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Harshini Mukundan
Physical Chemistry and Applied Spectroscopy, Chemistry Division, Los Alamos National Laboratory, Los Alamos, NM 87545, USA

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Review
Published: 12 May 2021 in Toxins
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Macrolides are a diverse class of hydrophobic compounds characterized by a macrocyclic lactone ring and distinguished by variable side chains/groups. Some of the most well characterized macrolides are toxins produced by marine bacteria, sea sponges, and other species. Many marine macrolide toxins act as biomimetic molecules to natural actin-binding proteins, affecting actin polymerization, while other toxins act on different cytoskeletal components. The disruption of natural cytoskeletal processes affects cell motility and cytokinesis, and can result in cellular death. While many macrolides are toxic in nature, others have been shown to display therapeutic properties. Indeed, some of the most well known antibiotic compounds, including erythromycin, are macrolides. In addition to antibiotic properties, macrolides have been shown to display antiviral, antiparasitic, antifungal, and immunosuppressive actions. Here, we review each functional class of macrolides for their common structures, mechanisms of action, pharmacology, and human cellular targets.

ACS Style

Kiersten Lenz; Katja Klosterman; Harshini Mukundan; Jessica Kubicek-Sutherland. Macrolides: From Toxins to Therapeutics. Toxins 2021, 13, 347 .

AMA Style

Kiersten Lenz, Katja Klosterman, Harshini Mukundan, Jessica Kubicek-Sutherland. Macrolides: From Toxins to Therapeutics. Toxins. 2021; 13 (5):347.

Chicago/Turabian Style

Kiersten Lenz; Katja Klosterman; Harshini Mukundan; Jessica Kubicek-Sutherland. 2021. "Macrolides: From Toxins to Therapeutics." Toxins 13, no. 5: 347.

Technical report
Published: 01 May 2021 in Agnostic Immunity
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Harshini Mukundan; Univ. Of New Mexico; Johns Hopkins Univ.; Univ. Of Nebraska; Medical Univ. Of South Carolina. Agnostic Immunity [Slides]. Agnostic Immunity 2021, 1 .

AMA Style

Harshini Mukundan, Univ. Of New Mexico, Johns Hopkins Univ., Univ. Of Nebraska, Medical Univ. Of South Carolina. Agnostic Immunity [Slides]. Agnostic Immunity. 2021; ():1.

Chicago/Turabian Style

Harshini Mukundan; Univ. Of New Mexico; Johns Hopkins Univ.; Univ. Of Nebraska; Medical Univ. Of South Carolina. 2021. "Agnostic Immunity [Slides]." Agnostic Immunity , no. : 1.

Research article
Published: 07 April 2021 in PLOS ONE
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Lipoarabinomannan (LAM), an amphiphilic lipoglycan of the Mycobacterium tuberculosis cell wall, is a diagnostic target for tuberculosis. Previous work from our laboratory and others suggests that LAM is associated with host serum lipoproteins, which may in turn have implications for diagnostic assays. Our team has developed two serum assays for amphiphile detection: lipoprotein capture and membrane insertion. The lipoprotein capture assay relies on capture of the host lipoproteins, exploiting the biological association of host lipoprotein with microbial amphiphilic biomarkers to “concentrate” LAM. In contrast, the membrane insertion assay is independent of the association between pathogen amphiphiles and host lipoprotein association, and directly captures LAM based on its thermodynamic propensity for association with a supported lipid membrane, which forms the functional surface of an optical biosensor. In this manuscript, we explored the use of these assays for the detection of LAM in sera from adults whose tuberculosis status had been well-characterized using conventional microbiological tests, and endemic controls. Using the lipoprotein capture assay, LAM signal/noise ratios were >1.0 in 29/35 (83%) individuals with culture-confirmed active tuberculosis, 8/13 (62%) individuals with tuberculosis symptoms, but no positive culture for M. tuberculosis, and 0/6 (0%) symptom-free endemic controls. To evaluate serum LAM levels without bias associated with potential differences in circulating host lipoprotein concentrations between individuals, we subsequently processed available samples to liberate LAM from associated host lipoprotein assemblies followed by direct detection of the pathogen biomarker using the membrane insertion approach. Using the membrane insertion assay, signal/noise for detection of serum LAM was greater than that observed using the lipoprotein capture method for culture-confirmed TB patients (6/6), yet remained negative for controls (2/2). Taken together, these results suggest that detection of serum LAM is a promising TB diagnostic approach, but that further work is required to optimize assay performance and to decipher the implications of LAM/host lipoprotein associations for diagnostic assay performance and TB pathogenesis.

ACS Style

Shailja Jakhar; Ramamurthy Sakamuri; Dung Vu; Priya Dighe; Loreen R. Stromberg; Laura Lilley; Nicolas Hengartner; Basil I. Swanson; Emmanuel Moreau; Susan E. Dorman; Harshini Mukundan. Interaction of amphiphilic lipoarabinomannan with host carrier lipoproteins in tuberculosis patients: Implications for blood-based diagnostics. PLOS ONE 2021, 16, e0243337 .

AMA Style

Shailja Jakhar, Ramamurthy Sakamuri, Dung Vu, Priya Dighe, Loreen R. Stromberg, Laura Lilley, Nicolas Hengartner, Basil I. Swanson, Emmanuel Moreau, Susan E. Dorman, Harshini Mukundan. Interaction of amphiphilic lipoarabinomannan with host carrier lipoproteins in tuberculosis patients: Implications for blood-based diagnostics. PLOS ONE. 2021; 16 (4):e0243337.

Chicago/Turabian Style

Shailja Jakhar; Ramamurthy Sakamuri; Dung Vu; Priya Dighe; Loreen R. Stromberg; Laura Lilley; Nicolas Hengartner; Basil I. Swanson; Emmanuel Moreau; Susan E. Dorman; Harshini Mukundan. 2021. "Interaction of amphiphilic lipoarabinomannan with host carrier lipoproteins in tuberculosis patients: Implications for blood-based diagnostics." PLOS ONE 16, no. 4: e0243337.

Journal article
Published: 05 March 2021 in Scientific Reports
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The separation of biomarkers from blood is straightforward in most molecular biology laboratories. However, separation in resource-limited settings, allowing for the successful removal of biomarkers for diagnostic applications, is not always possible. The situation is further complicated by the need to separate hydrophobic signatures such as lipids from blood. Herein, we present a microfluidic device capable of centrifugal separation of serum from blood at the point of need with a system that is compatible with biomarkers that are both hydrophilic and hydrophobic. The cross-flow filtration device separates serum from blood as efficiently as traditional methods and retains amphiphilic biomarkers in serum for detection.

ACS Style

Kiersten D. Lenz; Shailja Jakhar; Jing W. Chen; Aaron S. Anderson; Dylan C. Purcell; Mohammad O. Ishak; Jennifer F. Harris; Leyla E. Akhadov; Jessica Z. Kubicek-Sutherland; Pulak Nath; Harshini Mukundan. A centrifugal microfluidic cross-flow filtration platform to separate serum from whole blood for the detection of amphiphilic biomarkers. Scientific Reports 2021, 11, 1 -8.

AMA Style

Kiersten D. Lenz, Shailja Jakhar, Jing W. Chen, Aaron S. Anderson, Dylan C. Purcell, Mohammad O. Ishak, Jennifer F. Harris, Leyla E. Akhadov, Jessica Z. Kubicek-Sutherland, Pulak Nath, Harshini Mukundan. A centrifugal microfluidic cross-flow filtration platform to separate serum from whole blood for the detection of amphiphilic biomarkers. Scientific Reports. 2021; 11 (1):1-8.

Chicago/Turabian Style

Kiersten D. Lenz; Shailja Jakhar; Jing W. Chen; Aaron S. Anderson; Dylan C. Purcell; Mohammad O. Ishak; Jennifer F. Harris; Leyla E. Akhadov; Jessica Z. Kubicek-Sutherland; Pulak Nath; Harshini Mukundan. 2021. "A centrifugal microfluidic cross-flow filtration platform to separate serum from whole blood for the detection of amphiphilic biomarkers." Scientific Reports 11, no. 1: 1-8.

Research article
Published: 01 February 2021 in PLOS Neglected Tropical Diseases
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Non-typhoidal Salmonella (NTS) is a major global health concern that often causes bloodstream infections in areas of the world affected by malnutrition and comorbidities such as HIV and malaria. Developing a strategy to control the emergence and spread of highly invasive and antimicrobial resistant NTS isolates requires a comprehensive analysis of epidemiological factors and molecular pathogenesis. Here, we characterize 11 NTS isolates that caused bloodstream infections in pediatric patients in Siaya, Kenya from 2003–2010. Nine isolates were identified as S. Typhimurium sequence type 313 while the other two were S. Enteritidis. Comprehensive genotypic and phenotypic analyses were performed to compare these isolates to those previously identified in sub-Saharan Africa. We identified a S. Typhimurium isolate referred to as UGA14 that displayed novel plasmid, pseudogene and resistance features as compared to other isolates reported from Africa. Notably, UGA14 is able to ferment both lactose and sucrose due to the acquisition of insertion elements on the pKST313 plasmid. These findings show for the first time the co-evolution of plasmid-mediated lactose and sucrose metabolism along with cephalosporin resistance in NTS further elucidating the evolutionary mechanisms of invasive NTS phenotypes. These results further support the use of combined genomic and phenotypic approaches to detect and characterize atypical NTS isolates in order to advance biosurveillance efforts that inform countermeasures aimed at controlling invasive and antimicrobial resistant NTS.

ACS Style

Jessica Z. Kubicek-Sutherland; Gary Xie; Migun Shakya; Priya K. Dighe; Lindsey L. Jacobs; Hajnalka Daligault; Karen Davenport; Loreen R. Stromberg; Zachary R. Stromberg; Qiuying Cheng; Prakasha Kempaiah; John Michael Ong’Echa; Vincent Otieno; Evans Raballah; Samuel Anyona; Collins Ouma; Patrick S. G. Chain; Douglas J. Perkins; Harshini Mukundan; Benjamin H. McMahon; Norman A. Doggett. Comparative genomic and phenotypic characterization of invasive non-typhoidal Salmonella isolates from Siaya, Kenya. PLOS Neglected Tropical Diseases 2021, 15, e0008991 .

AMA Style

Jessica Z. Kubicek-Sutherland, Gary Xie, Migun Shakya, Priya K. Dighe, Lindsey L. Jacobs, Hajnalka Daligault, Karen Davenport, Loreen R. Stromberg, Zachary R. Stromberg, Qiuying Cheng, Prakasha Kempaiah, John Michael Ong’Echa, Vincent Otieno, Evans Raballah, Samuel Anyona, Collins Ouma, Patrick S. G. Chain, Douglas J. Perkins, Harshini Mukundan, Benjamin H. McMahon, Norman A. Doggett. Comparative genomic and phenotypic characterization of invasive non-typhoidal Salmonella isolates from Siaya, Kenya. PLOS Neglected Tropical Diseases. 2021; 15 (2):e0008991.

Chicago/Turabian Style

Jessica Z. Kubicek-Sutherland; Gary Xie; Migun Shakya; Priya K. Dighe; Lindsey L. Jacobs; Hajnalka Daligault; Karen Davenport; Loreen R. Stromberg; Zachary R. Stromberg; Qiuying Cheng; Prakasha Kempaiah; John Michael Ong’Echa; Vincent Otieno; Evans Raballah; Samuel Anyona; Collins Ouma; Patrick S. G. Chain; Douglas J. Perkins; Harshini Mukundan; Benjamin H. McMahon; Norman A. Doggett. 2021. "Comparative genomic and phenotypic characterization of invasive non-typhoidal Salmonella isolates from Siaya, Kenya." PLOS Neglected Tropical Diseases 15, no. 2: e0008991.

Preprint content
Published: 20 November 2020
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Lipoarabinomannan (LAM), an amphiphilic lipoglycan of the Mycobacterium tuberculosis cell wall, is a diagnostic target for tuberculosis. Previous work from our laboratory and others suggests that LAM is associated with host serum lipoproteins, which may in turn have implications for diagnostic assays. Our team has developed two serum assays for amphiphile detection: lipoprotein capture and membrane insertion. The lipoprotein capture assay relies on capture of the host lipoproteins, exploiting the biological association of host lipoprotein with microbial amphiphilic biomarkers to “concentrate” LAM. In contrast, the membrane insertion assay is independent of the association between pathogen amphiphiles and host lipoprotein association, and directly captures LAM based on its thermodynamic propensity for association with a supported lipid membrane, which forms the functional surface of an optical biosensor. In this manuscript, we explored the use of these assays for the detection of LAM in sera from adults whose tuberculosis status had been well-characterized using conventional microbiological tests, and endemic controls. Using the lipoprotein capture assay, LAM signal/noise ratios were >1.0 in 29/35 (83%) individuals with culture-confirmed active tuberculosis, 8/13 (62%) individuals with tuberculosis symptoms but no positive culture for M. tuberculosis, and 0/6 (0%) symptom-free endemic controls. To evaluate serum LAM levels without bias associated with potential differences in circulating host lipoprotein concentrations between individuals, we subsequently processed available samples to liberate LAM from associated host lipoprotein assemblies followed by direct detection of the pathogen biomarker using the membrane insertion approach. Using the membrane insertion assay, signal/noise for detection of serum LAM was greater than that observed using the lipoprotein capture method for culture-confirmed TB patients (6/6), yet remained negative for controls (2/2). Taken together, these results suggest that detection of serum LAM is a promising TB diagnostic approach. Further work is required to optimize assay performance and to decipher the implications of LAM/host lipoprotein associations for diagnostic assay performance and TB pathogenesis.

ACS Style

Shailja Jakhar; Ramamurthy Sakamuri; Dung Vu; Priya Dighe; Loreen R. Stromberg; Laura Lilley; Nicolas Hengartner; Basil I. Swanson; Emmanuel Moreau; Susan E. Dorman; Harshini Mukundan. Interaction of Amphiphilic Lipoarabinomannan with Host Carrier Lipoproteins in Tuberculosis Patients: Implications for Blood-based Diagnostics. 2020, 1 .

AMA Style

Shailja Jakhar, Ramamurthy Sakamuri, Dung Vu, Priya Dighe, Loreen R. Stromberg, Laura Lilley, Nicolas Hengartner, Basil I. Swanson, Emmanuel Moreau, Susan E. Dorman, Harshini Mukundan. Interaction of Amphiphilic Lipoarabinomannan with Host Carrier Lipoproteins in Tuberculosis Patients: Implications for Blood-based Diagnostics. . 2020; ():1.

Chicago/Turabian Style

Shailja Jakhar; Ramamurthy Sakamuri; Dung Vu; Priya Dighe; Loreen R. Stromberg; Laura Lilley; Nicolas Hengartner; Basil I. Swanson; Emmanuel Moreau; Susan E. Dorman; Harshini Mukundan. 2020. "Interaction of Amphiphilic Lipoarabinomannan with Host Carrier Lipoproteins in Tuberculosis Patients: Implications for Blood-based Diagnostics." , no. : 1.

Review
Published: 23 September 2020 in International Journal of Molecular Sciences
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Tuberculosis (TB) is a major public health concern for all ages. However, the disease presents a larger challenge in pediatric populations, partially owing to the lack of reliable diagnostic standards for the early identification of infection. Currently, there are no biomarkers that have been clinically validated for use in pediatric TB diagnosis. Identification and validation of biomarkers could provide critical information on prognosis of disease, and response to treatment. In this review, we discuss how the “omics” approach has influenced biomarker discovery and the advancement of a next generation rapid point-of-care diagnostic for TB, with special emphasis on pediatric disease. Limitations of current published studies and the barriers to their implementation into the field will be thoroughly reviewed within this article in hopes of highlighting future avenues and needs for combating the problem of pediatric tuberculosis.

ACS Style

Shailja Jakhar; Alexis A. Bitzer; Loreen R. Stromberg; Harshini Mukundan. Pediatric Tuberculosis: The Impact of “Omics” on Diagnostics Development. International Journal of Molecular Sciences 2020, 21, 6979 .

AMA Style

Shailja Jakhar, Alexis A. Bitzer, Loreen R. Stromberg, Harshini Mukundan. Pediatric Tuberculosis: The Impact of “Omics” on Diagnostics Development. International Journal of Molecular Sciences. 2020; 21 (19):6979.

Chicago/Turabian Style

Shailja Jakhar; Alexis A. Bitzer; Loreen R. Stromberg; Harshini Mukundan. 2020. "Pediatric Tuberculosis: The Impact of “Omics” on Diagnostics Development." International Journal of Molecular Sciences 21, no. 19: 6979.

Technical report
Published: 24 January 2020 in UNIVERSAL BACTERIAL SENSOR
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Harshini Mukundan. UNIVERSAL BACTERIAL SENSOR. UNIVERSAL BACTERIAL SENSOR 2020, 1 .

AMA Style

Harshini Mukundan. UNIVERSAL BACTERIAL SENSOR. UNIVERSAL BACTERIAL SENSOR. 2020; ():1.

Chicago/Turabian Style

Harshini Mukundan. 2020. "UNIVERSAL BACTERIAL SENSOR." UNIVERSAL BACTERIAL SENSOR , no. : 1.

Journal article
Published: 02 August 2019 in Scientific Reports
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Invasive non-typhoidal Salmonella (NTS) is among the leading causes of blood stream infections in sub-Saharan Africa and other developing regions, especially among pediatric populations. Invasive NTS can be difficult to treat and have high case-fatality rates, in part due to emergence of strains resistant to broad-spectrum antibiotics. Furthermore, improper treatment contributes to increased antibiotic resistance and death. Point of care (POC) diagnostic tests that rapidly identify invasive NTS infection, and differentiate between resistant and non-resistant strains, may greatly improve patient outcomes and decrease resistance at the community level. Here we present for the first time a model for NTS dynamics in high risk populations that can analyze the potential advantages and disadvantages of four strategies involving POC diagnostic deployment, and the resulting impact on antimicrobial treatment for patients. Our analysis strongly supports the use of POC diagnostics coupled with targeted antibiotic use for patients upon arrival in the clinic for optimal patient and public health outcomes. We show that even the use of imperfect POC diagnostics can significantly reduce total costs and number of deaths, provided that the diagnostic gives results quickly enough that patients are likely to return or stay to receive targeted treatment.

ACS Style

Carrie Manore; Todd Graham; Alexa Carr; Alicia Feryn; Shailja Jakhar; Harshini Mukundan; Hannah Callender Highlander. Modeling and Cost Benefit Analysis to Guide Deployment of POC Diagnostics for Non-typhoidal Salmonella Infections with Antimicrobial Resistance. Scientific Reports 2019, 9, 1 -17.

AMA Style

Carrie Manore, Todd Graham, Alexa Carr, Alicia Feryn, Shailja Jakhar, Harshini Mukundan, Hannah Callender Highlander. Modeling and Cost Benefit Analysis to Guide Deployment of POC Diagnostics for Non-typhoidal Salmonella Infections with Antimicrobial Resistance. Scientific Reports. 2019; 9 (1):1-17.

Chicago/Turabian Style

Carrie Manore; Todd Graham; Alexa Carr; Alicia Feryn; Shailja Jakhar; Harshini Mukundan; Hannah Callender Highlander. 2019. "Modeling and Cost Benefit Analysis to Guide Deployment of POC Diagnostics for Non-typhoidal Salmonella Infections with Antimicrobial Resistance." Scientific Reports 9, no. 1: 1-17.

Brief report
Published: 18 April 2019 in Microbiology Resource Announcements
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We report here the genome sequence of a Staphylococcus xylosus clinical isolate, strain SMA0341-04 (UGA5), which contains one chromosome and at least one plasmid. Notably, strain SMA0341-04 (UGA5) contains the tetracycline efflux major facilitator superfamily (MFS) transporter ( tetK ) gene.

ACS Style

Qiuying Cheng; Gary Xie; Hajnalka Daligault; Karen Davenport; Cheryl Gleasner; Lindsey Jacobs; Jessica Kubicek-Sutherland; Tessa LeCuyer; Vincent Otieno; Evans Raballah; Norman Doggett; Benjamin McMahon; Douglas J. Perkins; Harshini Mukundan. Genome Sequence of a Staphylococcus xylosus Clinical Isolate, Strain SMA0341-04 (UGA5), from Siaya County Referral Hospital in Siaya, Kenya. Microbiology Resource Announcements 2019, 8, e01625-18 .

AMA Style

Qiuying Cheng, Gary Xie, Hajnalka Daligault, Karen Davenport, Cheryl Gleasner, Lindsey Jacobs, Jessica Kubicek-Sutherland, Tessa LeCuyer, Vincent Otieno, Evans Raballah, Norman Doggett, Benjamin McMahon, Douglas J. Perkins, Harshini Mukundan. Genome Sequence of a Staphylococcus xylosus Clinical Isolate, Strain SMA0341-04 (UGA5), from Siaya County Referral Hospital in Siaya, Kenya. Microbiology Resource Announcements. 2019; 8 (16):e01625-18.

Chicago/Turabian Style

Qiuying Cheng; Gary Xie; Hajnalka Daligault; Karen Davenport; Cheryl Gleasner; Lindsey Jacobs; Jessica Kubicek-Sutherland; Tessa LeCuyer; Vincent Otieno; Evans Raballah; Norman Doggett; Benjamin McMahon; Douglas J. Perkins; Harshini Mukundan. 2019. "Genome Sequence of a Staphylococcus xylosus Clinical Isolate, Strain SMA0341-04 (UGA5), from Siaya County Referral Hospital in Siaya, Kenya." Microbiology Resource Announcements 8, no. 16: e01625-18.

Journal article
Published: 17 April 2019 in Scientific Reports
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Bacteremia is a leading cause of death in sub-Saharan Africa where childhood mortality rates are the highest in the world. The early diagnosis of bacteremia and initiation of treatment saves lives, especially in high-disease burden areas. However, diagnosing bacteremia is challenging for clinicians, especially in children presenting with co-infections such as malaria and HIV. There is an urgent need for a rapid method for detecting bacteremia in pediatric patients with co-morbidities to inform treatment. In this manuscript, we have developed and clinically validated a novel method for the direct detection of amphiphilic pathogen biomarkers indicative of bacteremia, directly in aqueous blood, by mimicking innate immune recognition. Specifically, we have exploited the interaction of amphiphilic pathogen biomarkers such as lipopolysaccharides (LPS) from Gram-negative bacteria and lipoteichoic acids (LTA) from Gram-positive bacteria with host lipoprotein carriers in blood, in order to develop two tailored assays – lipoprotein capture and membrane insertion – for their direct detection. Our assays demonstrate a sensitivity of detection of 4 ng/mL for LPS and 2 ng/mL for LTA using a waveguide-based optical biosensor platform that was developed at LANL. In this manuscript, we also demonstrate the application of these methods for the detection of LPS in serum from pediatric patients with invasive Salmonella Typhimurium bacteremia (n = 7) and those with Staphylococcal bacteremia (n = 7) with 100% correlation with confirmatory culture. Taken together, these results demonstrate the significance of biochemistry in both our understanding of host-pathogen biology, and development of assay methodology, as well as demonstrate a potential new approach for the rapid, sensitive and accurate diagnosis of bacteremia at the point of need.

ACS Style

Jessica Z. Kubicek-Sutherland; Dung M. Vu; Aneesa Noormohamed; Heather M. Mendez; Loreen R. Stromberg; Christine A. Pedersen; Astrid Hengartner; Katja E. Klosterman; Haley A. Bridgewater; Vincent Otieno; Qiuying Cheng; Samuel B. Anyona; Collins Ouma; Evans Raballah; Douglas J. Perkins; Benjamin H. McMahon; Harshini Mukundan. Direct detection of bacteremia by exploiting host-pathogen interactions of lipoteichoic acid and lipopolysaccharide. Scientific Reports 2019, 9, 1 -14.

AMA Style

Jessica Z. Kubicek-Sutherland, Dung M. Vu, Aneesa Noormohamed, Heather M. Mendez, Loreen R. Stromberg, Christine A. Pedersen, Astrid Hengartner, Katja E. Klosterman, Haley A. Bridgewater, Vincent Otieno, Qiuying Cheng, Samuel B. Anyona, Collins Ouma, Evans Raballah, Douglas J. Perkins, Benjamin H. McMahon, Harshini Mukundan. Direct detection of bacteremia by exploiting host-pathogen interactions of lipoteichoic acid and lipopolysaccharide. Scientific Reports. 2019; 9 (1):1-14.

Chicago/Turabian Style

Jessica Z. Kubicek-Sutherland; Dung M. Vu; Aneesa Noormohamed; Heather M. Mendez; Loreen R. Stromberg; Christine A. Pedersen; Astrid Hengartner; Katja E. Klosterman; Haley A. Bridgewater; Vincent Otieno; Qiuying Cheng; Samuel B. Anyona; Collins Ouma; Evans Raballah; Douglas J. Perkins; Benjamin H. McMahon; Harshini Mukundan. 2019. "Direct detection of bacteremia by exploiting host-pathogen interactions of lipoteichoic acid and lipopolysaccharide." Scientific Reports 9, no. 1: 1-14.

Journal article
Published: 04 April 2019 in Toxins
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Mycolactone, the amphiphilic macrolide toxin secreted by Mycobacterium ulcerans, plays a significant role in the pathology and manifestations of Buruli ulcer (BU). Consequently, it follows that the toxin is a suitable target for the development of diagnostics and therapeutics for this disease. Yet, several challenges have deterred such development. For one, the lipophilic nature of the toxin makes it difficult to handle and store and contributes to variability associated with laboratory experimentation and purification yields. In this manuscript, we have attempted to incorporate our understanding of the lipophilicity of mycolactone in order to define the optimal methods for the storage, handling, and purification of this toxin. We present a systematic correlation of variability associated with measurement techniques (thin-layer chromatography (TLC), mass spectrometry (MS), and UV-Vis spectrometry), storage conditions, choice of solvents, as well as the impact of each of these on toxin function as assessed by cellular cytotoxicity. We also compared natural mycolactone extracted from bacterial culture with synthesized toxins in laboratory (solvents, buffers) and physiologically relevant (serum) matrices. Our results point to the greater stability of mycolactone in organic, as well as detergent-containing, solvents, regardless of the container material (plastic, glass, or silanized tubes). They also highlight the presence of toxin in samples that may be undetectable by any one technique, suggesting that each detection approach captures different configurations of the molecule with varying specificity and sensitivity. Most importantly, our results demonstrate for the very first time that amphiphilic mycolactone associates with host lipoproteins in serum, and that this association will likely impact our ability to study, diagnose, and treat Buruli ulcers in patients.

ACS Style

Jessica Z. Kubicek-Sutherland; Dung M. Vu; Aaron S. Anderson; Timothy C. Sanchez; Paul J. Converse; Ricardo Martí-Arbona; Eric L. Nuermberger; Basil I. Swanson; Harshini Mukundan. Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone. Toxins 2019, 11, 202 .

AMA Style

Jessica Z. Kubicek-Sutherland, Dung M. Vu, Aaron S. Anderson, Timothy C. Sanchez, Paul J. Converse, Ricardo Martí-Arbona, Eric L. Nuermberger, Basil I. Swanson, Harshini Mukundan. Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone. Toxins. 2019; 11 (4):202.

Chicago/Turabian Style

Jessica Z. Kubicek-Sutherland; Dung M. Vu; Aaron S. Anderson; Timothy C. Sanchez; Paul J. Converse; Ricardo Martí-Arbona; Eric L. Nuermberger; Basil I. Swanson; Harshini Mukundan. 2019. "Understanding the Significance of Biochemistry in the Storage, Handling, Purification, and Sampling of Amphiphilic Mycolactone." Toxins 11, no. 4: 202.

Technical report
Published: 09 November 2018 in The Microbe Strikes Back: Emerging infectious Diseases and the need for point of care diagnostics
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ACS Style

Harshini Mukundan. The Microbe Strikes Back: Emerging infectious Diseases and the need for point of care diagnostics. The Microbe Strikes Back: Emerging infectious Diseases and the need for point of care diagnostics 2018, 1 .

AMA Style

Harshini Mukundan. The Microbe Strikes Back: Emerging infectious Diseases and the need for point of care diagnostics. The Microbe Strikes Back: Emerging infectious Diseases and the need for point of care diagnostics. 2018; ():1.

Chicago/Turabian Style

Harshini Mukundan. 2018. "The Microbe Strikes Back: Emerging infectious Diseases and the need for point of care diagnostics." The Microbe Strikes Back: Emerging infectious Diseases and the need for point of care diagnostics , no. : 1.

Preprint
Published: 06 August 2018
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Invasive non-typhoidal Salmonella (NTS) is among the leading causes of blood stream infections in sub-Saharan Africa and other developing regions, especially among pediatric populations. Invasive NTS can be difficult to treat and have high case-fatality rates, in part due to emergence of strains resistant to broad-spectrum antibiotics. Furthermore, improper treatment contributes to increased antibiotic resistance and death. Point of care (POC) diagnostic tests that rapidly identify invasive NTS infection, and differentiate between resistant and non-resistant strains, may greatly improve patient outcomes and decrease resistance at the community level. Here we present for the first time a model for NTS dynamics in high risk populations that can analyze the potential advantages and disadvantages of four strategies involving POC diagnostic deployment, and the resulting impact on antimicrobial treatment for patients. Our analysis strongly supports the use of POC diagnostics coupled with targeted antibiotic use for patients upon arrival in the clinic for optimal patient and public health outcomes. We show that even the use of imperfect POC diagnostics can significantly reduce total costs and number of deaths, provided that the diagnostic gives results quickly enough that patients are likely to return or stay to receive targeted treatment.

ACS Style

Carrie Manore; Todd Graham; Alexa Carr; Alicia Feryn; Shailja Jakhar; Harshini Mukundan; Hannah Callender Highlander; Hannah Callender. Modeling and Cost Benefit Analysis to Guide Deployment of POC Diagnostics for Non-typhoidal Salmonella Infections with Antimicrobial Resistance. 2018, 384933 .

AMA Style

Carrie Manore, Todd Graham, Alexa Carr, Alicia Feryn, Shailja Jakhar, Harshini Mukundan, Hannah Callender Highlander, Hannah Callender. Modeling and Cost Benefit Analysis to Guide Deployment of POC Diagnostics for Non-typhoidal Salmonella Infections with Antimicrobial Resistance. . 2018; ():384933.

Chicago/Turabian Style

Carrie Manore; Todd Graham; Alexa Carr; Alicia Feryn; Shailja Jakhar; Harshini Mukundan; Hannah Callender Highlander; Hannah Callender. 2018. "Modeling and Cost Benefit Analysis to Guide Deployment of POC Diagnostics for Non-typhoidal Salmonella Infections with Antimicrobial Resistance." , no. : 384933.

Research article
Published: 14 June 2018 in PLOS ONE
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Recognition of Pathogen-associated Molecular Patterns (PAMPs) by Toll-like receptors is central to innate immunity. Many bacterial PAMPs such as lipopolysaccharide (LPS) and lipoteichoic acid have amphiphilic properties. The hydrophobicity of amphiphilic PAMPs contributes to increasing entropy and causes these molecules to self-aggregate or bind host carrier proteins in aqueous physiological environments. The goal of this work was to determine how innate immune signaling is impacted by physical presentation and association of amphiphilic PAMPs with serum carrier proteins, using LPS as an example molecule. Specifically, we measured LPS-induced cytokine profiles in murine macrophages when the antigen was presented associated with the various serum carrier proteins in serum versus a serum-depleted system. Our study demonstrates that the observed cytokine profiles are dramatically different when LPS is presented in buffer, versus in serum when it is associated with proteins, specifically with respect to inhibition of pro-inflammatory cytokines in the latter. These studies suggest that LPS-mediated cytokine expression is dependent on its presentation in physiological systems. The amphiphilicity of bacterial PAMPs and consequent association with lipoproteins is a feature, which should be taken into account in the design of in vitro experiments. Further studies of the interdependencies of different serum carriers can identify pathways for drug delivery and diagnostics.

ACS Style

Loreen R. Stromberg; Heather M. Mendez; Jessica Z. Kubicek-Sutherland; Steven W. Graves; Nicolas W. Hengartner; Harshini Mukundan. Presentation matters: Impact of association of amphiphilic LPS with serum carrier proteins on innate immune signaling. PLOS ONE 2018, 13, e0198531 .

AMA Style

Loreen R. Stromberg, Heather M. Mendez, Jessica Z. Kubicek-Sutherland, Steven W. Graves, Nicolas W. Hengartner, Harshini Mukundan. Presentation matters: Impact of association of amphiphilic LPS with serum carrier proteins on innate immune signaling. PLOS ONE. 2018; 13 (6):e0198531.

Chicago/Turabian Style

Loreen R. Stromberg; Heather M. Mendez; Jessica Z. Kubicek-Sutherland; Steven W. Graves; Nicolas W. Hengartner; Harshini Mukundan. 2018. "Presentation matters: Impact of association of amphiphilic LPS with serum carrier proteins on innate immune signaling." PLOS ONE 13, no. 6: e0198531.

Review
Published: 30 May 2018 in BMC Infectious Diseases
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Background: Emerging pathogens such as Zika, chikungunya, Ebola, and dengue viruses are serious threats to national and global health security. Accurate forecasts of emerging epidemics and their severity are critical to minimizing subsequent mortality, morbidity, and economic loss. The recent introduction of chikungunya and Zika virus to the Americas underscores the need for better methods for disease surveillance and forecasting. Methods: To explore the suitability of current approaches to forecasting emerging diseases, the Defense Advanced Research Projects Agency (DARPA) launched the 2014–2015 DARPA Chikungunya Challenge to forecast the number of cases and spread of chikungunya disease in the Americas. Challenge participants (n=38 during final evaluation) provided predictions of chikungunya epidemics across the Americas for a six-month period, from September 1, 2014 to February 16, 2015, to be evaluated by comparison with incidence data reported to the Pan American Health Organization (PAHO). This manuscript presents an overview of the challenge and a summary of the approaches used by the winners. Results: Participant submissions were evaluated by a team of non-competing government subject matter experts based on numerical accuracy and methodology. Although this manuscript does not include in-depth analyses of the results, cursory analyses suggest that simpler models appear to outperform more complex approaches that included, for example, demographic information and transportation dynamics, due to the reporting biases, which can be implicitly captured in statistical models. Mosquito-dynamics, population specific information, and dengue-specific information correlated best with prediction accuracy. Conclusion: We conclude that with careful consideration and understanding of the relative advantages and disadvantages of particular methods, implementation of an effective prediction system is feasible. However, there is a need to improve the quality of the data in order to more accurately predict the course of epidemics.

ACS Style

Sara Y. Del Valle; Benjamin H. McMahon; Jason Asher; Richard Hatchett; Joceline C. Lega; Heidi E. Brown; Mark E. Leany; Yannis Pantazis; David J. Roberts; Sean Moore; A Townsend Peterson; Luis E. Escobar; Huijie Qiao; Nicholas W. Hengartner; Harshini Mukundan. Summary results of the 2014-2015 DARPA Chikungunya challenge. BMC Infectious Diseases 2018, 18, 1 -14.

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Sara Y. Del Valle, Benjamin H. McMahon, Jason Asher, Richard Hatchett, Joceline C. Lega, Heidi E. Brown, Mark E. Leany, Yannis Pantazis, David J. Roberts, Sean Moore, A Townsend Peterson, Luis E. Escobar, Huijie Qiao, Nicholas W. Hengartner, Harshini Mukundan. Summary results of the 2014-2015 DARPA Chikungunya challenge. BMC Infectious Diseases. 2018; 18 (1):1-14.

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Sara Y. Del Valle; Benjamin H. McMahon; Jason Asher; Richard Hatchett; Joceline C. Lega; Heidi E. Brown; Mark E. Leany; Yannis Pantazis; David J. Roberts; Sean Moore; A Townsend Peterson; Luis E. Escobar; Huijie Qiao; Nicholas W. Hengartner; Harshini Mukundan. 2018. "Summary results of the 2014-2015 DARPA Chikungunya challenge." BMC Infectious Diseases 18, no. 1: 1-14.

Journal article
Published: 22 May 2018 in Online Journal of Public Health Informatics
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ObjectiveOur goal is to develop deployable strategies for infectious disease diagnosis at the point-of-care that are applicable to multiple hosts of infection - conforming to the global One Health Strategy for diagnostics. We aim to develop methods that do not require prior knowledge of the pathogen in question, and can facilitate rapid and effective decision-making and situational awareness.IntroductionThere is an urgent need for diagnostic strategies for infections which are host-independent, so as to effectively track zoonotic spread, monitor animal carriers of pathogens, and evaluate transmission dynamics. Infection of a host - pathogen or human- by an animal results in recognition by the immune response, which consequently causes release of inflammatory mediators. Many scientists have explored the use of cytokines as diagnostic indicators of disease, but the conserved nature of the immune response in humans and animals results in cross-reactivity among many pathogens, making evaluation of the results difficult, especially in high disease burden populations. Measuring the pathogen-specific signature, however, is advantageous - as it offers discrete identification of active infection, and discrimination from exposure. It also offers a universal strategy that can be applied to human and animal hosts of infection - allowing for One Health Biosurveillance. Achieving this, however, requires the development of a) tailored strategies for the measurement of biochemically disparate pathogen signatures in clinical samples and b) ultra-sensitive detection of such signatures in the host. The sensor team at Los Alamos National Laboratory is working on both of these aspects, and the development of One Health Diagnostic platforms, the focus of the work presented here.MethodsMany of the biomarkers secreted by bacterial pathogens and recognized by innate immune receptors elicit host cytokine responses that are amphiphilic (largely glycolipids, lipoproteins or lipoglycans). Based on such, our team has developed tailored methods – membrane insertion and lipoprotein capture for the capture and detection of these greasy and stealthy molecules in infected blood. Sensitivity and specificity, assay optimization, alinearity associated with lipidic molecules and assay parameter development for biomarkers associated with bacterial pathogens will be presented. Considerations for clinical sampling for amphiphile detection in blood, and clinical study design will be demonstrated. Our team has developed an ultra-sensitive biosensor for detection of biomarkers in complex matrices based on the evanescent field properties of single mode planar optical waveguides. Detection of amphiphilic biomarkers in clinical samples, using the waveguide-based biosensor and tailored methods for the detection of such molecules has allowed for the diagnosis of infection in both human and animals hosts. Herein, we present two examples of the same; 1) diagnosis of tuberculosis in humans, cattle and badgers using pathogen-based diagnostic strategies; and 2) detection of Gram-negative Lipopolysacharides in beef and in patients with Salmonella-induced sepsis.ResultsWe have developed tailored ultra-sensitive waveguide-biosensor assays for the detection of Lipoarabinomannan (and other biomarkers) from Mycobacteria and demonstrated feasibility in blinded clinical studies in humans and cattle, demonstrating one -health compatibility.We have also demonstrated detection of lipopolysacharides from eight different serotypes of Shiga-toxin carrying E. coli in beef, and Salmonella Typhimurium in pediatric patients using the same approach.ConclusionsWe have demonstrated clinical feasibility of a One Health Strategy for point-of-care diagnostics of bacterial infections in blood for tuberculosis and Gram-negative pathogens in clinical samples. The results will be demonstrated with several discussion points on the consideration of unconventional biochemistry of pathogens for diagnostics, factors influencing point-of-care deployment and integration of diagnostic platforms for biosurveillance. One Health Considerations and challenges therein will also merit discussion.ReferencesSakamuri, R; et al. Accurate Tracking of Bovine Tuberculosis Biomarkers in Infected Cattle using a Novel Biomarker Capture Strategy, Analytical Sciences, 33(4): 457-460, 2017.Noormohammed A et al. Detection of Salmonella LPS in Patient blood by Lipoprotein Capture. Proc. SPIE 10072, Optical Diagnostics and Sensing XVII: Toward Point-of-Care Diagnostics, 100720A , 2017. doi:10.1117/12.2253506Stromberg L et al. Membrane Insertion for the Detection of Lipopolysaccharides: Exploring the Dynamics of Amphiphile-in-Lipid Assays, PloS One, April 2016.Sakamuri RM et al. Association of lipoarabinomannan with high density lipoprotein in blood: implications for diagnostics.Tuberculosis (Edinb). 2013 May;93(3):301-7. doi: 10.1016/j.tube.2013.02.015. Epub 2013 Mar 16Mukundan H et al. Understanding the interaction of Lipoarabinomannan with membrane mimetic architectures. Tuberculosis, 92(1) 32-47, 2012.Mukundan H et al., Waveguide-based Sensors for Pathogen Detection, Invited Review, Sensors, 9(7), 5783-5809.

ACS Style

Harshini Mukundan. Developing and Deploying Universal Diagnostic Platforms for One-Health Biosurveillance. Online Journal of Public Health Informatics 2018, 10, 1 .

AMA Style

Harshini Mukundan. Developing and Deploying Universal Diagnostic Platforms for One-Health Biosurveillance. Online Journal of Public Health Informatics. 2018; 10 (1):1.

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Harshini Mukundan. 2018. "Developing and Deploying Universal Diagnostic Platforms for One-Health Biosurveillance." Online Journal of Public Health Informatics 10, no. 1: 1.

Book chapter
Published: 12 July 2017 in Escherichia coli - Recent Advances on Physiology, Pathogenesis and Biotechnological Applications
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Detection Methods for Lipopolysaccharides: Past and Present | InTechOpen, Published on: 2017-07-12. Authors: Loreen R. Stromberg, Heather M. Mendez and Harshini Mukundan

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Loreen Stromberg; Heather M. Mendez; Harshini Mukundan. Detection Methods for Lipopolysaccharides: Past and Present. Escherichia coli - Recent Advances on Physiology, Pathogenesis and Biotechnological Applications 2017, 1 .

AMA Style

Loreen Stromberg, Heather M. Mendez, Harshini Mukundan. Detection Methods for Lipopolysaccharides: Past and Present. Escherichia coli - Recent Advances on Physiology, Pathogenesis and Biotechnological Applications. 2017; ():1.

Chicago/Turabian Style

Loreen Stromberg; Heather M. Mendez; Harshini Mukundan. 2017. "Detection Methods for Lipopolysaccharides: Past and Present." Escherichia coli - Recent Advances on Physiology, Pathogenesis and Biotechnological Applications , no. : 1.

Review
Published: 04 July 2017 in Biosensors
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Rapid diagnosis is crucial to effectively treating any disease. Biological markers, or biomarkers, have been widely used to diagnose a variety of infectious and non-infectious diseases. The detection of biomarkers in patient samples can also provide valuable information regarding progression and prognosis. Interestingly, many such biomarkers are composed of lipids, and are amphiphilic in biochemistry, which leads them to be often sequestered by host carriers. Such sequestration enhances the difficulty of developing sensitive and accurate sensors for these targets. Many of the physiologically relevant molecules involved in pathogenesis and disease are indeed amphiphilic. This chemical property is likely essential for their biological function, but also makes them challenging to detect and quantify in vitro. In order to understand pathogenesis and disease progression while developing effective diagnostics, it is important to account for the biochemistry of lipid and amphiphilic biomarkers when creating novel techniques for the quantitative measurement of these targets. Here, we review techniques and methods used to detect lipid and amphiphilic biomarkers associated with disease, as well as their feasibility for use as diagnostic targets, highlighting the significance of their biochemical properties in the design and execution of laboratory and diagnostic strategies. The biochemistry of biological molecules is clearly relevant to their physiological function, and calling out the need for consideration of this feature in their study, and use as vaccine, diagnostic and therapeutic targets is the overarching motivation for this review.

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Jessica Z. Kubicek-Sutherland; Dung M. Vu; Heather M. Mendez; Shailja Jakhar; Harshini Mukundan. Detection of Lipid and Amphiphilic Biomarkers for Disease Diagnostics. Biosensors 2017, 7, 25 .

AMA Style

Jessica Z. Kubicek-Sutherland, Dung M. Vu, Heather M. Mendez, Shailja Jakhar, Harshini Mukundan. Detection of Lipid and Amphiphilic Biomarkers for Disease Diagnostics. Biosensors. 2017; 7 (4):25.

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Jessica Z. Kubicek-Sutherland; Dung M. Vu; Heather M. Mendez; Shailja Jakhar; Harshini Mukundan. 2017. "Detection of Lipid and Amphiphilic Biomarkers for Disease Diagnostics." Biosensors 7, no. 4: 25.

Proceedings article
Published: 22 February 2017 in Biophysics, Biology and Biophotonics II: the Crossroads
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Lipopolysaccharide (LPS) is an amphiphilic lipoglycan that is the primary component of the outer membrane of Gramnegative bacteria. Classified as a pathogen associated molecular pattern (PAMPs), LPS is an essential biomarker for identifying pathogen serogroups. Structurally, LPS is comprised of a hydrophobic lipophilic domain that partitions into the outer membrane of Gram-negative bacteria. Previous work by our team explored biophysical interactions of LPS in supported lipid bilayer assemblies (sLBAs), and demonstrated LPS-induced hole formation in DOPC lipid bilayers. Here, we have incorporated cholesterol and sphingomyelin into sLBAs to evaluate the interaction of LPS in a more physiologically relevant system. The goal of this work was to determine whether increasing membrane complexity of sLBAs, and changing physiological parameters such as temperature, affects LPS-induced hole formation. Integrating cholesterol and sphingomyelin into sLBAs decreased LPS-induced hole formation at lower concentrations of LPS, and bacterial serotype contributed to differences in hole formation as a response to changes in temperature. We also investigated the possibility of LPS-induced hole formation in cellular systems using the cytokine response in both TLR4 (+)/(-) murine macrophages. LPS was presented to each cell line in murine serum, delipidated serum, and buffer (i.e. no serum), and the resulting cytokine levels were measured. Results indicate that the method of LPS presentation directly affects cellular cytokine expression. The two model systems presented in this study provide preliminary insight into the interactions of LPS in the host, and suggest the significance of amphiphile-carrier interactions in regulating host-pathogen biology during infection. © (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.

ACS Style

Heather M. Mendez; Loreen R. Stromberg; Kirstie Swingle; Steven W. Graves; Gabriel Montaño; Harshini Mukundan. Serogroup-specific interactions of lipopolysaccharides with supported lipid bilayer assemblies. Biophysics, Biology and Biophotonics II: the Crossroads 2017, 10075, 100750 .

AMA Style

Heather M. Mendez, Loreen R. Stromberg, Kirstie Swingle, Steven W. Graves, Gabriel Montaño, Harshini Mukundan. Serogroup-specific interactions of lipopolysaccharides with supported lipid bilayer assemblies. Biophysics, Biology and Biophotonics II: the Crossroads. 2017; 10075 ():100750.

Chicago/Turabian Style

Heather M. Mendez; Loreen R. Stromberg; Kirstie Swingle; Steven W. Graves; Gabriel Montaño; Harshini Mukundan. 2017. "Serogroup-specific interactions of lipopolysaccharides with supported lipid bilayer assemblies." Biophysics, Biology and Biophotonics II: the Crossroads 10075, no. : 100750.