Invasive Areas Research Articles

Behavioral preferences of Solenopsis invicta Buren to essential oils and active compounds from amiaceae plants

The red imported fire ant (RIFA) (Solenopsis invicta Buren) has caused serious ecological and public safety problems in invasive areas. While extensive use of pesticides to control RIFA may negatively affect human health and environment, essential oils (EOs) of Lamiaceae possessing aromatic smell and multiple biological activities are potential alternatives of chemical pesticides. In this study, we evaluated the behavioral preferences of different castes of RIFA towards the EOs from five species of Lamiaceae plants (Salvia miltiorrhiza, Perilla frutescens, Pogostemon cablin, Schizonepeta tenusfolia, Mentha canadensis), and then investigated the active compounds in these EOs that trigger insect behavioral preferences by gas chromatography-electroantennographic detection (GC-EAD). The results of behavioral experiment indicated that three EOs strongly altered the behavior of RIFA. Notably, all concentrations of P. frutescens EO showed a significant repellent activity on virgin queen ants, with a repellent rate of 62.50% at the lowest concentration of 2 μg/ml. The repellent effect of S. tenusfolia EO against small worker ants reached a very significant difference between control and treatment groups, with the highest repellent rate of 85.00% observed at 1000 μg/ml. Save for the lowest concentration 2 μg/ml, other doses of M. canadensis EO showed a remarkable repellent effect on small worker ants, demonstrating a positive dose-effect relationship (with repellent rates of 77.50% and 65.00% at 1000 and 8 μg/ml, respectively). A total of 39 EAD-active compounds were detected in these EOs, and 12 of them were subsequently evaluated for behavioral preferences of RIFA. Our findings indicated that six compounds (menthone, menthol, caryophyllene oxide, pulegone, 3-methylcyclopentanol, eugenol) consistently triggered behavioral preferences in worker ants. Meanwhile, six compounds, including borneol, eugenol, phenylethyl alcohol, perilla ketone, 3-methylcyclopentanol, α-guaiene, were found to significantly alter the behavioral selectivity of virgin queen ants. These results may help to develop application value of selected Lamiaceae plants EOs and contribute to the exploration of new green control methods for invasive pests.

Risks posed by invasive species to the provision of ecosystem services in Europe

Invasive species significantly impact biodiversity and ecosystem services, yet understanding these effects at large spatial scales remains a challenge. Our study addresses this gap by assessing the current and potential future risks posed by 94 invasive species to seven key ecosystem services in Europe. We demonstrate widespread potential impacts, particularly on outdoor recreation, habitat maintenance, crop provisioning, and soil and nitrogen retention. Exposure to invasive species was higher in areas with lower provision of ecosystem services, particularly for regulating and cultural services. Exposure was also high in areas where ecosystem contributions to crop provision and nitrogen retention were at their highest. Notably, regions vital for ecosystem services currently have low invasion suitability, but face an average 77% increase in potential invasion area. Here we show that, while high-value ecosystem service areas at the highest risk represent a small fraction of Europe (0-13%), they are disproportionally important for service conservation. Our study underscores the importance of monitoring and protecting these hotspots to align management strategies with international biodiversity targets, considering both invasion vulnerability and ecosystem service sustainability.

Correlation between tumor invasion and somatostatin receptor subtypes in acromegaly.

The low expression of somatostatin receptor (SSTR) subtypes in somatotropinoma is associated with a poor response to somatostatin analogs (SSAs). However, the correlation between SSTRs and tumor invasion has not yet been clarified. Therefore, the authors aimed to investigate the relationship between SSTRs and tumor invasion, as well as the correlation between tumor invasiveness and pharmacological response to SSAs. A total of 102 patients with acromegaly who underwent surgery between December 2016 and December 2021 at the largest pituitary tumor surgery center in southern China were included in this retrospective study. Patients were divided into the noninvasive tumor group (Knosp grades 0-2 and Hardy-Wilson grade I or II) and invasive group (either Knosp grade 3 or 4 or Hardy-Wilson grade III or IV). The positive response to SSAs was defined by the following criteria after at least 3 months of SSA treatment: 1) ≥ 50% reduction or age- and sex-adjusted normal range of insulin-like growth factor-1 (IGF-1) level; 2) ≥ 80% reduction in or normal range of growth hormone (GH) level; or 3) > 20% reduction in tumor volume. The reference for the normal range of age- and sex-adjusted serum IGF-1 levels was derived from a survey of 2791 healthy adults (1339 males and 1452 females) in China. Demographics and clinical characteristics including tumor size, biochemical assessment, expression levels of SSTRs, and response to preoperative SSAs were compared between the invasive group and noninvasive group. Receiver operating characteristic (ROC) curve analysis was performed to assess the association between SSTR2 and tumor invasion. Compared with the noninvasive group, the invasive group presented with a larger tumor size (9.99 ± 10.41 cm3 vs 3.50 ± 4.02 cm3, p < 0.001), relatively lower SSTR2 expression (p < 0.001), and poorer response to SSAs (36.4% vs 91.7%, p < 0.001). In addition, there was a significant negative correlation between SSTR2 mRNA level and tumor size (r = -0.214, p = 0.031). However, there were no statistically significant differences in the expression of SSTR1, SSTR3, and SSTR5 between the groups. ROC analysis revealed that the low SSTR2 mRNA level was closely associated with tumor invasion (area under the curve 0.805, p < 0.0001). Tumor invasion is negatively correlated with SSTR2 level but is not associated with other SSTR subtypes. Patients with invasive tumors have a poorer response to SSA therapy, which may be due to the low level of SSTR2 expression. Therefore, SSTR2 could be considered as a routine investigative marker for aiding management of postoperative residual tumors.

Abstract 553: Assessment of CNTN4 expression and its association with PD-L1 across 18 various cancers using an artificial intelligence-powered immunohistochemistry analyzer

Abstract Background: Contactin-4 (CNTN4) has been identified as an inhibitory immune checkpoint protein and mainly expressed on tumor cells. It interacts with APP present in T cells, leading to a reduction in T cell proliferation, activation, and responsiveness to tumors. In this study, we applied artificial intelligence (AI)-powered analyzers to examine the expression of CNTN4 and its association with PD-L1 in various solid tumors. Methods: The AI-powered immunohistochemistry (IHC) analyzers were developed using pan-cancer whole slide images (WSIs) stained with 18 different IHC types. Pathologists annotated cells and tissues on the WSIs. The universal model was trained by annotations from IHC WSI, which could detect tumor cells with four grade intensities (strong, moderate, weak, and negative) and segment invasive cancer area. Finally, the AI analyzer can count the tumor cells in an invasive cancer area and calculate the ratio based on intensity. The AI analyzer was applied to the pan-cancer tissue microarray dataset (n=795) consisting of 18 cancers to validate the analyzer. The dataset was stained with rabbit monoclonal antibody (clone EPR 8735) and PD-L1 22C3 antibody, respectively. The expression levels of CNTN4 and PD-L1 were calculated with H-score (range 0 to 300) or tumor proportion score (TPS). Results: When positivity was defined as moderate-to-strong staining intensity in ≥ 30%, 50%, 75%, and 90%, 42.4% (337/795), 25.5% (203/795), 11.4% (91/795), and 5.2% (41/795) of samples were CNTN4 positive, respectively. In case of moderate-to-strong staining in ≥ 50% tumor cells, CNTN4 expression were most frequently observed in hepatocellular carcinoma (63.9%), endometrial carcinoma (50.0%), stomach cancer (43.9%), pancreatic cancer (40.0%), and prostate cancer (35.4%). Notably, 10% of cases with an H-score &amp;lt; 200 (75/757) showed a range of PD-L1 expression (PD-L1 TPS ≥ 1%), while all of the samples with an H-score ≥ 200 (n=38) were PD-L1 TPS &amp;lt;1%. Conclusions: This study describes the CNTN4 expression levels in various tumors using an AI-powered IHC analyzer. CNTN4 tends to be expressed in opposition to PD-L1, making it a viable immune therapeutic target in cancers that do not conventionally express PD-L1. Citation Format: Seunghwan Shin, Jimin Moon, Soo Ick Cho, Chan-Young Ock, Bu-Nam Jeon, Gyeongyeon Kim, Hyunkyung Yu, Miyoung Cha. Assessment of CNTN4 expression and its association with PD-L1 across 18 various cancers using an artificial intelligence-powered immunohistochemistry analyzer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 553.

Abstract 1509: Microglial dominance in glioma dissemination: Unraveling TAMs' impact and therapeutic implications through modulating cathepsin activity

Abstract Introduction: It is well known that tumor-associated macrophages/microglia (TAMs) play a vital role in brain tumor progression. Our previous studies suggested that the activated microglia could reduce the cytotoxic effect of chemotherapeutic drugs on glioma cells in a 2D/3D model. This study delves into TAMs within diverse brain TME regions, emphasizing the distinction of resident microglia from infiltrating macrophages in invasion areas and their role in glioma migration and invasion. Material and Method: We employed the murine orthotopic astrocytoma model (ALTS1C1), dissecting brain tumor tissues into normal, invasion, edge, and core regions. Immunofluorescence staining, utilizing TMEM119 and CD68 markers to identify resident microglia and infiltrating macrophages, respectively. The IHC results were analyzed with QuPath software and validated via flow cytometry. Microglia (BV2) and macrophage (RAW264.7) cell lines were directly co-cultured with ALTS1C1 in 2D and 3D migration/invasion models. Bulk RNA sequencing analysis unraveled the differences between BV2 and RAW264.7. Bone marrow-derived macrophages (BMDM) were used for the TAMs activation test, and the cathepsin inhibitor, E64, was applied to elucidate cathepsin's role in microglia within co-culture models. Results: The IHC results reveal more TMEM119+ microglia than CD68+ infiltrating macrophages in the tumor invasion area. On the other hand, there are more CD68+ infiltrating macrophages than TMEM119+ microglia in the tumor core regions, signifying a specific role of microglia in tumor invasion. Flow cytometry further supported that more CD11b+CD45low microglia prevalence in tumor surrounding tissues than in tumor core. ALTS1C1 astrocytoma cells, in the presence of microglia, exhibited more increased invasion and migration ability than the presence of macrophages in 2D/3D models. RNA sequencing highlighted M2 activation and cathepsin gene upregulation in microglia over macrophages. M2-polarized TAMs enhanced 2D migration, while cathepsin inhibition significantly decreased glioma's 3D invasion potential. Conclusion: This study identifies microglia as primary TAMs in brain tumor invasion areas and their roles in glioma cell survival and dissemination. Notable differences in activation states and the critical impact of cathepsin in microglia for their roles on brain tumor invasion and survival offer promising therapeutic avenues. Targeting cathepsin in microglia emerges as a novel glioma prevention and treatment strategy. These findings contribute valuable insights to enhance clinical interventions for glioma patients. Citation Format: Sheng-Yan Wu, Chi-Shiun Chiang. Microglial dominance in glioma dissemination: Unraveling TAMs' impact and therapeutic implications through modulating cathepsin activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1509.

Abstract 238: Investigating size-dependent invasion behaviors and therapeutic responses of the 42MGBA glioblastoma spheroids

Abstract Introduction: Glioblastoma (GBM) is one of the most aggressive malignant brain tumors. Many studies have reported that the tumor microenvironment (TME) contributes greatly to invasion and drug resistance of GBM. Our lab has previously described a tri-culture model using methacrylamide-functionalized gelatin (GelMA) with brain microvascular endothelial cells (BMVEC), human normal astrocytes (NHA), and human brain vascular pericyte (HBVP) recapitulated the in vivo brain microenvironment. However, phenotypic drug response was only observed under supraphysiological dosage. To evaluate therapeutic response more precisely, we decided to combine the Tri-culture system with the IdenTx3 microfluidic device (AIM Biotech). By separating the immature perivascular network from the GBM spheroids, we are hoping to generate a model that can more realistically mimic the TME. While many studies have used cancer spheroid in GBM studies, little is known about the impact of spheroids size. Since spheroid size correlates with nutrient and oxygen gradients, we are also curious to find out whether an increase in spheroid size increases the necrotic population and affects drug response and invasion. Materials: Wild-type (WT) and TMZ resistant 42MGBA cell lines were seeded at different cell numbers to generate spheroids of different sizes. Spheroids were dissociated for flow analysis to quantify the necrotic population. In the invasion assay, spheroids were encapsulated in GelMA hydrogel with or without the addition of perivascular cells including BMVECs, HBVP, and NHA (3:1:1). Vasculature formation inside the chip was initiated 5 days prior to the introduction of GBM spheroids. Results: 42MGBA-WT and 42MGBA-TMZres spheroids demonstrated phenotypical and proliferative differences. Spheroids sizes showed a direct relationship with necrotic cell numbers. Small spheroids (1000 cells) and large spheroids (10,000 cells) were encapsulated inside the hydrogel and subjected to different treatments of temozolomide. A difference in invasion area and proliferation was found when subjected to bulk or metronomic dosages. The two 42MGBA cell lines displayed an increase in invading area when co-cultured with perivascular cells. This result coincided with our previous finding that perivascular network increases GBM proliferation and invasion in GBM6 spheroid (Ngo et al, 2022). In addition, a different invasion behavior was observed between the 42MGBA-WT and 42MGBA-TMZres spheroids when introduced inside the microfluidic device. Surprisingly, the wildtype showed a larger invading area compared to the TMZ resistant. Conclusion: We demonstrated a direct relationship between size and necrotic population in the 42MGBA spheroids. We also showed that bulk or metronomic treatment exert different effects on the cancer spheroids. Finally, we have verified that TME is critical to invasion and drug resistance. Citation Format: Sheridan Fok, Yoanna Ivanova, Victoria Kriuchkovskaia, Brendan Harley. Investigating size-dependent invasion behaviors and therapeutic responses of the 42MGBA glioblastoma spheroids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 238.

Abstract 566: The anticancer effects of polyisoprenylated cysteinyl amide inhibitors involves interaction with the KRAS chaperone calmodulin

Abstract Pancreatic cancer is the third leading cause of cancer-related deaths with a 5-year survival rate of less than 10%. Approximately 90% of reported cases are driven by KRAS mutations. The mutant KRAS acquires its functional localization through interactions with chaperone proteins via its polybasic polyisoprenylated tail. Polyisoprenylated cysteinyl amide inhibitors (PCAIs) mimic this region possibly disrupting the polyisoprenylation-dependent interactions with chaperone proteins. Here, we determine the effect of newly developed PCAIs that were designed to be more water-soluble on cell viability, 3D spheroid invasion, spheroid degeneration and F-actin filaments on MIA PaCa-2 and PANC-1 cells. Of the seven new PCAIs analogs tested, NSL-AB-51 and NSL-AB-45 were the most potent, with EC50 values of 2.5 and 5.6 µM against MIA PaCa-2, and 1.7 and 4.5 µM against PANC-1 cells, respectively. At 5 and 10 µM, NSL-AB-51 decreased the invasion area of 3D spheroids by 98 and 99%, respectively. MIA PaCa-2 spheroids degenerated by 61% after 72 h treatment with 10 µM NSL-AB-51. Treatment with 1.0 μM NSL-AB-51 caused cell rounding and reduction of mean cell areas by 69% and F-actin filaments by 96%. Moreover, to identify the pharmacological target of the PCAIs, NSL-AB-45 was coupled to an epoxy-activated Sepharose resin for affinity trapping of PCAIs-interacting proteins. Affinity-analysis of supernatants and detergent membrane extracts from MIA PaCa-2 and PANC-1 cells revealed single Coomassie band of about 17 kDa that was identified as calmodulin by western blotting analysis. The calmodulin-PCAIs affinity interaction was found to be strongest in the presence of calcium and weakest in the presence of EDTA. These findings reveal that calmodulin is the likely target of PCAIs. Since calmodulin is the chaperone protein responsible for KRAS trafficking and localization to the inner surface of the plasma membrane, this finding strongly suggests, at least in part, an anticancer role for the PCAIs that involve disruption of KRAS functions. Citation Format: Kweku Ofosu-Asante, Jassy Mary Lazarte, Amarender Burra, Nazarius S. Lamango. The anticancer effects of polyisoprenylated cysteinyl amide inhibitors involves interaction with the KRAS chaperone calmodulin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 566.

Abstract 2306: A deep learning framework for automated segmentation and analysis of 3D breast cancer spheroid invasion dynamics

Abstract Invasion of neighboring tissues is a cardinal feature of malignancy, notably observed in aggressive cancers like breast cancer, where it can lead to significant morbidity. To explore cancer invasion dynamics, the implementation of a three-dimensional (3D) tumor spheroid invasion assay offers a swift approach to mimicking a tumor micro-region or micro-metastasis. Differential Interference Contrast (DIC) time-lapse imaging was chosen for its fluorescence-free and non-destructive advantages in capturing live spheroid movement. However, the subsequent analysis posed challenges, including out-of-focus cells and a voluminous dataset with diverse microscope planes and time points. Our study introduces a segmentation framework featuring in-focus spatial stacking algorithm and novel training architecture that enabling in-depth automated analysis of 3D spheroid invasion behaviors within a microenvironment. Firstly, the MDA-MB-231 breast cancer spheroids were cultured in a microwell dish and then incorporated into a collagen type I matrix for the 3D spheroid invasion assay. Capturing spheroid invasion dynamics involved acquiring 15 z-plane images (spaced at 20 µm intervals) per timepoint for each spheroid using DIC microscopy at 15-minute intervals over a 28-hour period, employing a 20X objective lens. We refined the Focus Stacking framework into Multiple Slices Blurry Stacking (MSB-Stack) by pre-filtering blurred areas before stacking images. This process produced 6390 slices consolidated into 426 labeled stacked images, highlighting Invasive area, Spheroid core, and Invasive cells post-invasion. We then enhanced Mask R-CNN (Region based-Convolutional Neural Networks) with a weak-to-strong augmentation mechanism, leveraging well-segmented samples from MSB-Stack for training on both fully stacked and blurred images. Blurry Consistency Mask R-CNN (BCMask R-CNN) demonstrates overall improvement across feature extractors, outperforming Mask R-CNN. Using Mean average precision (mAP) as the main metric for instance segmentation and detection, our proposed model achieves 73.0% and 65.2% on mAP @[0.5, 0.95], showcasing robustness. Notably, our best results for mAP @0.5 reach 96.6% and 96.7% on both tasks, underscoring the effectiveness of consistency training in mitigating unclear boundary issues during preprocessing. In conclusion, our proposed method, MSB-Stack, effectively addresses the challenge of unclear boundaries in 3D breast cancer cell DIC microscopy datasets. The integration of weak-to-strong consistency training in our model, BCMask R-CNN, not only mitigates biases but also enhances performance across different backbones, setting the stage for future advancements in cell-tracking tasks and broader migratory analyses. Citation Format: Thi Kim Ngan Ngo, Thanh-Huy Nguyen, Fang-Yu Lin, Ting-Yuan Tu. A deep learning framework for automated segmentation and analysis of 3D breast cancer spheroid invasion dynamics [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2306.

GDF-8 improves in vitro implantation and cryo-tolerance by stimulating the ALK5-SMAD2/3 signaling in bovine IVF embryo development.

Transforming growth factor-beta (TGF-β) plays a critical role in regulating trophoblast invasion and proliferation. Growth differentiation factor-8 (GDF-8) is a member of the TGF-β superfamily and is categorized as a myostatin subtype. It is primarily a secreted protein synthesized in skeletal muscle cells. It is expressed in the placenta, reproductive tissues, and cells. In this study, we investigated the role of GDF-8 in the development and hatching rate of bovine embryos. We noted a notable elevation (p < 0.05) in the development and hatching rates compared to the control embryos. Furthermore, the GDF-8 group showed a significantly improved total cell number (p < 0.05) and an increase in trophectoderm ratio inner cell mass (trophectoderm: inner cell mass) cells (p < 0.001) compared to the control group. Additionally, blastocysts treated with GDF-8 exhibited significantly higher mRNA levels of caudal-type homeobox 2 (CDX2) (p < 0.05). The trophoblast invasion area was significantly larger in the GDF-8 group than in the control group (p < 0.01). Furthermore, qRT-PCR analysis revealed significantly higher mRNA levels (p < 0.05) of matrix metalloproteinases 9 (MMP9) and follistatin-like 3(FSTL3), both of which are associated with the ALK5-SMAD2/3 signaling pathway, in the GDF-8 group than those in the control group. The mRNA expression levels of genes related to tight junctions (TJ) and adherent junctions were higher in the GDF-8 group than those in the control group (p < 0.05). After 24h of thawing, blastocysts were analyzed using 4-kDa FITC-dextran, which revealed a higher TJ integrity in the GDF-8 group (p < 0.01). Thus, GDF-8 plays a crucial role in bovine embryonic development, in vitro implantation, and cryotolerance.

Can ecological niche models be used to accurately predict the distribution of invasive insects? A case study of Hyphantria cunea in China.

In recent decades, ecological niche models (ENMs) have been widely used to predict suitable habitats for species. However, for invasive organisms, the prediction accuracy is unclear. In this study, we employed the most widely used maximum entropy (MaxEnt) model and ensemble model (EM) Biomod2 and verified the practical effectiveness of the ENM in predicting the distribution areas of invasive insects based on the true occurrence of Hyphantria cunea in China. The results showed that when only limited data of invasive areas were used, the two ENMs could not effectively predict the distribution of suitable habitats of H. cunea, although the use of global data can greatly improve the prediction accuracy of ENMs. When analyzing the same data, Biomod2's prediction accuracy was significantly better than that of MaxEnt. For long-term predictions, the area of suitable habitat predicted by the ENMs was much greater than the occurrence area; for short-term predictions, the accuracy of the predicted area was significantly improved. Under the current conditions, the area of suitable habitat for H. cunea in China is 118 × 104 km2, of which 59.32% is moderately or highly suitable habitat. Future climate change could significantly increase the suitable habitat area of H. cunea in China, and the predicted area of suitable habitats in all climate scenarios exceeded 355 × 104 km2, accounting for 36.98% of the total land area in China. This study demonstrates the use of ENMs to study invasive insects and provides a reference for the management of H. cunea in China.

Tumor Budding, Poorly Differentiated Clusters, and Stroma Percentage in Uterine Endometrioid Carcinomas Are Morphologic Findings Associated With Adverse Clinical Outcomes

Uterine endometrioid carcinoma (UEC) has well-defined morphologic features that carry prognostic significance and guide treatment. In addition to the well-known features, tumor budding (TB), poorly differentiated clusters (PDCs), and amount of stroma within the invasive front are associated with a poor prognostic outcome in many carcinomas. Here, we evaluate TB, PDCs, and the stroma percentage in UEC and correlate these findings with morphologic features known to be associated with a poor clinical outcome and with recurrence-free survival (RFS). We performed a retrospective search of our institution’s EMR for cases of hysterectomy for UEC. We collected clinical data including disease recurrence, death, and data associated with poor patient outcomes. Cases were evaluated for TB, PDCs, and percent stroma by 2 pathologists. Ten 20× fields were examined, and the one with the most PDCs and TB was evaluated. Percent stroma was evaluated in a ×10 field of tumor at the area of deepest invasion. Less than 10% stroma was defined as stroma poor, and &gt;10% stroma was defined as stroma rich. Statistical testing and analysis were conducted, and P-value was set at 0.05. One hundred thirteen cases were evaluated. Decreased RFS was seen with TB (P=0.03), at least 5 PDCs (P=0.01), and stroma-rich tumors (P=0.043). This study shows a statistically significant association between TB presence, at least 5 PDCs, stroma-rich tumors, and decreased RFS in UEC. We conclude that TB, PDCs and stromal evaluation are strong independent prognostic indicators in UEC and provide additional value to the currently used morphologic assessment of UEC.

Suppression of Neovascularization by Topical and Subconjunctival Bevacizumab After High-Risk Corneal Transplantation

To assess the effectiveness of topical and subconjunctival bevacizumab in suppressing vascularization in graft and host bed after high-risk corneal transplantation. Secondary analysis of prospective, randomized, double-blind, placebo-controlled multicentric clinical trial. The study includes patients aged > 18 years who underwent high-risk penetrating keratoplasty, which was defined as corneal vascularization in ≥ 1 quadrants of the corneal graft and host bed, excluding the limbus. Patients were randomized to treatment and control groups. The patients in the treatment group received subconjunctival injection of bevacizumab (2.5 mg/0.1 ml) on the day of the procedure, followed by topical bevacizumab (10 mg/ml) 4 times per day for 4 weeks. The patients in control group received injection of vehicle (0.9% sodium chloride) on the day of procedure, followed by topical vehicle (carboxymethylcellulose sodium 1%) 4 times a day for 4 weeks. Vessel and invasion area of vessels in the corneal graft and host beds. This study included 56 eyes of 56 patients who underwent high-risk corneal transplantation, with equal numbers in the bevacizumab and vehicle (control) treatment groups. The mean age of patients who received bevacizumab was 61.2 ± 15.9 years, and the mean age of those treated with vehicle was 60.0 ± 16.1 years. The vessel area at baseline was comparable in the bevacizumab (16.72% ± 3.19%) and control groups (15.48% ± 3.12%; P= 0.72). Similarly, the invasion areas were also similar in the treatment (35.60% ± 2.47%) and control (34.23% ± 2.64%; P= 0.9) groups at baseline. The reduction in vessel area was significantly higher in the bevacizumab-treated group (83.7%) over a period of 52 weeks compared with the control group (61.5%; P < 0.0001). In the bevacizumab-treated group, invasion area was reduced by 75.8% as compared with 46.5% in the control group. The vessel area was similar at 52 weeks postprocedure in cases of first (3.54% ± 1.21%) and repeat (3.80% ± 0.40%) corneal transplantation in patients who received bevacizumab treatment. In the vehicle-treated patients, the vessel area was significantly higher in repeat (9.76% ± 0.32%) compared with first (8.06% ± 1.02%; P < 0.0001) penetrating keratoplasty. In the bevacizumab treatment group, invasion areas at week 52 were comparable in first (11.70% ± 3.38%) and repeat (11.64% ± 1.74%) procedures, whereas invasion area was significantly higher in repeat (27.87% ± 2.57%) as compared with first (24.11% ± 2.17%) penetrating keratoplasty in vehicle-treated patients. In patients undergoing vascularized high-risk corneal transplantation, bevacizumab is efficacious in reducing vascularization of corneal graft and host bed, thereby reducing the risk of corneal graft rejection in vascularized host beds. Proprietary or commercial disclosure may be found in the Footnotes and Disclosures at the end of this article.

Limited effects of culling on the behavior of invasive lionfish (Pterois miles) in the Mediterranean.

Invasive species pose serious threats to ecosystems. To reduce ecological and economic consequences of invasions, efforts are made to control invaders and evaluating the effects of such efforts is paramount. Lionfishes (Pterois volitans and Pterois miles) are native to the Indo-Pacific Ocean and pose a major threat to local ecosystems in the invaded Atlantic and Mediterranean. Culling via spearfishing is a widespread measure to limit lionfish population size in invaded ranges. However, like most hunted fishes, lionfish alter their behavior after repeated culling, potentially decreasing the effectiveness of future culls. Previous studies on lionfish in the Caribbean have shown that lionfish are less bold after repeated culling. However, the impact of culling on lionfish in their newest invasive range, the Mediterranean, remains enigmatic. To determine the behavioral changes in response to culling in this second area of invasion, we tested for effects of culling on the behavior of lionfish in Cyprus, a region heavily impacted by the lionfish invasion in the Mediterranean. We compared the response of lionfish to an approaching free diver holding a metal pole (imitating a spear fisher) between protected areas where spearfishing is restricted and areas where culls are frequently conducted. We also assessed whether activity, hiding pattern, and site fidelity differed between these culled and unculled sites. Overall, we found limited effects of culling on the traits measured, indicating surprising resistance to culling-induced behavioral changes in Mediterranean lionfish. Future studies should monitor invasive lionfish population densities and the effects of culling in more detail to tailor management plans and reduce the negative effects of these fish in specific invaded ranges.

Invasive carcinoma segmentation in whole slide images using MS-ResMTUNet

Identifying the invasive cancer area is a crucial step in the automated diagnosis of digital pathology slices of the breast. When examining the pathological sections of patients with invasive ductal carcinoma, several evaluations are required specifically for the invasive cancer area. However, currently there is little work that can effectively distinguish the invasive cancer area from the ductal carcinoma in situ in whole slide images. To address this issue, we propose a novel architecture named ResMTUnet that combines the strengths of vision transformer and CNN, and uses multi-task learning to achieve accurate invasive carcinoma recognition and segmentation in breast cancer. Furthermore, we introduce a multi-scale input model based on ResMTUnet with conditional random field, named MS-ResMTUNet, to perform segmentation on WSIs. Our systematic experimentation has shown that the proposed network outperforms other competitive methods and effectively segments invasive carcinoma regions in WSIs. This lays a solid foundation for subsequent analysis of breast pathological slides in the future. The code is available at: https://github.com/liuyiqing2018/MS-ResMTUNet

Global spatial distribution of Prosopis juliflora -one of the world's worst 100 invasive alien species under changing climate using multiple machine learning models.

Climate change is one of the factors contributing to the spread of invasive alien species. As a result, it is critical to investigate potential invasion dynamics on a global scale in the face of climate change. We used updated occurrence data, bioclimatic variables, and Köppen-Geiger climatic zones to better understand the climatic niche dynamics of Prosopis juliflora L. (Fabaceae). In this study, we first compared several algorithms-MaxEnt, generalized linear model (GLM), artificial neural network (ANN), generalized boosted model (GBM), generalized additive model (GAM), and random forest (RF)-to investigate the relationships between species-environment and climate for mesquite. We identified the global climate niche similarity sites (NSSs) using the coalesce approach. This study focused on the current and future climatic suitability of P. juliflora under two global circulation models (GCMs) and two climatic scenarios, i.e., Representative Concentration Pathways (RCPs), 4.5 and 8.5, for 2050 and 2070, respectively. Sensitivity, specificity, true skill statistic (TSS), kappa coefficient, and correlation were used to evaluate model performance. Among the tested models, the machine learning algorithm random forest (RF) demonstrated the highest accuracy. The vast swaths of currently uninvaded land on multiple continents are ideal habitats for invasion. Approximately 9.65% of the area is highly suitable for the establishment of P. juliflora. Consequently, certain regions in the Americas, Africa, Asia, Europe, and Oceania have become particularly vulnerable to invasion. In relation to RCPs, we identified suitable area changes (expansion, loss, and stability). The findings of this study show that NSSs and RCPs increase the risk of invasion in specific parts of the world. Our findings contribute to a cross-border continental conservation effort to combat P. juliflora expansion into new potential invasion areas.

Mitral Annulus Calcium Score in Patients With Calcific Mitral Stenosis Undergoing Invasive Hemodynamic Assessment.

Aortic valve calcium score is associated with hemodynamic severity of aortic stenosis. Whether this association is present in calcific mitral stenosis remains unknown. This study was a retrospective analysis of consecutive patients with mitral stenosis secondary to mitral annular calcification (MAC) undergoing transseptal catheterization. All patients underwent invasive mitral valve assessment via direct left atrial and left ventricular pressure measurement. Computed tomography within 1 year of cardiac catheterization and with adequate visualization of the mitral annulus was included. MAC calcium score quantification by Agatston method was obtained offline using dedicated software (Aquarius, TeraRecon, V.4). Median patient age was 66.9±11.2 years, 47% of patients were women, 50% had coronary artery disease, 40% had atrial fibrillation, 47% had prior cardiac surgery, and 33% had prior chest radiation. Median diastolic mitral valve gradient was 9.4±3.4 mm Hg on echocardiography and 8.5±4 mm Hg invasively. Invasive median mitral valve area using the Gorlin formula was 1.87±0.9 cm2. Median MAC calcium score for the cohort was 7280±7937 Hounsfield units. MAC calcium score correlated with the presence of atrial fibrillation (P=0.02) but was not associated with other comorbidities. There was no correlation between MAC calcium score and mitral valve area (r=0.07; P=0.6) or mitral valve gradient (r=-0.03; P=0.8). MAC calcium score did not correlate with invasively measured mitral valve gradient and mitral valve area in patients with MAC-related mitral stenosis, suggesting that calcium score should not be used as a surrogate for invasive hemodynamic parameters.

Climate Change Facilitates the Potentially Suitable Habitats of the Invasive Crop Insect Ectomyelois ceratoniae (Zeller)

Invasive alien insects directly or indirectly driven by climate change threaten crop production and increase economic costs worldwide. Ectomyelois ceratoniae (Zeller) is a highly reproductive invasive crop insect that can severely damage fruit commodities and cause significant economic losses globally. Estimating the global potentially suitable habitats (PSH) of E. ceratoniae is an important aspect of its invasive risk assessment and early warning. Here, we constructed an optimized MaxEnt model based on the global distribution records of E. ceratoniae, and nine environmental variables (EVs), to predict its global PSH under current and future climates. Our results showed that the RM value was 2.0 and the mean area under receiver operating characteristic curve (AUC) value was 0.972, indicating the high accuracy of the optimal MaxEnt model. The mean temperature of driest quarter (bio9, 50.2%), mean temperature of wettest quarter (bio8, 16.9%), temperature seasonality (bio4, 9.7%), and precipitation of coldest quarter (bio19, 9.1%) were the significant EVs affecting its distribution patterns. The global PSH of E. ceratoniae are mainly located in western Asia under current climate scenarios (687.57 × 104 km2), which showed an increasing trend under future climate scenarios. The PSH of E. ceratoniae achieved the maximum under the shared socioeconomic pathway (SSP) 1–2.6 in the 2030s and under the SSP2-4.5 in the 2050s. The increased PSH of E. ceratoniae are mainly located in southwestern Asia, northwestern Europe, northwestern South America, northwestern North America, southern Oceania, and northwestern Africa. Our findings suggest that quarantine officials and governmental departments in the above high-risk invasion areas should strengthen monitoring and early warning to control E. ceratoniae; in particular, cultural measures should be taken in areas where its further expansion is expected in the future.

Identification of invasive subpopulations using spatial transcriptome analysis in thyroid follicular tumors.

Follicular tumors include follicular thyroid adenomas and carcinomas; however, it is difficult to distinguish between the two when the cytology or biopsy material is obtained from a portion of the tumor. The presence or absence of invasion in the resected material is used to differentiate between adenomas and carcinomas, which often results in the unnecessary removal of the adenomas. If nodules that may be follicular thyroid carcinomas are identified preoperatively, active surveillance of other nodules as adenomas is possible, which reduces the risk of surgical complications and the expenses incurred during medical treatment. Therefore, we aimed to identify biomarkers in the invasive subpopulation of follicular tumor cells. We performed a spatial transcriptome analysis of a case of follicular thyroid carcinoma and examined the dynamics of CD74 expression in 36 cases. We identified a subpopulation in a region close to the invasive area, and this subpopulation expressed high levels of CD74. Immunohistochemically, CD74 was highly expressed in the invasive and peripheral areas of the tumor. Although high CD74 expression has been reported in papillary and anaplastic thyroid carcinomas, it has not been analyzed in follicular thyroid carcinomas. Furthermore, the heterogeneity of CD74 expression in thyroid tumors has not yet been reported. The CD74-positive subpopulation identified in this study may be useful in predicting invasion of follicular thyroid carcinomas.

Assessing the Current and Future Potential Distribution of Solanum rostratum Dunal in China Using Multisource Remote Sensing Data and Principal Component Analysis

Accurate information concerning the spatial distribution of invasive alien species’ habitats is essential for invasive species prevention and management, and ecological sustainability. Currently, nationwide identification of suitable habitats for the highly destructive and potentially invasive weed, Solanum rostratum Dunal (S. rostratum), poses a series of challenges. Simultaneously, research on potential future invasion areas and likely directions of spread has not received adequate attention. This study, based on species occurrence data and multi-dimensional environmental variables constructed from multi-source remote sensing data, utilized Principal Component Analysis (PCA) in combination with the Maxent model to effectively model the current and future potential habitat distribution of S. rostratum in China, while quantitatively assessing the various factors influencing its distribution. Research findings indicate that the current suitable habitat area of S. rostratum covers 1.3952 million km2, all of which is located in northern China. As the trend of climate warming persists, the potential habitat suitability range of S. rostratum is projected to shift southward and expand in the future; while still predominantly located in northern China, it will have varying degrees of expansion at different time frames. Notably, during the period from 2040 to 2061, under the SSP1-2.6 scenario, the habitat area exhibits the most significant increase, surpassing the current scenario by 19.23%. Furthermore, attribution analysis based on PCA inverse transformation reveals that a combination of soil, climate, spatial, humanistic, and topographic variables collectively influence the suitability of S. rostratum habitats, with soil factors, in particular, playing a dominant role and contributing up to 75.85%. This study identifies target areas for the management and control of S. rostratum, providing valuable insights into factor selection and variable screening methods in species distribution modeling (SDM).

Characteristics and immune checkpoint status of radioiodine-refractory recurrent papillary thyroid carcinomas from Ukrainian Chornobyl Tissue Bank donors.

The radioiodine-refractory (RAI-R) recurrent papillary thyroid carcinomas (PTCs) are more frequent in elderly patients and have an unfavorable prognosis. Data on the prevalence and characteristics of RAI-R recurrent PTCs in patients of young and middle age with or without a history of radiation exposure in childhood are poorly described. The aim of the current study was: i) to determine the frequency of RAI-R recurrent PTCs among donors of the Chornobyl Tissue Bank (CTB) and analyze the clinicopathological features of primary tumors (PTs), primary metastases (PMTSs), recurrent metastases (RMTSs) and risk factors for RMTS, and ii) to determine the immune checkpoint status (ICS) of the RAI-R recurrent PTCs and to assess the factors associated with ICS positivity. Sixty RAI-R recurrent PTCs (46 exposed to radiation and 14 non-exposed, 2.5% of all cases registered with the CTB) from the Ukrainian patients aged up to 48 years were identified. The clinicopathological characteristics of the PTs moderately to weakly resembled those of the PMTS and RMTS from the same patients while the metastatic tissues were highly similar. The multivariate model of RMTS included the dominant solid-trabecular growth pattern of the PT, cystic changes, N1b metastases, and the probability of a causation (POC) of PTC by radiation as risk factors. Among these factors, the lateral PMTS (N1b) had the strongest effect. The longer period of latency (a POC component) was the second statistically significant characteristic. ICS percent agreement between the PT and RAI-R RMTS was 91.5%; 23.7% of PTs and 28.8% of RMTSs had positive ICS (positive PD-L1 tumor epithelial cells (TECs) and positive PD-L1/PD1 tumor-associated immune cells). ICS positivity of PTs was associated with pronounced oncocytic changes and high density of the p16INK4A-positive TECs in the invasive areas of PTs. In RMTSs, ICS positivity was associated with pronounced oncocytic changes and Ki-67 labeling index ≥ 4.5% of PTs, and the dominant solid-trabecular growth pattern, Ki-67 labeling index ≥ 7.6% and p16INK4A-positivity of RMTS. The findings are of clinical relevance and may be useful for developing individual treatment approaches for patients with RAI-R recurrent PTCs possibly involving immunotherapy.