Exosc9 Initiates SUMO-Dependent lncRNA TERRA Degradation to Impact Telomeric Integrity in Endocrine Therapy Insensitive Hormone Receptor-Positive Breast Cancer.

Long, noncoding RNAs (lncRNAs) are indispensable for normal cell physiology and, consequently, are tightly regulated in human cells. Yet, unlike mRNA, substantially less is known about the mechanisms for lncRNA degradation. It is important to delineate the regulatory control of lncRNA degradation, particularly for lncRNA telomeric repeat-containing RNA (TERRA), as the TERRA-telomere R-loops dictate cell cycle progression and genomic stability. We now report that the exosome complex component Exosc9 degrades lncRNA TERRA in human mammary epithelial cells. Heterochromatin protein 1 alpha (HP1α) recruits Exosc9 to the telomeres; specifically, the SUMO-modified form of HP1α supports interaction with Exosc9 and, as previously reported, lncRNA TERRA. The telomeric enrichment of Exosc9 is cell cycle-dependent and consistent with the loss of telomeric TERRA in the S/G2 phase. Elevated Exosc9 is frequently observed and drives the growth of endocrine therapy-resistant (ET-R) HR+ breast cancer (BCa) cells. Specifically, the knockdown of Exosc9 inversely impacts telomeric R-loops and the integrity of the chromosome ends of ET-R cells. Consistently, Exosc9 levels dictate DNA damage and the sensitivity of ET-R BCa cells to PARP inhibitors. In this regard, Exosc9 may serve as a promising biomarker for predicting the response to PARP inhibitors as a targeted monotherapy for ET-R HR+ BCa.

Objective Analysis of Breast Symmetry in Female Patients Undergoing Breast Reconstruction After Total Mastectomy.

Background: Satisfaction with the breast aesthetic outcome is an expectation of breast reconstruction surgery, which is an integral part of cancer treatment for many patients. We evaluated postreconstruction breast symmetry in 82 female patients using distance and volume measurements. Objectives: Clinical factors, such as reconstruction type (implant-based and autologous reconstruction), laterality, timing of reconstruction (immediate, delayed, and sequential), radiation therapy (RT), and demographic factors (age, BMI, race, and ethnicity), were evaluated as predictors of postoperative symmetry. Matched preoperative and postoperative measurements for a subset of 46 patients were used to assess correlation between preoperative and postoperative symmetry. Methods: We used standardized differences between the left and right breasts for the sternal notch to lowest visible point distance and breast volume as metrics for breast, positional symmetry, and volume symmetry, respectively. We performed statistical tests to compare symmetry between subgroups of patients based on reconstruction type, laterality, timing, RT, and demographics. Results: Overall, reconstruction type, reconstruction timing, and RT were observed to be factors significantly associated with postoperative symmetry, with implant reconstructions and immediate reconstruction procedures, and no RT showing better postoperative breast volume symmetry. Subgroup analyses, for both reconstruction type and laterality, showed superior volume symmetry for the bilateral implant reconstructions. No correlation was observed between preoperative and postoperative breast symmetry. Demographic factors were not significant predictors of postreconstruction symmetry. Conclusions: This comprehensive analysis examines multiple clinical factors in a single study and will help both patients and surgeons make informed decisions about reconstruction options at their disposal.

Impact of implant-based breast reconstruction on bra fit.

Comfortable and well-fitting bras are necessary for good quality of life but hard to find for women who undergo reconstruction after breast cancer treatment. This study aimed to provide data to inform bra designs for breast cancer survivors. We measured anatomical distances used in bra design on 3D clinical photographs of patients who underwent unilateral and bilateral implant-based reconstruction to quantify changes after reconstruction relative to the measured values before the person underwent surgery. We performed additional assessments of symmetry before surgery and after reconstruction, and we used regression analyses to identify associations between the measurements and patient characteristics, such as BMI. Overall, almost all measurements changed significantly in implant-based reconstructed breasts relative to native breasts. We highlight several aspects of ergonomic bra design that will be impacted by the changes in anatomical distances. Practitioner summary: Implant-based breast reconstruction surgery changes the breast so that off-the-rack bras are inadequate. This study provides designers with measurement data from women who underwent implant-based reconstruction to inform bra designs for this population. The key factor designers need to account for is the semi-spherical shape of the reconstructed breast.

Research Infrastructure Core Facilities at Research Centers in Minority Institutions: Part I-Research Resources Management, Operation, and Best Practices.

Funded by the National Institutes of Health (NIH), the Research Centers in Minority Institutions (RCMI) Program fosters the development and implementation of innovative research aimed at improving minority health and reducing or eliminating health disparities. Currently, there are 21 RCMI Specialized (U54) Centers that share the same framework, comprising four required core components, namely the Administrative, Research Infrastructure, Investigator Development, and Community Engagement Cores. The Research Infrastructure Core (RIC) is fundamentally important for biomedical and health disparities research as a critical function domain. This paper aims to assess the research resources and services provided and evaluate the best practices in research resources management and networking across the RCMI Consortium. We conducted a REDCap-based survey and collected responses from 57 RIC Directors and Co-Directors from 98 core leaders. Our findings indicated that the RIC facilities across the 21 RCMI Centers provide access to major research equipment and are managed by experienced faculty and staff who provide expert consultative and technical services. However, several impediments to RIC facilities operation and management have been identified, and these are currently being addressed through implementation of cost-effective strategies and best practices of laboratory management and operation.

Breast Decisions: Recommender System for Appearance Counseling about Breast Reconstruction.

Appearance counseling is an important component of the consent process for breast reconstruction. The purpose of appearance counseling is to help the patient form realistic expectations of what she might look like after breast reconstruction. In this article, we introduce a recommender system, "BreastDecisions," for appearance counseling that suggests photographs of previous patients that are tailored to a specific patient to help her form realistic expectations of her own reconstruction. Methods: We present user specifications and algorithm parameters needed to incorporate the recommender system into the appearance counseling workflow. We demonstrate the system for a common counseling scenario using a knowledgebase of previous breast reconstruction patients. The medical appropriateness of the recommended photographs for use in appearance counseling was evaluated by experts using a four-point rating system. Results: The recommender system presents photographs that are medically appropriate for counseling a specific patient, depicts typical outcomes, and adapts to a variety of clinical workflows. For each of 33 patients taken as examples of breast reconstruction patients, we used the system to identify photographs for appearance counseling. The baseline average medical appropriateness of the recommended photographs was between mostly appropriate (some explanation needed) and medically appropriate (minimal explanation needed). We demonstrate filtering and ranking steps to reduce the number of recommended photographs and increase the average medical appropriateness. Conclusions: Our recommender system automatically suggests photographs of previous breast reconstruction patients for use in counseling a patient about appearance outcomes. The system is patient-specific and customizable to a particular surgeon's practice.

Potential of Intraoperative 3D Photography and 3D Visualization in Breast Reconstruction.

Although pre- and postoperative three-dimensional (3D) photography are well-established in breast reconstruction, intraoperative 3D photography is not. We demonstrate the process of intraoperative acquisition and visualization of 3D photographs for breast reconstruction and present clinicians' opinions about intraoperative visualization tools. METHODS: Mastectomy specimens were scanned with a handheld 3D scanner during breast surgery. The 3D photographs were processed to compute morphological measurements of the specimen. Three visualization modalities (screen-based viewing, augmented reality viewing, and 3D printed models) were created to show different representations of the 3D photographs to plastic surgeons. We interviewed seven surgeons about the usefulness of the visualization methods. RESULTS: The average time for intraoperative acquisition of 3D photographs of the mastectomy specimen was 4 minutes, 8 seconds ± 44 seconds. The average time for image processing to compute morphological measurements of the specimen was 54.26 ± 40.39 seconds. All of the interviewed surgeons would be more inclined to use intraoperative visualization if it displayed information that they are currently missing (eg, the target shape of the reconstructed breast mound). Additionally, the surgeons preferred high-fidelity visualization tools (such as 3D printing) that are easy-to-use and have minimal disruption to their current workflow. CONCLUSIONS: This study demonstrates that 3D photographs can be collected intraoperatively within acceptable time limits, and quantitative measurements can be computed timely to be utilized within the same procedure. We also report surgeons' comments on usability of visualization methods and of measurements of the mastectomy specimen, which can be used to guide future surgical practice.

Goldilocks Principle: Preference for Change in Breast Size in Breast Cancer Reconstruction Patients.

Patients' preferences regarding changing or maintaining their breast size after mastectomy and reconstruction are important but understudied determinants of post-surgical satisfaction and quality of life. The goal of this study was to identify factors associated with preferences for changing or maintaining breast size for women undergoing breast reconstruction at The University of Texas MD Anderson Cancer Center in the United States from 2011 to 2014. The average age of participants was 45.7 ± 9.1 years. At baseline, mean average breast volumes were 755.7 ± 328.4 mL for all women (n = 48), 492.3 mL ± 209.3 for 13 women who preferred to be "bigger than now," 799.2 mL ± 320.9 for 25 women who preferred to remain "about the same," and 989.3 mL ± 253.1 for 10 women who preferred "smaller than now." Among the 23 women who preferred to change their breast size, 19 desired to shift toward the mean. Women with the smallest and largest 20% of baseline breast size were more likely to desire a change toward the mean (p = 0.006). Multinomial logistic regression models found average breast volume and satisfaction with breast size to be the most important factors associated with preferences for changing or maintaining breast size for women undergoing breast reconstruction. This study provides preliminary evidence for a "Goldilocks principle" in women's preferences for breast size change in the context of breast reconstruction, and identifies hypotheses for future studies of the associations among preference for change in breast size, preference achievement, and post-reconstruction body image.

Review of Quantitative Imaging for Objective Assessment of Fat Grafting Outcomes in Breast Surgery.

This article presents a comprehensive review of the use of quantitative measurements based on breast images to assess fat retention, breast aesthetics, and fat necrosis after autologous fat grafting. Breast volume measured from MRI and 3-dimensional surface images is widely used as a measure of fat retention. Breast aesthetics are assessed from a combination of anthropometric (distance and contour) measurements on breast surface images. Examination of radiologic images (ultrasonography, mammography, and MRI) is utilized to assess fat necrosis. The article discusses implementation guidelines for objective outcome assessment to support robust quantification and enable investigations of fat grafting efficacy. Level of Evidence: 4.

High-performance computing for SARS-CoV-2 RNAs clustering: a data science‒based genomics approach.

Nowadays, Genomic data constitutes one of the fastest growing datasets in the world. As of 2025, it is supposed to become the fourth largest source of Big Data, and thus mandating adequate high-performance computing (HPC) platform for processing. With the latest unprecedented and unpredictable mutations in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the research community is in crucial need for ICT tools to process SARS-CoV-2 RNA data, e.g., by classifying it (i.e., clustering) and thus assisting in tracking virus mutations and predict future ones. In this paper, we are presenting an HPC-based SARS-CoV-2 RNAs clustering tool. We are adopting a data science approach, from data collection, through analysis, to visualization. In the analysis step, we present how our clustering approach leverages on HPC and the longest common subsequence (LCS) algorithm. The approach uses the Hadoop MapReduce programming paradigm and adapts the LCS algorithm in order to efficiently compute the length of the LCS for each pair of SARS-CoV-2 RNA sequences. The latter are extracted from the U.S. National Center for Biotechnology Information (NCBI) Virus repository. The computed LCS lengths are used to measure the dissimilarities between RNA sequences in order to work out existing clusters. In addition to that, we present a comparative study of the LCS algorithm performance based on variable workloads and different numbers of Hadoop worker nodes.

Quantitative 3-Dimensional Photographic Assessment of Breast Cosmesis After Whole Breast Irradiation for Early Stage Breast Cancer: A Secondary Analysis of a Randomized Clinical Trial.

PURPOSE: Our purpose was to use 3-dimensional (3D) surface photography to quantitatively measure breast cosmesis within the framework of a randomized clinical trial of conventionally fractionated (CF) and hypofractionated (HF) whole breast irradiation (WBI); to identify how 3D measurements are associated with patient- and physician-reported cosmesis; and to determine whether objective measures of breast symmetry varied by WBI treatment arm or transforming growth factor ß 1 (TGFß1) status. METHODS AND MATERIALS: From 2011 to 2014, 287 women age ≥40 with ductal carcinoma in situ or early-stage invasive breast cancer were enrolled in a multicenter trial and randomized to HF-WBI or CF-WBI with a boost. Three-dimensional surface photography was performed at 3 years posttreatment. Patient-reported cosmetic outcomes were recorded with the Breast Cancer Treatment Outcome Scale. Physician-reported cosmetic outcomes were assessed by the Radiation Therapy Oncology Group scale. Volume ratios and 6 quantitative measures of breast symmetry, termed F1-6C, were calculated using the breast contour and fiducial points assessed on 3D surface images. Associations between all metrics, patient- and physician-reported cosmesis, treatment arm, and TGFß1 genotype were performed using the Kruskal-Wallis test and multivariable logistic regression models. RESULTS: Among 77 (39 CF-WBI and 38 HF-WBI) evaluable patients, both patient- and physician-reported cosmetic outcomes were significantly associated with the F1C vertical symmetry measure (both P < .05). Higher dichotomized F1C and volumetric symmetry measures were associated with improved patient- and physician-reported cosmesis on multivariable logistic regression (both P ≤ .05). There were no statistically significant differences in vertical symmetry or volume measures between treatment arms. Increased F6C horizontal symmetry was observed in the CF-WBI arm (P = .05). Patients with the TGFß1 C-509T variant allele had lower F2C vertical symmetry measures (P = .02). CONCLUSIONS: Quantitative 3D image-derived measures revealed comparable cosmetic outcomes with HF-WBI compared with CF-WBI. Our findings suggest that 3D surface imaging may be a more sensitive method for measuring subtle cosmetic changes than global patient- or physician-reported assessments.

Multifunctional Bio-Nanocomposite Coatings for Perishable Fruits.

Hunger and chronic undernourishment impact over 800 million people, which translates to ≈10.7% of the world's population. While countries are increasingly making efforts to reduce poverty and hunger by pursuing sustainable energy and agricultural practices, a third of the food produced around the globe still is wasted and never consumed. Reducing food shortages is vital in this effort and is often addressed by the development of genetically modified produce or chemical additives and inedible coatings, which create additional health and environmental concerns. Herein, a multifunctional bio-nanocomposite comprised largely of egg-derived polymers and cellulose nanomaterials as a conformal coating onto fresh produce that slows down food decay by retarding ripening, dehydration, and microbial invasion is reported. The coating is edible, washable, and made from readily available inexpensive or waste materials, which makes it a promising economic alternative to commercially available fruit coatings and a solution to combat food wastage that is rampant in the world.

Novel Silicon Titanium Diboride Micropatterned Substrates for Cellular Patterning.

Both hard material photolithography and soft lithography are widely used for patterned cell culture. Soft lithography techniques enable bioactive molecule incorporation, however complex surface modifications are required to introduce specific ligands or proteins in conventional photolithography. In this study, we demonstrate human umbilical vein cell (HUVEC) and adult bone marrow derived mesenchymal stem cell (MSC) patterning on titanium diboride (TiB2) layers deposited on silicon (Si) substrates by electron-beam evaporation and micropatterned using photolithography. Micropatterned cell growth specificity on geometric shapes of circle and/or lines is achieved via differential growth factors adsorption in the presence of heparin. Specifically, the deposited films of TiB2 showed increased stiffness, hardness, hydrophilicity and surface charge when compared to background Si. These substrates were found to be compatible with HUVEC and MSC viability, based on biomarker expression and RNA-sequence transcriptome analysis. Cell-type dependent, micropattern selective cell growth, such as contact guidance, alignment, and durotaxis, were observed. In addition, MSC clustering was achieved, enabling a three-dimensional (3D) aggregate based microenvironment during culture. This study clearly demonstrates the potential of microfabricated Si and TiB2 biomaterials for patterned cell culture in vitro, independent of any additional surface modification.

Natural Breast Symmetry in Preoperative Breast Cancer Patients.

Plastic surgeons aim to achieve breast symmetry during cosmetic and reconstructive breast surgery. They rely on measures of breast size, position, and projection to determine and achieve breast symmetry, but normative data on symmetry in preoperative breast reconstruction patients are scarce. METHODS: A statistical evaluation was performed to examine the relationship of breast symmetry to demographic and clinical factors such as age, body mass index (BMI), race, and cancer status in a sample population of 87 patients who were scheduled to undergo mastectomy and breast reconstruction. The sternal notch to nipple (SN-N) distance and breast volume were measured on three-dimensional images, and distance and volume ratios across the left and right breasts were compared to determine symmetry. Ptosis grades were recorded and grade agreement (match) across the left and right breasts was assessed to determine shape symmetry. RESULTS: A substantial portion of women (41.4%) showed SN-N distance differences >5 mm and 50.6% exhibited a volume difference >50 mL between their right and left breasts. Multiple linear regression modeling did not show any association between age, BMI, and cancer status and the SN-N and volume ratios. Race showed an association with volume symmetry but not with SN-N symmetry. A higher BMI increased the likelihood of ptosis disagreement. Additionally, tumor size did not impact overall breast symmetry. CONCLUSION: This study provides normative data on the extent of breast asymmetry in preoperative patients that can guide physicians in setting realistic goals for reconstruction procedures and manage patients' expectations related to outcomes.

Body image dissatisfaction in patients undergoing breast reconstruction: Examining the roles of breast symmetry and appearance investment.

OBJECTIVE: Reconstruction as part of treatment for breast cancer is aimed at mitigating body image concerns after mastectomy. Although algorithms have been developed to objectively assess breast reconstruction outcomes, associations between objectively quantified breast aesthetic appearance and patient-reported body image outcomes have not been examined. Further, the role of appearance investment in explaining a patient's body image is not well understood. We investigated the extent to which objectively quantified breast symmetry and patient-reported appearance investment were associated with body image dissatisfaction in patients undergoing cancer-related breast reconstruction. METHODS: Breast cancer patients in different stages of reconstruction (n = 190) completed self-report measures of appearance investment and body image dissatisfaction. Vertical extent and horizontal extent symmetry values, which are indicators of breast symmetry, were calculated from clinical photographs. Associations among breast symmetry, appearance investment, body image dissatisfaction, and patient clinical factors were examined. Multi-variable regression was used to evaluate the extent to which symmetry and appearance investment were associated with body image dissatisfaction. RESULTS: Vertical extent symmetry, but not horizontal extent symmetry, was associated with body image dissatisfaction. Decreased vertical extent symmetry (ß = -.19, P < .05) and increased appearance investment (ß = .45, P < .001) were significantly associated with greater body image dissatisfaction while controlling for clinical factors. CONCLUSIONS: Breast symmetry and patient appearance investment both significantly contribute to an understanding of patient-reported body image satisfaction during breast reconstruction treatment.

Quantitative Assessment of Breast Cosmetic Outcome After Whole-Breast Irradiation.

PURPOSE: To measure, by quantitative analysis of digital photographs, breast cosmetic outcome within the setting of a randomized trial of conventionally fractionated (CF) and hypofractionated (HF) whole-breast irradiation (WBI), to identify how quantitative cosmesis metrics were associated with patient- and physician-reported cosmesis and whether they differed by treatment arm. METHODS AND MATERIALS: From 2011 to 2014, 287 women aged ≥40 with ductal carcinoma in situ or early invasive breast cancer were randomized to HF-WBI (42.56 Gy/16 fractions [fx] + 10-12.5 Gy/4-5 fx boost) or CF-WBI (50 Gy/25 fx + 10-14 Gy/5-7 fx). At 1 year after treatment we collected digital photographs, patient-reported cosmesis using the Breast Cancer Treatment and Outcomes Scale, and physician-reported cosmesis using the Radiation Therapy Oncology Group scale. Six quantitative measures of breast symmetry, labeled M1-M6, were calculated from anteroposterior digital photographs. For each measure, values closer to 1 imply greater symmetry, and values closer to 0 imply greater asymmetry. Associations between M1-M6 and patient- and physician-reported cosmesis and treatment arm were evaluated using the Kruskal-Wallis test. RESULTS: Among 245 evaluable patients, patient-reported cosmesis was strongly associated with M1 (vertical symmetry measure) (P<.01). Physician-reported cosmesis was similarly correlated with M1 (P<.01) and also with M2 (vertical symmetry, P=.01) and M4 (horizontal symmetry, P=.03). At 1 year after treatment, HF-WBI resulted in better values of M2 (P=.02) and M3 (P<.01) than CF-WBI; treatment arm was not significantly associated with M1, M4, M5, or M6 (P≥.12). CONCLUSIONS: Quantitative assessment of breast photographs reveals similar to improved cosmetic outcome with HF-WBI compared with CF-WBI 1 year after treatment. Assessing cosmetic outcome using these measures could be useful for future comparative effectiveness studies and outcome reporting.

Transcriptional regulation of core autophagy and lysosomal genes by the androgen receptor promotes prostate cancer progression.

AR (androgen receptor) signaling is crucial for the development and maintenance of the prostate as well as the initiation and progression of prostate cancer. Despite the AR's central role in prostate cancer progression, it is still unclear which AR-mediated processes drive the disease. Here, we identified 4 core autophagy genes: ATG4B, ATG4D, ULK1, and ULK2, in addition to the transcription factor TFEB, a master regulator of lysosomal biogenesis and function, as transcriptional targets of AR in prostate cancer. These findings were significant in light of our recent observation that androgens promoted prostate cancer cell growth in part through the induction of autophagy. Expression of these 5 genes was essential for maximal androgen-mediated autophagy and cell proliferation. In addition, expression of each of these 5 genes alone or in combination was sufficient to increase prostate cancer cell growth independent of AR activity. Further, bioinformatic analysis demonstrated that the expression of these genes correlated with disease progression in 3 separate clinical cohorts. Collectively, these findings demonstrate a functional role for increased autophagy in prostate cancer progression, provide a mechanism for how autophagy is augmented, and highlight the potential of targeting this process for the treatment of advanced prostate cancer.

Identification of vascular disruptor compounds by analysis in zebrafish embryos and mouse embryonic endothelial cells.

To identify vascular disruptor compounds (VDCs), this study utilized an in vivo zebrafish embryo vascular model in conjunction with a mouse endothelial cell model to screen a subset of the U.S. Environmental Protection Agency (EPA) ToxCast Phase I chemical inventory. In zebrafish, 161 compounds were screened and 34 were identified by visual inspection as VDCs, of which 28 were confirmed as VDCs by quantitative image analysis. Testing of the zebrafish VDCs for their capacity to inhibit endothelial tube formation in the murine yolk-sac-derived endothelial cell line C166 identified 22 compounds that both disrupted zebrafish vascular development and murine endothelial in vitro tubulogenesis. Putative molecular targets for the VDCs were predicted using EPA's Toxicological Prioritization Index tool and a VDC signature based on a proposed adverse outcome pathway for developmental vascular toxicity. In conclusion, our screening approach identified 22 novel VDCs, some of which were active at nanomolar concentrations.

Computation of breast ptosis from 3D surface scans of the female torso.

Stereophotography is now finding a niche in clinical breast surgery, and several methods for quantitatively measuring breast morphology from 3D surface images have been developed. Breast ptosis (sagging of the breast), which refers to the extent by which the nipple is lower than the inframammary fold (the contour along which the inferior part of the breast attaches to the chest wall), is an important morphological parameter that is frequently used for assessing the outcome of breast surgery. This study presents a novel algorithm that utilizes three-dimensional (3D) features such as surface curvature and orientation for the assessment of breast ptosis from 3D scans of the female torso. The performance of the computational approach proposed was compared against the consensus of manual ptosis ratings by nine plastic surgeons, and that of current 2D photogrammetric methods. Compared to the 2D methods, the average accuracy for 3D features was ~13% higher, with an increase in precision, recall, and F-score of 37%, 29%, and 33%, respectively. The computational approach proposed provides an improved and unbiased objective method for rating ptosis when compared to qualitative visualization by observers, and distance based 2D photogrammetry approaches.

Inferior Breast-Chest Contour Detection in 3-D Images of the Female Torso.

Stereophotogrammetry is finding increased use in clinical breast surgery, both for breast reconstruction after oncological procedures and cosmetic augmentation and reduction. The ability to visualize and quantify morphological features of the breast facilitates pre-operative planning and post-operative outcome assessment. The contour outlining the lower half of the breast is important for the quantitative assessment of breast aesthetics. Based on this inferior breast contour, relevant morphological measures, such as breast symmetry, volume, and ptosis, can be determined. In this paper, we present an approach for automatically detecting the inferior contour of the breast in 3D images. Our approach employs surface curvature analysis and is able to detect the breast contour with high accuracy, achieving an average error of 1.64 mm and a dice coefficient in the range of 0.72-0.87 when compared with the manually annotated contour (ground truth). In addition, the detected contour is used to facilitate the detection of the lowest visible point on the breast, which is an important landmark for breast morphometric analysis.