Identification of a novel ER-NFĸB-driven stem-like cell population associated with relapse of ER+ breast tumors.

BACKGROUND: Up to 40% of patients with estrogen receptor-positive (ER+) breast cancer experience relapse. This can be attributed to breast cancer stem cells (BCSCs), which are known to be involved in therapy resistance, relapse, and metastasis. Therefore, there is an urgent need to identify genes/pathways that drive stem-like cell properties in ER+ breast tumors. METHODS: Using single-cell RNA sequencing and various bioinformatics approaches, we identified a unique stem-like population and established its clinical relevance. With follow-up studies, we validated our bioinformatics findings and confirmed the role of ER and NFĸB in the promotion of stem-like properties in breast cancer cell lines and patient-derived models. RESULTS: We identified a novel quiescent stem-like cell population that is driven by ER and NFĸB in multiple ER+ breast cancer models. Moreover, we found that a gene signature derived from this stem-like population is expressed in primary ER+ breast tumors, endocrine therapy-resistant and metastatic cell populations and predictive of poor patient outcome. CONCLUSIONS: These findings indicate a novel role for ER and NFĸB crosstalk in BCSCs biology and understanding the mechanism by which these pathways promote stem properties can be exploited to improve outcomes for ER+ breast cancer patients at risk of relapse.

Estrogen Receptor-Regulated Gene Signatures in Invasive Breast Cancer Cells and Aggressive Breast Tumors.

Most metastatic breast cancers arise from estrogen receptor α (ER)-positive disease, and yet the role of ER in promoting metastasis is unclear. Here, we used an ER+ breast cancer cell line that is highly invasive in an ER- and IKKß-dependent manner. We defined two ER-regulated gene signatures that are specifically regulated in the subpopulations of invasive cells. The first consists of proliferation-associated genes, which is a known function of ER, which actually suppress rather than enhance invasion. The second signature consists of genes involved in essential biological processes, such as organelle assembly and vesicle trafficking. Importantly, the second subpopulation-specific signature is associated with aggressive disease and poor patient outcome, independently of proliferation. These findings indicate a complex interplay between ER-driven proliferation and invasion, and they define new ER-regulated gene signatures that are predictive of aggressive ER+ breast cancer.

ASSESSMENT OF OCCUPATIONAL EXPOSURE IN THE MAIN PAEDIATRIC INTERVENTIONAL RADIOLOGY PROCEDURES.

The aim of this study is to evaluate the personal dose equivalent Hp(10) in the most frequent (non-cardiac) paediatric interventional radiology (PIR) procedures: central venous catheters (CVC), hepatic/biliary and sclerotherapy interventions. i2 active solid-state dosemeters placed over the lead apron were used to monitor the exposure of three interventional radiologists over 18 months. A database was created to register all procedures performed by each radiologist (including the type of procedure and the kerma-area product, PKA). The mean Hp(10) per procedure for CVC, sclerotherapy and hepatic/biliary interventions was respectively 0.01 ± 0.01 mSv, 0.18 ± 0.13 mSv and 0.12 ± 0.06 mSv (k = 2). A similar value of Hp(10)/PKA was found despite the type of procedure or the patient weight (~10 µSv/Gy·cm2). There was high variability among individual interventions, probably due to the variable level of complexity, which led to uncertainties in the measurements' mean higher than those associated with the dosemeter's angular and energy dependence. i2 therefore proved suitable for monitoring Hp(10) in PIR procedures.

Role of correlated noise in textural features extraction.

Predictive models of tumor response based on heterogeneity metrics in medical images, such as textural features, are highly suggestive. However, the demonstrated sensitivity of these features to noise does affect the model being developed. An in-depth analysis of the noise influence on the extraction of texture features was performed based on the assumption that an improvement in information quality can also enhance the predictive model. A heuristic approach was used that recognizes from the beginning that the noise has its own texture and it was analysed how it affects the quantitative signal data. A simple procedure to obtain noise image estimation is shown; one which makes it possible to extract the noise-texture features at each observation. The distance measured between the textural features in signal and estimated noise images allows us to determine the features affected in each observation by the noise and, for example, to exclude some of them from the model. A demonstration was carried out using synthetic images applying realistic noise models found in medical images. Drawn conclusions were applied to a public cohort of clinical images obtained using FDG-PET to show how the predictive model could be improved. A gain in the area under the receiver operating characteristic curve between 10 and 20% when noise texture information is used was shown. An improvement between 20 and 30% can be appreciated in the estimated model quality.

Occupational doses to the eye lens in pediatric and adult noncardiac interventional radiology procedures.

PURPOSE: To assess occupational lens exposure in a mixed interventional radiology department, comparing pediatric and adult procedures. To analyze the correlation between the lens dose and the doses measured at the chest and collar level and the kerma-area product (PKA ). METHODS: For 17 months, three radiologists performing both pediatric and adult interventions were monitored by means of 14 dosimeters per worker: 12 single-point optically stimulated luminescent (OSL) dosimeters calibrated in terms of Hp (0.07) were placed on the inside and outside of two pairs of lead glasses, one for pediatric procedures and one for adult interventions; another whole-body OSL dosimeter calibrated in terms of Hp (10) was placed over the thyroid shield; finally, an additional active solid-state dosimeter, also calibrated for Hp (10), was worn on the chest, over the apron. Furthermore, a database was created to register the demographic and dosimetric data of the procedures, as well as the name of the radiologist acting as first operator. RESULTS: For the three radiologists, who performed 276-338 procedures/year (20% pediatric), cumulative annual doses to the left bare eye exceeded 20 mSv (21-61 mSv). Considering the glasses' protection, annual doses exceeded 6 mSv (13-48 mSv) for both eyes. No important differences were observed in lens dose per procedure between pediatric and adult interventions (0.16 vs 0.18, 0.12 vs 0.09, and 0.07 vs 0.07 mSv), although lens dose per PKA was 4.1-4.5 times higher in pediatrics (5.8 vs 1.3, 3.3 vs 0.8, and 2.6 vs 0.6 µSv/Gy·cm2 ) despite a similar use of the ceiling-suspended screen. Lens doses were highly correlated with collar readings (with Pearson coefficients [r] ranging from 0.86 to 0.98) and with chest readings (with r ranging from 0.75 to 0.93). However, slopes of the linear regressions varied greatly among radiologists. CONCLUSIONS: There is real risk of exceeding the occupational dose limit to the eye lens in mixed interventional radiology rooms if radiation protection tools are not used properly. Regular monitoring of the lens dose is recommended, given lens exposure might easily exceed 6 mSv/yr. Using a collar dosimeter for this purpose might be suitable if it is preceded by an individualized regression analysis. The same radiation protection measures should be applied to interventional radiologists regardless of whether they are treating pediatric or adult patients.

The NF-κB Pathway Promotes Tamoxifen Tolerance and Disease Recurrence in Estrogen Receptor-Positive Breast Cancers.

The purpose of this study was to identify critical pathways promoting survival of tamoxifen-tolerant, estrogen receptor α positive (ER+) breast cancer cells, which contribute to therapy resistance and disease recurrence. Gene expression profiling and pathway analysis were performed in ER+ breast tumors of patients before and after neoadjuvant tamoxifen treatment and demonstrated activation of the NF-κB pathway and an enrichment of epithelial-to mesenchymal transition (EMT)/stemness features. Exposure of ER+ breast cancer cell lines to tamoxifen, in vitro and in vivo, gives rise to a tamoxifen-tolerant population with similar NF-κB activity and EMT/stemness characteristics. Small-molecule inhibitors and CRISPR/Cas9 knockout were used to assess the role of the NF-κB pathway and demonstrated that survival of tamoxifen-tolerant cells requires NF-κB activity. Moreover, this pathway was essential for tumor recurrence following tamoxifen withdrawal. These findings establish that elevated NF-κB activity is observed in breast cancer cell lines under selective pressure with tamoxifen in vitro and in vivo, as well as in patient tumors treated with neoadjuvant tamoxifen therapy. This pathway is essential for survival and regrowth of tamoxifen-tolerant cells, and, as such, NF-κB inhibition offers a promising approach to prevent recurrence of ER+ tumors following tamoxifen exposure. IMPLICATIONS: Understanding initial changes that enable survival of tamoxifen-tolerant cells, as mediated by NF-κB pathway, may translate into therapeutic interventions to prevent resistance and relapse, which remain major causes of breast cancer lethality.

Local diagnostic reference levels for paediatric non-cardiac interventional radiology procedures.

PURPOSE: To establish local diagnostic reference levels (DRLs) for non-cardiac interventional procedures in paediatrics. METHODS: The type of procedure, the patient's weight and age and dose-related data from 279 interventions was recorded in a database completed by interventional radiologists, radiographers and technicians of the Medical Physics department. These procedures were classified into 14 categories and 6 weight ranges. Local DRLs were proposed for those ranges in which a sample of at least 15 patients could be gathered and were calculated as the third quartile (Q3) of the air kerma-area product (PKA) values. The Q3 of the fluoroscopy time (FT) and number of digital subtraction angiography (DSA) images were also obtained. Finally, the correlation between PKA and weight was analysed. RESULTS: Local DRLs are proposed for three types of procedures: hepatic/biliary interventions (5-15 kg, 1304 cGy·cm2; 15-30 kg, 2121 cGy·cm2), sclerotherapy procedures (15-30 kg, 704 cGy·cm2; 30-50 kg, 4049 cGy·cm2; 50-80 kg, 3734 cGy·cm2) and central venous catheter (CVC) procedures (5-15 kg, 84 cGy·cm2). Hepatic/biliary interventions showed a moderate correlation (r = 0.61), while sclerotherapy procedures presented a poor correlation (r = 0.34) between PKA and weight, possibly due to the PKA dependence on the complexity level. Regarding CVC procedures, a clearly higher correlation was found when the fluoroscopy PKA value was normalised to the FT (r = 0.85 vs r = 0.35). CONCLUSIONS: The results support the feasibility of establishing DRLs for the most common procedures (sclerotherapy, hepatic/biliary and CVC interventions) despite the small number of paediatric interventions.

Cytoplasmic ERα and NFκB Promote Cell Survival in Mouse Mammary Cancer Cell Lines.

There is a desperate need in the field for mouse mammary tumors and cell lines that faithfully mimic estrogen receptor (ER) expression and activity found in human breast cancers. We found that several mouse mammary cancer cell lines express ER but fail to demonstrate classical estrogen-driven proliferation or transcriptional activity. We investigated whether these cell lines may be used to model tamoxifen resistance by using small molecule inhibitors to signaling pathways known to contribute to resistance. We found that the combination of NFκB inhibition and ER antagonists significantly reduced cell proliferation in vitro, as well as growth of syngeneic tumors. Surprisingly, we found that ER was localized to the cytoplasm, regardless of any type of treatment. Based on this, we probed extra-nuclear functions of ER and found that co-inhibition of ER and NFκB led to an increase in oxidative stress and apoptosis. Together, these findings suggest that cytoplasmic ER and NFκB may play redundant roles in protecting mammary cancer cells from oxidative stress and cell death. Although this study has not identified a mouse model with classical ER activity, cytoplasmic ER has been described in a small subset of human breast tumors, suggesting that these findings may be relevant for some breast cancer patients.

Effect of salinity on the synthesis and concentration of glycine betaine in osmoregulatory tissues from juvenile shrimps Litopenaeus vannamei.

In marine animals, glycine betaine is one of the main osmolytes accumulated under osmotic stress conditions; nevertheless, in penaeids, shrimps little is known about the pathways involved in glycine betaine biosynthesis. In animal cells, glycine betaine is synthesized by the enzyme betaine aldehyde dehydrogenase (BADH). We herein investigated the salinity effect on the synthesis and concentration of glycine betaine on white shrimp Litopenaeus vannamei. Shrimps were subjected to 10, 20, 35, 40, 50, and 60 ppt salinity conditions for seven days. BADH activity increased in hepatopancreas and gills of shrimps subjected to salinities above 35 ppt salinity. In muscle, the BADH activity decreased at 35 ppt salinity. In hepatopancreas from shrimps subjected to 50 and 60 ppt salinities, BADH activity increased 1.1 and 1.7-fold. At 60 ppt salinity, BADH activity increased 1.5-fold respect to 35 ppt in gills. Glycine betaine concentration increased in hepatopancreas, gills, muscle, and hemolymph in shrimps subjected to salinities above 35 ppt. Glycine betaine concentration also increased at 20 ppt salinity, while at 10 ppt, not detected significant differences. The catch of glycine betaine from hemolymph by the cell likely is carried out to avoid protein denaturalization. Ammonia concentration in the aquarium's water only increased at salinities of 20 ppt and 10 ppt (1.1-fold relative to 35 ppt). Our data demonstrated that in L. vannamei, salinity regulates BADH activity and glycine betaine content in a tissue-specific manner.

Segmentation improvement through denoising of PET images with 3D-context modelling in wavelet domain.

Positron emission tomography (PET) images have been incorporated into the radiotherapy process as a powerful tool to assist in the contouring of lesions, leading to the emergence of a broad spectrum of automatic segmentation schemes for PET images (PET-AS). However, not all proposed PET-AS algorithms take into consideration the previous steps of image preparation. PET image noise has been shown to be one of the most relevant affecting factors in segmentation tasks. This study demonstrates a nonlinear filtering method based on spatially adaptive wavelet shrinkage using three-dimensional context modelling that considers the correlation of each voxel with its neighbours. Using this noise reduction method, excellent edge conservation properties are obtained. To evaluate the influence in the segmentation schemes of this filter, it was compared with a set of Gaussian filters (the most conventional) and with two previously optimised edge-preserving filters. Five segmentation schemes were used (most commonly implemented in commercial software): fixed thresholding, adaptive thresholding, watershed, adaptive region growing and affinity propagation clustering. Segmentation results were evaluated using the Dice similarity coefficient and classification error. A simple metric was also included to improve the characterisation of the filters used for induced blurring evaluation, based on the measurement of the average edge width. The proposed noise reduction procedure improves the results of segmentation throughout the performed settings and was shown to be more stable in low-contrast and high-noise conditions. Thus, the capacity of the segmentation method is reinforced by the denoising plan used.

Biplane interventional pediatric system with cone-beam CT: dose and image quality characterization for the default protocols.

The aim of this study was to assess image quality and radiation dose of a biplane angiographic system with cone-beam CT (CBCT) capability tuned for pediatric cardiac procedures. The results of this study can be used to explore dose reduction techniques. For pulsed fluoroscopy and cine modes, polymethyl methacrylate phantoms of various thicknesses and a Leeds TOR 18-FG test object were employed. Various fields of view (FOV) were selected. For CBCT, the study employed head and body dose phantoms, Catphan 504, and an anthropomorphic cardiology phantom. The study also compared two 3D rotational angiography protocols. The entrance surface air kerma per frame increases by a factor of 3-12 when comparing cine and fluoroscopy frames. The biggest difference in the signal-to- noise ratio between fluoroscopy and cine modes occurs at FOV 32 cm because fluoroscopy is acquired at a 1440 × 1440 pixel matrix size and in unbinned mode, whereas cine is acquired at 720 × 720 pixels and in binned mode. The high-contrast spatial resolution of cine is better than that of fluoroscopy, except for FOV 32 cm, because fluoroscopy mode with 32 cm FOV is unbinned. Acquiring CBCT series with a 16 cm head phantom using the standard dose protocol results in a threefold dose increase compared with the low-dose protocol. Although the amount of noise present in the images acquired with the low-dose protocol is much higher than that obtained with the standard mode, the images present better spatial resolution. A 1 mm diameter rod with 250 Hounsfield units can be distinguished in reconstructed images with an 8 mm slice width. Pediatric-specific protocols provide lower doses while maintaining sufficient image quality. The system offers a novel 3D imaging mode. The acquisition of CBCT images results in increased doses administered to the patients, but also provides further diagnostic information contained in the volumetric images. The assessed CBCT protocols provide images that are noisy, but with very good spatial resolution.

Comparative Study of the Optical and Textural Properties of Tetrapyrrole Macrocycles Trapped Within ZrO2, TiO2, and SiO2 Translucent Xerogels.

The entrapping of physicochemical active molecules inside mesoporous networks is an appealing field of research due to the myriad of potential applications in optics, photocatalysis, chemical sensing, and medicine. One of the most important reasons for this success is the possibility of optimizing the properties that a free active species displays in solution but now trapped inside a solid substrate. Additionally it is possible to modulate the textural characteristics of substrates, such as pore size, specific surface area, polarity and chemical affinity of the surface, toward the physical or chemical adhesion of a variety of adsorbates. In the present document, two kinds of non-silicon metal alkoxides, Zr and Ti, are employed to prepare xerogels containing entrapped tetrapyrrolic species that could be inserted beforehand in analogue silica systems. The main goal is to develop efficient methods for trapping or binding tetrapyrrole macrocycles inside TiO2 and ZrO2 xerogels, while comparing the properties of these systems against those of the SiO2 analogues. Once the optimal synthesis conditions for obtaining translucent monolithic xerogels of ZrO2 and TiO2 networks were determined, it was confirmed that these substrates allowed the entrapment, in monomeric form, of macrocycles that commonly appear as aggregates within the SiO2 network. From these experiments, it could be determined that the average pore diameters, specific surface areas, and water sorption capacities depicted by each one of these substrates, are a consequence of their own nature combined with the particular structure of the entrapped tetrapyrrole macrocycle. Furthermore, the establishment of covalent bonds between the intruding species and the pore walls leads to the obtainment of very similar pore sizes in the three different metal oxide (Ti, Zr, and Si) substrates as a consequence of the templating effect of the encapsulated species.

High-Quality Draft Genomes of Two Vibrio parahaemolyticus Strains Aid in Understanding Acute Hepatopancreatic Necrosis Disease of Cultured Shrimps in Mexico.

The high-quality draft genomes of two Vibrio parahaemolyticus strains, one that causes the acute hepatopancreatic necrosis disease (AHPND) in cultured shrimps (FIM-S1708(+)), and another that does not (FIM-S1392(-)) are reported. A chromosome-scale assembly for the FIM-S1392(-) genome is reported here. The analysis of the two genomes gives some clues regarding the genomic differences between the strains.

Crossed and linked histories of tetrapyrrolic macrocycles and their use for engineering pores within sol-gel matrices.

The crossed and linked histories of tetrapyrrolic macrocycles, interwoven with new research discoveries, suggest that Nature has found in these structures a way to ensure the continuity of life. For diverse applications porphyrins or phthalocyanines must be trapped inside solid networks, but due to their nature, these compounds cannot be introduced by thermal diffusion; the sol-gel method makes possible this insertion through a soft chemical process. The methodologies for trapping or bonding macrocycles inside pristine or organo-modified silica or inside ZrO2 xerogels were developed by using phthalocyanines and porphyrins as molecular probes. The sizes of the pores formed depend on the structure, the cation nature, and the identities and positions of peripheral substituents of the macrocycle. The interactions of the macrocyclic molecule and surface Si-OH groups inhibit the efficient displaying of the macrocycle properties and to avoid this undesirable event, strategies such as situating the macrocycle far from the pore walls or to exchange the Si-OH species by alkyl or aryl groups have been proposed. Spectroscopic properties are better preserved when long unions are established between the macrocycle and the pore walls, or when oligomeric macrocyclic species are trapped inside each pore. When macrocycles are trapped inside organo-modified silica, their properties result similar to those displayed in solution and their intensities depend on the length of the alkyl chain attached to the matrix. These results support the prospect of tuning up the pore size, surface area, and polarity inside the pore cavities in order to prepare efficient catalytic, optical, sensoring, and medical systems. The most important feature is that research would confirm again that tetrapyrrolic macrocycles can help in the development of the authentic pore engineering in materials science.