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3. A Synthetic Polymicrobial Community Biofilm Model Demonstrates Spatial Partitioning, Tolerance to Antimicrobial Treatment, Reduced Metabolism, and Small Colony Variants Typical of Chronic Wound Biofilms

Author

Ammara Khalid, Alan R. Cookson, David E. Whitworth, Michael L. Beeton, Lori I. Robins, and Sarah E. Maddocks

Subjects
polymicrobial, antimicrobial, wound infection, Medicine
Abstract

Understanding chronic wound infection is key for successful treatment and requires accurate laboratory models. We describe a modified biofilm flow device that effectively mimics the chronic wound environment, including simulated wound fluid, a collagen-based 3D biofilm matrix, and a five-species mixture of clinically relevant bacteria (Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Citrobacter freundii). Mixed biofilms were cultured for between 3 and 14 days with consistent numbers of bacteria that exhibited reduced metabolic activity, which increased with a high dose of glucose. S. aureus was recovered from biofilms as a small colony variant, but as a normal colony variant if P. aeruginosa was excluded from the system. Bacteria within the biofilm did not co-aggregate but formed discrete, species-specific clusters. Biofilms demonstrated differential tolerance to the topical antimicrobials Neosporin and HOCl, consistent with protection due to the biofilm lifestyle. The characteristics exhibited within this model match those of real-world wound biofilms, reflecting the clinical scenario and yielding a powerful in vitro tool that is versatile, inexpensive, and pivotal for understanding chronic wound infection.

Published
2023
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5. Long-Term Stability and Efficacy of NCT Solutions.

Author

Staudinger, Gabriel J., Thomas, Zach M., Hooper, Sarah E., Williams, Jeffrey F., and Robins, Lori I.

Subjects
CHRONIC wounds & injuries, ULTRAVIOLET-visible spectroscopy, NUCLEAR magnetic resonance spectroscopy, HIGH temperatures, PSEUDOMONAS aeruginosa
Abstract

To realize the potential for the use of N-chlorotaurine (NCT) in healthcare, a better understanding of the long-term stability of the compound in water is needed. An array of analytical procedures is required that can measure changes in NCT concentration over time and allow for the detection and identification of contaminants and likely degradation end products. We used UV-Vis and NMR spectroscopy, HPLC, and LCMS to establish the stability of NCT in solutions subjected to prolonged ambient and elevated temperatures. Stability proved to be dependent on concentration with half-lives of ~120 days and ~236 days for 1% and 0.5% solutions of NCT at ~20 °C. Regardless of initial pH, all solutions shifted toward and maintained a pH of ~8.3 at 20 °C and 40 °C. NCT at 500 µg/mL and 250 µg /mL inhibited biofilm formation by Pseudomonas aeruginosa and Staphylococcus aureus but did not disperse established biofilms. NCT exposure to the biofilms had profound effects on the viability of both bacteria, reducing live organisms by >90%. Exposure of Interleukin-6 (IL-6) to 11 µM NCT reduced the binding of IL-6 to an immobilized specific antibody by ~48%, which is 5× the amount required for HOCl to bring about the same effect in this test system. Our data demonstrate the potency of the compound as an antimicrobial agent with potential benefits in the management of infected chronic wounds and suggest that NCT may contribute to anti-inflammatory processes in vivo by direct modification of cytokine mediators. [ABSTRACT FROM AUTHOR]

Published
2024
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10. Hypochlorous acid as a disinfectant for high‐risk HPV: Insight into the mechanism of action

Author

Lori I. Robins, Andrew Clark, Philip R. Gafken, Samina Alam, Janice Milici, Reem Hassan, Che‐Yen Wang, Jeffrey Williams, and Craig Meyers

Subjects
Human papillomavirus 16, Infectious Diseases, Virology, Papillomavirus Infections, Humans, Capsid Proteins, Disinfectants, Hypochlorous Acid
Abstract

Medical instruments that are not autoclavable but may become contaminated with high-risk human papillomaviruses (HPVs) during use must be thoroughly disinfected to avoid the possibility of iatrogenic transmission of infection. There is an expectation that prolonged soaking of instruments in the United States Food and Drug Administration-cleared chemical disinfectant solutions will result in high-level decontamination, but HPV16 and HPV18 are known to be resistant to commonly used formulations. However, they are susceptible to a variety of oxidative agents, including those based on chlorine. Here, we tested the efficacy of homogeneous hypochlorous acid (HOCl) solutions against mature infectious virions of HPV16 and HPV18 dried onto butadiene styrene coupons and ultrasonic probes. Both viruses were inactivated to4 log reduction value (LRV) after 15 s on coupons and 5 min on ultrasonic probes. Morphologic changes became evident within those contact times by transmission electron microscopy when HPV16 virus-like particles were exposed to HOCl under identical conditions. Mass spectrometry analysis of trypsin-digested products of L1 capsid proteins exposed to HOCl showed that mostly conserved residues were modified by oxidation and that these changes rapidly lead to instability of the protein demonstrable on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Modifications to these residues may contribute to rapid virus inactivation. The use of homogeneous HOCl solutions for HPV decontamination provides a highly effective means of assuring the safety of nonautoclavable medical instruments.

Published
2022
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12. The Effect of a Geriatric Simulation-enhanced Interprofessional Education on Health Profession Students

Author

Sheau-Huey Chiu, Lori I. Kidd, Diane K. Brown, James A. Grand, and Christine Heifner Graor

Subjects
Nursing (miscellaneous), Social work, Disease, Interprofessional education, Health professions, medicine.disease, Speech therapy, Education, Nursing, Modeling and Simulation, medicine, Dementia, Psychology, Healthcare providers
Abstract

Background Alzheimer's disease is the most common form of dementia, making it urgent to prepare health providers in interprofessional teams to care for those affected. Methods An unfolding geriatric simulation-enhanced interprofessional education program was designed using an ACE.S case and breakout activities for an interprofessional group of nursing, social work, speech therapy and nutrition students. Results Pre post education measures revealed a decrease in perceived challenges for interprofessional collaboration, with no change in readiness for interprofessional learning. Satisfaction with the education design was rated positively, and individual education components were rated as valuable. Conclusions This study offers educators an effective example of an unfolding active interprofessional geriatric education related to Alzheimer's care.

Published
2021
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13. Development of a Mental Health Nursing Simulation: Challenges and Solutions

Author

Kidd, Lori I., Morgan, Karyn I., and Savery, John R.

Abstract

Nursing education programs are proliferating rapidly in the United States in an effort to meet demand for nurse professionals. Multiple challenges arise from this rapid expansion. One challenge is finding sufficient clinical sites to accommodate students. Increased competition for scarce resources requires creativity in clinical contracting. This paper examines the challenges associated with providing virtual clinical--experiences and environments rich in diversity and exposure, yet safe for experimentation and learning of mental health nursing students. (Contains 2 figures and 1 table.)

Published
2012

14. Testing the efficacy of topical antimicrobial treatments using a two‐ and five‐species chronic wound biofilm model

Author

Rebecca L Plant, Lori I. Robins, Andreea-Gabriela Nedelea, and Sarah E Maddocks

Subjects
Chronic wound, Staphylococcus aureus, biology, Pseudomonas aeruginosa, Chemistry, Biofilm, General Medicine, medicine.disease_cause, biology.organism_classification, Antimicrobial, Applied Microbiology and Biotechnology, Enterococcus faecalis, Microbiology, Anti-Infective Agents, Biofilms, Streptococcus pyogenes, Wound Infection, medicine, Humans, medicine.symptom, Bacteria, Biotechnology
Abstract

Aims The effectiveness of commercially available wound dressings and a HOCl gel formulation was tested against two- and five-species biofilms in a dynamic in vitro chronic wound infection model. Method Two-species biofilms (Pseudomonas aeruginosa and Staphylococcus aureus) were cultured using a biofilm flow device and treated with wound dressings containing silver, iodine, polyhexamethylene biguanide, crystal violet or HOCl gel at 5 h. Five-species biofilms (P. aeruginosa, S. aureus, Enterococcus faecalis, Streptococcus pyogenes and Escherichia coli) were similarly cultured and treated with HOCl gel at 5 and 24 h. Multidose experiments used two- and five-species biofilms with HOCl applied at 24, 48 and 72 h. Results None of the treatments completely disrupted the biofilms and, with the exception of silver, bacteria recovered in number post-treatment. HOCl was most effective when applied to 24 h established biofilms with most activity against P. aeruginosa. Recovery post-treatment was negligible with HOCl applied at 24 h and multiple doses indicated that bacteria were not becoming tolerant to treatment. Conclusions Realistic models are necessary to test the effectiveness of antimicrobial wound treatments to ensure findings are clinically translatable. HOCl gel shows promise as a new topical antimicrobial for wounds, especially due to its ability to inhibit P. aeruginosa. Significance and impact of the study This study highlights a need for robust in vitro data to support development and use of wound treatments that can only be obtained from the refinement of realistic infection models. Furthermore, it indicates the potential use of HOCl gel for chronic wound management.

Published
2021
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16. Challenging Our Assumptions: An Investigation into Student Understanding of the Gas Laws

Author

Robins, Lori I., Villagomez, Gisela, Dockter, Derek, Christopher, Elizabeth, Ortiz, Christine, Passmore, Cynthia, and Smith, Martin H.

Abstract

Teacher research--often called "action research"--is an intentional and systematic inquiry into one's own classroom practice with the goal of improved student learning (Cochran-Smith and Lytle 1993). In this article, the authors present a teacher research project undertaken to improve student understanding of the gas laws in a high school chemistry class. It addresses both the product of this teacher research project--insights into the teaching of gas laws--and the process and potential power of this approach. (Contains 7 figures.)

Published
2009
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18. Dialectical Behavior Therapy for Substance Abusers Adapted for Persons Living with HIV/AIDS with Substance Use Diagnoses and Borderline Personality Disorder

Author

Wagner, Elizabeth E., Miller, Alec L., and Greene, Lori I.

Abstract

The primary aim of this article is to describe modifications made to Dialectical Behavior Therapy (DBT) for a predominantly ethnic minority population of persons living with HIV/AIDS with substance-use diagnoses and borderline personality disorder (BPD) or three features of BPD plus suicidality (i.e., the triply diagnosed). Despite the myriad psychosocial needs of the triply diagnosed, there remains a dearth of treatments available that can adequately address the challenges presented by these individuals' dual diagnostic and HIV status. The key modifications we developed and describe in this article are (a) modification of Stage One, Target 2, behaviors to include HIV treatment adherence targets, (b) new and modified standard skills that address needs of the triply diagnosed client including methadone clinic-relevant skills and adherence skills; (c) expansion of the role of the therapist consultation group to include the DBT Therapist Path to HIV/AIDS Competence and HIV-related psychotherapy themes; and (d) use of a Consumer Advisory Board to provide consultation and feedback on treatment adaptations. A case study is presented that illustrates these modifications.

Published
2004
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19. Modification of Major Contributors Responsible for Latrine Malodor on Exposure to Hypochlorous Acid: The Potential for Simultaneously Impacting Odor and Infection Hazards to Encourage Latrine Use

Author

Erika K. Keim, Tim E. Dennler-Church, Mary C. Hall, JoAnne M. Mulligan, Jeremy C. Butz, John Scott Meschke, Joseph E. McKinley, Jeffrey F. Williams, and Lori I. Robins

Subjects
Hypochlorous acid, chemistry.chemical_compound, Virology, Escherichia coli, Medicine, Humans, Dimethyl disulfide, Food science, Sanitation, Toilet Facilities, Levivirus, Toilet, business.industry, Significant difference, Pit latrine, Articles, Oxidants, Hypochlorous Acid, Infectious Diseases, chemistry, Odor, Communicable Disease Control, Odorants, Latrine, Parasitology, Dimethyl trisulfide, business
Abstract

Open defecation remains a common practice in developing countries and leads to high incidence and prevalence of acute gastroenteritis, which is most often caused by human noroviruses (human NoV). Encouraging the use of toilets and pit latrines is one method of improving sanitation; however, it is often hindered by not only cultural traditions but also from a reluctance to use latrines and toilets due to their odor and impression of uncleanliness. In an effort to establish new means to encourage toilet and latrine use, laboratory experiments tested the ability of hypochlorous acid (HOCl) to modify the malodorous compounds identified in the air in latrines in developing countries (indole, p-cresol, dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS), and butyric acid) and inactivate MS2 bacteriophage, a surrogate for human NoV. After 5 minutes, > 94% of indole, p-cresol, DMDS, and DMTS was modified as determined by high-pressure liquid chromatography in the presence of 100 ppm HOCl. A log10 reduction value (LRV) greater than 6 was seen for MS2 bacteriophage after 5 minutes of exposure to 100 ppm HOCl in solution. Sensory studies indicated that there was a significant difference (P ≤ 0.05) between the untreated and HOCl-treated samples for all five malodorous compounds tested. The findings suggest that introduction of HOCl into the headspace air could encourage latrine and toilet use. Optimization of HOCl dosing in air to accomplish both odor control and reduction of infectious hazards is worthy of further study.

Published
2020

24. An Authentic Poverty Simulation for Health Care Profession Students Using Community Volunteers Experiencing Poverty

Author

Lori I. Kidd, Rose M Resler, Greta A Lax, and Sheri A Hartman

Subjects
Adult, Male, Volunteers, Students, Health Occupations, Resource (biology), 020205 medical informatics, Attitude of Health Personnel, Health Personnel, media_common.quotation_subject, education, Population, Empathy, 02 engineering and technology, Education, Young Adult, 03 medical and health sciences, Health care, 0202 electrical engineering, electronic engineering, information engineering, Humans, Education, Nursing, Poverty, Simulation Training, Curriculum, media_common, education.field_of_study, Medical education, 030504 nursing, business.industry, LPN and LVN, United States, Nursing Education Research, Incentive, Review and Exam Preparation, Female, Fundamentals and skills, 0305 other medical science, business, Psychology, Realism
Abstract

Background Poverty is a harsh reality for more than 40 million Americans, which can lead to detrimental health outcomes. Problem Considering health professionals encounter clients of low-income status, increasing awareness and empathy toward this population is essential. Approach This article describes a novel approach to conducting poverty simulations by using community volunteers that are currently living or have recently lived in poverty, lending to a more authentic experience for students. The community volunteers acted as resource workers during the simulation. Outcomes Five lessons were learned as part of this authentic approach including the importance of orientation, consistent volunteer recruitment, volunteer appreciation, simulation role ownership, and importance of incentives. Conclusion This approach is an effective initial step in increasing awareness and empathy among health care profession students toward individuals living in poverty. Use of community volunteers with personal experience with poverty enhances the realism of this experience for students.

Published
2019
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26. Modification of IL-6 by hypochlorous acid: effects on receptor binding and possible role in treatment of COVID-19

Author

Jeffrey F. Williams, John Scott Meschke, Deborah Robins, Lori I. Robins, Scott Edgar, Philip R. Gafken, and Erika K. Keim

Subjects
chemistry.chemical_compound, Methionine, Cytokine, Hypochlorous acid, chemistry, Biochemistry, In vivo, medicine.medical_treatment, Hypobromous acid, medicine, Tryptophan, Receptor, In vitro
Abstract

Interleukin-6 (IL-6) has been implicated in the pathogenesis of acute inflammatory events in COVID-19 patients. We exposed IL-6 to hypochlorous acid (HOCl) in vitro at concentrations reported to develop in vivo. After HOCl treatment the cytokine failed to bind to IL-6 receptors in a bioassay using engineered human cells. Similar results followed exposure of IL-6 to N-chlorotaurine (NCT) and hypobromous acid (HOBr). SDS-PAGE analysis of HOCl-treated IL-6 showed neither fragmentation nor aggregation, suggesting that the modifications induced by these agents occurred on the intact protein. Mass spectrometry of intact and trypsin-digested fragments identified oxidative changes limited to two amino acid residues, methionine 161 and tryptophan 157, both of which have been implicated in receptor binding of the cytokine. Further studies on the effects of hypohalous acids and their halogenated amine derivatives on IL-6 and related cytokines is needed.

Published
2021
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27. Control of felinine-derived malodor in cat litter

Author

Charles D Mackenzie, Xin Gao, Carol Flegler, Carolina Seek, Lori I. Robins, and Stephanie Napier

Subjects
Litter (animal), 040301 veterinary sciences, 0402 animal and dairy science, 04 agricultural and veterinary sciences, Biology, 040201 dairy & animal science, Gas Chromatography-Mass Spectrometry, 0403 veterinary science, chemistry.chemical_compound, Animal science, chemistry, Anti-Infective Agents, Odor control, Odorants, Cats, Animals, Felinine, Cysteine, Small Animals
Abstract

Objectives Malodors stemming from soiled cat litter are a major frustration for cat owners, despite the widespread use of absorbent litters with claims of odor control. Technologies for effective litter odor control have not been rigorously evaluated. Here, we report on the effectiveness of a novel litter formulation of 1-monochlorodimethylhydantoin (MCDMH)-modified clinoptilolite zeolite (MCDMH-Z) to control the odors of 3-mercapto-3-methylbutanol (3M3MB) and ammonia, the principal products generated by the enzymatic breakdown of felinine and urea, respectively. Methods The efficacy of MCDMH-Z for the odor control of 3M3MB was determined by solid-phase microextraction and gas chromatography mass spectrometry analysis, colorimetric analysis and a sensory panel. Enzyme inhibition was monitored by a colorimetric coupled assay for ammonia. The antimicrobial properties were measured by a reduction in colony-forming units (CFUs). Results 3M3MB proved highly susceptible to modification by MCDMH-Z granules. Headspace above litter exposed to MCDMH-Z showed no detectable 3M3MB; levels >59 ng were detected in commercially available products. Urease activity decreased by >97% after incubation with MCDMH-Z to 0.14 mg/ml. Cat litter F showed comparable inhibition (0.13 mg/ml); others showed less inhibition, producing up to 4.8 mg/ml of ammonia. MCDMH-Z reduced the CFUs of Proteus vulgaris by six log reduction values in 30 mins; in the same amount of time, no reduction was seen with commercial products tested. The odor control capability of the MCDMH-Z granules was further supported by a sensory panel scoring 3M3MB-spiked litters. Conclusions and relevance Samples of commercially available litter products showed an effect on malodor, or inhibition of urease, or contained antimicrobial activity; no samples were capable of accomplishing these concurrently. In contrast, MCDMH-Z granules were effective in all three test categories. Control of felinine-derived odors, in particular, has the potential to improve cat owner satisfaction, and may beneficially affect cat behaviors provoked by pheromonally active sulfurous metabolites deposited in the litter.

Published
2021

34. Modification of Major Contributors Responsible for Latrine Malodor on Exposure to Hypochlorous Acid: The Potential for Simultaneously Impacting Odor and Infection Hazards to Encourage Latrine Use

Author

Dennler-Church, Tim E., primary, Butz, Jeremy C., additional, McKinley, Joseph E., additional, Keim, Erika K., additional, Hall, Mary C., additional, Meschke, John S., additional, Mulligan, JoAnne M., additional, Williams, Jeffrey F., additional, and Robins, Lori I., additional

Published
2020
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38. Control of felinine-derived malodor in cat litter.

Author

Robins, Lori I, Napier, Stephanie, Seek, Carolina M, Gao, Xin, Flegler, Carol, and Mackenzie, Charles D

Abstract

Objectives: Malodors stemming from soiled cat litter are a major frustration for cat owners, despite the widespread use of absorbent litters with claims of odor control. Technologies for effective litter odor control have not been rigorously evaluated. Here, we report on the effectiveness of a novel litter formulation of 1-monochlorodimethylhydantoin (MCDMH)-modified clinoptilolite zeolite (MCDMH-Z) to control the odors of 3-mercapto-3-methylbutanol (3M3MB) and ammonia, the principal products generated by the enzymatic breakdown of felinine and urea, respectively. Methods: The efficacy of MCDMH-Z for the odor control of 3M3MB was determined by solid-phase microextraction and gas chromatography mass spectrometry analysis, colorimetric analysis and a sensory panel. Enzyme inhibition was monitored by a colorimetric coupled assay for ammonia. The antimicrobial properties were measured by a reduction in colony-forming units (CFUs). Results: 3M3MB proved highly susceptible to modification by MCDMH-Z granules. Headspace above litter exposed to MCDMH-Z showed no detectable 3M3MB; levels >59 ng were detected in commercially available products. Urease activity decreased by >97% after incubation with MCDMH-Z to 0.14 mg/ml. Cat litter F showed comparable inhibition (0.13 mg/ml); others showed less inhibition, producing up to 4.8 mg/ml of ammonia. MCDMH-Z reduced the CFUs of Proteus vulgaris by six log reduction values in 30 mins; in the same amount of time, no reduction was seen with commercial products tested. The odor control capability of the MCDMH-Z granules was further supported by a sensory panel scoring 3M3MB-spiked litters. Conclusions and relevance: Samples of commercially available litter products showed an effect on malodor, or inhibition of urease, or contained antimicrobial activity; no samples were capable of accomplishing these concurrently. In contrast, MCDMH-Z granules were effective in all three test categories. Control of felinine-derived odors, in particular, has the potential to improve cat owner satisfaction, and may beneficially affect cat behaviors provoked by pheromonally active sulfurous metabolites deposited in the litter. [ABSTRACT FROM AUTHOR]

Published
2022
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39. Testing the efficacy of topical antimicrobial treatments using a two‐ and five‐species chronic wound biofilm model.

Author

Nedelea, Andreea‐Gabriela, Plant, Rebecca L., Robins, Lori I., and Maddocks, Sarah E.

Subjects
CHRONIC wounds & injuries, STREPTOCOCCUS pyogenes, BIOFILMS, GENTIAN violet, ENTEROCOCCUS faecalis, PSEUDOMONAS aeruginosa
Abstract

Aims: The effectiveness of commercially available wound dressings and a HOCl gel formulation was tested against two‐ and five‐species biofilms in a dynamic in vitro chronic wound infection model. Method: Two‐species biofilms (Pseudomonas aeruginosa and Staphylococcus aureus) were cultured using a biofilm flow device and treated with wound dressings containing silver, iodine, polyhexamethylene biguanide, crystal violet or HOCl gel at 5 h. Five‐species biofilms (P. aeruginosa, S. aureus, Enterococcus faecalis, Streptococcus pyogenes and Escherichia coli) were similarly cultured and treated with HOCl gel at 5 and 24 h. Multidose experiments used two‐ and five‐species biofilms with HOCl applied at 24, 48 and 72 h. Results: None of the treatments completely disrupted the biofilms and, with the exception of silver, bacteria recovered in number post‐treatment. HOCl was most effective when applied to 24 h established biofilms with most activity against P. aeruginosa. Recovery post‐treatment was negligible with HOCl applied at 24 h and multiple doses indicated that bacteria were not becoming tolerant to treatment. Conclusions: Realistic models are necessary to test the effectiveness of antimicrobial wound treatments to ensure findings are clinically translatable. HOCl gel shows promise as a new topical antimicrobial for wounds, especially due to its ability to inhibit P. aeruginosa. Significance and Impact of the Study: This study highlights a need for robust in vitro data to support development and use of wound treatments that can only be obtained from the refinement of realistic infection models. Furthermore, it indicates the potential use of HOCl gel for chronic wound management. [ABSTRACT FROM AUTHOR]

Published
2022
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42. The Changing Landscape for Stroke Prevention in AF: Findings From the GLORIA-AF Registry Phase 2

Author

Huisman, M. V., Rothman, K. J., Paquette, M., Teutsch, C., Diener, H. -C., Dubner, S. J., Halperin, J. L., C. S., Ma, Zint, K., Elsaesser, A., Bartels, D. B., Lip, G. Y. H., Abban, D., Abdul, N., Abelson, M., Ackermann, A., Adams, F., Adams, L., Adragao, P., Ageno, W., Aggarwal, R., Agosti, S., Marin, J. A., Aguilar, F., Aguilar Linares, J. A., Aguinaga, L., Ahmad, Z., Ainsworth, P., Al Ghalayini, K., Al Ismail, S., Alasfar, A., Alawwa, A., Al-Dallow, R., Alderson, L., Alexopoulos, D., Ali, A., Ali, M., Aliyar, P., Al-Joundi, T., Al Mahameed, S., Almassi, H., Almuti, K., Al-Obaidi, M., Alshehri, M., Altmann, U., Alves, A. R., Al-Zoebi, A., Amara, W., Amelot, M., Amjadi, N., Ammirati, F., Andrawis, N., Angoulvant, D., Annoni, G., Ansalone, G., Antonescu, S. A., Ariani, M., Arias, J. C., Armero, S., Arora, R., Arora, C., Ashcraft, W., Aslam, M. S., Astesiano, A., Audouin, P., Augenbraun, C., Aydin, S., Azar, R., Azim, A., Aziz, S., Backes, L. M., Baig, M., Bains, S., Bakbak, A., Baker, S., Bakhtiar, K., Bala, R., Banayan, J., Bandh, S., Bando, S., Banerjee, S., Bank, A., Barbarash, O., Baron, G., Barr, C., Barrera, C., Barton, J., Kes, V. B., Baula, G., Bayeh, H., Bazargani, N., Behrens, S., Bell, A., Benezet-Mazuecos, J., Benhalima, B., Berdague, P., Berg van den, B. J., Bergen van, P. F. M. M., Berngard, E., Bernstein, R., Berrospi, P., Berti, S., Bertomeu, V., Berz, A., Bettencourt, P., Betzu, R., Beyer-Westendorf, J., Bhagwat, R., Black, T., Blanco Ibaceta, J. H., Bloom, S., Blumberg, E., Bo, M., Bockisch, V., Bohmer, E., Bongiorni, M. G., Boriani, G., Bosch, R., Boswijk, D. J., Bott, J., Bottacchi, E., Kalan, M. B., Brandes, A., Bratland, B., Brautigam, D., Breton, N., Brouwers, P. J. A. M., Browne, K., Bruguera, J., Brunehaut, M., Brunschwig, C., Buathier, H., Buhl, A., Bullinga, J., Butcher, K., Cabrera Honorio, J. W., Caccavo, A., Cadinot, D., Cai, S., Calvi, V., Camm, J., Candeias, R., Capo, J., Capucci, A., Cardoso, J. N., Duarte Vera, Y. C., Carlson, B., Carvalho, P., Cary, S., Casanova, R., Casu, G., Cattan, S., Cavallini, C., Cayla, G., Cha, T. J., Cha, K. S., Chaaban, S., Chae, J. K., Challappa, K., Chand, S., Chandrashekar, H., Chang, M., Charbel, P., Chartier, L., Chatterjee, K., Cheema, A., Chen, S. -A., Chevallereau, P., Chiang, F. -T., Chiarella, F., Chih-Chan, L., Cho, Y. K., Choi, D. J., Chouinard, G., Danny, Chow, H. F., Chrysos, D., Chumakova, G., Jose Roberto Chuquiure Valenzuela, E. J., Cieza-Lara, T., Nica, V. C., Ciobotaru, V., Cislowski, D., Citerne, O., Claus, M., Clay, A., Clifford, P., Cohen, S., Cohen, A., Colivicchi, F., Collins, R., Compton, S., Connors, S., Conti, A., Buenostro, G. C., Coodley, G., Cooper, M., Corbett, L., Corey, O., Coronel, J., Corrigan, J., Cotrina Pereyra, R. Y., Cottin, Y., Coutu, B., Cracan, A., Crean, P., Crenshaw, J., Crijns, H. J. G. M., Crump, C., Cucher, F., Cudmore, D., Cui, L., Culp, J., Darius, H., Dary, P., Dascotte, O., Dauber, I., Davee, T., Davies, R., Davis, G., Davy, J. -M., Dayer, M., De La Briolle, A., de Mora, M., De Teresa, E., De Wolf, L., Decoulx, E., Deepak, S., Defaye, P., Del-Carpio Munoz, F., Brkljacic, D. D., Deluche, L., Destrac, S., Deumite, N. J., Di Legge, S., Dibon, O., Diemberger, I., Dillinger, J., Dionisio, P., Naydenov, S., Dotani, I., Dotcheva, E., D'Souza, A., Dubrey, S., Ducrocq, X., Dupljakov, D., Duthinh, V., Dutra, O. P., Dutta, D., Duvilla, N., Dy, J., Dziewas, R., Eaton, C., Eaves, W., Ebinger, M., Eck van, J. W. M., Edwards, T., Egocheaga, I., Ehrlich, C., Eisenberg, S., El Hallak, A., El Jabali, A., El Mahmoud, R., El Shahawy, M., Eldadah, Z., Elghelbazouri, F., Elhag, O., El-Hamdani, M., Elias, D., Ellery, A., El-Sayed, H., Elvan, A., Erickson, B., Espaliat, E., Essandoh, L., Everington, T., Evonich, R., Ezhov, A., Facila, L., Farsad, R., Fayard, M., Fedele, F., Gomes Ferreira, L. G., Ferreira, D., Santos, J. F., Ferrier, A., Finsen, A., First, B., Fisher, R., Floyd, J., Folk, T., Fonseca, C., Fonseca, L., Forman, S., Forsgren, M., Foster, M., Foster, N., Frais, M., Frandsen, B., Frappe, T., Freixa, R., French, W., Freydlin, M., Frickel, S., Fruntelata, A. G., Fujii, S., Fujino, Y., Fukunaga, H., Furukawa, Y., Gabelmann, M., Gabris, M., Gadsboll, N., Galin, P., Galinier, M., Ganim, R., Garcia, R., Quintana, A. 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O, Iglesias, I, Ikpoh, M, Imberti, D, Ince, H, Indolfi, C, Ionova, T, Ip, J, Irles, D, Iseki, H, Ismail, Y, Israel, N, Isserman, S, Iteld, B, Ivanchura, G, Iyer, R, Iyer, V, Iza Villanueva, R, Jackson Voyzey, E, Jaffrani, N, Jäger, F, Jain, M, James, M, Jamon, Y, Jang, S, Pereira Jardim, C, Jarmukli, N, Jeanfreau, R, Jenkins, R, Jiang, X, Jiang, H, Jiang, T, Jiang, N, Jimenez, J, Jobe, R, Joffe, I, Johansson, B, Jones, N, Moura Jorge, J, Jouve, B, Jundi, M, Jung, W, Jung, B, Jung, K, Kabbani, S, Kabour, A, Kafkala, C, Kajiwara, K, Kalinina, L, Kampus, P, Kanda, J, Kapadia, S, Karim, A, Karolyi, L, Kashou, H, Kastrup, A, Katsivas, A, Kaufman, E, Kawai, K, Kawajiri, K, Kazmierski, J, Keeling, P, Kerfes, G, Kerr Saraiva, J, Ketova, G, Khaira, A, Khalid, M, Khludeeva, E, Khripun, A, Kim, D, Kim, N, Kim, K, Kim, Y, Kim, J, Kinova, E, Klein, A, Kleinschnitz, C, Kmetzo, J, Kneller, G, Knezevic, A, Koch, S, Koenig, K, Angela Koh, S, Köhrmann, M, Koons, J, Korabathina, R, Korennova, O, Koschutnik, M, Kosinski, E, Kovacic, D, Kowalczyk, J, Koziolova, N, Kragten, J, Krause, L, Kreidieh, I, Krenning, B, Krishnaswamy, K, Krysiak, W, Kuck, K, Kumar, S, Kümler, T, Kuniss, M, Kuo, J, Küppers, A, Kurrelmeyer, K, Kwan, T, Kyo, E, Labovitz, A, Lacroix, A, Lam, A, Lanas Zanetti, F, Landau, C, Landini, G, Lang, W, Larsen, T, Laske, V, Lavandier, K, Law, N, Lee, M, Lee, D, Leitão, A, Lejay, D, Lelonek, M, Lenarczyk, R, Leprince, P, Lequeux, B, Leschke, M, Ley, N, Li, Z, Li, Y, Li, X, Li, W, Liang, J, Lieber, I, Lillestol, M, Limon Rodriguez, R, Lin, H, Litchfield, J, Liu, Z, Liu, X, Liu, Y, Liu, F, Liu, W, Llamas Esperon, G, Llisterri, J, Lo, T, Lo, E, Lobos, J, Lodde, B, Loiselet, P, López Sendón, J, Lorga Filho, A, Lori, I, Luo, M, Lupovitch, S, Lyrer, P, Zuhairy, H, Ma, G, Ma, H, Madariaga, I, Maeno, K, Magnin, D, Mahmood, S, Mahood, K, Maid, G, Mainigi, S, Makaritsis, K, Maldonado Villalon, J, Malhotra, R, Malik, A, Mallecourt, C, Mallik, R, Manning, R, Manolis, A, Mantas, I, Manzur Jattin, F, Marcionni, N, Marín, F, Santana, A, Martinez, J, Martinez, L, Maskova, P, Hernández, N, Matskeplishvili, S, Matsuda, K, Mavri, A, May, E, Mayer, N, Mazon, P, Mcclure, J, Mccormack, T, Mcgarity, W, Mcguire, M, Mcintyre, H, Mclaughlin, P, Mclaurin, B, Medina Palomino, F, Mehta, P, Mehzad, R, Meinel, A, Melandri, F, Mena, A, Meno, H, Menzies, D, Metcalf, K, Meyer, B, Miarka, J, Mibach, F, Michalski, D, Michel, P, Chreih, R, Mikdadi, G, Mikhail, M, Mikus, M, Milicic, D, Militaru, C, Miller, G, Milonas, C, Minescu, B, Mintale, I, Miralles, A, Mirault, T, Mistry, D, Mitchell, G, Miu, N, Miyamoto, N, Moccetti, T, Mohammed, A, Nor, A, Molina de Salazar, D, Molon, G, Molony, D, Mondillo, S, Mont, L, Moodley, R, Moore, R, Ribeiro Moreira, D, Mori, K, Moriarty, A, Morka, J, Moschos, N, Mota Gomes, M, Mousallem, N, Moya, A, Mügge, A, Mulhearn, T, Muller, J, Muresan, C, Muse, D, Musial, W, Musumeci, F, Nadar, V, Nageh, T, Nair, P, Nakagawa, H, Nakamura, Y, Nakayama, T, Nam, K, Napalkov, D, Natarajan, I, Nayak, H, Nechvatal, L, Neiman, J, Nerheim, P, Neuenschwander, F, Nishida, K, Nizov, A, Novikova, T, Novo, S, Nowalany Kozielska, E, Nsah, E, Nunez Fragoso, J, Nyvad, O, de Los Rios Ibarra, M, O'Donnell, M, O'Donnell, P, Oh, D, Oh, Y, Daniel Oh, C, O'Hara, G, Oikonomou, K, Olalla, J, Olivari, Z, Oliver, R, Olympios, C, Osborne, J, Osca, J, Osman, R, Osunkoya, A, Padanilam, B, Panchenko, E, Pandey, A, Vicenzo de Paola, A, Paraschos, A, Pardell, H, Park, H, Park, J, Parkash, R, Parker, I, Parrens, E, Parris, R, Passamonti, E, Patel, J, Patel, R, Pentz, W, Persic, V, Perticone, F, Peters, P, Petkar, S, Pezo, L, Pham, D, Cao Phai, G, Phlaum, S, Pineau, J, Pineda Velez, A, Pini, R, Pinter, A, Pinto, F, Pirelli, S, Pivac, N, Pizzini, A, Pocanic, D, Calin Podoleanu, C, Polanczyk, C, Polasek, P, Poljakovic, Z, Pollock, S, Polo, J, Poock, J, Poppert, H, Porro, Y, Pose, A, Poulain, F, Poulard, J, Pouzar, J, Povolny, P, Pozzer, D, Pras, A, Prasad, N, Prevot, S, Protasov, K, Prunier, L, Puleo, J, Pye, M, Qaddoura, F, Quedillac, J, Raev, D, Rahimi, S, Raisaro, A, Rama, B, Ranadive, N, Randall, K, Ranjith, N, Raposo, N, Rashid, H, Raters, C, Rauch Kroehnert, U, Rebane, T, Regner, S, Renzi, M, Reyes Rocha, M, Reza, S, Ria, L, Richter, D, Rickli, H, Rickner, K, Rieker, W, Rigo, F, Ripoll, T, Fonteles Ritt, L, Roberts, D, Pascual, C, Briones, I, Reyes, H, Roelke, M, Roman, M, Romeo, F, Ronner, E, Ronziere, T, Rooyer, F, Rosenbaum, D, Roth, S, Rozkova, N, Rubacek, M, Rubalcava, F, Rubanenko, O, Rubin, A, Borret, M, Rybak, K, Sabbour, H, Morales, O, Sakai, T, Salacata, A, Salecker, I, Salem, A, Salfity, M, Salguero, R, Salvioni, A, Samson, M, Sanchez, G, Sandesara, C, Saporito, W, Sasaoka, T, Sattar, P, Savard, D, Scala, P, Scemama, J, Schaupp, T, Schellinger, P, Scherr, C, Schmitz, K, Schmitz, B, Schmitz, L, Schnitzler, R, Schnupp, S, Schoeniger, P, Schön, N, Schuster, S, Schwimmbeck, P, Seamark, C, Seebass, R, Seidl, K, Seidman, B, Sek, J, Sekaran, L, Seko, Y, Sepulveda Varela, P, Sevilla, B, Shah, V, Shah, A, Shah, N, Shanes, J, Sharareh, A, Sharma, V, Shaw, L, Shimizu, Y, Shimomura, H, Shin, D, Shin, E, Shite, J, Shoukfeh, M, Shoultz, C, Silver, F, Sime, I, Simmers, T, Singal, D, Singh, N, Siostrzonek, P, Sirajuddin, M, Skeppholm, M, Smadja, D, Smith, R, Smith, D, Soda, H, Sofley, C, Sokal, A, Sotolongo, R, de Souza, O, Sparby, J, Spinar, J, Sprigings, D, Spyropoulos, A, Stakos, D, Steinberg, A, Steinwender, C, Stergiou, G, Stites, H, Stoikov, A, Strasser, R, Streb, W, Styliadis, I, Su, G, Su, X, Suarez, R, Sudnik, W, Sueyoshi, A, Sukles, K, Sun, L, Suneja, R, Svensson, P, Ziekenhuis, A, Szavits Nossan, J, Taggeselle, J, Takagi, Y, Takhar, A, Tallet, J, Tamm, A, Tanaka, S, Tanaka, K, Tang, A, Tang, S, Tassinari, T, Tayama, S, Tayebjee, M, Tebbe, U, Teixeira, J, Tesloianu, D, Tessier, P, The, S, Thevenin, J, Thomas, H, Timsit, S, Topkis, R, Torosoff, M, Touze, E, Traissac, T, Trendafilova, E, Troyan, B, Tsai, W, Tse, H, Tsutsui, H, Tsutsui, T, Tuininga, Y, Turakhia, M, Turk, S, Turner, W, Tveit, A, Twiddy, S, Tytus, R, Ukrainski, G, Valdovinos Chavez, S, Van De Graaff, E, Vanacker, P, Vardas, P, Vargas, M, Vassilikos, V, Vazquez, J, Venkataraman, A, Verdecchia, P, Vester, E, Vial, H, Vinereanu, D, Vlastaris, A, Vogel, C, vom Dahl, J, von Mering, M, Vora, K, Wakefield, P, Walia, J, Walter, T, Wang, M, Wang, N, Wang, F, Wang, X, Wang, Z, Wang, K, Watanabe, K, Wei, J, Weimar, C, Weinrich, R, Wen, M, Wheelan, K, Wicke, J, Wiemer, M, Wild, B, Wilke, A, Willems, S, Williams, M, Williams, D, Winkler, A, Wirtz, J, Witzenbichler, B, Wong, D, Lawrence Wong, K, Wong, B, Wozakowska Kaplon, B, Wu, Z, Wu, S, Wyatt, N, Xu, Y, Xu, X, Yamada, A, Yamamoto, K, Yamanoue, H, Yamashita, T, Bryan Yan, P, Yang, Y, Yang, T, Yao, J, Yarlagadda, C, Yeh, K, Yotov, Y, Yvorra, S, Zahn, R, Zamorano, J, Zanini, R, Zarich, S, Zebrack, J, Zenin, S, Zeuthen, E, Zhang, X, Zhang, Q, Zhang, D, Zhang, H, Zhao, S, Zhao, X, Zheng, Y, Zheng, Q, Zhou, J, Zimmermann, S, Zimmermann, R, Zukerman, L, and Zwaan van der, C

Subjects
Male, oral anticoagulation, Internationality, Middle Aged, registry, Antithrombins, Dabigatran, Stroke, Cross-Sectional Studies, Fibrinolytic Agents, Humans, Female, atrial fibrillation, Prospective Studies, Registries, Cardiology and Cardiovascular Medicine, Aged, Atrial Fibrillation
Abstract

Background GLORIA-AF (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients with Atrial Fibrillation) is a prospective, global registry program describing antithrombotic treatment patterns in patients with newly diagnosed nonvalvular atrial fibrillation at risk of stroke. Phase 2 began when dabigatran, the first non–vitamin K antagonist oral anticoagulant (NOAC), became available. Objectives This study sought to describe phase 2 baseline data and compare these with the pre-NOAC era collected during phase 1. Methods During phase 2, 15,641 consenting patients were enrolled (November 2011 to December 2014); 15,092 were eligible. This pre-specified cross-sectional analysis describes eligible patients’ baseline characteristics. Atrial fibrillation disease characteristics, medical outcomes, and concomitant diseases and medications were collected. Data were analyzed using descriptive statistics. Results Of the total patients, 45.5% were female; median age was 71 (interquartile range: 64, 78) years. Patients were from Europe (47.1%), North America (22.5%), Asia (20.3%), Latin America (6.0%), and the Middle East/Africa (4.0%). Most had high stroke risk (CHA2DS2-VASc [Congestive heart failure, Hypertension, Age ≥75 years, Diabetes mellitus, previous Stroke, Vascular disease, Age 65 to 74 years, Sex category] score ≥2; 86.1%); 13.9% had moderate risk (CHA2DS2-VASc = 1). Overall, 79.9% received oral anticoagulants, of whom 47.6% received NOAC and 32.3% vitamin K antagonists (VKA); 12.1% received antiplatelet agents; 7.8% received no antithrombotic treatment. For comparison, the proportion of phase 1 patients (of N = 1,063 all eligible) prescribed VKA was 32.8%, acetylsalicylic acid 41.7%, and no therapy 20.2%. In Europe in phase 2, treatment with NOAC was more common than VKA (52.3% and 37.8%, respectively); 6.0% of patients received antiplatelet treatment; and 3.8% received no antithrombotic treatment. In North America, 52.1%, 26.2%, and 14.0% of patients received NOAC, VKA, and antiplatelet drugs, respectively; 7.5% received no antithrombotic treatment. NOAC use was less common in Asia (27.7%), where 27.5% of patients received VKA, 25.0% antiplatelet drugs, and 19.8% no antithrombotic treatment. Conclusions The baseline data from GLORIA-AF phase 2 demonstrate that in newly diagnosed nonvalvular atrial fibrillation patients, NOAC have been highly adopted into practice, becoming more frequently prescribed than VKA in Europe and North America. Worldwide, however, a large proportion of patients remain undertreated, particularly in Asia and North America. (Global Registry on Long-Term Oral Antithrombotic Treatment in Patients With Atrial Fibrillation [GLORIA-AF]; NCT01468701)

Published
2017

43. A Mechanistic Study of Thioester Hydrolysis with Heavy Atom Kinetic Isotope Effects

Author

John F. Marlier, Emily J. Fogle, Lori I. Robins, Richard L. Redman, Anthony D. Stillman, and Matthew A. Denison

Subjects
chemistry.chemical_classification, Thiocholine, Molecular Structure, Sulfur Compounds, Hydrolysis, Organic Chemistry, Inorganic chemistry, Formylthiocholine, Thioester, Kinetic energy, Bond order, Kinetics, State structure, Isotopes, chemistry, Computational chemistry, Kinetic isotope effect, Atom
Abstract

The carbonyl-C, carbonyl-O, and leaving-S kinetic isotope effects (KIEs) were determined for the hydrolysis of formylthiocholine. Under acidic conditions, (13)k(obs) = 1.0312, (18)k(obs) = 0.997, and (34)k(obs) = 0.995; for neutral conditions, (13)k(obs) = 1.022, (18)k(obs) = 1.010, and (34)k(obs) = 0.996; and for alkaline conditions, (13)k(obs) = 1.0263, (18)k(obs) = 0.992, and (34)k(obs) = 1.000. The observed KIEs provided helpful insights into a qualitative description of the bond orders in the transition state structure.

Published
2015
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45. A kinetic and isotope effect investigation of the urease-catalyzed hydrolysis of hydroxyurea

Author

Marlier, John F., Robins, Lori I., Tucker, Kathryn A., Rawlings, Jill, Anderson, Mark A., and Cleland, W.W.

Subjects
Nitrogen -- Chemical properties, Ammonia -- Chemical properties, Hydrolases -- Structure, Hydrolases -- Chemical properties, Enzymes -- Structure, Enzymes -- Chemical properties, Biological sciences, Chemistry
Abstract

Kinetic isotope effects for both phases (rapid burst phase and slow plateau phase) of the urease-catalyzed hydrolysis of hydroxyurea were measured at pH 6.0 and 25 degree [Celsius] to understand the observed nitrogen isotope effects for the ammonia leaving group and hydroxylamine leaving group. The observed similarity of the magnitude of the carbon isotope effects argues for formation of a common intermediate during both phases.

Published
2010

46. Inactivation of Prions and Amyloid Seeds with Hypochlorous Acid

Author

Allison Kraus, Byron Caughey, Laura Sangaré, Bradley R. Groveman, Jeffrey Francis Williams, Luis Contreras, Katie Phillips, Brent Race, Andrew G. Hughson, Eri Saijo, Daniel S. Terry, Gianluigi Zanusso, Matteo Manca, Lori I. Robins, and Virkamal K. Dhaliwal

Subjects
0301 basic medicine, Bleach, Scrapie, Biochemistry, protein amyloid seeds, Animal Diseases, Prion Diseases, chemistry.chemical_compound, Zoonoses, Medicine and Health Sciences, Bioassay, prion decontamination, lcsh:QH301-705.5, Mammals, Hypochlorites, Hypochlorous acid, prions, prion inactivation, Neurodegenerative diseases, 3. Good health, Animal Prion Diseases, Chemistry, Infectious Diseases, Neurology, Sodium hypochlorite, Vertebrates, Physical Sciences, Metallurgy, Hamsters, Research Article, lcsh:Immunologic diseases. Allergy, Amyloid, Prions, Bovine spongiform encephalopathy, Immunology, Materials Science, Biology, Microbiology, Rodents, 03 medical and health sciences, In vivo, Virology, Genetics, medicine, Alloys, Animals, Molecular Biology, Organisms, Chemical Compounds, Biology and Life Sciences, Proteins, medicine.disease, Stainless Steel, Creutzfeldt-Jakob disease, 030104 developmental biology, lcsh:Biology (General), chemistry, Steel, Amniotes, Amyloid Proteins, Parasitology, Salts, lcsh:RC581-607, Zoology
Abstract

Hypochlorous acid (HOCl) is produced naturally by neutrophils and other cells to kill conventional microbes in vivo. Synthetic preparations containing HOCl can also be effective as microbial disinfectants. Here we have tested whether HOCl can also inactivate prions and other self-propagating protein amyloid seeds. Prions are deadly pathogens that are notoriously difficult to inactivate, and standard microbial disinfection protocols are often inadequate. Recommended treatments for prion decontamination include strongly basic (pH ≥~12) sodium hypochlorite bleach, ≥1 N sodium hydroxide, and/or prolonged autoclaving. These treatments are damaging and/or unsuitable for many clinical, agricultural and environmental applications. We have tested the anti-prion activity of a weakly acidic aqueous formulation of HOCl (BrioHOCl) that poses no apparent hazard to either users or many surfaces. For example, BrioHOCl can be applied directly to skin and mucous membranes and has been aerosolized to treat entire rooms without apparent deleterious effects. Here, we demonstrate that immersion in BrioHOCl can inactivate not only a range of target microbes, including spores of Bacillus subtilis, but also prions in tissue suspensions and on stainless steel. Real-time quaking-induced conversion (RT-QuIC) assays showed that BrioHOCl treatments eliminated all detectable prion seeding activity of human Creutzfeldt-Jakob disease, bovine spongiform encephalopathy, cervine chronic wasting disease, sheep scrapie and hamster scrapie; these findings indicated reductions of ≥103- to 106-fold. Transgenic mouse bioassays showed that all detectable hamster-adapted scrapie infectivity in brain homogenates or on steel wires was eliminated, representing reductions of ≥~105.75-fold and >104-fold, respectively. Inactivation of RT-QuIC seeding activity correlated with free chlorine concentration and higher order aggregation or destruction of proteins generally, including prion protein. BrioHOCl treatments had similar effects on amyloids composed of human α-synuclein and a fragment of human tau. These results indicate that HOCl can block the self-propagating activity of prions and other amyloids., Author Summary Many serious diseases have been linked to pathogenic states of various proteins. These naturally occurring proteins can be corrupted to form aggregates such as prions and amyloids that propagate in and between tissues by acting as seeds that convert the normal form of the protein into more of the pathological form. For example, corrupted prion protein can cause fatal transmissible neurodegenerative diseases such as Creutzfeldt-Jakob disease in humans, chronic wasting disease in cervids and bovine spongiform encephalopathy. Other amyloid-forming protein aggregates are pathogenic in Parkinson’s, Alzheimer’s, and other diseases. The fact that prions and amyloids are composed predominantly of tough, tightly packed proteins makes them unusually resistant to conventional microbial disinfection procedures. Infectious prions can persist indefinitely in, or on, a variety of materials such as tissues, fluids, tools, instruments, and environmental surfaces, making it important to identify decontaminants that are effective without being dangerous or damaging. Here we show that hypochlorous acid, a disinfectant that is produced naturally by certain cells within the body, has strong anti-prion and anti-amyloid activity. We find that a non-irritating and broadly applicable hypochlorous acid preparation can disinfect prions in tissue homogenates and on stainless steel wires serving as surrogates for surgical instruments.

Published
2016

47. Uridine-based inhibitors as new leads for antibiotics targeting Escherichia coli LpxC

Author

Barb, Adam W., Leavy, Tanya M., Robins, Lori I., Ziqiang Guan, Six, David A., Pei Zhou, Bertozzi, Carolyn R., and Raetz, Christian R. H.

Subjects
Drug resistance in microorganisms -- Analysis, Escherichia coli -- Genetic aspects, Escherichia coli -- Physiological aspects, Enzyme inhibitors -- Chemical properties, Enzyme inhibitors -- Structure, Uridine -- Chemical properties, Biological sciences, Chemistry
Abstract

Several studies are conducted for the identification of a series of uridine-based inhibitors that can be used in the antibiotics targeting Escherichia coli LpxC. The article proves that these new inhibitors are extremely beneficial, as they can be integrated in antibiotics for targeting the UDP binding pocket.

Published
2009

48. A facile synthesis of new 5H-indazolo[3,2-b]benzo[d]-1,3-oxazines via one-pot intramolecular bis-heterocyclizations

Author

Butler, Jeffery D., Solano, Danielle M., Robins, Lori I., Haddadin, Makhluf J., and Kurth, Mark J.

Subjects
Antiviral agents -- Usage, Nuclear magnetic resonance spectroscopy -- Usage, Oxazoles -- Chemical properties, Oxazoles -- Structure, Ring formation (Chemistry) -- Analysis, Biological sciences, Chemistry
Abstract

Intramolecular bis-heterocyclization reaction is used to synthesize 5H-indazolo-[3,2-b]benzo[d]-1,3-oxazine. The derivatives obtained by reacting with different substrates showed a wide range of biological activities.

Published
2008

49. Diazocinones: Synthesis and conformational analysis

Author

Robins, Lori I., Carpenter, Richard D., Fettinger, James C., Haddadin, Makhluf J., Tinti, Dino S., and Kurth, Mark J.

Subjects
Methanol -- Chemical properties, Conformational analysis, Biological sciences, Chemistry
Abstract

A general route to novel isoxazole-substituted dihydrodiazocinones is achieved, which proceeds by the condensation of tetrazines with cyclobutanones in methanolic KOH at room temperature. These eight-membered ring diaza heterocycles exist in relatively few, low energy, conformationally accessible states and experience limited conformational flexibility and conformational analysis confirms the twist-boat-chair (TBC) conformation as the major contributor to the thermodynamic isomer.

Published
2006

50. Structural Basis for the Sugar Nucleotide and Acyl-Chain Selectivity of Leptospira interrogans LpxA

Author

Christian R. H. Raetz, Lori I. Robins, and Allison H. Williams

Subjects
Models, Molecular, Protein Conformation, Stereochemistry, Crystallography, X-Ray, medicine.disease_cause, Biochemistry, Article, Protein Structure, Secondary, Substrate Specificity, Lipid A, chemistry.chemical_compound, Protein structure, Catalytic Domain, medicine, Transferase, Escherichia coli, Uridine Diphosphate N-Acetylglucosamine, biology, Fatty Acids, Lauric Acids, Active site, Hydrogen Bonding, biology.organism_classification, Recombinant Proteins, Kinetics, Uridine diphosphate N-acetylglucosamine, chemistry, Pantetheine, Biocatalysis, biology.protein, Leptospira interrogans, Oxyanion hole, Acyltransferases
Abstract

Leptospira interrogans, a spirochete that inhabits areas with warm, humid climates, thrives in fresh water sources contaminated by animal urine (1). L. interrogans infect humans who come in contact with the contaminated water or soils, causing Weil's disease, which damages the liver, kidneys, heart and lungs (1), and may be fatal if left untreated (2). L. interrogans is a spirochete that is able to survive outside of its host and one of the few that synthesizes lipopolysaccharide (LPS) (3, 4). Accordingly, all L. interrogans genomes encode orthologues of the Escherichia coli Lpx enzymes (3, 4), which are responsible for the assembly of the Kdo2-lipid A anchor of LPS (Fig. 1). Figure 1 Substrate selectivity of LiLpxA versus EcLpxA and the origin of UDP-GlcNAc3N Some Gram-negative bacteria, including L. interrogans, synthesize variants of lipid A in which a 2,3-diamino-2,3-dideoxy-d-glucopyranose (GlcN3N) unit replaces one or both glucosamine residues (5, 6). L. interrogans lipid A contains two such GlcN3N units (6) (Fig. 1A). The GlcN3N residues are derived from UDP-2-acetamido-3-amino-2,3-dideoxy-α-d-glucopyranose (UDP-GlcNAc3N) (Fig. 1A) (7, 8). This material is synthesized from UDP-GlcNAc by the sequential actions of the dehydrogenase GnnA and the transaminase GnnB; these proteins are not present in E. coli (Fig. 1A) (7, 8). L. interrogans LpxA (LiLpxA) transfers the R-3-hydroxylauroyl chain from R-3-hydroxylauroyl-ACP to UDP-GlcNAc3N (6, 7) (Fig. 1A). LiLpxA shows an absolute selectivity for UDP-GlcNAc3N (Fig. 1A) (7). EcLpxA acylates both UDP-GlcNAc and UDP-GlcNAc3N in vitro with equal efficiency (7) (not shown in Fig. 1A), but UDP-GlcNAc3N is not synthesized by wild-type E. coli. LiLpxA also shows a striking preference for R-3-hydroxylauroyl-ACP over other hydroxyacyl-ACPs (7) (Fig. 1A). E. coli LpxA (EcLpxA) is highly selective for R-3-hydroxymyristoyl-ACP (Fig. 1B), but it can also slowly utilize R-3-hydroxylauroyl- and R-3-hydroxypalmitoyl-ACP (7, 9, 10). Initial biochemical studies of EcLpxA suggested a mechanism in which H125 is the catalytic base (11) (Scheme 1). Recent structural studies have confirmed that the NE2 atom of H125 is within hydrogen-bonding distance of the 3-OH group of the glucosamine ring in complexes of EcLpxA with UDP-GlcNAc (12). These structures also elucidated the roles of the conserved EcLpxA residues Q73, L75, H99, H122, H144, Q161, N198, and R205 in substrate binding (Scheme 1) (12). The NH group of G143 is the proposed oxyanion hole (Scheme 1). The preference of EcLpxA for R-3-hydroxymyristoyl-ACP is explained by the positioning of G173 and H191 (12). The reaction catalyzed by EcLpxA with UDP-GlcNAc is reversible and thermodynamically unfavorable in the forward direction (11, 13). Scheme 1 Proposed catalytic mechanism of EcLpxA and roles of key residues in substrate binding. The structure of an LpxA orthologue selective for UDP-GlcNAc3N has not been investigated. Herein, we report the crystal structures of LiLpxA in the free form at 2.10 A, with bound UDP-3-N-(R-3-hydroxylauroyl)-GlcNAc3N at 2.10 A, and with bound R-3-hydroxylauroyl-methylphosphopantetheine at 2.12 A. The latter compound is an effective model substrate for LiLpxA but not EcLpxA. The space group is trigonal (P321). The asymmetric unit of LiLpxA contains three protein molecules, whereas that of EcLpxA contains only one (12, 14). However, the active site cleft of the biologically relevant LiLpxA homotrimer (which is not the same as three monomers in the asymmetric unit) is similar to that of EcLpxA (12), and its structure explains the R-3-hydroxylauroyl-ACP selectivity of LiLpxA. The altered position of the backbone carbonyl moiety of Q68 in LiLpxA provides a favorable H-bond acceptor for the NH2 moiety of UDP-GlcNAc3N but discriminates against the 3-OH group of UDP-GlcNAc. The UDP-GlcNAc3N selectivity of LiLpxA is reminiscent of the mechanism of bacterial resistance to vancomycin.

Published
2009
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