Oncology Commons^™

Lpa, Hgf, And Egf Utilize Distinct Combinations Of Signaling Pathways To Promote Migration And Invasion Of Mda-Mb-231 Breast Carcinoma Cells, Susan M. W. Harrison, Teresa Knifley, Min Chen, Kathleen L. O'Connor

Markey Cancer Center Faculty Publications

BACKGROUND: Various pathways impinge on the actin-myosin pathway to facilitate cell migration and invasion including members of the Rho family of small GTPases and MAPK. However, the signaling components that are considered important for these processes vary substantially within the literature with certain pathways being favored. These distinctions in signaling pathways utilized are often attributed to differences in cell type or physiological conditions; however, these attributes have not been systematically assessed.

METHODS: To address this question, we analyzed the migration and invasion of MDA-MB-231 breast carcinoma cell line in response to various stimuli including lysophosphatidic acid (LPA), hepatocyte growth factor …

Measuring The Sixth Vital Sign: A Descriptive Analysis Of Distress In Individuals With Head And Neck Cancer And Their Caregivers, Catherine C. Bornbaum

Electronic Thesis and Dissertation Repository

Introduction: Distress has become so problematic in oncology that it has been recognized as the “sixth vital sign” implying that distress monitoring should occur as routinely as the monitoring of one’s temperature or blood pressure. The research reported herein investigated the impact of head and neck cancer on levels of distress, commonly reported problems, and perceptions of quality of life in individuals with head and neck cancer and their caregivers.

Method: Two distinct studies were conducted; the first explored the patient experience of distress and quality of life while the second assessed the caregiver experience of these same constructs. A …

Igf-Ir Promotes Prostate Cancer Growth By Stabilizing Α5Β1 Integrin Protein Levels., Aejaz Sayeed, Carmine Fedele, Marco Trerotola, Kirat K Ganguly, Lucia R Languino

Department of Cancer Biology Faculty Papers

Dynamic crosstalk between growth factor receptors, cell adhesion molecules and extracellular matrix is essential for cancer cell migration and invasion. Integrins are transmembrane receptors that bind extracellular matrix proteins and enable cell adhesion and cytoskeletal organization. They also mediate signal transduction to regulate cell proliferation and survival. The type 1 insulin-like growth factor receptor (IGF-IR) mediates tumor cell growth, adhesion and inhibition of apoptosis in several types of cancer. We have previously demonstrated that β1 integrins regulate anchorage-independent growth of prostate cancer (PrCa) cells by regulating IGF-IR expression and androgen receptor-mediated transcriptional functions. Furthermore, we have recently reported that IGF-IR …

Recommendations For High-Priority Research On Cancer-Related Fatigue In Children And Adults., Andrea M. Barsevick, Michael R. Irwin, Pamela Hinds, Andrew Miller, Ann Berger, Paul Jacobsen, Sonia Ancoli-Israel, Bryce B Reeve, Karen Mustian, Ann O'Mara, Jin-Shei Lai, Michael Fisch, David Cella

Department of Medical Oncology Faculty Papers

Over the past decades, some scientific progress has been made in understanding and treating cancer-related fatigue (CRF). However, three major problems have limited further progress: lack of agreement about measurement, inadequate understanding of the underlying biology, and problems in the conduct of clinical trials for CRF. This commentary reports the recommendations of a National Cancer Institute Clinical Trials Planning Meeting and an ongoing National Cancer Institute working group to address these problems so that high-priority research and clinical trials can be conducted to advance the science of CRF and its treatment. Recommendations to address measurement issues included revising the current …

Altered Cholesterol Metabolism In Human Cancers Unraveled By Label-Free Spectroscopic Imaging, Shuhua Yue

Open Access Dissertations

Despite tremendous scientific achievements, cancer remains the second leading cause of death in the United States. Metabolic reprogramming has been increasingly recognized as a core hallmark of cancer. My dissertation work identified novel diagnostic markers and therapeutic targets for human cancers through the study of cholesterol in cancer cells.

Enabled by label-free Raman spectromicroscopy, we performed the first quantitative analysis of lipogenesis at single cell level in human patient cancerous tissues. Our imaging data revealed an unexpected, aberrant accumulation of esterified cholesterol in lipid droplets of high-grade prostate cancer and metastases, but not in normal prostate, benign prostatic hyperplasia, or …

Synthesis And Antiproliferative Activities Of Quebecol And Its Analogs, Kasiviswanadharaju Pericherla, Amir Nasrolahi Shirazi, V. Kameshwara Rao, Rakesh Tiwari, Nicholas Dasilva, Kellen Mccaffrey, Yousef A. Beni, Antonio González- Sarrías, Navindra P. Seeram, Keykavous Parang, Anil Kumar

Pharmacy Faculty Articles and Research

Simple and efficient synthesis of quebecol and a number of its analogs was accomplished in five steps. The synthesized compounds were evaluated for antiproliferative activities against human cervix adenocarcinoma (HeLa), human ovarian carcinoma (SK-OV-3), human colon carcinoma (HT-29), and human breast adenocarcinoma (MCF-7) cancer cell lines. Among all the compounds, 7c, 7d, 7f, and 8f exhibited antiproliferative activities against four tested cell lines with inhibition over 80% at 75 mu M after 72 h, whereas, compound 7b and 7g were more selective towards MCF-7 cell line. The IC50 values for compounds 7c, 7d, and 7f were 85.1 mu M, 78.7 …

Oncolog, Volume 58, Number 10, October 2013, Bryan Tutt, Sunni Hosemann, C. Wilcox

OncoLog MD Anderson's Report to Physicians (All issues)

• Sentinel Lymph Node Dissection After Neoadjuvant Chemotherapy for Node-Positive Breast Cancer May replace Axillary Dissection for Some Patients: Sentinel lymph node dissection has become a standard staging tool for breast cancer patients with no clinical eveidence of disease in the lymph nodes
• INBRIEF: miR-200 MicroRNAs Slow Tumor Growth
• Compass: Resectable or Borderline Resectable Pancreatic Adenocarcinoma: Initial Treatment Options-Neoadjuvent chemoradiation may benefit patients with resectable disease
• HOUSE CALL: Hereditary Cancers: Genetic screening can give valuable information

I Don't Want To Save Second Base, Chelsea E. Broe

SURGE

Tomorrow kicks off Breast Cancer Awareness Month, and I have one request: This October, let’s not save second base.

I know, I know, you probably think this month is a good thing. If not for all of those T-shirts demanding every female-bodied person to feel their boobies, or the bracelets simply proclaiming “I love boobies,” people with breasts might forget that they even have them, or at the very least might start to think that their breasts are their own business. But the female body seems to be an object owned by the public, so we must always be reminded …

Resistance Of Human Cytomegalovirus To Cyclopropavir Maps To A Base Pair Deletion In The Open Reading Frame Of Ul97, Brian G. Gentry, Laura E. Vollmer, Ellie D. Hall, Katherine Z. Borysko, Jiri Zemlicka, Jeremy P. Kamil, John C. Drach

Oncology Faculty Publications

Human cytomegalovirus (HCMV) is a widespread pathogen in the human population, affecting many immunologically immature and immunocompromised patients, and can result in severe complications, such as interstitial pneumonia and mental retardation. Current chemotherapies for the treatment of HCMV infections include ganciclovir (GCV), foscarnet, and cidofovir. However, the high incidences of adverse effects (neutropenia and nephrotoxicity) limit the use of these drugs. Cyclopropavir (CPV), a guanosine nucleoside analog, is 10-fold more active against HCMV than GCV (50% effective concentrations [EC₅₀s] = 0.46 and 4.1 μM, respectively). We hypothesize that the mechanism of action of CPV is similar to that …

Oncolog, Volume 58, Number 09, September 2013, Jill Deisigne, Markeda Wade

OncoLog MD Anderson's Report to Physicians (All issues)

• Advances in Neuroimaging Help Unveil the Mechanisms of Chemobrain: Chemobrain, a catchall term for the cognitive dysfunction and neurophysiological changes produced bt chemotherapy toxicity, is an important quality-of-life issue for cancer survivors
• Opioid Rotation Can Improve Pain Control and Reduce Neurotoxicity for Cancer Patients: Opioids are a main stay in the relief of pain-especially the chronic cancer-related pain often encountered in the palliative care setting. However, extended opioid use can result in tolerance, dependence, and opioid-induced neurotoxicity
• INBRIEF: Researchers Identity Inhibitor of Cancer-Promoting Protein SKp2/ Change in CA-125 Level Over Time Shows Promise for Ovarian cancer Screening
• HOUSE CALL: Myths …

Clinical And Dosimetric Implications Of Air Gaps Between Bolus And Skin Surface During Radiation Therapy, Yousaf Khan, J Eduardo Villarreal-Barajas, Mona Udowicz, Richie Sinha, Wazir Muhammad, Ahmed N. Abbasi, Amjad Hussain

Department of Radiation Oncology

Purpose: The main objective of the study was to evaluate the effect of air gaps of 0 - 5.0 cm between bolus and skin for 1.0 cm Superflab bolus on surface dose (DSurf) and depth of maximum dose (dmax) in solid water and Rando® phantoms.

Methods: In this work, the effects of bolus to surface distance on DSurf and variation in dmax were analyzed in a solid water phantom and in an anthropomorphic Rando® phantom for different field sizes, using Gafchromic® EBT films and farmer chamber.

Results: For field sizes of 5 × 5 cm2 the DSurf …

Cyclin D1 Determines Estrogen Signaling In The Mammary Gland In Vivo., Mathew C Casimiro, Chenguang Wang, Z Li, Gabriele Disante, Nicole E Willmart, Sankar Addya, Lei Chen, Yang Liu, Michael P. Lisanti, Richard Pestell

Department of Cancer Biology Faculty Papers

The CCND1 gene, which is frequently overexpressed in cancers, encodes the regulatory subunit of a holoenzyme that phosphorylates the retinoblastoma protein. Although it is known that cyclin D1 regulates estrogen receptor (ER)α transactivation using heterologous reporter systems, the in vivo biological significance of cyclin D1 to estrogen-dependent signaling, and the molecular mechanisms by which cyclin D1 is involved, are yet to be elucidated. Herein, genome-wide expression profiling conducted of 17β-estradiol-treated castrated virgin mice deleted of the Ccnd1 gene demonstrated that cyclin D1 determines estrogen-dependent gene expression for 88% of estrogen-responsive genes in vivo. In addition, expression profiling of 17β-estradiol-stimulated cyclin …

Chronic Cadmium Exposure Stimulates Sdf-1 Expression In An Erα Dependent Manner, Esmeralda Ponce, Natalie B. Aquino, Maggie Louie

Collected Faculty and Staff Scholarship

Cadmium is an omnipotent environmental contaminant associated with the development of breast cancer. Studies suggest that cadmium functions as an endocrine disruptor, mimicking the actions of estrogen in breast cancer cells and activating the receptor to promote cell growth. Although acute cadmium exposure is known to promote estrogen receptor-mediated gene expression associated with growth, the consequence of chronic cadmium exposure is unclear. Since heavy metals are known to bioaccumulate, it is necessary to understand the effects of prolonged cadmium exposure. This study aims to investigate the effects of chronic cadmium exposure on breast cancer progression. A MCF7 breast cancer cell …

The Impact Of Lateral Electron Disequilibrium On Stereotactic Body Radiation Therapy Of Lung Cancer, Brandon Disher

Electronic Thesis and Dissertation Repository

Stereotactic Body Radiation Therapy (SBRT) is an effective treatment option for patients with inoperable early-stage lung cancer. SBRT uses online image-guidance technology [e.g. cone-beam CT (CBCT)] to focus small-fields of high energy x-rays onto a tumour to deliver ablative levels of radiation dose (e.g. 54 Gy) in a few treatment fractions (e.g. 3). For the combination of these treatment parameters and a low density lung, lateral electron disequilibrium (LED) can potentially occur, reducing lung and tumour doses. The goal of this thesis was to determine the impact of LED on stereotactic body radiation therapy for lung cancer.

The effect of …

A Survey Of The Barriers Associated With Academic-Based Cancer Research Commercialization, Nathan L. Vanderford, L. Todd Weiss, Heidi L. Weiss

Markey Cancer Center Faculty Publications

Commercialization within the academic setting is associated with many challenges and barriers. Previous studies investigating these challenges/barriers have, in general, broadly focused on multiple disciplines and, oftentimes, several institutions simultaneously. The goal of the study presented here was to analyze a range of barriers that may be broadly associated with commercializing academic-based cancer research. This goal was addressed via a study of the barriers associated with cancer research commercialization at the University of Kentucky (UK). To this end, a research instrument in the form of an electronic survey was developed. General demographic information was collected on study participants and two …

Oncogenes And Inflammation Rewire Host Energy Metabolism In The Tumor Microenvironment: Ras And Nfκb Target Stromal Mct4., Ubaldo E. Martinez-Outshoorn, Md, Joseph Curry, Md, Ying-Hui Ko, Md, Zhao Lin, Md, Madalina Tuluc, Md, David Cognetti, Md, Ruth Birbe, Md, Edmund A. Pribitkin, Md, Alessandro Bombonati, Richard Pestell, Md, Anthony Howell, Federica Sotgia, Michael P. Lisanti, Md

Kimmel Cancer Center Faculty Papers

Here, we developed a model system to evaluate the metabolic effects of oncogene(s) on the host microenvironment. A matched set of "normal" and oncogenically transformed epithelial cell lines were co-cultured with human fibroblasts, to determine the "bystander" effects of oncogenes on stromal cells. ROS production and glucose uptake were measured by FACS analysis. In addition, expression of a panel of metabolic protein biomarkers (Caveolin-1, MCT1, and MCT4) was analyzed in parallel. Interestingly, oncogene activation in cancer cells was sufficient to induce the metabolic reprogramming of cancer-associated fibroblasts toward glycolysis, via oxidative stress. Evidence for "metabolic symbiosis" between oxidative cancer cells …

Adoptive T Cell Therapy For Metastatic Colorectal Cancer, Shauna Ebanks

Summer Training Program in Cancer Immunotherapy

Colorectal cancer is the second leading cause of cancer-related death in US (1). Most of the mortality reflects local, regional and distant metastatic tumors. Cancers that are confined within the wall of the colon are often curable with surgery while tumor cells that have spread into other organs like liver and lung has low curable possibility, even though the chemotherapies and monoclonal antibodies can extend the person's life and improve quality of life(2, 3). Surgery is the main treatment for primary colorectal tumors but recurrence can happen and develop into metastasis. In addition, colorectal cancer …

Unraveling The Mechanism Of 2,4-Dinitrofluorobenzene-Associated Cell Death, Franklin Thelmo

Summer Training Program in Cancer Immunotherapy

The mechanism by which the hapten 2,4-dinitrophenyl (DNP) mediates cell death is unclear according to current literature. To determine how DNP mediates death in the B16F10 murine melanoma cell line in vitro, cells were modified with 2,4-dinitro-1-fluorobenzene (DNFB). Following modification, cell lysates were collected and analyzed via Western blottings with known apoptotic or necrotic markers. Our results indicated that DNFB treated cells do not express the apoptotic marker Cleaved Caspase-3 and have no change in levels of the necrotic marker Cyclophilin A. We conclude that DNFB is not inducing apoptosis in B16F10 cells. Future research will further examine the …

Coupling S100a4 To Rhotekin Alters Rho Signaling Output In Breast Cancer Cells, Min Chen, Anne R. Bresnick, Kathleen L. O'Connor

Markey Cancer Center Faculty Publications

Rho signaling is increasingly recognized to contribute to invasion and metastasis. In this study, we discovered that metastasis-associated protein S100A4 interacts with the Rho-binding domain (RBD) of Rhotekin, thus connecting S100A4 to the Rho pathway. Glutathione S-transferase pull-down and immunoprecipitation assays demonstrated that S100A4 specifically and directly binds to Rhotekin RBD, but not the other Rho effector RBDs. S100A4 binding to Rhotekin is calcium-dependent and uses residues distinct from those bound by active Rho. Interestingly, we found that S100A4 and Rhotekin can form a complex with active RhoA. Using RNA interference, we determined that suppression of both S100A4 and …

Metastatic Castration-Resistant Prostate Cancer: Critical Review Of Enzalutamide, Joelle El-Amm, Nihar Patel, Ashley Freeman, Jeanny B. Aragon-Ching

Medicine Faculty Publications

Enzalutamide, previously known as MDV300, is an oral, second-generation androgen receptor (AR) signaling inhibitor or antagonist that was approved by the Food and Drug Administration in 2012 for the treatment of metastatic castrate-resistant prostate cancer (mCRPC) postdocetaxel. Preclinical studies have demonstrated impressive affinity to the AR compared to the first-generation AR inhibitors. The landmark Phase III AFFIRM trial demonstrated improved overall survival benefit compared to placebo in addition to improvement in all tested parameters. Enzalutamide is currently being studied in several trials prechemotherapy and in earlier settings of prostate cancer. This review will discuss the mechanism of action of enzalutamide, …

Cancer Metabolism: New Validated Targets For Drug Discovery., Federica Sotgia, Ubaldo E. Martinez-Outshoorn, Md, Michael P. Lisanti

Kimmel Cancer Center Faculty Papers

Recent studies in cancer metabolism directly implicate catabolic fibroblasts as a new rich source of i) energy and ii) biomass, for the growth and survival of anabolic cancer cells. Conversely, anabolic cancer cells upregulate oxidative mitochondrial metabolism, to take advantage of the abundant fibroblast fuel supply. This simple model of "metabolic-symbiosis" has now been independently validated in several different types of human cancers, including breast, ovarian, and prostate tumors. Biomarkers of metabolic-symbiosis are excellent predictors of tumor recurrence, metastasis, and drug resistance, as well as poor patient survival. New pre-clinical models of metabolic-symbiosis have been generated and they genetically validate …

Evaluating The Utility Of Clinical Criteria For The Identification Of Lynch Syndrome Among Endometrial Cancer Patients, Amanda S. Bruegl

Dissertations & Theses (Open Access)

Background: Lynch Syndrome (LS) is a familial cancer syndrome with a high prevalence of colorectal and endometrial carcinomas among affected family members. Clinical criteria, developed from information obtained from familial colorectal cancer registries, have been generated to identify individuals at elevated risk for having LS. In 2007, the Society of Gynecologic Oncology (SGO) codified criteria to assist in identifying women presenting with gynecologic cancers at elevated risk for having LS. These criteria have not been validated in a population-based setting.

Materials and Methods: We retrospectively identified 412, unselected endometrial cancer cases. Clinical and pathologic information were obtained from the electronic …

A Statement On Vemurafenib-Resistant Melanoma., Edward J Hartsough, A E Aplin

Department of Cancer Biology Faculty Papers

Despite recent advancements in the treatment of late-stage mutant BRAF (V600E/K) melanomas, a major hurdle continues to be acquired resistance to BRAF inhibitors such as vemurafenib. The mechanisms for resistance have proven to be heterogeneous, emphasizing the need to use broad therapeutic approaches. In this issue, the study "Stat3-targeted therapies overcome the acquired resistance to vemurafenib in melanomas" by Liu et al. proposes that signal transducer and activator of transcription 3 (STAT3)-paired box 3 (PAX3) signaling may be a mechanism that is used by melanomas to resist RAF inhibitors.

Coping With Pediatric Cancer: Conversational Methods Utilized By Parents And Children When Dealing With Pediatric Cancer, Chelsi Morgan Walls

UNLV Theses, Dissertations, Professional Papers, and Capstones

This study analyzed how people perceived parents should communicate with their child regarding pediatric cancer treatments. When dealing with pediatric cancer, it is vital that parents and their child communicate about the illness in order to effectively cope with the cancer. Using Uncertainty Management Theory, appraisals, inferences, and illusions, are examined in this study to discover how much affect-management and buffering would be used to manage the illness. Under UMT, the coping mechanisms of affect-management (i.e., religious coping and behavioral disengagement), and buffering (avoidance and child distraction) depend upon how individuals appraise the uncertain situation (positive vs. negative), the inferences …

Oncolog, Volume 58, Number 08, August 2013, Bryan Tutt, Sarah Bronson

OncoLog MD Anderson's Report to Physicians (All issues)

• Specialized Care Improves Lives of patients With Head and Neck Lymphedema: A unique program is producing dramatic results for patients with head and neck lymphedema, a side effect of cancer treatment that can interfere with patient's ability to speak, swallow, or breath
• Targeted Cancer Therapies May Help Overcome Resistance to Radiation Therapy: Some cancer cells are resilient enough to withstand and recover from the damage to their DNA caused by radiation therapy. But recent studies have shown that adding molecularly targeted agents to radiation therapy can prevent the repair of this radiation-induced damage and therapy improve the treatment response of …

Efficacy Of Gold Silica Nanoshells And Gold Nanorods For Photothermal Therapy Of Human Glioma Spheroids, Suyog Jung Chhetri

UNLV Theses, Dissertations, Professional Papers, and Capstones

Gold-based nanoparticles including gold-silica nano-spheres and gold nano-rods have been investigated for a number of therapeutic and diagnostic applications. The ability of these nanoparticles to convert light into heat energy makes them particularly appealing for photothermal therapy in which cancer cells are destroyed via light-induced heat generation. The overall objective of the study is to compare the efficacy of gold-silica nano-spheres and gold nano-rods in an in vitro system consisting of human brain tumor (glioma) spheroids.

Delivery of the nanoparticles to the spheroids was accomplished using murine macrophages. Nanoparticles (spheres or rods) were incubated with macrophages for 24 hours. Thereafter, …

Failure Patterns In Resected Pancreas Adenocarcinoma: Lack Of Predicted Benefit To Smad4 Expression., Jordan M. Winter, Laura H. Tang, David S. Klimstra, Weiguo Liu, Irena Linkov, Murray F. Brennan, Michael I. DʼAngelica, Ronald P. Dematteo, Yuman Fong, William R. Jarnagin, Eileen M. OʼReilly, Peter J. Allen

Department of Surgery Faculty Papers

OBJECTIVE: To determine whether SMAD4 expression is associated with recurrence pattern after resection for pancreatic ductal adenocarcinoma (PDA).

BACKGROUND: SMAD4 expression status has been reported to be associated with patterns of failure in PDA, but studies have not examined recurrence patterns after resection.

METHODS: A tissue microarray was constructed including 127 patients with resected PDA and either short-term (<12 >months) or long-term (>30 months) survival. SMAD4 expression was evaluated by immunohistochemistry and categorized as present or lost in tumor cells. Conventional pathologic features (lymph node metastases, positive resection margin, poor grade, and tumor size) were recorded, and disease-specific outcomes …

Health Literacy And Cancer Prevention: It’S Not What You Say It’S What They Hear, Kathleen M. Mazor, Douglas W. Roblin, Andrew E. Williams, Paul J. K. Han, Mary E. Costanza, Sarah L. Cutrona, Terry S. Field, Bridget Gaglio, Joann L. Wagner, Brandi E. Robinson, Vinutha Vijayadeva

Mary E. Costanza

Background: A growing body of literature documents the relationship between health literacy and important health behaviors and outcomes. Most research to date has focused on print literacy–few studies have examined literacy with respect to spoken information (“spoken health literacy”). We sought to examine the extent to which responses to physician advice about cancer prevention and screening were associated with spoken health literacy.

Methods: Participants listened to 3 simulated physician-patient discussions addressing: 1) Prostate Specific Antigen (PSA) testing; 2) tamoxifen for breast cancer prevention; and 3) colorectal cancer (CRC) screening. The physician provided information on risks and benefits but did not …

Outcomes Of Peginterferon Alfa-2a And Ribavirin Combination Therapy In A Resident-Initiated, Multidisciplinary, Hepatitis C Clinic, Nicole M. Agostino Do, Erini Vasiliadis Do, K Nadeem Ahmed Md, Suzanne J. Templer Do, Edward R. Norris Md, Fapa, Fapm, Charles M. Brooks Md, Eric J. Gertner Md, Mph, Joseph L. Yozviak Do, Facp

Joseph L Yozviak DO, FACP

No abstract provided.

Combination Of Sirna-Directed Gene Silencing With Cisplatin Reverses Drug Resistance In Human Non-Small Cell Lung Cancer, Shanthi Ganesh, Arun K. Iyer, Jan Weller, David V. Morrissey, Mansoor M. Amiji

Pharmaceutical Sciences Faculty Publications

One of the most challenging aspects of lung cancer therapy is the rapid acquisition of multidrug-resistant (MDR) phenotype. One effective approach would be to identify and downregulate resistance-causing genes in tumors using small interfering RNAs (siRNAs) to increase the sensitivity of tumor cells to chemotherapeutic challenge. After identifying the overexpressed resistance-related antiapoptotic genes (survivin and bcl-2) in cisplatin-resistant cells, the siRNA sequences were designed and screened to select the most efficacious candidates. Modifications were introduced in them to minimize off-target effects. Subsequently, the combination of siRNA and cisplatin that gave the maximum synergy was identified in resistant cells. We then …