall highly correlated with the ground-observed stream discharges, with a Pearson’s correlation coefficient of 0.98. The detected inundation extent was found to depend strongly on hurricane rainfall and topography in lowlands within watersheds. Normal climate, which connects to mean soil moisture, also contributed to the differentiated flooding extent among watersheds. The higher the accumulated Fiona rain and the lower the mean elevation in the flat lowlands, the larger the detected surface flooding extent at the watershed scale. Additionally, the drier the climate, which might indicate drier soils, the smaller the surface flooding areas. The approach used in this study is limited by the penetration capability of C-band SAR; further application of L-band images would expand the detection to flooding under dense vegetation. Detecting flooding by applying machine learning techniques to SAR satellite images provides an effective, efficient, and reliable approach to flood assessment in coastal regions on a large scale, hence helping to guide emergency responses and policy making and to mitigate flooding disasters.

Reforestation often occurs when the economy shifts from agriculture to industry and services such as tourism. However, there is a lack of coherent knowledge and investigation about the impact of reforestation in the tropics on hydrological variability as well as flood risks. It is unclear how changes in forest cover and pattern will affect flood risks and watershed response to future altered rainfall intensity. This study uses the Soil Water Assessment Tool (SWAT+) to simulate the impact of reforestation, the locality of forest, and the concentrated rainfall on the hydrology of the largest watershed in Puerto Rico. SWAT+ is a computer model simulating watershed hydrology driven by meteorological input and the characteristics of soils and land use. We hypothesized that increased forest cover, especially at low elevation range, would reduce flood risk and that reduced rain days while maintaining the mean annual rainfall invariant would increase stream discharge variability. We found that reforestation significantly reduced large discharges but increased small discharges; that forest at low elevation tended to reduce large and extreme discharges in comparison with forest at high elevation; and that more concentrated rainfall not only increased the rainfall variability but also increased the discharge variability. However, both the impact of shifting forest locality and the response of watershed to altered rainfall intensity strongly depended on geophysical factors such as ranges of elevation and slope. Moving forests to lower elevation in subbasins with steeper slopes showed a stronger reduction in extreme discharges than in subbasins with flatter slopes. On the other hand, subbasins with steeper slopes tended to response more strongly to more concentrated rainfall with greater increase in discharge variability than subbasins with flatter slopes. To cope with future increased climate variability, our results favor reforestation at lower elevation for watershed with large elevation ranges and steep slopes.

Abstract Background Growing evidence has proved that stromal cells, as the critical component of tumor microenvironment (TME), are closely associated with tumor's progression. However, the model based on stromal score to predict progression‐free survival (PFS) in papillary thyroid carcinoma (PTC) has not been developed. The study aimed at exploring the relation between stromal score and prognosis, then establishing a nomogram to predict PFS of patients with PTC. Method We obtained the stromal score and clinicopathological characteristics of PTC patients from The Cancer Genome Atlas (TCGA) database. Cox regression analysis assisted in selecting prognosis‐related factors. A stromal score‐based nomogram was built and verified in the training and validation cohorts, respectively. The calibration curve, concordance index (C‐index), decision curve analysis (DCA) as well as receiver operating characteristic (ROC) curve assisted in measuring the performance exhibited by the nomogram. Results We divided 381 PTC patients into the training cohort (n = 269) and the validation cohort (n = 112) randomly. Compared with patients who had a low stromal score, patients with a high stromal score appeared with significantly better PFS [Hazard ratio (HR) and 95% confidence interval (CI): 0.294, 0.130–0.664]. The C‐index of the PFS nomogram was 0.764 (0.662–0.866) in the training cohort and 0.717 (0.603–0.831) in the validation cohort. The calibration curves for PFS prediction in the nomogram were remarkably consistent with the actual observation. DCA indicated superior performance of the nomogram to predict PFS than the American Joint Committee on Cancer (AJCC) Tumor Node Metastasis (TNM) staging system. The ROC curves showed the favorable sensitivity and specificity of the novel nomogram. Conclusion High stromal score was significantly associated with improved PFS in patients with PTC. The nomogram based on the stromal score and clinicopathological patterns yielded a reliable performance to predict the prognosis of PTC.

Endo180, a collagen binding receptor, is highly expressed in a subset of cancer-associated fibroblasts. The authors show, using knockout mice and 3D in vitro assays, that Endo180 depletion impairs tumour fibroblast contractility and viability resulting in reduced tumour growth and metastasis.

Despite the many strategies employed to slow the spread of cancer, the development of new anti-tumor drugs and the minimization of side effects have been major research hotspots in the anti-tumor field. Natural drugs are a huge treasure trove of drug development, and they have been widely used in the clinic as anti-tumor drugs. Selaginella species in the family Selaginellaceae are widely distributed worldwide, and they have been well-documented in clinical practice for the prevention and treatment of cancer. Biflavonoids are the main active ingredients in Selaginella, and they have good biological and anti-tumor activities, which warrant extensive research. The promise of biflavonoids from Selaginella (SFB) in the field of cancer therapy is being realized thanks to new research that offers insights into the multi-targeting therapeutic mechanisms and key signaling pathways. The pharmacological effects of SFB against various cancers in vitro and in vivo are reviewed in this review. In addition, the types and characteristics of biflavonoid structures are described in detail; we also provide a brief summary of the efforts to develop drug delivery systems or combinations to enhance the bioavailability of SFB monomers. In conclusion, SFB species have great potential to be developed as adjuvant or even primary therapeutic agents for cancer, with promising applications.

Mode instability (MI) has become a major factor that restricts the brightness enhancement of fiber laser. This paper studies the MI threshold characteristics of the fiber oscillators with refrigerated ytterbium-doped fiber (YDF). In the experiments, two fiber lasers that can operate at low temperature were built. We rigorously tested the MI threshold of the lasers with the YDF placed in two different environments. At room temperature environment, the MI threshold of the oscillator was measured with the temperature of the YDF water cooling plate ranging from 25 °C to 5 °C. When the cooling temperature of the YDF drops from 20 °C to 5 °C, the MI threshold of the laser rises gradually, but the increase is small. In another experiment, the YDF is assembled in a constant temperature test chamber, in which the cooling of the YDF is more efficient. It is found that as the cooling temperature drops, the MI threshold of the laser has an obvious upward trend. In the process of the cooling temperature of the YDF dropped from 20 °C to −10 °C, the MI threshold of the laser was increased from 867 W to above 931 W, an increase of nearly 10%. It is the first detailed experimental demonstration which shows that decreasing the cooling temperature of the gain fiber can indeed increase the MI threshold of the fiber oscillator. This work can clarify the influence of cooling temperature on the laser thermal effect, which is conducive to perfecting the theoretical model of the MI effect.

ObjectiveThe objectives of this study were to prospectively 1) explore the characteristics and enhanced patterns of carotid body tumors (CBTs) at color Doppler ultrasound (CDU) and contrast-enhanced ultrasonography (CEUS) qualitatively and quantitatively and 2) compare CDU and CEUS for their morphology and vascularity signature.MethodsCDU and CEUS with Sonovue® were used to evaluate 25 CBT lesions. The comparison between these ultrasonic modalities included the size, Shamblin type, vascularity, and feeding vessels of the lesion areas. The time–intensity curve (TIC) analysis was used to obtain the dynamics of the contrast-enhancement features of CBTs.ResultsThe TIC analysis presented a fast wash-in [wash-in time: 3.00 ± 1.10 s, mean ± SD] and slow wash-out [wash-out time: 58.79 ± 24.21 s, mean ± SD] pattern in the CBT lesions, with a high area under the curve (AUC) of 669.68 ± 143.46 mm2 (mean ± SD). In comparison with CDU, CEUS was superior in identifying Shamblin type I or III CBT lesions (χ2 = 17.389, p=0.002). It detected a significant difference in the AUC between moderate and marked vascularity groups (563.33 ± 102.63 vs. 707.22 ± 138.81, t=-2.311, p=0.031.), while CDU observed no significant difference between these two groups. Although CDU was more sensitive than CEUS in detecting feeding vessels (100% vs. 88%), CEUS better visualized the origins of feeding vessels (χ2 = 9.162, p=0.010).ConclusionCEUS can better investigate the Shamblin type and vascularity of CBT lesions than CDU. CBTs displayed a fast wash-in, slow wash-out pattern with high AUC in the TIC analysis in the CEUS mode. CDU is more sensitive in detecting feeding vessels than CEUS, while CEUS can better visualize the origins of feeding vessels.

Brown cotton fiber (BCF) is a unique raw material of naturally colored cotton (NCC). But characteristics of the regulatory gene network and metabolic components related to the proanthocyanidins biosynthesis pathway at various stages of its fiber development remain unclear. Here, the dynamic changes in proanthocyanidins biosynthesis components and transcripts in the BCF variety “Zong 1-61” and its white near-isogenic lines (NILs) “RT” were characterized at five fiber developmental stages (0, 5, 10, 15, and 20 days post-anthesis; DPA). Enrichment analysis of differentially expressed genes (DEGs), comparison of metabolome differences, and pathway enrichment analysis of a weighted gene correlation network analysis together revealed the dominant gene expression of flavonoid biosynthesis (FB), phenylpropanoid metabolisms, and some carbohydrate metabolisms at 15 or 20 DPA than white cotton. Eventually, 63 genes were identified from five modules putatively related to FB. Three R2R3-MYB and two bHLH transcription factors were predicted as the core genes. Further, GhANS, GhANR1, and GhUFGT2 were preliminarily regulated by GhMYB46, GhMYB6, and GhMYB3, respectively, according to yeast one-hybrid assays in vitro. Our findings provide an important transcriptional regulatory network of proanthocyanidins biosynthesis pathway and dynamic flavonoid metabolism profiles.

Background Schizophrenia is a serious mental illness affecting approximately 20 million individuals globally. Both genetic and environmental factors contribute to the illness. If left undiagnosed and untreated, schizophrenia results in impaired social function, repeated hospital admissions, reduced quality of life and decreased life expectancy. Clinical diagnosis largely relies on subjective evidence, including self-reported experiences, and reported behavioural abnormalities followed by psychiatric evaluation. In addition, psychoses may occur along with other conditions, and the symptoms are often episodic and transient, posing a significant challenge to the precision of diagnosis. Therefore, objective, specific tests using biomarkers are urgently needed for differential diagnosis of schizophrenia in clinical practice.Aims We aimed to provide evidence-based and consensus-based recommendations, with a summary of laboratory measurements that could potentially be used as biomarkers for schizophrenia, and to discuss directions for future research.Methods We searched publications within the last 10 years with the following keywords: ‘schizophrenia’, ‘gene’, ‘inflammation’, ‘neurotransmitter’, ‘protein marker’, ‘gut microbiota’, ‘pharmacogenomics’ and ‘biomarker’. A draft of the consensus was discussed and agreed on by all authors at a round table session.Results We summarised the characteristics of candidate diagnostic markers for schizophrenia, including genetic, inflammatory, neurotransmitter, peripheral protein, pharmacogenomic and gut microbiota markers. We also proposed a novel laboratory process for diagnosing schizophrenia in clinical practice based on the evidence summarised in this paper.Conclusions Further efforts are needed to identify schizophrenia-specific genetic and epigenetic markers for precise diagnosis, differential diagnosis and ethnicity-specific markers for the Chinese population. The development of novel laboratory techniques is making it possible to use these biomarkers clinically to diagnose disease.

Abstract Background Root systems are critical for plant growth and development. The Casparian strip in root systems is involved in stress resistance and maintaining homeostasis. Casparian strip membrane domain proteins (CASPs) are responsible for the formation of Casparian strips. Results To investigate the function of CASPs in cotton, we identified and characterized 48, 54, 91 and 94 CASPs from Gossypium arboreum, Gossypium raimondii, Gossypium barbadense and Gossypium hirsutum, respectively, at the genome-wide level. However, only 29 common homologous CASP genes were detected in the four Gossypium species. A collinearity analysis revealed that whole genome duplication (WGD) was the primary reason for the expansion of the genes of the CASP family in the four cotton species. However, dispersed duplication could also contribute to the expansion of the GaCASPs gene family in the ancestors of G. arboreum. Phylogenetic analysis was used to cluster a total of 85 CASP genes from G. arboreum and Arabidopsis into six distinct groups, while the genetic structure and motifs of CASPs were conserved in the same group. Most GaCASPs were expressed in diverse tissues, with the exception of that five GaCASPs (Ga08G0113, Ga08G0114, Ga08G0116, Ga08G0117 and Ga08G0118) that were highly expressed in root tissues. Analyses of the tissue and subcellular localization suggested that GaCASP27 genes (Ga08G0117) are membrane protein genes located in the root. In the GaCASP27 silenced plants and the Arabidopsis mutants, the lateral root number significantly increased. Furthermore, GaMYB36, which is related to root development was found to regulate lateral root growth by targeting GaCASP27. Conclusions This study provides a fundamental understanding of the CASP gene family in cotton and demonstrates the regulatory role of GaCASP27 on lateral root growth and development.

Purpose: To evaluate the long-term survival and functional outcomes of patients with prolonged disorders of consciousness (pDoC) 1–8 years after brain injuries. Methods: Retrospective study to assess the long-term survival and functional outcomes of patients with pDoC was conducted. We performed Cox regression and multivariate logistic regression to calculate hazard ratios (HRs) for the outcome of survival and to identify risk factors of the functional outcome. Results: We recruited 154 patients with pDoC. The duration of follow-up from disease onset was 1–8 years. The median age was 46 years (IQR, 32–59), and 65.6% (n = 101) of them were men. During the follow-up period, one hundred and ten patients (71.4%) survived; among them, 52 patients had a good outcome. From the overall survival curve, the 1-, 3-, and 8-year survival rates of patients were about 80.5%, 72.0%, and 69.7%, respectively. Cox regression analysis revealed a significant association between the lower APACHE II score (p = 0.005) (cut-off score ≥ 18) and the presence of sleep spindles (p = 0.001) with survival. Logistic regression analysis demonstrated a higher CRS-R score (cut-off score ≥ 7), and presence of sleep spindles were related to a favorable outcome among patients with pDoC. Conclusions: Sleep spindles are correlated with both long-term survival and long-term functional outcome in pDoC patients.

Abstract Background Endocrine therapy reduces breast cancer mortality by 40%, but resistance remains a major clinical problem. In this study, we sought to investigate the impact of aromatase inhibitor (AI) therapy on gene expression and identify gene modules representing key biological pathways that relate to early AI therapy resistance. Methods Global gene expression was measured on pairs of core-cut biopsies taken at baseline and at surgery from 254 patients with ER-positive primary breast cancer randomised to receive 2-week presurgical AI (n = 198) or no presurgical treatment (control n = 56) from the POETIC trial. Data from the AI group was adjusted to eliminate artefactual process-related changes identified in the control group. The response was assessed by changes in the proliferation marker, Ki67. Results High baseline ESR1 expression associated with better AI response in HER2+ tumours but not HER2− tumours. In HER2− tumours, baseline expression of 48 genes associated with poor antiproliferative response (p

Abstract Background Endocrine therapies are still the main strategy for the treatment of oestrogen receptor-positive (ER+) breast cancers (BC), but resistance remains problematic. Cross-talk between ER and PI3K/AKT/mTORC has been associated with ligand-independent transcription of ER. We have previously reported the anti-proliferative effects of the combination of everolimus (an mTORC1 inhibitor) with endocrine therapy in resistance models, but potential routes of escape via AKT signalling can lead to resistance; therefore, the use of dual mTORC1/2 inhibitors has met with significant interest. Methods To address this, we tested the effect of vistusertib, a dual mTORC1 and mTORC2 inhibitor, in a panel of endocrine-resistant and endocrine-sensitive ER+ BC cell lines, with varying PTEN, PIK3CA and ESR1 mutation status. End-points included proliferation, cell signalling, cell cycle and effect on ER-mediated transcription. Two patient-derived xenografts (PDX) modelling endocrine resistance were used to assess the efficacy of vistusertib, fulvestrant or the combination on tumour progression, and biomarker studies were conducted using immunohistochemistry and RNA-seq technologies. Results Vistusertib caused a dose-dependent decrease in proliferation of all the cell lines tested and reduced abundance of mTORC1, mTORC2 and cell cycle markers, but caused an increase in abundance of EGFR, IGF1R and ERBB3 in a context-dependent manner. ER-mediated transcription showed minimal effect of vistusertib. Combined therapy of vistusertib with fulvestrant showed synergy in two ER+ PDX models of resistance to endocrine therapy and delayed tumour progression after cessation of therapy. Conclusions These data support the notion that models of acquired endocrine resistance may have a different sensitivity to mTOR inhibitor/endocrine therapy combinations.

Cancer cells must develop distinct metabolic adaptations to survive in challenging metastatic environments. Here, the authors find, via an in vivo RNAi screen, that the aldo-keto reductase AKR1B10 limits the toxic side effects of oxidative stress to sustain fatty acid oxidation and promote metastatic colonisation.

Abstract Background Cupressus gigantea, a rare and endangered tree species with remarkable medicinal value, is endemic to the Tibetan Plateau. Yet, little is known about the underlying genetics of the unique ecological adaptability of this extremely long-lived conifer with a large genome size. Here, we present its first de novo and multi-tissue transcriptome in-depth characterization. Results We performed Illumina paired-end sequencing and RNA libraries assembly derived from terminal buds, male and female strobili, biennial leaves, and cambium tissues taken from adult C. gigantea. In total, large-scale high-quality reads were assembled into 101,092 unigenes, with an average sequence length of 1029 bp, and 6848 unigenes (6.77%) were mapped against the KEGG databases to identify 292 pathways. A core set of 41,373 genes belonging to 2412 orthologous gene families shared between C. gigantea and nine other plants was revealed. In addition, we identified 2515 small to larger-size gene families containing in total 9223 genes specific to C. gigantea, and enriched for gene ontologies relating to biotic interactions. We identified an important terpene synthases gene family expansion with its 121 putative members. Conclusions This study presents the first comprehensive transcriptome characterization of C. gigantea. Our results will facilitate functional genomic studies to support genetic improvement and conservation programs for this endangered conifer.

Abstract Background Dissemination of breast cancers to the brain is associated with poor patient outcome and limited therapeutic options. In this study we sought to identify novel regulators of brain metastasis by profiling mouse mammary carcinoma cells spontaneously metastasising from the primary tumour in an immunocompetent syngeneic host. Methods 4T1 mouse mammary carcinoma sublines derived from primary tumours and spontaneous brain and lung metastases in BALB/c mice were subject to genome-wide expression profiling. Two differentially expressed genes, Id2 and Aldh3a1, were validated in in-vivo models using mouse and human cancer cell lines. Clinical relevance was investigated in datasets of breast cancer patients with regards to distant metastasis-free survival and brain metastasis relapse-free survival. The role of bone morphogenetic protein (BMP)7 in regulating Id2 expression and promoting cell survival was investigated in two-dimensional and three-dimensional in-vitro assays. Results In the spontaneous metastasis model, expression of Id2 and Aldh3a1 was significantly higher in 4T1 brain-derived sublines compared with sublines from lung metastases or primary tumour. Downregulation of expression impairs the ability of cells to colonise the brain parenchyma whereas ectopic expression in 4T1 and human MDA-MB-231 cells promotes dissemination to the brain following intracardiac inoculation but has no impact on the efficiency of lung colonisation. Both genes are highly expressed in oestrogen receptor (ER)-negative breast cancers and, within this poor prognosis sub-group, increased expression correlates with reduced distant metastasis-free survival. ID2 expression also associates with reduced brain metastasis relapse-free survival. Mechanistically, BMP7, which is present at significantly higher levels in brain tissue compared with the lungs, upregulates ID2 expression and, after BMP7 withdrawal, this elevated expression is retained. Finally, we demonstrate that either ectopic expression of ID2 or BMP7-induced ID2 expression protects tumour cells from anoikis. Conclusions This study identifies ID2 as a key regulator of breast cancer metastasis to the brain. Our data support a model in which breast cancer cells that have disseminated to the brain upregulate ID2 expression in response to astrocyte-secreted BMP7 and this serves to support metastatic expansion. Moreover, elevated ID2 expression identifies breast cancer patients at increased risk of developing metastatic relapse in the brain.

Contrast-enhanced ultrasound (CEUS) can be used to evaluate microcirculation in cancers, which in turn is associated with the biologic features and ultimately patient prognosis. We conducted a retrospective analysis to examine potential association between CEUS parameters and prognosis in patients with papillary thyroid cancer (PTC). The analysis included 306 patients who underwent CEUS prior to thyroidectomy at our center during a period from 2012 to 2019. Subjects with excellent response (ER) were compared to the non-ER group (including indeterminate response, biochemical incomplete response and structural incomplete response). During the median follow-up of 34 months, ER was observed in 195 (63.7%) subjects. The remaining 111 (36.3%) patients developed non-ER events, with distant metastasis in five (1.6%) cases. In a multivariate COX regression, non-ER event was associated with the male sex (OR = 1.83, 95%CI: 1.21–2.76) and blood-rich enhancement in CEUS (OR = 1.69, 95%CI: 1.04–2.75). Based on this finding, we developed a predictive model: high risk for developing non-ER events was defined as having both risk factors; low risk was defined as having none or only one risk. In receiver operating characteristic (ROC) analysis, the area under the curve was 0.59 (95%CI: 0.52–0.66). The sensitivity and specificity were 17.1 and 95.4%, respectively. The positive and negative predictive values were 67.9 and 66.9%, respectively. In conclusion, blood-rich enhancement in CEUS is associated with non-ER events after thyroidectomy in patients with PTC.

Objectives: The purpose of this study was to verify the veracity and reliability of the INCNS score for prediction of neurological ICU (NICU) mortality and 3-month functional outcome and mortality in comatose patients.Methods: In this prospective study, data of the patients admitted to NICU from January 2013 to January 2019 were collected for validation. The 3-month functional outcomes were evaluated using modified Rankin Scale (mRS). By using the receiver operating characteristics curve (ROC) analysis, we compared the INCNS score with Glasgow Coma Scale (GCS), Full Outline of Un-Responsiveness Score (FOUR) and Acute Physiology and Chronic Health Evaluation II (APACHE II) for assessment of the predictive performance of these scales for 3-month functional outcome and mortality and NICU mortality performed at 24- and 72-h after admission to the NICU.Results: Totally 271 patients were used for evaluation; the INCNS score achieved an AUC (area under the receiver operating characteristic curve) of 0.766 (95% CI: 0.711–0.815) and 0.824 (95% CI: 0.774–0.868) for unfavorable functional outcomes, an AUC of 0.848 (95% CI: 0.800–0.889) and 0.892 (95% CI: 0.848–0.926) for NICU mortality, and an AUC of 0.811 (95% CI: 0.760–0.856) and 0.832 (95% CI: 0.782–0.874) for the 3-month mortality after discharge from the NICU at 24- and 72-h. The INCNS score exhibited a significantly better predictive performance of mortality and 3-month functional outcomes than FOUR and GCS. There was no significant difference in predicting NICU mortality and 3-month functional outcomes between INCNS and APACHE II, but INCNS had better predictive performance of 3-month mortality than APACHE II.Conclusions: The INCNS score could be used for predicting the functional outcomes and mortality rate of comatose patients.

The clinical utility of genetic testing for epilepsy has been enhanced with the advancement of next-generation sequencing (NGS) technology along with the rapid updating of publicly available databases. The aim of this study was to evaluate the diagnostic yield of NGS and assess the value of reinterpreting genetic test results in children and adults with epilepsy. We performed genetic testing on 200 patients, including 82 children and 118 adults. The results were classified into three categories: positive, inconclusive, or negative. The reinterpretation of inconclusive results was conducted in April 2020. Overall, we identified disease-causing variants in 12% of the patients in the original analysis, and 14.5% at reinterpretation. The diagnostic yield for adults with epilepsy was similar to that for children (11 vs. 19.5%, p = 0.145). After reinterpretation, 9 of the 86 patients who initially had inconclusive results obtained a clinically significant change in diagnosis. Among these nine revised cases, five obtained positive diagnoses, representing a diagnosis rate of 5.8% (5/86). Manual searches for additional evidence of pathogenicity for candidate variants and updated patient clinical information were the main reasons for diagnostic reclassification. This study emphasizes the diagnostic potential of combining NGS and reinterpretation of inconclusive genetic test reports in children and adults with epilepsy.

Mangrove forests have unique ecosystem functions and services, yet the coastal mangroves in tropics are often disturbed by tropical cyclones. Hurricane Maria swept Puerto Rico and nearby Caribbean islands in September 2017 and caused tremendous damage to the coastal mangrove systems. Understanding the vulnerability and resistance of mangrove forests to disturbances is pivotal for future restoration and conservation. In this study, we used LiDAR point clouds to derive the canopy height of five major mangrove forests, including true mangroves and mangrove associates, along the coast of Puerto Rico before and after the hurricanes, which allowed us to detect the spatial variations of canopy height reduction. We then spatially regressed the pre-hurricane canopy height and the canopy height reduction on biophysical factors such as the elevation, the distance to rivers/canals within and nearby, the distance to coast, tree density, and canopy unevenness. The analyses resulted in the following findings. The pre-hurricane canopy height increased with elevation when elevation was low and moderate but decreased with elevation when elevation was high. The canopy height reduction increased quadratically with the pre-hurricane canopy height, but decreased with elevation for the four sites dominated by true mangroves. The site of Palma del Mar dominated by Pterocarpus, a mangrove associate, experienced the strongest wind, and the canopy height reduction increased with elevation. The canopy height reduction decreased with the distance to rivers/canals only for sites with low to moderate mean elevation of 0.36–0.39 m. In addition to the hurricane winds, the rainfall during hurricanes is an important factor causing canopy damage by inundating the aerial roots. In summary, the pre-hurricane canopy structures, physical environment, and external forces brought by hurricanes interplayed to affect the vulnerability of coastal mangroves to major hurricanes.

Type 2 diabetes (T2D) is a rapidly growing epidemic, which leads to increased mortality rates and health care costs. Nutrients (namely, carbohydrates, fat, protein, mineral substances, and vitamin), sensing, and management are central to metabolic homeostasis, therefore presenting a leading factor contributing to T2D. Understanding the comprehensive effects and the underlying mechanisms of nutrition in regulating glucose metabolism and the interactions of diet with genetics, epigenetics, and gut microbiota is helpful for developing new strategies to prevent and treat T2D. In this review, we discuss different mechanistic pathways contributing to T2D and then summarize the current researches concerning associations between different nutrients intake and glucose homeostasis. We also explore the possible relationship between nutrients and genetic background, epigenetics, and metagenomics in terms of the susceptibility and treatment of T2D. For the specificity of individual, precision nutrition depends on the person’s genotype, and microbiota is vital to the prevention and intervention of T2D.

Objective The deficient placental blood perfusion caused by the attenuated infiltration of trophoblast cells is a key factor in the occurrence of preeclampsia (PE). Furthermore, the long noncoding (lnc)RNA SNHG12 (small nucleolar RNA host gene 12) can promote the proliferation and metastasis of multiple tumor cells. However, whether lncRNA SNHG12 affects proliferation and metastasis of trophoblast cells is unclear. Methods We examined the level of lncRNA SNHG12 in plasma and placenta of patients with PE and constructed trophoblast cells with overexpressed or knocked down SNHG12. CCK-8, wound healing, and Transwell assays were used to detect alterations in proliferation, migration, and invasion of trophoblast cells. Western blotting was used to detect proteins related to the epithelial–mesenchymal transition (EMT), and cell cycle assays clarified cell cycle distribution. Results LncRNA SNHG12 promoted the proliferation, migration, and invasion of trophoblast cells. The expression of matrix metalloproteinase-2 (MMP-2) and MMP-9, β-catenin, and vimentin were positively correlated with SNHG12, and expression of E-cadherin was negatively correlated with SNHG12. SNHG12 also promoted the transition of trophoblast cells from G 0 /G 1 to S phase. Conclusion Overall, lncRNA SNHG12 promoted the migration and invasion of trophoblast cells by inducing the progression of EMT.

To evaluate the effectiveness of the number of central compartment lymph nodes (CLNs) on ultrasound (US) in predicting CLN metastasis (CLNM). We prospectively studied 309 papillary thyroid cancer (PTC) patients who underwent thyroidectomy with CLN dissection at our center from May 2017 to July 2017. The number and features of CLNs were evaluated preoperatively via US. All US examinations were performed using a Philips iU 22 or a GE Logiq 9 machine. Correlations between CLNs observed via preoperative US and amount of CLNM were evaluated. We found that ≥2 CLNs on the preoperative US was associated with CLNM (P5 metastatic CLNs) (P

Abstract Background Several prognostic signatures for early oestrogen receptor-positive (ER+) breast cancer have been established with a 10-year follow-up. We tested the hypothesis that signatures optimised for 0–5-year and 5–10-year follow-up separately are more prognostic than a single signature optimised for 10 years. Methods Genes previously identified as prognostic or associated with endocrine resistance were tested in publicly available microarray data set using Cox regression of 747 ER+/HER2− samples from post-menopausal patients treated with 5 years of endocrine therapy. RNA expression of the selected genes was assayed in primary ER+/HER2− tumours from 948 post-menopausal patients treated with 5 years of anastrozole or tamoxifen in the TransATAC cohort. Prognostic signatures for 0–10, 0–5 and 5–10 years were derived using a penalised Cox regression (elastic net). Signature comparison was performed with likelihood ratio statistics. Validation was done by a case-control (POLAR) study in 422 samples derived from a cohort of 1449. Results Ninety-three genes were selected by the modelling of microarray data; 63 of these were significantly prognostic in TransATAC, most similarly across each time period. Contrary to our hypothesis, the derived early and late signatures were not significantly more prognostic than the 18-gene 10-year signature. The 18-gene 10-year signature was internally validated in the TransATAC validation set, showing prognostic information similar to that of Oncotype DX Recurrence Score, PAM50 risk of recurrence score, Breast Cancer Index and IHC4 (score based on four IHC markers), as well as in the external POLAR case-control set. Conclusions The derived 10-year signature predicts risk of metastasis in patients with ER+/HER2− breast cancer similar to commercial signatures. The hypothesis that early and late prognostic signatures are significantly more informative than a single signature was rejected.

ESR1 mutations occur in endocrine-resistant patients but have not yet been reported in in vitro models of breast cancer. Here, the authors report the discovery of naturally occurring ESR1 Y537Cand ESR1 Y537S mutations in two breast cancer cell lines after acquisition of resistance to long-term-estrogen-deprivation.

The coastal mangrove forest bears important ecosystem functions and services, including the protection of shorelines and coastal communities. While coastal mangroves often suffer severe damage during storms, understanding the vulnerability and resistance of mangroves to the damage at a landscape scale is crucial for coastal mangrove management and conservation. In September 2017, two consecutive major hurricanes caused tremendous damage to the coastal mangroves in the Caribbean. By utilizing LiDAR data taken before and after the hurricanes in a basin mangrove forest in Northeast Puerto Rico, we analyzed the spatial variation of a canopy structure before the hurricanes and hurricane-induced canopy height reduction and explored possible drivers by means of spatial regressions. Regarding the canopy structure, we found that the pre-hurricane canopy height of the mangrove forest decreased with elevation and distance to the freshwater/sewage canals within the forest, and these two drivers explained 82% of variations in the mangrove canopy height. The model, thus, implies that freshwater and nutrient inputs brought by the canals tend to promote the canopy height, and mangrove trees at lower elevation are especially more advantageous. Similarly, tree densities decreased with the canopy height but increased with the elevation and the distance to the canals. We also found that this mangrove forest suffered on average a 53% canopy height reduction, reflecting mostly heavy crown defoliation and the rupture of branches. The regression, which explains 88% of spatial variation in the canopy height reduction, showed that mangroves with a higher canopy or lower density, or growing in lower elevation, or being closer to the canals suffered more damage. Our findings indicate that delivered freshwater/sewage by means of human-made canals has a strong impact on the canopy structure as well as its resistance to tropical storms. Freshwater and sewage tend to release the salinity stress and nutrient deficit and, thus, to promote the mangrove canopy height. However, the addition of freshwater and nutrients might also increase the risk of mangrove damage during the storms probably because of an altered allometry of assimilates.

Tropical dry forest is vulnerable to increased climate variability with more frequent and severe storms. Studies of hurricane impact on tropical dry forest often focused on individual tree traits. How trees in tropical dry forests work together to combat wind damage is still unclear. To address this, we integrated ground-observed ecosystem structure from National Ecological Observation Network (NEON) with airborne-LiDAR images and analyzed resistance in forest structure of Guánica dry forest in Puerto Rico to major hurricanes in 2017 at the forest-stand level. Using each plot instead of the individual tree as the base unit, we regressed mean changes in stem height and fractions of lost or damaged stems at 15 plots on mean stem diameter, mean and standard deviation of stem height, stem density, and topography. Meanwhile, using the LiDAR-derived canopy heights, we compared the changes in canopy height before and after the hurricanes and regressed spatially the canopy height change on prior-hurricane tree cover, canopy height, and rugosity. We found that the damage was small in places with high stem density or high tree cover. Ground-observed damage in terms of height reduction significantly increased with the standard deviation of stem height, an index of roughness, but decreased with the mean stem diameter of the plots. LiDAR-detected damage in terms of reduction in canopy height was also found to decrease with tree cover and mean canopy height when the canopy height was small or moderate but increase with the rugosity. The fraction of lost stems significantly decreased with the stem density, and the fraction of damaged stems significantly increased with the roughness and the plot elevation. The collective parameters of forest stand quantified from ground-observation and LiDAR, such as stem density, tree cover, and canopy roughness or rugosity, highlighted mutual supports of trees and played important roles in resisting damages to the tropical dry forest during major hurricanes.

Changes in vegetation phenology due to global climate change directly impact the dynamic balance of terrestrial carbon and nutrients and the biodiversity pattern, and send feedbacks to climate system. It is crucial to understand spatiotemporal patterns and mechanisms of vegetation phenology change and their relationship with climatic change. In this paper, we quantified the spatiotemporal patterns of start and end of growing season for seven vegetation types in Northeast China Transect (NECT) and North–South Transect of East China (NSTEC) during 2001–2013 using MODIS Enhanced Vegetation Index (EVI). The integral of EVI during growing season was also calculated as an additional phenological index. We then proceeded to analyze the relationship between the three phenological metrics and climate variables including daytime and nighttime temperature and precipitation. To study the temporal phenological trend of deciduous broadleaved forest in northeast China, we identified stands of deciduous broadleaved forest without land cover changes during 2001–2013 and derived the slopes of temporal changes in phenological metrics. Regression was applied to quantify the relationship between phenological trends and trends of climate variables. The vegetation types along the two transects in eastern China have distinctly different phenological characteristics and the strength of climate modulations on phenology varies among vegetation types. The study of temporal trend of phenological changes of the deciduous broadleaved forest in northeast China revealed that nighttime temperature is the most important driver. Increasing in nighttime temperature in spring tends to advance the start of growing season, while increasing in nighttime temperature during growing season tends to delay the growing season. Cold daytime prior to growing season also favors advanced start of growing season. On the other hand, summer nighttime temperature tends to bring down, whereas precipitation tends to enhance, the maximum EVI and hence the EVI integral as the latter is mostly determined by maximum EVI and to a less extent by length of growing season.

Aromatase inhibitors are used to treat oestrogen receptor positive breast cancers but the molecular basis for the response of patients is unclear. Here, the authors use samples from an aromatase inhibitor clinical trial and show that tumours from poor responders have more mutations than good responders and also more frequently harbour p53 mutations.

Summary: Endocrine therapy (ET) is the standard of care for estrogen receptor-positive (ER+) breast cancers. Despite its efficacy, ∼40% of women relapse with ET-resistant (ETR) disease. A global transcription analysis in ETR cells reveals a downregulation of the neutral and basic amino acid transporter SLC6A14 governed by enhanced miR-23b-3p expression, resulting in impaired amino acid metabolism. This altered amino acid metabolism in ETR cells is supported by the activation of autophagy and the enhanced import of acidic amino acids (aspartate and glutamate) mediated by the SLC1A2 transporter. The clinical significance of these findings is validated by multiple orthogonal approaches in a large cohort of ET-treated patients, in patient-derived xenografts, and in in vivo experiments. Targeting these amino acid metabolic dependencies resensitizes ETR cells to therapy and impairs the aggressive features of ETR cells, offering predictive biomarkers and potential targetable pathways to be exploited to combat or delay ETR in ER+ breast cancers. : Bacci et al. find that endocrine-resistant ER+ breast cancers are characterized by enhanced miR-23b-3p, autophagy activation, and import of aspartate and glutamate that fuel catabolic and anabolic pathways, which are essential for their aggressive features. The molecular players involved in this metabolic scenario are of clinical significance and have prognostic and predictive value. Keywords: endocrine therapy, resistance, metabolic reprogramming, estrogen receptor, amino acid transporters, aspartate, glutamate, miRNA, SLCs

Rapamycin (Rap) and its derivatives, called rapalogs, are being explored in clinical trials targeting cancer and neurodegeneration. The underlying mechanisms of Rap actions, however, are not well understood. Mechanistic target of rapamycin (mTOR), a lysosome-localized protein kinase that acts as a critical regulator of cellular growth, is believed to mediate most Rap actions. Here, we identified mucolipin 1 (transient receptor potential channel mucolipin 1 [TRPML1], also known as MCOLN1), the principle Ca2+ release channel in the lysosome, as another direct target of Rap. Patch-clamping of isolated lysosomal membranes showed that micromolar concentrations of Rap and some rapalogs activated lysosomal TRPML1 directly and specifically. Pharmacological inhibition or genetic inactivation of mTOR failed to mimic the Rap effect. In vitro binding assays revealed that Rap bound directly to purified TRPML1 proteins with a micromolar affinity. In both healthy and disease human fibroblasts, Rap and rapalogs induced autophagic flux via nuclear translocation of transcription factor EB (TFEB). However, such effects were abolished in TRPML1-deficient cells or by TRPML1 inhibitors. Hence, Rap and rapalogs promote autophagy via a TRPML1-dependent mechanism. Given the demonstrated roles of TRPML1 and TFEB in cellular clearance, we propose that lysosomal TRPML1 may contribute a significant portion to the in vivo neuroprotective and anti-aging effects of Rap via an augmentation of autophagy and lysosomal biogenesis.

Extranodal nasal-type natural killer/T-cell lymphoma (NKTCL) is an Epstein-Barr virus (EBV)-associated lymphoma with a strong tendency relapse or be refractory in response to chemotherapy. Development of a new strategy for NKTCL treatment is still quite necessary. In this study, we found that aspirin treatment suppresses VEGF expression in NKTCL SNK-6 cells. Further investigation showed that aspirin treatment increases histone methylation in the range of −100~0 that is proximal to the transcription start site on the VEGF promoter, subsequently decreasing the binding ability of Sp1 to the VEGF promoter with VEGF suppression. Furthermore, aspirin treatment modulates mitochondrial function with increased ROS formation and apoptosis in NKTCL cells. Aspirin treatment alone slightly inhibits NKTCL SNK-6 tumor growth and EBV replication; while in the presence of histone deacetylase inhibitor (HDACi) chidamide (CDM), aspirin significantly suppresses the VEGF signaling pathway with increased ROS overgeneration and EBV inhibition. We also showed that with the addition of chidamide, aspirin significantly suppresses NKTCL tumor growth in both in vitro cell culture and in vivo mouse model with prolonged mouse survival. This is the first time that the potential mechanism for aspirin-mediated VEGF suppression and anti-tumor effect has been discovered, and this study provides a new strategy for anti-tumor drug development for NKTCL treatment based on aspirin-mediated targeting of the VEGF signaling pathway and ROS formation.

Prolactin (PRL) production in mammals has been demonstrated in extrapituitary gland, which can activate autocrine/paracrine signaling pathways to regulate physiological activity. In the current study, we characterized the gene expression profiles of PRL, prolactin receptor (PRLR) and signal transducers and activators of transcription 5 (STAT5) in the scented glandular tissues of the muskrats, to further elucidate the relationship between PRL and the scented glandular functions of the muskrats. The weight and volume of the scented glands in the breeding season were significantly higher than those of the non-breeding season. Immunohistochemical data showed that PRL, PRLR and STAT5/phospho-STAT5 (pSTAT5) were found in the glandular and epithelial cells of the scented glands in both seasons. Furthermore, we found that PRL, PRLR and STAT5 had higher immunoreactivities in the scented glands during the breeding season when compared to those of the non-breeding season. In parallel, the gene expressions of PRL, PRLR and STAT5 were significantly higher in the scented glands during the breeding season than those of the non-breeding season. The concentrations of PRL in scented glandular tissues and sera were measured by enzyme-linked immunosorbent assay (ELISA), and their levels were both notably higher in the breeding season than those of the non-breeding season. These findings suggested that the scented glands of the muskrats were capable of extrapituitary synthesis of PRL, which might attribute PRL a specific function to an endocrine or autocrine/paracrine mediator.

Reactive oxygen species (ROS) damage cell components, necessitating their clearance through autophagy. Here, the authors show that ROS can induce autophagy by triggering TRPML1 to release Ca2+from the lysosomal lumen, in turn activating the autophagy and lysosomal biogenesis regulator TFEB.

How cancer-associated fibroblasts become activated to support metastasis is not well understood. Here, Avgustinova et al. show that tumour cell-derived Wnt7a activates TGFβ signaling in fibroblasts, inducing them to remodel the microenvironment to the state, which promotes tumour invasiveness and metastasis.

Objective To explore the role of 18F-fluoro-2-deoxy-D-glucose (18F-FDG) PET imaging for evaluation of the prognosis of patients with chronic disorders of consciousness. Methods 18F-FDG PET was performed to analyze glucose metabolism of external and internal awareness networks in 20 patients with chronic disorders of consciousness, and to calculate standardized uptake value (SUV) of each region. Compare the SUV of each region between conscious [Glasgow Outcome Scale-Extended (GOS-E) > 2] and unconscious (GOS-E ≤ 2) patients after 6 months follow-up. Results After 6 months, one case was lost, 8 cases recovered consciousness during the follow-up, 11 were unconscious (2 cases) or dead (9 cases). The age of unconscious or dead patients was significantly higher than that of conscious (P = 0.001), while the Coma Recovery Scale-Revised (CRS-R) score (P = 0.030) and the SUV of bilateral prefrontal lobes (left P = 0.000, right P = 0.002), bilateral parietal lobes (left P = 0.003, right P = 0.007), bilateral precuneus (left P = 0.000, right P = 0.003), anterior (P = 0.001) and posterior (P = 0.006) cingulate cortices of unconscious or dead patients were significantly lower than that of conscious patients. Conclusions The SUV of externaland internal awareness networks in 18F-FDG PET imaging is related to the prognosis of patients with chronic disorders of consciousness. The higher the SUV is, the better prognosis will be achieved. DOI: 10.3969/j.issn.1672-6731.2015.12.003

Temperate forests and grasslands carry key ecosystem functions and provide essential services. Remote-sensing derived greenness has been widely used to assess the response of ecosystem function to climate and land-cover changes. Although reforestation and grassland restoration have been proposed to enhance the regional greenness in Northern China, the independent contribution of climate without the interference of land-cover change at meso and large scales has rarely been explored. To separate the impacts of climate change on vegetation greenness from those of land-cover/use change, we identified large patches of forests and grasslands in Northern China without land-cover/use changes in 2001–2015 and derived their greenness using MODIS enhanced vegetation index (EVI). We found that most deciduous-broadleaved forest patches showed greening, and the significant slope of the annual mean and maximum EVI are 3.97 ± 0.062 × 10−3 and 4.8 ± 0.116 × 10−3 yr−1, respectively. On the contrary, grassland patches showed great spatial heterogeneity and only those in the east showed greening. The partial correlation analysis between EVI and climate showed that the greening of grassland patches is primarily supported by the increased growing-season precipitation with mean significant coefficient of 0.72 ± 0.01. While wet-year (0.57 ± 0.01) and nongrowing-season precipitation (0.68 ± 0.01) significantly benefit greening of deciduous-broadleaved forests, the altered temperature seasonality modulates their greening spatial-heterogeneously. The increased growing-season minimum temperature might lengthen the growing season and contribute to the greening for the temperature-limited north as shown by positive partial correlation coefficient of 0.66 ± 0.01, but might elevate respiration and reduce greening of the forests in the south as shown by negative coefficient of −0.70 ± 0.01. Daytime warming in growing season is found to favor the drought-tolerant oak dominated forest in the south due to enhanced photosynthesis, but may not favor the forests dominated by less-drought-tolerant birch in the north due to potential water stress. Therefore, grassland greening was essentially promoted by the growing-season precipitation, however, in addition to being driven by precipitation, greening of deciduous forests was regulated spatial-heterogeneously by asymmetrical diurnal and seasonal warming which could be attributed to species composition.

High-carbon sequestrations of tropical montane forests and coastal mangroves have been greatly disturbed by intensified extreme climate events such as alternating hurricanes and droughts. However, few studies of the hurricane impact have taken into consideration the legacy of past climate events and analyzed the heterogeneity of hurricane impacts between montane forests and coastal mangroves. Here, we studied the impact of Hurricanes Irma and Maria in 2017 on coastal mangroves and upland forests in Puerto Rico after a severe drought during 2015–2016. We investigated the island-wide immediate impact on greenness using fused vegetation index from Sentinel-2 and Landsat-8, and the impact on and the recovery of 62 homogeneous vegetation patches by deriving an impact index and one-year recovery ratio (RR). A linear mixed-effect model was applied to explore roles of hurricane wind, rainfall, topography, and biological components in the impact and the recovery. Island-wide, the immediate impact is highly spatial-heterogeneous. Although most of the island was browned, a green-up strip in the dry south showed benefits from the hurricane rain which relieved the prior drought stress. Coastal mangroves experienced the greatest impact and slowest recovery with relative recovery of 0.44 compared to recovery greater than 0.70 for upland forests, and evergreen forests sustained significantly more damage than deciduous forests. The recovery of evergreen forests was on average 11 days earlier and faster than that of mangroves. Mangrove recovery was mostly limited by inundation-related factors such as elevation, slope, and drainage capacity. While higher elevation relates to slower recovery for upland forests, it favors mangrove recovery. Particularly, mangrove recovery is facilitated by river presence, explaining 65% variation in RR. The differentiated response, recovery, and underlying mechanisms highlighted a complicated array of external forces, geophysical/ biological modulators, and legacy of past climate events in determining and understanding hurricanes’ impact on tropical ecosystems.

After publication of the original article [1], we were notified that an author’s surname has been erroneously spelled. Elisabetta Maragoni’s family name should be replaced with Marangoni.

Studying the complex mechanisms underlying breast cancer metastasis and therapy response necessitates relevant in vivo models, particularly syngeneic models with an intact immune system. Two syngeneic spontaneously metastatic sublines, D2A1-m1 and D2A1-m2, were generated from the poorly metastasising BALB/c-derived D2A1 cell line by serial in vivo passaging. In vivo and in vitro analyses revealed distinct and shared characteristics of the metastatic D2A1-m1 and D2A1-m2 sublines. In particular, D2A1-m1 cells are more aggressive in experimental metastasis assays, while D2A1-m2 cells are more efficient at disseminating from the primary tumour in spontaneous metastasis assays. Surprisingly, classical metastasis-associated in vitro phenotypes, such as enhanced proliferation, migration and invasion, are reduced in the sublines compared to the parental cell line. Further, evasion of immune control cannot fully explain their enhanced metastatic properties. By contrast, both sublines show increased resistance to apoptosis when cultured in non-adherent conditions and, for the D2A1-m2 subline, increased 3D tumour spheroid growth. Moreover, the enhanced spontaneous metastatic phenotype of the D2A1-m2 subline is associated with an increased ability to recruit an activated tumour stroma. The metastatic D2A1-m1 and D2A1-m2 cell lines provide additional syngeneic models for investigating the different steps of the metastatic cascade and thereby represent valuable tools for breast cancer researchers. Finally, this study highlights that morphology and cell behaviour in 2D cell-based assays cannot be used as a reliable predictor of metastatic behaviour in vivo.