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Week Of January 7, 2019, New York Medical College

InTouch

• Center for Disaster Medicine Hosts Mass Casualty Triage Training at Local Hospital
• Touro College of Dental Medicine Names Howard Israel, D.D.S., Vice Dean for Academic Affairs
• Touro College of Dental Medicine and the Westchester Institute for Human Development Sign Academic Affiliation Agreement

Intranasal Peptide-Based Fpva-Klh Conjugate Vaccine Protects Mice From Pseudomonas Aeruginosa Acute Murine Pneumonia, Emel Sen-Kilic, Catherine B. Blackwood, Dylan T. Boehm, Wiliam T. Witt, Aaron C. Malkowski, Justin R. Bevere, Ting Y. Wong, Jesse M. Hall, Shelby D. Bradford, Melinda E. Varney, Fredrick Heath Damron, Mariette Barbier

Faculty & Staff Scholarship

Pseudomonas aeruginosa is an opportunistic pathogen causing acute and chronic respiratory infections associated with morbidity and mortality, especially in patients with cystic fibrosis. Vaccination against P. aeruginosa before colonization may be a solution against these infections and improve the quality of life of at-risk patients. To develop a vaccine against P. aeruginosa, we formulated a novel peptide-based P. aeruginosa subunit vaccine based on the extracellular regions of one of its major siderophore receptors, FpvA. We evaluated the effectiveness and immunogenicity of the FpvA peptides conjugated to keyhole limpet hemocyanin (KLH) with the adjuvant curdlan in a murine vaccination and challenge …

Mechanistic Study Of Diatrizoic Acid Induced Proximal Tubule Cytotoxicity, Dakota Blake Ward

Theses, Dissertations and Capstones

Radiocontrast media (RCM) are necessary for many diagnostic procedures such as arteriography, percutaneous coronary intervention (PCI), and computed tomography. Contrast-induced acute kidney injury (CI-AKI) is the third most common cause of hospital-associated kidney damage accounting for 10-25% of cases worldwide. The mechanisms of contrast-induced renal impairment are not entirely known, but diminished renal hemodynamics, inflammatory responses, and direct cytotoxicity have been hypothesized. The purpose of this study was to investigate the mechanisms of direct cytotoxicity observed in HK-2 cells following treatment with diatrizoic acid (DA) and to determine the source of this damage. Mitochondrial function, endoplasmic reticulum (ER) stress, oxidative …

Janus Kinase 1 Is Required For Transcriptional Reprograming Of Murine Astrocytes In Response To Endoplasmic Reticulum Stress, Savannah G. Sims, Gordon P. Meares

Faculty & Staff Scholarship

Neurodegenerative diseases are associated with the accumulation of misfolded proteins in the endoplasmic reticulum (ER), leading to ER stress. To adapt, cells initiate the unfolded protein response (UPR). However, severe or unresolved UPR activation leads to cell death and inflammation. The UPR is initiated, in part, by the transER membrane kinase PKR-like ER kinase (PERK). Recent evidence indicates ER stress and inflammation are linked, and we have shown that this involves PERKdependent signaling via Janus Kinase (JAK) 1. This signaling provokes the production of soluble inflammatory mediators such as interleukin-6 (IL-6) and chemokine C-C motif ligand 2 (CCL2). We, therefore, …

Endoplasmic Reticulum Stress Differentially Modulates The Il-6 Family Of Cytokines In Murine Astrocytes And Macrophages, Cristina L. Sanchez, Savnnah G. Sims, John D. Nowery, Gordon P. Meares

Faculty & Staff Scholarship

In many diseases, misfolded proteins accumulate within the endoplasmic reticulum (ER), leading to ER stress. In response, the cell initiates the unfolded protein response (UPR) to reestablish homeostasis. Additionally, in response to ER stress, various cell types mount an inflammatory response involving interleukin (IL)-6. While IL-6 has been widely studied, the impact of ER stress on other members of the IL-6 cytokine family, including oncostatin (OSM), IL-11, ciliary neurotrophic factor (CNTF), and leukemia inhibitor factor (LIF) remains to be elucidated. Here, we have examined the expression of the IL-6 family cytokines in response to pharmacologically-induced ER stress in astrocytes and …

A Role For Regulator Of G Protein Signaling-12 (Rgs12) In The Balance Between Myoblast Proliferation And Differentiation, Adam B. Schroer, Junaith S. Mohamed, Melinda D. Wilard, Vincent Setola, Emily Oesteich, David P. Siderovski

Faculty & Staff Scholarship

• Regulators of G Protein Signaling (RGS proteins) inhibit G protein-coupled receptor (GPCR) signaling by accelerating the GTP hydrolysis rate of activated Gα subunits. Some RGS proteins exert additional signal modulatory functions, and RGS12 is one such protein, with five additional, functional domains: a PDZ domain, a phosphotyrosine-binding domain, two Ras-binding domains, and a Gα·GDP-binding GoLoco motif. RGS12 expression is temporospatially regulated in developing mouse embryos, with notable expression in somites and developing skeletal muscle. We therefore examined whether RGS12 is involved in the skeletal muscle myogenic program. In the adult mouse, RGS12 is expressed in the tibialis anterior (TA) muscle, …

In Vivo Gene Essentiality And Metabolism In Bordetella Pertussis, Laura A. Gonyar, Patrick E. Gelbach, Dennis G. Mcduffe, Alexander F. Koeppel, Qing Chen, Gloria Lee, Louise M. Temple, Scott Stibitz, Erik L. Hewlwtt, Jason A. Papin, F. Heath Damron, Joshua C. Eby

Faculty & Staff Scholarship

Bordetella pertussis is the causative agent of whooping cough, a serious respiratory illness affecting children and adults, associated with prolonged cough and potential mortality. Whooping cough has reemerged in recent years, emphasizing a need for increased knowledge of basic mechanisms of B. pertussis growth and pathogenicity. While previous studies have provided insight into in vitro gene essentiality of this organism, very little is known about in vivo gene essentiality, a critical gap in knowledge, since B. pertussis has no previously identified environmental reservoir and is isolated from human respiratory tract samples. We hypothesize that the metabolic capabilities of B. pertussis …

Caspase-11 Mediates Neutrophil Chemotaxis And Extracellular Trap Formation During Acute Gouty Arthritis Through Alteration Of Cofilin Phosphorylation, Kyle Caution, Nicholas Young, Frank Robledo-Avila, Kathrin Krause, Arwa Abu Khweek, Kaitlin Hamilton, Asmaa Badr, Anup Vaidya, Kylene Daily, Hawin Gosu, Midhun N. K. Anne, Mostafa Eltobgy, Duaa Dakhlallah, Sudha Argwal, Shady Estfanous, Xiaoli Zhang, Santiago Partida-Sanchez, Mikhail A. Gavrilin, Wael N. Jarjour, Amal O. Amer

Faculty & Staff Scholarship

Gout is characterized by attacks of arthritis with hyperuricemia and monosodium urate (MSU) crystal-induced inflammation within joints. Innate immune responses are the primary drivers for tissue destruction and inflammation in gout. MSU crystals engage the Nlrp3 inflammasome, leading to the activation of caspase-1 and production of IL-1β and IL-18 within gout-affected joints, promoting the influx of neutrophils and monocytes. Here, we show that caspase-11−/− mice and their derived macrophages produce significantly reduced levels of gout-specific cytokines including IL-1β, TNFα, IL-6, and KC, while others like IFNγ and IL-12p70 are not altered. IL-1β induces the expression of caspase-11 in an IL-1 …

Defining Inflammatory Cell States In Rheumatoid Arthritis Joint Synovial Tissues By Integrating Single-Cell Transcriptomics And Mass Cytometry, F. Zhang, K. Wei, K. Slowikowski, C. Y. Fonseka, A. Rao, Kelly, S. M. Goodman, D. Tabechian, P. K. Gregersen, S. Raychaudhuri, +55 Additional Authors

Journal Articles

© 2019, The Author(s), under exclusive licence to Springer Nature America, Inc. To define the cell populations that drive joint inflammation in rheumatoid arthritis (RA), we applied single-cell RNA sequencing (scRNA-seq), mass cytometry, bulk RNA sequencing (RNA-seq) and flow cytometry to T cells, B cells, monocytes, and fibroblasts from 51 samples of synovial tissue from patients with RA or osteoarthritis (OA). Utilizing an integrated strategy based on canonical correlation analysis of 5,265 scRNA-seq profiles, we identified 18 unique cell populations. Combining mass cytometry and transcriptomics revealed cell states expanded in RA synovia: THY1(CD90) + HLA-DRA hi sublining fibroblasts, IL1B + …

Rnai Transfection Optimized In Primary Naïve B Cells For The Targeted Analysis Of Human Plasma Cell Differentiation, T. Shih, S. De, B. J. Barnes

Journal Articles

Upon antigen recognition, naïve B cells undergo rapid proliferation followed by differentiation to specialized antibody secreting cells (ASCs), called plasma cells. Increased circulating plasma cells are reported in patients with B cell-associated malignancies, chronic graft-vs.-host disease, and autoimmune disorders. Our aim was to optimize an RNAi-based method that efficiently and reproducibly knocks-down genes of interest in human primary peripheral B cells for the targeted analysis of ASC differentiation. The unique contributions of transcriptional diversity in species-specific regulatory networks and the mechanisms of gene function need to be approached directly in human B cells with tools to hone our basic inferences …

Identification Of Extracellular Dna-Binding Proteins In The Biofilm Matrix., Jeffrey S. Kavanaugh, Caralyn E. Flack, Jessica Lister, Erica B. Ricker, Carolyn B. Ibberson, Christian Jenul, Derek E. Moormeier, Elizabeth A. Delmain, Kenneth W. Bayles, Alexander R. Horswill

Journal Articles: Pathology and Microbiology

We developed a new approach that couples Southwestern blotting and mass spectrometry to discover proteins that bind extracellular DNA (eDNA) in bacterial biofilms. Using Staphylococcus aureus as a model pathogen, we identified proteins with known DNA-binding activity and uncovered a series of lipoproteins with previously unrecognized DNA-binding activity. We demonstrated that expression of these lipoproteins results in an eDNA-dependent biofilm enhancement. Additionally, we found that while deletion of lipoproteins had a minimal impact on biofilm accumulation, these lipoprotein mutations increased biofilm porosity, suggesting that lipoproteins and their associated interactions contribute to biofilm structure. For one of the lipoproteins, SaeP, we …

Naktide Targeted To Adipocytes Ameliorates Western Diet Induced Obesity, Rebecca D. Pratt

Theses, Dissertations and Capstones

Obesity has become a worldwide epidemic and is a major risk factor for metabolic syndrome. It is believed that cellular oxidant stress plays a key role in both the development and maintenance of obesity as well as its associated comorbidities such as diabetes, cardiovascular disease, and nonalcoholic steatohepatitis (NASH). We have previously reported that systemic administration of pNaKtide, which targets the Na/K-ATPase oxidant amplification loop was able to decrease oxidative stress and adiposity in mice fed a high fat and fructose supplemented western diet (WD). As adipocytes are believed to play an active role in the development of obesity and …

Structural And Functional Determinants Of Rodent And Human Surfactant Protein A: A Synthesis Of Binding And Computational Data, Armen Nalian, Todd M. Umstead, Ching-Hui Yang, Patricia Silveyra, Neal J. Thomas, Joanna Floros, Francis X. Mccormack, Zissis C. Chroneos

Faculty Publications

Surfactant protein A (SP-A) provides surfactant stability, first line host defense, and lung homeostasis by binding surfactant phospholipids, pathogens, alveolar macrophages (AMs), and epithelial cells. Non-primates express one SP-A protein whereas humans express two: SP-A1 and SP-A2 with core intra- and inter-species differences in the collagen-like domain. Here, we used macrophages and solid phase binding assays to discern structural correlates of rat (r) and human (h) SP-A function. Binding assays using recombinant rSP-A expressed in insect cells showed that lack of proline hydroxylation, truncations of amino-terminal oligomerization domains, and site-directed serine (S) or alanine (A) mutagenesis of cysteine 6 (C6S), …

Modernizing Biomedical Training: Replacing Live Animal Laboratories With Human Simulation, John Pawlowski, David Feinstein, Marie L. Crandall, Shalin Gala

Biomedicine and Animal Models in Research Collection

This chapter reviews the global trend towards a modernization of biomedical education in favor of simulation-based training methods, which studies confirm improve student learning and transference of applied skills to clinical practice, reduce laboratory costs, and spare animals from harmful procedures.

Dna Damage And Cytokine Production In Non-Target Irradiated Lymphocytes, Jane Bryant, Laura Shields, Christopher Hynes, Orla L. Howe, Brendan Mcclean, Fiona Lyng

Articles

In advanced radiotherapy, treatment of the tumor with high-intensity modulated fields is balanced with normal tissue sparing. However, the non-target dose delivered to surrounding healthy tissue within the irradiated volume is a potential cause for concern. Whether the effects observed are caused after exposure to out-of-field radiation or bystander effects through neighboring irradiated cells is not fully understood. The goal of this study was to determine the effect of exposure to out-of-field radiation in lymphocyte cell lines and primary blood cells. The role of cellular radiosensitivity in altering bystander responses in out-of-field exposed cells was also investigated. Target cells were …

Understanding Pseudomonas Aeruginosa Alginate Regulation And Its Link To Chronic Lung Infections In Cystic Fibrosis Patients, Roy Al Ahmar

Theses, Dissertations and Capstones

Cystic Fibrosis (CF) is a genetic disorder caused by mutations in the gene encoding Cystic Fibrosis Transmembrane Conductance Regulator (CFTR). This disorder results in thick lung secretions which compromise the patient’s immune system. Chronic lung infections with Pseudomonas aeruginosa biofilms is a major factor that contributes to poor outcome for clinical treatment. The overall hypothesis of this dissertation is that small colony variant and pyrimidine biosynthesis are linked with biofilm formation, alginate production is dependent on MucA proteolysis, rifaximin can be repurposed to treat biofilm infection and multiplex PCR can be used to rapidly measure the bacterial cell density in …

Urease Is An Essential Component Of The Acid Response Network Of Staphylococcus Aureus And Is Required For A Persistent Murine Kidney Infection, Chunyi Zhou, Fatema Bhinderwala, Mckenzie K. Lehman, Vinai Chittezham Thomas, Sujata S. Chaudhari, Kelsey J. Yamada, Kirk W. Foster, Robert Powers, Tammy Kielian, Paul D. Fey

Journal Articles: Pathology and Microbiology

Staphylococcus aureus causes acute and chronic infections resulting in significant morbidity. Urease, an enzyme that generates NH3 and CO2 from urea, is key to pH homeostasis in bacterial pathogens under acidic stress and nitrogen limitation. However, the function of urease in S. aureus niche colonization and nitrogen metabolism has not been extensively studied. We discovered that urease is essential for pH homeostasis and viability in urea-rich environments under weak acid stress. The regulation of urease transcription by CcpA, Agr, and CodY was identified in this study, implying a complex network that controls urease expression in response to changes in metabolic …

Impact Of Work-Life Balance On Health-Related Quality Of Life Among College Students, Emily Vernet

Honors Undergraduate Theses

Work-life balance involves the management of work and social responsibilities. Specifically, it describes the ability to meet the demands of multiple roles involving academics, social, personal, and professional life. Literature has shown that there is an increasing number of students who are working while taking classes at a university. Previous research has described how young adults balance working and going to school and how work hours influence student's mental, physical, and health behaviors. The purpose of this study is to examine the association between a student's ability to balance work demands, student and life responsibilities, and health outcomes. Using a …

Unraveling Pdi And Its Interaction With Ab Toxins, Jessica Guyette

Electronic Theses and Dissertations

Protein disulfide isomerase (PDI) is an essential endoplasmic reticulum (ER) protein that acts as both an oxidoreductase and chaperone. It exhibits substantial flexibility and undergoes cycles of unfolding and refolding in its interaction with cholera toxin (Ctx), which is a unique property of PDI. This unfolding allows PDI to disassemble the Ctx holotoxin, which is required for Ctx activity. Here, we investigated the unfolding and refolding property of PDI and how this affects its interaction with bacterial toxins. PDI showed remarkable redox-linked conformational resilience that allows it to refold after being thermally stressed. Deletion constructs of PDI showed that both …

Detection Of Active Caspase-3 In Mouse Models Of Stroke And Alzheimer's Disease With A Novel Dual Positron Emission Tomography/Fluorescent Tracer [ 68 Ga]Ga-Tc3-Ogdota, Valeriy G. Ostapchenko, Jonatan Snir, Mojmir Suchy, Jue Fan, M. Rebecca Cobb, Blaine A. Chronik, Michael Kovacs, Vania F. Prado, Robert H.E. Hudson, Stephen H. Pasternak, Marco A.M. Prado, Robert Bartha

Medical Biophysics Publications

© 2019 Valeriy G. Ostapchenko et al. Apoptosis is a feature of stroke and Alzheimer's disease (AD), yet there is no accepted method to detect or follow apoptosis in the brain in vivo. We developed a bifunctional tracer [ 68 Ga]Ga-TC3-OGDOTA containing a cell-penetrating peptide separated from fluorescent Oregon Green and 68 Ga-bound labels by the caspase-3 recognition peptide DEVD. We hypothesized that this design would allow [ 68 Ga]Ga-TC3-OGDOTA to accumulate in apoptotic cells. In vitro, Ga-TC3-OGDOTA labeled apoptotic neurons following exposure to camptothecin, oxygen-glucose deprivation, and β-amyloid oligomers. In vivo, PET showed accumulation of [ 68 Ga]Ga-TC3-OGDOTA in …

Overview Of Direct Thrombin Inhibitors For Use In Staphylococcus Aereus Infections, Joseph C. Risler

Honors Undergraduate Theses

The pathogenicity and intractable nature of the microorganism Staphylococcus aureus (SA) has been long documented and highlighted by many health care agencies, with emphasis on its ability to exploit the human coagulation system to deadly effect. Two drugs from a class of inhibitors known as Direct Thrombin Inhibitors (DTI) have been shown to have a substantial effect on the enzyme secreted by SA known as Staphylocoagulase (SC), but up until now the application of this potential treatment has been limited. This paper strives to supply an overview of these clinical studies and propose a novel protocol for testing DTI's on …

Can Nanotechnology Be The Leading Method In Detecting And Treating Cerebral Tumors?, Daniel D. Singh, Zuri Jules-Culver

OUR Journal: ODU Undergraduate Research Journal

Nanotechnology refers to the manipulation or design of materials and structures with desired features in the 1nm–1000 nm size range. The blood brain barrier (BBB) is a major obstacle that drugs must overcome in order to reach tumor cells. The role of this barrier is to transport essential nutrients while protecting and regulating the internal environment. Nanoparticles have been shown to transport drugs through this barrier and accumulate in tumor cells. This is significant since nanoparticles are drug carriers allowing chemotherapeutic drugs to accumulate in target areas (Sun et al., 2017). This is possible because they are able to be …

Mir-9 Promotes The Phenotypic Switch Of Vascular Smooth Muscle Cells By Targeting Klf5, Xiaochun Lu, Shitang Ma, Bo Zhou, Tieling Li

Turkish Journal of Medical Sciences

Background/aim: Diabetic vascular smooth muscle cells (VSMCs) are characterized by increased proliferation and migration. Small noncoding microRNAs (miRNAs) have been considered critical modulators of the VSMC phenotypic switch after an environmental stimulus. However, microRNA in high glucose-induced proinflammation and its atherogenic effect is still ambiguous. Materials and methods: The technique of qRT-PCR was used to examine the expression of miR-9 in VSMCs. The downstream signaling protein relative to miR-9 regulation, Krüppel-like factor 5, and some marker genes of contractile VSMCs were analyzed by western blotting and qRT-PCR. Luciferase reporter assay was used to detect the expression of KLF5, which is …

Pathologic And Hemodynamic Changes Of Common Carotid Artery In Obstructive Sleep Apnea Hypopnea Syndrome In A Porcine Model, Yongyi Liu, Lu Gao, Weinong Lv, Lin Lin, Yi Wang, Fan Jiang, Fan Feng

Turkish Journal of Medical Sciences

Background/aim: To prepare a porcine model of obstructive sleep apnea-hypopnea syndrome (OSAHS) and observe the pathological and hemodynamic changes in the common carotid artery.Materials and methods: Twelve male miniature pigs were randomly divided into the model and control group (n = 6). Pigs in the model group were kept in an air-flow negative pressure chamber at 0.96 ± 0.01 kPa, and the air oxygen content, temperature, and humidity were kept at normal culture conditions in both groups. After pigs in the model group presented symptoms of OSAHS, changes in the hemodynamics and morphology of the carotid artery were analyzed using …

External-Internal Ureteral Catheterization Technique In Treatment Of Ureteral Injuries, Hasanali̇ Durmaz

Turkish Journal of Medical Sciences

Background/aim: It was aimed to describe the external-internal ureteral catheterization technique and evaluate its safety, efficacy, and reliability in iatrogenic and traumatic ureteral injuries.Materials and methods: A retrospective review was performed on patients with iatrogenic and traumatic ureteral injury, treated using the external-internal ureteral catheterization technique between May 2012 and January 2018 in our hospital. A total of 14 patients were investigated with clinical, postoperative, and follow-up findings, as well as technical outcomes.Results: The urology, gynecology, and general surgery departments referred patients for treatment at a rate of 57% (n = 8), 36% (n = 5), and 7% (n = …

Obesity Perception Survey Among Youth In Turkey: Instrument Development And Test-Retest Reliability, Wasantha Jayawardene, Sali̇h Pinar, Mohammad Torabi, Pengcheng Xun, Mustafa Kami̇l Özer

Turkish Journal of Medical Sciences

Background/aim: We aimed to develop an instrument that can assess the perceptions and opinions of young people regarding the causes and consequences of obesity and the role of individuals, families, communities, and government in addressing obesity.Materials and methods: A 36-question (101-item) survey was developed by adopting, translating, and revising multiple-choice or Likert-scale questions from existing surveys to assure construct cross-cultural validity. A two-factor mixed-effects model estimated the intraclass correlation coefficient (ICC) to measure the test-retest reliability of questions administered 2 weeks apart to a convenient sample of İstanbul high school and university students, aged 15?25 years (n = 122).Results: The …

Mechanism Of Nanogp8 In Glioblastoma Multiforme, Jonhoi Smith

Electronic Theses and Dissertations

Glioblastoma multiforme (GBM) is an incurable brain tumor, with patients only expecting to live 15 to 16 months post-diagnosis with the most current treatments options including surgery, chemotherapy, and radiation. How GBM resist treatment is still not very well known. However, cancer stem cells (CSCs), a subset of cells in GBM tissue considered responsible for therapeutic resistance and the poor patient prognosis. NANOG, a homeobox gene, is responsible for maintaining pluripotency of embryonic stem cells and is observed in CSCs. NANOGP8, a retrogene in the NANOG family is expressed in CSCs and provides cells with stem-like characteristics previously observed in …

Development Of A Functional Testing Platform For The Sensory Segment Of The Neuromuscular Reflex Arc, Alisha Colon

Electronic Theses and Dissertations

Investigations of human biology and disease have been hindered by the use of animal models. The information obtained from such studies often results in clinically irrelevant results and drug trial failures. Additionally, several governing bodies have been formulating legislation to move away from animal models and toward more ethical and efficient testing platforms for drug discovery and cosmetic research. As an answer to these issues, "body-on-a-chip" systems have been a rapidly developing field which easily recapitulates in vivo functionality, providing a more relevant, repeatable, and ethical testing platform to better predict biology. These systems can be used as human-based testing …

Structural Studies Of Some Proteins Responsible For Bacterial Pathogenesis, Kartik Manne

All ETDs from UAB

A wide range of infections is attributed to the pathogenesis of Gram-positive bacteria. The emergence of multi-drug-resistant Gram-positive pathogens has been a significant health concern and present serious therapeutic challenges globally. Some of the common Gram-positive bacterial species like Streptococcus pneumonia, Group B Streptococcus, and Staphylococcus aureus are infectious by employing several virulence factors for initial adhesion, colonization, biofilm initiation and dispersion, complement evasion, nutrient acquisition, and infection. The structural insights into these virulence factors are essential for understanding their mode of action, and also for developing a new class of antibiotics; a significant interest of many researchers. Pneumococcal adhesive …

Targeting The Nad Salvage Pathway For The Treatment Of The Parasitic Disease Schistosomiasis, Michael David Schultz

All ETDs from UAB

Schistosomiasis, also known as bilharzia, is the third most common parasitic infection worldwide and a major source of morbidity and mortality in developing countries. Caused by trematodes in the genus Schistosoma, this parasitic disease has no vaccine therapy and resources for drug development are scarce. Despite an unknown mechanism of action, praziquantal has become the primary source of treatment due to its efficacy against all species of Schistosoma. However, with increased drug resistance in endemic areas and low activity against immature parasites, relying on this single drug is unsustainable and risky. Hence, there is a growing need for new, safe …