3,830 results on '"Kevin V"'
Search Results
2. Plastic pollution and health metrics in wild juvenile green sea turtles (Chelonia mydas) from two Ecuadorian national parks: Galápagos and Machalilla
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Juan Pablo Muñoz-Pérez, Gregory A. Lewbart, Tania Toapanta, Helen Chadwick, Elvis D. Okoffo, Daniela Alarcón-Ruales, Leo Zurita-Arthos, Jen S. Jones, Fernando Cisneros, Andres Moreira-Mendieta, Cristina Vintimilla-Palacios, Cristina Miranda, Felipe Vallejo, Emma Houck, Rubén Alemán, Kamila Escobar-Flores, Alice Skehel, Jason Castañeda, Patricia Secoura, Shelly Vaden, Ceri Lewis, Tamara Galloway, Bryan Wallace, Brendan J. Godley, Matthew Cole, Penelope Lindeque, Kevin V. Thomas, Dominique A. Potvin, Carlos A. Valle, and Kathy A. Townsend
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juvenile green sea turtle ,wildlife ,plastic pollution ,health ,pyr-GC/MS ,FT-IR ,Zoology ,QL1-991 - Abstract
Marine vertebrates, particularly green sea turtles, are especially vulnerable to plastic pollution through ingestion or entanglement. This study investigated wild juvenile green sea turtles (Chelonia mydas) from two Ecuadorian national parks (Galápagos and Machallilla) to assess the prevalence of plastic pollution in their feces and its potential impact on various health metrics. We analyzed fecal samples from 46 juvenile green sea turtles using Fourier transform infrared spectroscopy (FT-IR) to quantify microplastics (MPs). A complementary methodology using pressurized liquid extraction with double-shot pyrolysis-mass spectrometry gas chromatography (Pyr-GC/MS) was also employed to quantify synthetic polymer mass concentrations. The results from these analyses were compared with blood analytes. FT-IR analysis revealed a mean of 4.4±5.2 MPs/g in fecal samples, with the highest quantities found in the Galápagos Marine Reserve (GMR). The most common MPs shape identified were fibers (x̄= 3.8±4.5 MPs/g), and the predominant synthetic polymers were polyvinyl alcohol (PVOH) and polyacrylates (PMMA). The daily intake of MPs by the sampled turtles ranged from a minimum of 312±409 MPs/day to a maximum of 430±563 MPs/day. Pyr-GC/MS analysis detected polyethylene (PE) with a mean of 367±1158 µg/g and polypropylene (PP) with a mean of 155±434 µg/g in fecal samples, with the highest pollution levels observed in the GMR. Both FT-IR and Pyr-GC/MS techniques detected plastic pollution in 98% of the sampled population. Although both FT-IR and Pyr-GC/MS are reliable methods, they produced slightly different results due to methodological variations. However, both supported the finding that turtles in the GMR were exposed to higher rates of plastic ingestion. Despite the turtles appearing clinically healthy based on blood analysis, significant differences in eleven health metrics were observed between turtles classified as less at risk and those most at risk for plastic pollution. Further research is necessary to understand the potential health implications of these findings.
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- 2024
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3. Addressing the climate crisis through engineering biology
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Emily R. Aurand, Tae Seok Moon, Nicole R. Buan, Kevin V. Solomon, Michael Köpke, and EBRC Technical Roadmapping Working Group
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Meteorology. Climatology ,QC851-999 ,Environmental sciences ,GE1-350 - Abstract
As the climate crisis deepens and the impacts are felt more often and more acutely worldwide, scientific, engineering, and policy communities need more tools and opportunities to make a difference in tackling climate challenges. The Engineering Biology Research Consortium (EBRC) has recently published a technical research roadmap, Engineering Biology for Climate & Sustainability, that describes and details short-, medium-, and long-term milestones for engineering biology tool and technology advancements that can be applied to mitigate, prevent, and adapt to climate change. These ambitious technical achievements can only be realized in the context of complementary research, policy, and investment and in combination with efforts from many other disciplines and approaches. Herein we illustrate the opportunities, as described by the roadmap, in engineering biology research and development to impact climate change and long-term environmental sustainability, and why and how engineering biology and subsequent biotechnologies should be among the most prominent of approaches to overcoming the climate crisis.
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- 2024
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4. Beta-2 adrenergic receptor agonism alters astrocyte phagocytic activity and has potential applications to psychiatric disease
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Ellen R. Bowen, Phillip DiGiacomo, Hannah P. Fraser, Kevin Guttenplan, Benjamin A. H. Smith, Marlene L. Heberling, Laura Vidano, Nigam Shah, Mehrdad Shamloo, Jennifer L. Wilson, and Kevin V. Grimes
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
Abstract Schizophrenia is a debilitating condition necessitating more efficacious therapies. Previous studies suggested that schizophrenia development is associated with aberrant synaptic pruning by glial cells. We pursued an interdisciplinary approach to understand whether therapeutic reduction in glial cell—specifically astrocytic—phagocytosis might benefit neuropsychiatric patients. We discovered that beta-2 adrenergic receptor (ADRB2) agonists reduced phagocytosis using a high-throughput, phenotypic screen of over 3200 compounds in primary human fetal astrocytes. We used protein interaction pathways analysis to associate ADRB2, to schizophrenia and endocytosis. We demonstrated that patients with a pediatric exposure to salmeterol, an ADRB2 agonist, had reduced in-patient psychiatry visits using a novel observational study in the electronic health record. We used a mouse model of inflammatory neurodegenerative disease and measured changes in proteins associated with endocytosis and vesicle-mediated transport after ADRB2 agonism. These results provide substantial rationale for clinical consideration of ADRB2 agonists as possible therapies for patients with schizophrenia.
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- 2023
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5. Genetic variation among elite inbred lines suggests potential to breed for BNI-capacity in maize
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César D. Petroli, Guntur V. Subbarao, Juan A. Burgueño, Tadashi Yoshihashi, Huihui Li, Jorge Franco Duran, and Kevin V. Pixley
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Medicine ,Science - Abstract
Abstract Biological nitrification inhibition (BNI) is a plant function where root systems release antibiotic compounds (BNIs) specifically aimed at suppressing nitrifiers to limit soil-nitrate formation in the root zone. Little is known about BNI-activity in maize (Zea mays L.), the most important food, feed, and energy crop. Two categories of BNIs are released from maize roots; hydrophobic and hydrophilic BNIs, that determine BNI-capacity in root systems. Zeanone is a recently discovered hydrophobic compound with BNI-activity, released from maize roots. The objectives of this study were to understand/quantify the relationship between zeanone activity and hydrophobic BNI-capacity. We assessed genetic variability among 250 CIMMYT maize lines (CMLs) characterized for hydrophobic BNI-capacity and zeanone activity, towards developing genetic markers linked to this trait in maize. CMLs with high BNI-capacity and ability to release zeanone from roots were identified. GWAS was performed using 27,085 SNPs (with unique positions on the B73v.4 reference genome, and false discovery rate = 10), and phenotypic information for BNI-capacity and zeanone production from root systems. Eighteen significant markers were identified; three associated with specific BNI-activity (SBNI), four with BNI-activity per plant (BNIPP), another ten were common between SBNI and BNIPP, and one with zeanone release. Further, 30 annotated genes were associated with the significant SNPs; most of these genes are involved in pathways of “biological process”, and one (AMT5) in ammonium regulation in maize roots. Although the inbred lines in this study were not developed for BNI-traits, the identification of markers associated with BNI-capacity suggests the possibility of using these genomic tools in marker-assisted selection to improve hydrophobic BNI-capacity in maize.
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- 2023
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6. Surface-Enhanced Raman Spectroscopy Combined with Multivariate Analysis for Fingerprinting Clinically Similar Fibromyalgia and Long COVID Syndromes
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Shreya Madhav Nuguri, Kevin V. Hackshaw, Silvia de Lamo Castellvi, Yalan Wu, Celeste Matos Gonzalez, Chelsea M. Goetzman, Zachary D. Schultz, Lianbo Yu, Rija Aziz, Michelle M. Osuna-Diaz, Katherine R. Sebastian, W. Michael Brode, Monica M. Giusti, and Luis Rodriguez-Saona
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long COVID ,fibromyalgia ,surface-enhanced Raman spectroscopy ,volumetric absorptive micro-sampling ,dried bloodspot cards ,Biology (General) ,QH301-705.5 - Abstract
Fibromyalgia (FM) is a chronic central sensitivity syndrome characterized by augmented pain processing at diffuse body sites and presents as a multimorbid clinical condition. Long COVID (LC) is a heterogenous clinical syndrome that affects 10–20% of individuals following COVID-19 infection. FM and LC share similarities with regard to the pain and other clinical symptoms experienced, thereby posing a challenge for accurate diagnosis. This research explores the feasibility of using surface-enhanced Raman spectroscopy (SERS) combined with soft independent modelling of class analogies (SIMCAs) to develop classification models differentiating LC and FM. Venous blood samples were collected using two supports, dried bloodspot cards (DBS, n = 48 FM and n = 46 LC) and volumetric absorptive micro-sampling tips (VAMS, n = 39 FM and n = 39 LC). A semi-permeable membrane (10 kDa) was used to extract low molecular fraction (LMF) from the blood samples, and Raman spectra were acquired using SERS with gold nanoparticles (AuNPs). Soft independent modelling of class analogy (SIMCA) models developed with spectral data of blood samples collected in VAMS tips showed superior performance with a validation performance of 100% accuracy, sensitivity, and specificity, achieving an excellent classification accuracy of 0.86 area under the curve (AUC). Amide groups, aromatic and acidic amino acids were responsible for the discrimination patterns among FM and LC syndromes, emphasizing the findings from our previous studies. Overall, our results demonstrate the ability of AuNP SERS to identify unique metabolites that can be potentially used as spectral biomarkers to differentiate FM and LC.
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- 2024
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7. The impact of COVID-19 pandemic on diagnosis and management of gastrointestinal cancers
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Byung Soo Yoo, Ankit Patel, Kevin V. Houston, Alejandra Vargas, Ana Rosa Vilela Sangay, Steve M. D’Souza, and David A. Johnson
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covid-19 pandemic ,gastrointestinal cancer ,gastric cancer ,cancer screening ,mortality ,prognosis ,Other systems of medicine ,RZ201-999 - Abstract
Gastrointestinal (GI) cancer is one of the leading causes of death that affect many patients around the world. The coronavirus disease 2019 (COVID-19) pandemic significantly impacted our healthcare system in large that diagnosis and management of GI cancer have suffered with a reduction in cancer screening. This review will describe the current practices of cancer screening during COVID-19 pandemic and summarize how each GI cancer (esophageal, gastric, colorectal, and hepatocellular cancers) has been affected by COVID-19. World widely there has been a decreasing trend in screening, diagnosis, and management of GI cancers during the COVID-19 pandemic. Many healthcare institutions are now observing the effect of this change and implementing practice variations to adapt to the pandemic.
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- 2023
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8. Characterization and quantification of oxidative stress induced particle debris from polypropylene surgical mesh
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Nicholas T. H. Farr, Cassandra Rauert, Alexander J. Knight, Alexander I. Tartakovskii, and Kevin V. Thomas
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debris ,degradation ,materials characterization ,particles ,pelvic organ prolapse ,polypropylene mesh ,Materials of engineering and construction. Mechanics of materials ,TA401-492 - Abstract
Abstract Explanted polypropylene (PP) surgical mesh has frequently been reported to show surface alterations, such as cracks and flaking. However, to date the consequence of PP mesh degradation is not clearly understood, particularly its potential to influence the biological host response of surrounding tissues. Of particular concern is a possible host reaction to polypropylene particles released through degradation of surgical mesh in vivo. This concern is driven by previous studies which have postulated that an oxidative stress environment has the potential to etch away particles from the surface of a PP fibers. The release of such particles is of considerable significance as particles in the nano‐ to micro range have been shown to have the capacity to irritate cells and stimulate the immune system. The authors are not aware of any previous studies that have attempted to characterize, quantify or identify any particles released from PP mesh after exposure to an oxidative stress environment. Characterization of the PP mesh, post oxidative stress exposure, including identification of particles was achieved through application of a range of techniques: low voltage‐scanning electron microscopy (LV‐SEM), pyrolysis gas chromatography mass spectrometry (Pyr‐GCMS), nano‐Fourier transform infrared spectroscopy (nano‐FTIR), scattering‐type, scanning near‐field optical microscopy (s‐SNOM), atomic force microscopy (AFM), attenuated total reflectance‐Fourier transform infrared spectroscopy (ATR‐FTIR) and uniaxial tensile testing. The findings of this study indicate that oxidative stress alone is a major factor in the production of PP particle debris. PP debris identified within solution, using Pyr‐GCMS, was shown to be in order of the micron scale.
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- 2023
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9. Prophylactic intranasal administration of lipid nanoparticle formulated siRNAs reduce SARS-CoV-2 and RSV lung infection
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Aroon Supramaniam, Yaman Tayyar, Daniel T.W. Clarke, Gabrielle Kelly, Dhruba Acharya, Kevin V. Morris, Nigel A.J. McMillan, and Adi Idris
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SARS-CoV-2 ,siRNA ,RSV ,LNP ,COVID-19 ,Intranasal ,Microbiology ,QR1-502 - Abstract
RNA interference (RNAi) is an emerging and promising therapy for a wide range of respiratory viral infections. This highly specific suppression can be achieved by the introduction of short-interfering RNA (siRNA) into mammalian systems, resulting in the effective reduction of viral load. Unfortunately, this has been hindered by the lack of a good delivery system, especially via the intranasal (IN) route. Here, we have developed an IN siRNA encapsulated lipid nanoparticle (LNP) in vivo delivery system that is highly efficient at targeting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and respiratory syncytial virus (RSV) lung infection in vivo. Importantly, IN siRNA delivery without the aid of LNPs abolishes anti-SARS-CoV-2 activity in vivo. Our approach using LNPs as the delivery vehicle overcomes the significant barriers seen with IN delivery of siRNA therapeutics and is a significant advancement in our ability to delivery siRNAs. The study presented here demonstrates an attractive alternate delivery strategy for the prophylactic treatment of both future and emerging respiratory viral diseases.
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- 2023
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10. Gastrointestinal microbiome and coronavirus disease: evidence of a bidirectional association
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Kevin V. Houston, Ankit Patel, Michael Saadeh, Alejandra Vargas, Ana Rosa Vilela Sangay, Steve M. D'Souza, Byung Soo Yoo, and David A. Johnson
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gastrointestinal microbiome ,dysbiosis ,coronavirus disease 2019 ,post-acute coronavirus disease 2019 syndrome ,long-haul coronavirus disease 2019 ,post infection irritable bowel syndrome ,Other systems of medicine ,RZ201-999 - Abstract
The gastrointestinal (GI) microbiome remains an emerging topic of study and the characterization and impact on human health and disease continue to be an area of great interest. Similarly, the coronavirus disease 2019 (COVID-19) pandemic has significantly impacted the healthcare system with active disease, lasting effects, and complications with the full impact yet to be determined. The most current evidence of the interaction between COVID-19 and the GI microbiome is reviewed, with a focus on key mediators and the microbiome changes associated with acute disease and post-acute COVID-19 syndrome (PACS).
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- 2023
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11. Immunomodulation strategies against COVID-19 evidence: key nutrients and dietary approaches
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Lindsey B. Cundra, Manasa Vallabhaneni, Michael Saadeh, Kevin V. Houston, Byung Soo Yoo, Steve D'Souza, and David A. Johnson
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coronavirus disease-2019 ,immunity ,diet ,fasting ,dietary supplements ,probiotics ,Other systems of medicine ,RZ201-999 - Abstract
The novel coronavirus disease-2019 (COVID-19) has created a major public health crisis. Various dietary factors may enhance immunological activity against COVID-19 and serve as a method to combat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The dietary factors that are responsible for boosting immunity may provide a therapeutic advantage in patients with COVID-19. Investigators have demonstrated that vitamins B6, B12, C, D, E, and K, and trace elements like zinc, copper, selenium, and iron may serve as important tools for immunomodulation. Herein this is a review the peer-reviewed literature pertaining to dietary immunomodulation strategies against COVID-19. This review is intended to better define the evidence that dietary modifications and supplementation could positively influence the proinflammatory state in patients with COVID-19 and improve clinical outcomes. With appropriate insight, therapeutic interventions are discussed and directed to potentially modulate host immunity to mitigate the disease mechanisms of COVID-19.
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- 2023
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12. Prevalence, risk and severity of SARS-CoV-2 infections in psoriasis patients receiving conventional systemic, biologic or topical treatment during the COVID-19 pandemic: a cross-sectional cohort study (PsoCOVID)
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Kevin V. Kwee, Jean-Luc Murk, Qiqi Yin, M. Birgitte Visch, Linda Davidson, Elke M. G. J. de Jong, Juul M. P. A. van den Reek, and Milan Tjioe
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sars-cov-2 ,covid-19 ,psoriasis ,biologics ,vaccines ,immunosuppression ,Dermatology ,RL1-803 - Abstract
Background The risk of SARS-CoV-2 infection does not appear to be increased for psoriasis patients using biologics compared to those on other treatments, but evidence is still limited. Objectives (1) to estimate the prevalence of SARS-CoV-2 infection in patients with psoriasis, (2) to compare SARS-CoV-2 infection rates for different psoriasis treatments groups (biologic vs. systemic conventional vs. topical therapy) corrected for confounders and (3) to describe patients with severe COVID-19 for all treatment groups. Methods In this cross-sectional cohort study all patients received a questionnaire to gather data on psoriasis treatment, SARS-CoV-2 infections and related risk factors. Simultaneously, they underwent a blood test to screen for antibodies to SARS-CoV-2 N-antigen. Prevalence of SARS-CoV-2 infections was calculated and logistic regression and Cox proportional-hazards models were performed to determine the association between treatment group and SARS-CoV-2 infection risk, corrected for confounders. Patients with severe COVID-19 disease were described and the mortality rate per treatment group was calculated for the target population. Results Patients were included between April 12 2021 and October 31 2021. Of 551 patients, 59 (10.7% (CI95% 8.3–13.6)) had experienced a SARS-CoV-2 infection, based on questionnaire data combined with serological data. In our study cohort, corrected for confounders, biologic or non-biologic systemic therapy users did not appear to have increased SARS-CoV-2 infection risk compared to patients using other treatment. Only 4 hospitalizations (0.7% (CI95% 0.2–1.0) were reported in our study population and no ICU admissions were reported. The rough mortality rate in the target cohort was 0.32% (CI95% 0.13–0.66) in all treatment groups. Conclusions Corrected for risk-mitigating behavior and vaccination status, a higher SARS-CoV-2 incidence for biologics or non-biologics systemics compared to other treatments could not be proven. Severe cases were infrequent in all treatment groups. This finding further strengthens treatment recommendations that systemic therapies for patients with psoriasis do not require preventive cessation for reduction of SARS-CoV-2 infection risk.
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- 2023
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13. Per- and polyfluoroalkyl substances (PFAS) in consumer products: Current knowledge and research gaps
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Pradeep Dewapriya, Lachlan Chadwick, Sara Ghorbani Gorji, Bastian Schulze, Sara Valsecchi, Saer Samanipour, Kevin V. Thomas, and Sarit L. Kaserzon
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Consumer products ,Textiles ,Food packaging ,PFASs ,Cosmetics ,Hazardous substances and their disposal ,TD1020-1066 - Abstract
While several sources of per- and polyfluoroalkyl substances (PFAS) are known, their use in consumer household products is far less explored. The aim of this study was to provide comprehensive bottom-up analysis of the types and concentrations of PFAS reported in the literature over the past decade. A total of 52 studies revealed 107 PFAS belonging to 15 different categories in 1040 consumer products. The highest number of products tested were from the USA (n = 389) followed by the Czech Republic (n = 111). Mean PFAS concentrations were highest in household firefighting products, followed by textile finishing agents and household chemicals. The highest diversity of PFAS was reported in textiles (72 PFAS). Fluorotelomer alcohol (FTOH), polyfluoroalkyl phosphate esters (PAPs), perfluorocarboxylic acid (PFCA) and perfluorosulfonic acid (PFSA) are the classes of PFAS of high interest. Eight out of 52 studies used High-Resolution Mass Spectrometry techniques. Highlighted knowledge gaps included (i) the development of analytical methods for detecting a range of PFAS in consumer products, (ii) method validation and QA/QC approaches, (iii) application of suspect and non-target analysis, and (iv) an understanding of human exposure risk. This review highlights that the presence of PFAS in consumer products is of concern and remains underexplored.
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- 2023
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14. Portable Mid-Infrared Spectroscopy Combined with Chemometrics to Diagnose Fibromyalgia and Other Rheumatologic Syndromes Using Rapid Volumetric Absorptive Microsampling
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Shreya Madhav Nuguri, Kevin V. Hackshaw, Silvia de Lamo Castellvi, Haona Bao, Siyu Yao, Rija Aziz, Scott Selinger, Zhanna Mikulik, Lianbo Yu, Michelle M. Osuna-Diaz, Katherine R. Sebastian, M. Monica Giusti, and Luis Rodriguez-Saona
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fibromyalgia ,rheumatic diseases ,central sensitization ,FT-MIR ,OPLS-DA ,point-of-care device ,Organic chemistry ,QD241-441 - Abstract
The diagnostic criteria for fibromyalgia (FM) have relied heavily on subjective reports of experienced symptoms coupled with examination-based evidence of diffuse tenderness due to the lack of reliable biomarkers. Rheumatic disorders that are common causes of chronic pain such as rheumatoid arthritis, systemic lupus erythematosus, osteoarthritis, and chronic low back pain are frequently found to be comorbid with FM. As a result, this can make the diagnosis of FM more challenging. We aim to develop a reliable classification algorithm using unique spectral profiles of portable FT-MIR that can be used as a real-time point-of-care device for the screening of FM. A novel volumetric absorptive microsampling (VAMS) technique ensured sample volume accuracies and minimized the variation introduced due to hematocrit-based bias. Blood samples from 337 subjects with different disorders (179 FM, 158 non-FM) collected with VAMS were analyzed. A semi-permeable membrane filtration approach was used to extract the blood samples, and spectral data were collected using a portable FT-MIR spectrometer. The OPLS-DA algorithm enabled the classification of the spectra into their corresponding classes with 84% accuracy, 83% sensitivity, and 85% specificity. The OPLS-DA regression plot indicated that spectral regions associated with amide bands and amino acids were responsible for discrimination patterns and can be potentially used as spectral biomarkers to differentiate FM and other rheumatic diseases.
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- 2024
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15. Early Diagnosis of Fibromyalgia Using Surface-Enhanced Raman Spectroscopy Combined with Chemometrics
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Haona Bao, Kevin V. Hackshaw, Silvia de Lamo Castellvi, Yalan Wu, Celeste Matos Gonzalez, Shreya Madhav Nuguri, Siyu Yao, Chelsea M. Goetzman, Zachary D. Schultz, Lianbo Yu, Rija Aziz, Michelle M. Osuna-Diaz, Katherine R. Sebastian, Monica M. Giusti, and Luis Rodriguez-Saona
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fibromyalgia ,surface-enhanced Raman spectroscopy ,central sensitization syndrome ,metabolic fingerprinting ,in-clinic disease diagnostics ,chemometrics ,Biology (General) ,QH301-705.5 - Abstract
Fibromyalgia (FM) is a chronic muscle pain disorder that shares several clinical features with other related rheumatologic disorders. This study investigates the feasibility of using surface-enhanced Raman spectroscopy (SERS) with gold nanoparticles (AuNPs) as a fingerprinting approach to diagnose FM and other rheumatic diseases such as rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), osteoarthritis (OA), and chronic low back pain (CLBP). Blood samples were obtained on protein saver cards from FM (n = 83), non-FM (n = 54), and healthy (NC, n = 9) subjects. A semi-permeable membrane filtration method was used to obtain low-molecular-weight fraction (LMF) serum of the blood samples. SERS measurement conditions were standardized to enhance the LMF signal. An OPLS-DA algorithm created using the spectral region 750 to 1720 cm−1 enabled the classification of the spectra into their corresponding FM and non-FM classes (Rcv > 0.99) with 100% accuracy, sensitivity, and specificity. The OPLS-DA regression plot indicated that spectral regions associated with amino acids were responsible for discrimination patterns and can be potentially used as spectral biomarkers to differentiate FM and other rheumatic diseases. This exploratory work suggests that the AuNP SERS method in combination with OPLS-DA analysis has great potential for the label-free diagnosis of FM.
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- 2024
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16. Ternary Heterostructures Based on BaTiO3/MoO3/Ag for Highly Efficient and Reusable Photocatalytic Applications
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Kevin V. Alex, José P. B. Silva, Koppole Kamakshi, and Koppole C. Sekhar
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ferroelectric ,heterostructures ,photocatalysis ,plasmonic ,semiconductors ,Physics ,QC1-999 ,Technology - Abstract
Abstract This work shows the fabrication of an efficient ternary heterostructure photocatalyst by integrating ferroelectric BaTiO3 (BTO) as the bottom layer, semiconductor MoO3 as the middle layer and plasmonic silver nanoparticles (Ag NPs) as the top layer, respectively. The BaTiO3/MoO3/Ag (BMA) heterostructure exhibits a higher photodegradation and photocatalytic efficiency of 100% for rhodamine B (RhB) dye under a UV–Visible light illumination of 60 min when compared with its binary heterostructure counterparts BaTiO3/Ag (BA) and MoO3/Ag (MA). The increased photocatalytic activity in BMA heterostructure is attributed to its enhanced interfacial electric field due to the electric double layer formation at BTO‐MoO3 and MoO3‐Ag interfaces. The higher blueshift in the surface plasmon resonance (SPR) peak observed for the BMA heterostructure clearly indicates an increased electron transfer toward the top Ag NPs layer under optical illumination. The higher resistive switching (RS) ratio, the increased difference in voltage minima, and the improved photocurrent generation, as evident from the I–V characteristics, illustrate the enhanced charge carrier generation and separation in BMA heterostructure. A smaller arc radius observed for the Nyquist plot of BMA heterostructure clearly showcases its increased interfacial charge transfer (CT). The CT mechanism and reusability of the BMA heterostructure are also studied.
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- 2023
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17. Festivals following the easing of COVID-19 restrictions: Prevalence of new psychoactive substances and illicit drugs
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Nikolaos Rousis, Richard Bade, Iván Romero-Sánchez, Jochen F. Mueller, Nikolaos S. Thomaidis, Kevin V. Thomas, and Emma Gracia-Lor
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Wastewater-based epidemiology ,Synthetic cathinones ,3-chloromethcathinone (3-CMC) ,N,N-dimethylpentylone ,2F-deschloroketamine (2F-DCK) ,Substance abuse ,Environmental sciences ,GE1-350 - Abstract
The market for illicit drugs and new psychoactive substances (NPS) has grown significantly and people attending festivals have been identified as being at high risk (high extent and frequency of substance use). Traditional public health surveillance data sources have limitations (high costs, long implementation times, and ethical issues) and wastewater-based epidemiology (WBE) can cost-effectively support surveillance efforts. Influent wastewater samples were analyzed for NPS and illicit drug consumption collected during New Year period (from 29-Dec-2021 to 4-Jan-2022) and a summer Festival (from 29-June-2022 to 12-July-2022) in a large city in Spain. Samples were analyzed for phenethylamines, cathinones, opioids, benzodiazepines, plant-based NPS, dissociatives, and the illicit drugs methamphetamine, MDA, MDMA, ketamine, heroin, cocaine, and pseudoephedrine by liquid chromatography mass spectrometry. High consumption rates of specific NPS and established illicit drugs were identified at the peak of each event. Furthermore, a dynamic change in NPS use (presence and absence of substances) was detected over a period of six months. Eleven NPS, including synthetic cathinones, benzodiazepines, plant-based NPS and dissociatives, and seven illicit drugs were found across both the New Year and summer Festival. Statistically significant differences (p
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- 2023
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18. Effect of the thickness on the photocatalytic and the photocurrent properties of ZnO films deposited by spray pyrolysis
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A. Sulthan Ibrahim, Kevin V. Alex, M. Bhakya Latha, K. Kamakshi, S. Sathish, J. P. B. Silva, and K. C. Sekhar
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Ultrasonic spray pyrolysis ,ZnO ,Photoluminescence ,I-V characteristics ,Photocatalytic ,Materials of engineering and construction. Mechanics of materials ,TA401-492 - Abstract
Abstract In this work, we have investigated the structure, morphology, photoluminescence, photocatalytic and photocurrent properties of ZnO thin films as a function of their film thickness (tZnO) fabricated via ultrasonic spray pyrolysis technique. The X-Ray diffraction patterns exhibited the formation of polycrystalline wurtzite phase of ZnO. Scanning electron microscopy images showed the uniform morphology with nanorod structure. The photosensitivity and photocatalytic efficiency are found to be optimum at tZnO = 1200 nm and are attributed to the improved photogeneration of charge carriers and higher concentration of oxygen vacancies. A direct correlation is established between the photosensitivity and photodegradation process. The incident photon-to-electron conversion efficiency (IPCE) and photocatalytic efficiency for the ZnO film at tZnO = 1200 nm are estimated to be 31.5% and 100% respectively. The obtained result suggests that ZnO thin films are potential candidates for applications in various optoelectronic devices.
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- 2022
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19. Metabolic Fingerprinting for the Diagnosis of Clinically Similar Long COVID and Fibromyalgia Using a Portable FT-MIR Spectroscopic Combined with Chemometrics
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Kevin V. Hackshaw, Siyu Yao, Haona Bao, Silvia de Lamo Castellvi, Rija Aziz, Shreya Madhav Nuguri, Lianbo Yu, Michelle M. Osuna-Diaz, W. Michael Brode, Katherine R. Sebastian, M. Monica Giusti, and Luis Rodriguez-Saona
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fibromyalgia ,post-acute sequelae of SARS-CoV-2 (PASC)/long COVID ,metabolic fingerprinting ,in-clinic disease diagnostics ,chemometrics ,blood ,Biology (General) ,QH301-705.5 - Abstract
Post Acute Sequelae of SARS-CoV-2 infection (PASC or Long COVID) is characterized by lingering symptomatology post-initial COVID-19 illness that is often debilitating. It is seen in up to 30–40% of individuals post-infection. Patients with Long COVID (LC) suffer from dysautonomia, malaise, fatigue, and pain, amongst a multitude of other symptoms. Fibromyalgia (FM) is a chronic musculoskeletal pain disorder that often leads to functional disability and severe impairment of quality of life. LC and FM share several clinical features, including pain that often makes them indistinguishable. The aim of this study is to develop a metabolic fingerprinting approach using portable Fourier-transform mid-infrared (FT-MIR) spectroscopic techniques to diagnose clinically similar LC and FM. Blood samples were obtained from LC (n = 50) and FM (n = 50) patients and stored on conventional bloodspot protein saver cards. A semi-permeable membrane filtration approach was used to extract the blood samples, and spectral data were collected using a portable FT-MIR spectrometer. Through the deconvolution analysis of the spectral data, a distinct spectral marker at 1565 cm−1 was identified based on a statistically significant analysis, only present in FM patients. This IR band has been linked to the presence of side chains of glutamate. An OPLS-DA algorithm created using the spectral region 1500 to 1700 cm−1 enabled the classification of the spectra into their corresponding classes (Rcv > 0.96) with 100% accuracy and specificity. This high-throughput approach allows unique metabolic signatures associated with LC and FM to be identified, allowing these conditions to be distinguished and implemented for in-clinic diagnostics, which is crucial to guide future therapeutic approaches.
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- 2023
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20. Pre-clinical data supporting immunotherapy for HIV using CMV-HIV-specific CAR T cells with CMV vaccine
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Min Guan, Laura Lim, Leo Holguin, Tianxu Han, Vibhuti Vyas, Ryan Urak, Aaron Miller, Diana L. Browning, Liliana Echavarria, Shasha Li, Shirley Li, Wen-Chung Chang, Tristan Scott, Paul Yazaki, Kevin V. Morris, Angelo A. Cardoso, M. Suzette Blanchard, Virginia Le Verche, Stephen J. Forman, John A. Zaia, John C. Burnett, and Xiuli Wang
- Subjects
chimeric antigen receptor T cell (CAR T) ,cytomegalovirus (CMV) vaccine ,broadly neutralizing antibody (bNAb) ,N6 ,HIV/AIDS ,immunotherapy ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
T cells engineered to express HIV-specific chimeric antigen receptors (CARs) represent a promising strategy to clear HIV-infected cells, but to date have not achieved clinical benefits. A likely hurdle is the limited T cell activation and persistence when HIV antigenemia is low, particularly during antiretroviral therapy (ART). To overcome this issue, we propose to use a cytomegalovirus (CMV) vaccine to stimulate CMV-specific T cells that express CARs directed against the HIV-1 envelope protein gp120. In this study, we use a GMP-compliant platform to engineer CMV-specific T cells to express a second-generation CAR derived from the N6 broadly neutralizing antibody, one of the broadest anti-gp120 neutralizing antibodies. These CMV-HIV CAR T cells exhibit dual effector functions upon in vitro stimulation through their endogenous CMV-specific T cell receptors or the introduced CARs. Using a humanized HIV mouse model, we show that CMV vaccination during ART accelerates CMV-HIV CAR T cell expansion in the peripheral blood and that higher numbers of CMV-HIV CAR T cells were associated with a better control of HIV viral load and fewer HIV antigen p24+ cells in the bone marrow upon ART interruption. Collectively, these data support the clinical development of CMV-HIV CAR T cells in combination with a CMV vaccine in HIV-infected individuals.
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- 2022
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21. Hypoxia-directed tumor targeting of CRISPR-Cas9 and HSV-TK suicide gene therapy using lipid nanoparticles
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Alicia Davis, Kevin V. Morris, and Galina Shevchenko
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CRISPR-Cas9 ,HSV-tk ,lipid nanoparticle ,cancer ,hypoxia ,hypoxia regulation ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Hypoxia is a characteristic feature of solid tumors that contributes to tumor aggressiveness and is associated with resistance to cancer therapy. The hypoxia inducible factor-1 (HIF-1) transcription factor complex mediates hypoxia-specific gene expression by binding to hypoxia-responsive element (HRE) sequences within the promoter of target genes. HRE-driven expression of therapeutic cargo has been widely explored as a strategy to achieve cancer-specific gene expression. By utilizing this system, we achieve hypoxia-specific expression of two therapeutically relevant cargo elements: the herpes simplex virus thymidine kinase (HSV-tk) suicide gene and the CRISPR-Cas9 nuclease. Using an expression vector containing five copies of the HRE derived from the vascular endothelial growth factor gene, we are able to show high transgene expression in cells in a hypoxic environment, similar to levels achieved using the cytomegalovirus (CMV) and CBh promoters. Furthermore, we are able to deliver our therapeutic cargo to tumor cells with high efficiency using plasmid-packaged lipid nanoparticles (LNPs) to achieve specific killing of tumor cells in hypoxic conditions while maintaining tight regulation with no significant changes to cell viability in normoxia.
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- 2022
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22. Gluten-Free Diet in Childhood Difficult-to-Treat Nephrotic Syndrome: A Pilot Feasibility Study
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Tarak Srivastava, Katherine M. Dell, Kevin V. Lemley, Debbie S. Gipson, Frederick J. Kaskel, Kevin Edward Meyers, Christian Faul, Ayelet Goldhaber, LauraJane Pehrson, and Howard Trachtman
- Subjects
glomerular disease ,nephrotic syndrome ,proteinuria ,gluten-free diet ,pilot study ,childhood ,difficult-to-treat disease ,zonulin ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Introduction: Minimal change disease in childhood can follow a frequently relapsing or steroid-dependent course in up to 40% of cases. Second-line immunosuppressive medications that are used to manage these patients are associated with significant adverse effects. There is a need for safer alternative treatments for difficult-to-treat nephrotic syndrome. Therefore, we conducted an open-label feasibility study to assess the safety and efficacy of a gluten-free diet as treatment for pediatric patients with difficult-to-treat nephrotic syndrome. As a second aim, we sought to determine if the plasma zonulin concentration can identify those who are more likely to respond to this intervention. Methods: Seventeen patients were placed on a gluten-free diet for 6 months. A positive response was defined as a 50% reduction in the relapse rate compared to the preceding 6 months or the ability to discontinue 1 immunosuppressive drug. Results: Five (29%) participants had a positive response to the dietary intervention. The gluten-free diet was well tolerated with no clinical or laboratory adverse events. Plasma zonulin concentration was elevated in patients who failed to benefit from the gluten-free diet. Discussion/Conclusion: A gluten-free diet may be a useful adjunctive intervention for patients with difficult-to-treat nephrotic syndrome that can be implemented prior to resorting to second-line immunosuppressive therapy. Development of the plasma zonulin level as a biomarker to predict efficacy would facilitate rational use of a gluten-free diet in the management of nephrotic syndrome.
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- 2022
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23. Engineered extracellular vesicles directed to the spike protein inhibit SARS-CoV-2
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Tristan A. Scott, Aroon Supramaniam, Adi Idris, Angelo A. Cardoso, Surya Shrivastava, Gabrielle Kelly, Nicole A. Grepo, Citradewi Soemardy, Roslyn M. Ray, Nigel A.J. McMillan, and Kevin V. Morris
- Subjects
extracellular vesicles ,nanobody ,SARS-CoV-2 ,COVID-19 ,therapeutic ,neutralization ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
SARS-CoV-2 (CoV-2) viral infection results in COVID-19 disease, which has caused significant morbidity and mortality worldwide. A vaccine is crucial to curtail the spread of SARS-CoV-2, while therapeutics will be required to treat ongoing and reemerging infections of SARS-CoV-2 and COVID-19 disease. There are currently no commercially available effective anti-viral therapies for COVID-19, urging the development of novel modalities. Here, we describe a molecular therapy specifically targeted to neutralize SARS-CoV-2, which consists of extracellular vesicles (EVs) containing a novel fusion tetraspanin protein, CD63, embedded within an anti-CoV-2 nanobody. These anti-CoV-2-enriched EVs bind SARS-CoV-2 spike protein at the receptor-binding domain (RBD) site and can functionally neutralize SARS-CoV-2. This work demonstrates an innovative EV-targeting platform that can be employed to target and inhibit the early stages of SARS-CoV-2 infection.
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- 2022
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24. Exosome-mediated stable epigenetic repression of HIV-1
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Surya Shrivastava, Roslyn M. Ray, Leo Holguin, Lilliana Echavarria, Nicole Grepo, Tristan A. Scott, John Burnett, and Kevin V. Morris
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Science - Abstract
A strategy to control HIV-1 infection is to stably repress HIV-1 and induce “deep latency”. Here the authors show that a recombinant anti-HIV-1-1 protein can be packaged as mRNA into exosomes and delivered systemically to repress HIV-1-1 within the context of virus infected mice and achieve long term silencing of HIV-1-1 expression.
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- 2021
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25. Socioeconomic status and public health in Australia: A wastewater-based study
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Nikolaos I. Rousis, Zhe Li, Richard Bade, Michael S. McLachlan, Jochen F. Mueller, Jake W. O'Brien, Saer Samanipour, Benjamin J. Tscharke, Nikolaos S. Thomaidis, and Kevin V. Thomas
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Wastewater-based epidemiology ,Metoprolol ,Atenolol acid ,Venlafaxine ,Sotalol ,Sitagliptin ,Environmental sciences ,GE1-350 - Abstract
Analysis of untreated municipal wastewater is recognized as an innovative approach to assess population exposure to or consumption of various substances. Currently, there are no published wastewater-based studies investigating the relationships between catchment social, demographic, and economic characteristics with chemicals using advanced non-targeted techniques. In this study, fifteen wastewater samples covering 27% of the Australian population were collected during a population Census. The samples were analysed with a workflow employing liquid chromatography high-resolution mass spectrometry and chemometric tools for non-target analysis. Socioeconomic characteristics of catchment areas were generated using Geospatial Information Systems software. Potential correlations were explored between pseudo-mass loads of the identified compounds and socioeconomic and demographic descriptors of the wastewater catchments derived from Census data. Markers of public health (e.g., cardiac arrhythmia, cardiovascular disease, anxiety disorder and type 2 diabetes) were identified in the wastewater samples by the proposed workflow. They were positively correlated with descriptors of disadvantage in education, occupation, marital status and income, and negatively correlated with descriptors of advantage in education and occupation. In addition, markers of polypropylene glycol (PPG) and polyethylene glycol (PEG) related compounds were positively correlated with housing and occupation disadvantage. High positive correlations were found between separated and divorced people and specific drugs used to treat cardiac arrhythmia, cardiovascular disease, and depression. Our robust non-targeted methodology in combination with Census data can identify relationships between biomarkers of public health, human behaviour and lifestyle and socio-demographics of whole populations. Furthermore, it can identify specific areas and socioeconomic groups that may need more assistance than others for public health issues. This approach complements important public health information and enables large-scale national coverage with a relatively small number of samples.
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- 2022
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26. A nationwide wastewater-based assessment of metformin consumption across Australia
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Dan Yang, Qiuda Zheng, Phong K. Thai, Fahad Ahmed, Jake W. O'Brien, Jochen F. Mueller, Kevin V. Thomas, and Ben Tscharke
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Wastewater ,Metformin ,Type 2 diabetes ,Socio-economic factors ,Spatial pattern ,Environmental sciences ,GE1-350 - Abstract
Metformin is the most widely used drug to treat type 2 diabetes. Monitoring spatial patterns of metformin use could provide new insights into treatment of type 2 diabetes and the distribution among populations. This study applied a wastewater-based epidemiological (WBE) approach to estimate metformin use in different populations across Australia and compared these estimates with traditional approaches of surveys and prescription data. Twenty-four-hour influent samples were collected from 75 wastewater treatment plants (WWTPs) across Australia in 2016 and analysed for metformin. Metformin was detected in all samples ranging in concentration from 8.2 to 191 µg/L (median 58 µg/L). Concentrations were converted to population-weighted average consumption at the national level, resulting in an average consumption of 28.6 g/day/1000 people across Australia, which was within 7% of estimates from national prescription statistics. In addition, results for five out of seven states had an estimated prevalence of type 2 diabetes within 20% compared to the traditional epidemiology surveys. Spatial patterns were also observed between urban and rural settings, with higher consumption rates of metformin found in Major Cities (22.5 ± 10.9 g/d/1000 people) and Inner Regional cities (25.4 ± 13.4 g/d/1000 people) than in Outer Regional (17.0 ± 8.1 g/d/1000 people) and Remote areas (15.1 ± 7.4 g/d/1000 people). Consumption estimates were also correlated against socioeconomic factors of the specific catchment areas. Greater metformin use was correlated with populations of lower education and income levels, while positive correlations were found between metformin consumption and consumption of allopurinol, caffeine and venlafaxine. Our study provides more evidence on the distribution of metformin use across Australia, which can be used to develop public health strategies to reduce the overall burden of type 2 diabetes in specific regions.
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- 2022
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27. Evaluation of the Elements of Short Hairpin RNAs in Developing shRNA-Containing CAR T Cells
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Ryan Urak, Brenna Gittins, Citradewi Soemardy, Nicole Grepo, Lior Goldberg, Madeleine Maker, Galina Shevchenko, Alicia Davis, Shirley Li, Tristan Scott, Kevin V. Morris, Stephen J. Forman, and Xiuli Wang
- Subjects
chimeric antigen receptor (CAR) ,short hairpin RNA (shRNA) ,hypoxanthine phosphoribosyltransferase 1 (HPRT) ,C-C chemokine receptor type 5 (CCR5) ,Tat ,Rev ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,RC254-282 - Abstract
Short hairpin RNAs (shRNAs) have emerged as a powerful tool for gene knockdown in various cellular systems, including chimeric antigen receptor (CAR) T cells. However, the elements of shRNAs that are crucial for their efficacy in developing shRNA-containing CAR T cells remain unclear. In this study, we evaluated the impact of different shRNA elements, including promoter strength, orientation, multiple shRNAs, self-targeting, and sense and antisense sequence composition on the knockdown efficiency of the target gene in CAR T cells. Our findings highlight the importance of considering multiple shRNAs and their orientation to achieve effective knockdown. Moreover, we demonstrate that using a strong promoter and avoiding self-targeting can enhance CAR T cell functionality. These results provide a framework for the rational design of CAR T cells with shRNA-mediated knockdown capabilities, which could improve the therapeutic efficacy of CAR T cell-based immunotherapy.
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- 2023
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28. Enhanced target cell specificity and uptake of lipid nanoparticles using RNA aptamers and peptides
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Roslyn M. Ray, Anders Højgaard Hansen, Maria Taskova, Bernhard Jandl, Jonas Hansen, Citra Soemardy, Kevin V. Morris, and Kira Astakhova
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aptamer ,blood–brain barrier ,gene therapy ,hiv-1 ,lipid nanoparticle ,Science ,Organic chemistry ,QD241-441 - Abstract
Lipid nanoparticles (LNPs) constitute a facile and scalable approach for delivery of payloads to human cells. LNPs are relatively immunologically inert and can be produced in a cost effective and scalable manner. However, targeting and delivery of LNPs across the blood–brain barrier (BBB) has proven challenging. In an effort to target LNPs composed of an ionizable cationic lipid (DLin-MC3-DMA), cholesterol, the phospholipid 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG 2000) to particular cell types, as well as to generate LNPs that can cross the BBB, we developed and assessed two approaches. The first was centered on the BBB-penetrating trans-activator of transcription (Tat) peptide or the peptide T7, and the other on RNA aptamers targeted to glycoprotein gp160 from human immunodeficiency virus (HIV) or C-C chemokine receptor type 5 (CCR5), a HIV-1 coreceptor. We report herein a CCR5-selective RNA aptamer that acts to facilitate entry through a simplified BBB model and that drives the uptake of LNPs into CCR5-expressing cells, while the gp160 aptamer did not. We further observed that the addition of cell-penetrating peptides, Tat and T7, did not increase BBB penetration above the aptamer-loaded LNPs alone. Moreover, we found that these targeted LNPs exhibit low immunogenic and low toxic profiles and that targeted LNPs can traverse the BBB to potentially deliver drugs into the target tissue. This approach highlights the usefulness of aptamer-loaded LNPs to increase target cell specificity and potentially deliverability of central-nervous-system-active RNAi therapeutics across the BBB.
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- 2021
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29. A role for a novel natural antisense-BDNF in the maintenance of nicotine-seeking
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Neil A. Youngson, Matthew R. Castino, Angela Stuart, Kelly A. Kershaw, Nathan M. Holmes, Eilish C. Heffernan, Paul D. Waters, Kevin V. Morris, and Kelly J. Clemens
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Infralimbic ,Non-coding RNA ,Addiction ,Rat ,Extinction ,Antisense ,Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
Brain derived neurotrophic factor (BDNF) is critical for the extinction of drug-seeking. Expression of the Bdnf gene is highly epigenetically regulated, including via interactions with non-coding RNA. Here we investigate whether a long non-coding RNA antisense to Bdnf exon IV is involved in extinction of drug-seeking. Strand-specific RNA sequencing confirmed the presence of a novel long non-coding RNA antisense to exon IV of the Bdnf gene in the ventromedial prefrontal cortex of adult male rats (Bdnf-IV-AS). Bdnf-IV-AS expression was validated using strand-specific reverse transcription and quantitative polymerase chain reaction following acquisition, extinction or abstinence from intravenous nicotine self-administration. Expression of Bdnf-IV-AS was elevated following intravenous nicotine self-administration but not experimenter-administered nicotine. Elevated Bdnf-IV-AS persisted across abstinence and to a greater extent following extinction training, suggesting an interaction between Bdnf-IV-AS, nicotine and extinction learning. A functional role of the Bdnf-IV-AS in extinction of nicotine-seeking was established by infusing gapmer oligonucleotides into the infralimbic cortex prior to extinction and testing for the effect of these infusions on reinstatement and reacquisition of nicotine-seeking. Knockdown of the Bdnf-IV-AS across cue-extinction, but not abstinence, significantly attenuated nicotine-primed reinstatement of nicotine-seeking. Bdnf-IV-AS accumulates in the infralimbic cortex across self-administration training, interferes with the inhibitory learning that underpins extinction of drug-seeking, and predisposes animals to drug relapse.
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- 2022
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30. RNAi to treat SARS‐CoV‐2—variant proofing the next generation of therapies
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Nigel A J McMillan, Kevin V Morris, and Adi Idris
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Medicine (General) ,R5-920 ,Genetics ,QH426-470 - Abstract
There is an urgent need to bring new antivirals to SARS‐CoV‐2 to the market. Indeed, in the last 3 months, we have seen at least two new antivirals approved, molnupiravir and paxlovid. Both are older established antivirals that show some efficacy against SARS‐CoV‐2. The work by Chang et al (2022) in the current issue of EMBO Molecular Medicine explores the use of short interfering RNAs to directly target SARS‐CoV‐2 and shows that RNAi is an effective approach to reducing, or even eliminating viral replication, depending on the experimental setting. This antiviral effect results in significant prevention of infection‐related pathology in animals. The key feature of this approach, besides its simplicity as naked siRNAs, is that all current variants are covered by this treatment.
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- 2022
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31. Broadly active zinc finger protein-guided transcriptional activation of HIV-1
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Tristan A. Scott, Denis O’Meally, Nicole Anne Grepo, Citradewi Soemardy, Daniel C. Lazar, Yue Zheng, Marc S. Weinberg, Vicente Planelles, and Kevin V. Morris
- Subjects
HIV-1 ,transcription ,zinc finger protein ,ZFN-362-VPR ,activation ,latency ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Human immunodeficiency virus type 1 (HIV-1) causes a persistent viral infection resulting in the demise of immune regulatory cells. Clearance of HIV-1 infection results in integration of proviral DNA into the genome of host cells, which provides a means for evasion and long-term persistence. A therapeutic compound that specifically targets and sustainably activates a latent HIV-1 provirus could be transformative and is the goal for the “shock-and-kill” approach to a functional cure for HIV-1. Substantial progress has been made toward the development of recombinant proteins that target specific genomic loci for gene activation, repression, or inactivation by directed mutations. However, most of these modalities are too large or too complex for efficient therapeutic application. We describe here the development and testing of a novel recombinant zinc finger protein transactivator, ZFP-362-VPR, which specifically and potently enhances proviral HIV-1 transcription both in established latency models and activity across different viral clades. Additionally, ZFP-362-VPR-activated HIV-1 reporter gene expression in a well-established primary human CD4+ T cell latency model and off-target pathways were determined by transcriptome analyses. This study provides clear proof of concept for the application of a novel, therapeutically relevant, protein transactivator to purge cellular reservoirs of HIV-1.
- Published
- 2021
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32. Designer nucleases to treat malignant cancers driven by viral oncogenes
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Tristan A. Scott and Kevin V. Morris
- Subjects
Oncogenic viruses ,ZFN ,TALEN ,CRISPR/Cas ,HPV ,Gammaherpesvirus ,Infectious and parasitic diseases ,RC109-216 - Abstract
Abstract Viral oncogenic transformation of healthy cells into a malignant state is a well-established phenomenon but took decades from the discovery of tumor-associated viruses to their accepted and established roles in oncogenesis. Viruses cause ~ 15% of know cancers and represents a significant global health burden. Beyond simply causing cellular transformation into a malignant form, a number of these cancers are augmented by a subset of viral factors that significantly enhance the tumor phenotype and, in some cases, are locked in a state of oncogenic addiction, and substantial research has elucidated the mechanisms in these cancers providing a rationale for targeted inactivation of the viral components as a treatment strategy. In many of these virus-associated cancers, the prognosis remains extremely poor, and novel drug approaches are urgently needed. Unlike non-specific small-molecule drug screens or the broad-acting toxic effects of chemo- and radiation therapy, the age of designer nucleases permits a rational approach to inactivating disease-causing targets, allowing for permanent inactivation of viral elements to inhibit tumorigenesis with growing evidence to support their efficacy in this role. Although many challenges remain for the clinical application of designer nucleases towards viral oncogenes; the uniqueness and clear molecular mechanism of these targets, combined with the distinct advantages of specific and permanent inactivation by nucleases, argues for their development as next-generation treatments for this aggressive group of cancers.
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- 2021
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33. Wastewater Surveillance Can Function as an Early Warning System for COVID-19 in Low-Incidence Settings
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Mohamad Assoum, Colleen L. Lau, Phong K. Thai, Warish Ahmed, Jochen F. Mueller, Kevin V. Thomas, Phil Min Choi, Greg Jackson, and Linda A. Selvey
- Subjects
wastewater surveillance ,SARS-CoV-2 ,early warning ,surveillance ,Medicine - Abstract
Introduction: During the first two years of the COVID-19 pandemic, Australia implemented a series of international and interstate border restrictions. The state of Queensland experienced limited COVID-19 transmission and relied on lockdowns to stem any emerging COVID-19 outbreaks. However, early detection of new outbreaks was difficult. In this paper, we describe the wastewater surveillance program for SARS-CoV-2 in Queensland, Australia, and report two case studies in which we aimed to assess the potential for this program to provide early warning of new community transmission of COVID-19. Both case studies involved clusters of localised transmission, one originating in a Brisbane suburb (Brisbane Inner West) in July–August 2021, and the other originating in Cairns, North Queensland in February–March 2021. Materials and Methods: Publicly available COVID-19 case data derived from the notifiable conditions (NoCs) registry from the Queensland Health data portal were cleaned and merged spatially with the wastewater surveillance data using statistical area 2 (SA2) codes. The positive predictive value and negative predictive value of wastewater detection for predicting the presence of COVID-19 reported cases were calculated for the two case study sites. Results: Early warnings for local transmission of SARS-CoV-2 through wastewater surveillance were noted in both the Brisbane Inner West cluster and the Cairns cluster. The positive predictive value of wastewater detection for the presence of notified cases of COVID-19 in Brisbane Inner West and Cairns were 71.4% and 50%, respectively. The negative predictive value for Brisbane Inner West and Cairns were 94.7% and 100%, respectively. Conclusions: Our findings highlight the utility of wastewater surveillance as an early warning tool in low COVID-19 transmission settings.
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- 2023
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34. Rapid Biomarker-Based Diagnosis of Fibromyalgia Syndrome and Related Rheumatologic Disorders by Portable FT-IR Spectroscopic Techniques
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Siyu Yao, Haona Bao, Shreya Madhav Nuguri, Lianbo Yu, Zhanna Mikulik, Michelle M. Osuna-Diaz, Katherine R. Sebastian, Kevin V. Hackshaw, and Luis Rodriguez-Saona
- Subjects
fibromyalgia ,biomarker ,disease diagnostics ,portable FT-IR spectroscopy ,chemometrics ,blood ,Biology (General) ,QH301-705.5 - Abstract
Fibromyalgia syndrome (FM), one of the most common illnesses that cause chronic widespread pain, continues to present significant diagnostic challenges. The objective of this study was to develop a rapid vibrational biomarker-based method for diagnosing fibromyalgia syndrome and related rheumatologic disorders (systemic lupus erythematosus (SLE), osteoarthritis (OA) and rheumatoid arthritis (RA)) through portable FT-IR techniques. Bloodspot samples were collected from patients diagnosed with FM (n = 122) and related rheumatologic disorders (n = 70), including SLE (n = 17), RA (n = 43), and OA (n = 10), and stored in conventional protein saver bloodspot cards. The blood samples were prepared by four different methods (blood aliquots, protein-precipitated extraction, and non-washed and water-washed semi-permeable membrane filtration extractions), and spectral data were collected with a portable FT-IR spectrometer. Pattern recognition analysis, OPLS-DA, was able to identify the signature profile and classify the spectra into corresponding classes (Rcv > 0.93) with excellent sensitivity and specificity. Peptide backbones and aromatic amino acids were predominant for the differentiation and might serve as candidate biomarkers for syndromes such as FM. This research evaluated the feasibility of portable FT-IR combined with chemometrics as an accurate and high-throughput tool for distinct spectral signatures of biomarkers related to the human syndrome (FM), which could allow for real-time and in-clinic diagnostics of FM.
- Published
- 2023
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35. Conditionally Replicating Vectors Mobilize Chimeric Antigen Receptors against HIV
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Ryan Z. Urak, Citradewi Soemardy, Roslyn Ray, Shirley Li, Galina Shevchenko, Tristan Scott, Laura Lim, Xiuli Wang, and Kevin V. Morris
- Subjects
HIV ,CAR T-cell ,45-46 ,Conditionally replicating vectors ,Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Human immunodeficiency virus (HIV) is an attractive target for chimeric antigen receptor (CAR) therapy. CAR T cells have proved remarkably potent in targeted killing of cancer cells, and we surmised that CAR T cells could prove useful in eradicating HIV-infected cells. Toward this goal, we interrogate several neutralizing single-chain variable fragments (scFvs) that target different regions of the HIV envelope glycoprotein, gp120. We find here that CAR T cells with scFv from NIH45-46 antibody demonstrated the highest cytotoxicity. Although NIH45-46 CAR T cells are capable of eliminating antigen-expressing cells, we wanted to address HIV reactivation from ex vivo culture of HIV patient-derived CAR T cells. In order to capitalize on the HIV reactivation, we developed a conditionally replicating lentiviral vector (crLV). The crLV can hijack HIV machinery, forming a chimeric lentivirus (LV) instead of HIV and delivered to uninfected cells. We find that CAR T cells generated with crLVs have similar CAR-mediated functionality as traditional CARs. We also demonstrate crLVs’ capability of expanding CAR percentage and protecting CD4 CAR T cell in HIV donors. Collectively, we demonstrate here that the novel crLV NIH45-46 CAR can serve as a strategy to combat HIV, as well as overcome HIV reactivation in CD4+ CAR T cells.
- Published
- 2020
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36. Longitudinal Changes in Health-Related Quality of Life in Primary Glomerular Disease: Results From the CureGN Study
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Shannon L. Murphy, John D. Mahan, Jonathan P. Troost, Tarak Srivastava, Amy J. Kogon, Yi Cai, T. Keefe Davis, Hilda Fernandez, Alessia Fornoni, Rasheed A. Gbadegesin, Emily Herreshoff, Pietro A. Canetta, Patrick H. Nachman, Bryce B. Reeve, David T. Selewski, Christine B. Sethna, Chia-shi Wang, Sharon M. Bartosh, Debbie S. Gipson, Katherine R. Tuttle, Ali Gharavi, Wooin Ahn, Gerald B. Appel, Rupali S. Avasare, Revekka Babayev, Ibrahim Batal, Andrew S. Bomback, Eric Brown, Eric S. Campenot, Pietro Canetta, Brenda Chan, Vivette D. D’Agati, Bartosz Foroncewicz, Gian Marco Ghiggeri, William H. Hines, Namrata G. Jain, Krzysztof Kiryluk, Fangming Lin, Francesca Lugani, Maddalena Marasa, Glen Markowitz, Sumit Mohan, Krzysztof Mucha, Thomas L. Nickolas, Jai Radhakrishnan, Maya K. Rao, Renu Regunathan-Shenk, Simone Sanna-Cherchi, Dominick Santoriello, Michael B. Stokes, Natalie Yu, Anthony M. Valeri, Ronald Zviti, Larry A. Greenbaum, William E. Smoyer, Amira Al-Uzri, Isa Ashoor, Diego Aviles, Rossana Baracco, John Barcia, Sharon Bartosh, Craig Belsha, Michael C. Braun, Aftab Chishti, Donna Claes, Carl Cramer, Keefe Davis, Elif Erkan, Daniel Feig, Michael Freundlich, Melisha Hanna, Guillermo Hidalgo, Amrish Jain, Myda Khalid, Mahmoud Kallash, MD, Jerome C. Lane, John Mahan, Nisha Mathews, Carla Nester, Cynthia Pan, Hiren Patel, Adelaide Revell, Rajasree Sreedharan, Julia Steinke, Scott E. Wenderfer, Craig S. Wong, Ronald Falk, William Cook, Vimal Derebail, Agnes Fogo, Adil Gasim, Todd Gehr, Raymond Harris, Jason Kidd, Louis-Philippe Laurin, Will Pendergraft, Vincent Pichette, Thomas Brian Powell, Matthew B. Renfrow, Virginie Royal, Lawrence B. Holzman, Sharon Adler, Charles Alpers, Raed Bou Matar, Elizabeth Brown, Michael Choi, Katherine M. Dell, Ram Dukkipati, Fernando C. Fervenza, Crystal Gadegbeku, Patrick Gipson, Leah Hasely, Sangeeta Hingorani, Michelle A. Hladunewich, Jonathan Hogan, J. Ashley Jefferson, Kenar Jhaveri, Duncan B. Johnstone, Frederick Kaskel, Amy Kogan, Jeffrey Kopp, Kevin V. Lemley, Laura Malaga- Dieguez, Kevin Meyers, Alicia Neu, Michelle Marie O'Shaughnessy, John F. O’Toole, Rulan Parekh, Heather Reich, Kimberly Reidy, Helbert Rondon, Kamalanathan K. Sambandam, John R. Sedor, Jeffrey Schelling, John C. Sperati, Agnes Swiatecka-Urban, Howard Trachtman, Joseph Weisstuch, Olga Zhdanova, Brenda Gillespie, Matthias Kretzler, Bruce M. Robinson, Laura Mariani, Matthew Wladkowski, and Lisa M. Guay-Woodford
- Subjects
edema ,health-related quality of life ,patient-reported outcomes ,primary glomerular disease ,Diseases of the genitourinary system. Urology ,RC870-923 - Abstract
Introduction: Prior cross-sectional studies suggest that health-related quality of life (HRQOL) worsens with more severe glomerular disease. This longitudinal analysis was conducted to assess changes in HRQOL with changing disease status. Methods: Cure Glomerulonephropathy (CureGN) is a cohort of patients with minimal change disease, focal segmental glomerulosclerosis, membranous nephropathy, IgA vasculitis, or IgA nephropathy. HRQOL was assessed at enrollment and follow-up visits 1 to 3 times annually for up to 5 years with the Patient-Reported Outcomes Measurement Information System (PROMIS). Global health, anxiety, and fatigue domains were measured in all; mobility was measured in children; and sleep-related impairment was measured in adults. Linear mixed effects models were used to evaluate HRQOL responsiveness to changes in disease status. Results: A total of 469 children and 1146 adults with PROMIS scores were included in the analysis. HRQOL improved over time in nearly all domains, though group-level changes were modest. Edema was most consistently associated with worse HRQOL across domains among children and adults. A greater number of symptoms also predicted worse HRQOL in all domains. Sex, age, obesity, and serum albumin were associated with some HRQOL domains. The estimated glomerular filtration rate (eGFR) was only associated with fatigue and adult physical health; proteinuria was not associated with any HRQOL domain in adjusted models. Conclusion: HRQOL measures were responsive to changes in disease activity, as indicated by edema. HRQOL over time was not predicted by laboratory-based markers of disease. Patient-reported edema and number of symptoms were the strongest predictors of HRQOL, highlighting the importance of the patient experience in glomerular disease. HRQOL outcomes inform understanding of the patient experience for children and adults with glomerular diseases.
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- 2020
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37. Preventive drugs for Huntington’s disease: A choice-based conjoint survey of patient preferences
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Marcus C. Parrish, Andrea Hanson-Kahn, V. Srinivasan, and Kevin V. Grimes
- Subjects
Huntington’s disease ,patient preferences ,choice-based conjoint survey ,preventive treatment ,regulatory policy ,Medicine - Abstract
Abstract Introduction: This research examined the perspective of the Huntington’s disease (HD) community regarding the use of predictive biomarkers as endpoints for regulatory approval of therapeutics to prevent or delay the onset of clinical HD in asymptomatic mutation carriers. Methods: An online, choice-based conjoint survey was shared with HD community members including untested at-risk individuals, presymptomatic mutation carriers, and symptomatic individuals. Across 15 scenarios, participants chose among two proposed therapies with differing degrees of biomarker improvement and side effects or a third option of no treatment. Results: Two hundred and thirty-eight responses were received. Attributes reflecting biomarker efficacy (e.g., prevention of brain atrophy on magnetic resonance imaging, reduced mutant huntingtin, or reduced inflammation biomarkers) had 3- to 7-fold greater importance than attributes representing side effects (e.g., increased risk of heart disease, cancer, and stroke over 20 years) and were more influential in directing choice of treatments. Reduction in mutant huntingtin protein was the most valued attribute overall. Multinomial logit model simulations based on survey responses demonstrated high interest among respondents (87–99% of the population) for drugs that might prevent or delay HD solely based upon biomarker evidence, even at the risk of serious side effects. Conclusion: These results indicate a strong desire among members of the HD community for preventive therapeutics and a willingness to accept significant side effects, even before the drug has been shown to definitively delay disease onset if the drug improves biomarker evidence of HD progression. Preferences of the HD community should inform regulatory policies for approving preventive therapies.
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- 2022
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38. Intravital imaging reveals glomerular capillary distension and endothelial and immune cell activation early in Alport syndrome
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Georgina Gyarmati, Urvi Nikhil Shroff, Audrey Izuhara, Xiaogang Hou, Stefano Da Sacco, Sargis Sedrakyan, Kevin V. Lemley, Kerstin Amann, Laura Perin, and János Peti-Peterdi
- Subjects
Nephrology ,Medicine - Abstract
Alport syndrome (AS) is a genetic disorder caused by mutations in type IV collagen that lead to defective glomerular basement membrane, glomerular filtration barrier (GFB) damage, and progressive chronic kidney disease. While the genetic basis of AS is well known, the molecular and cellular mechanistic details of disease pathogenesis have been elusive, hindering the development of mechanism-based therapies. Here, we performed intravital multiphoton imaging of the local kidney tissue microenvironment in a X-linked AS mouse model to directly visualize the major drivers of AS pathology. Severely distended glomerular capillaries and aneurysms were found accompanied by numerous microthrombi, increased glomerular endothelial surface layer (glycocalyx) and immune cell homing, GFB albumin leakage, glomerulosclerosis, and interstitial fibrosis by 5 months of age, with an intermediate phenotype at 2 months. Renal histology in mouse or patient tissues largely failed to detect capillary aberrations. Treatment of AS mice with hyaluronidase or the ACE inhibitor enalapril reduced the excess glomerular endothelial glycocalyx and blocked immune cell homing and GFB albumin leakage. This study identified central roles of glomerular mechanical forces and endothelial and immune cell activation early in AS, which could be therapeutically targeted to reduce mechanical strain and local tissue inflammation and improve kidney function.
- Published
- 2022
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39. Wastewater surveillance demonstrates high predictive value for COVID-19 infection on board repatriation flights to Australia
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Warish Ahmed, Aaron Bivins, Stuart L. Simpson, Paul M. Bertsch, John Ehret, Ian Hosegood, Suzanne S. Metcalfe, Wendy J.M. Smith, Kevin V. Thomas, Josh Tynan, and Jochen F. Mueller
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SARS-CoV-2 ,COVID-19 ,Wastewater surveillance ,Human health risks ,Enveloped viruses ,Aircraft ,Environmental sciences ,GE1-350 - Abstract
Controlling importation and transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from overseas travelers is essential for countries, such as Australia, New Zealand, and other island nations, that have adopted a suppression strategy to manage very low community transmission. Wastewater surveillance of SARS-CoV-2 RNA has emerged as a promising tool employed in public health response in many countries globally. This study aimed to establish whether the surveillance of aircraft wastewater can be used to provide an additional layer of information to augment individual clinical testing. Wastewater from 37 long-haul flights chartered to repatriate Australians was tested for the presence of SARS-CoV-2 RNA. Children 5 years or older on these flights tested negative for coronavirus disease 19 (COVID-19) (deep nasal and oropharyngeal reverse-transcription (RT)-PCR swab) 48 h before departure. All passengers underwent mandatory quarantine for 14-day post arrival in Howard Springs, NT, Australia. Wastewater from 24 (64.9 %) of the 37 flights tested positive for SARS-CoV-2 RNA. During the 14 day mandatory quarantine, clinical testing identified 112 cases of COVID-19. Surveillance for SARS-CoV-2 RNA in repatriation flight wastewater using pooled results from three RT-qPCR assays demonstrated a positive predictive value (PPV) of 87.5 %, a negative predictive value (NPV) of 76.9 % and 83.7% accuracy for COVID-19 cases during the post-arrival 14-day quarantine period. The study successfully demonstrates that the surveillance of wastewater from aircraft for SARS-CoV-2 can provide an additional and effective tool for informing the management of returning overseas travelers and for monitoring the importation of SARS CoV-2 and other clinically significant pathogens.
- Published
- 2022
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40. Nuclear microRNA-466c regulates Vegfa expression in response to hypoxia.
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Pia Laitinen, Mari-Anna Väänänen, Ida-Liisa Kolari, Petri I Mäkinen, Minna U Kaikkonen, Marc S Weinberg, Kevin V Morris, Paula Korhonen, Tarja Malm, Seppo Ylä-Herttuala, Thomas C Roberts, Mikko P Turunen, and Tiia A Turunen
- Subjects
Medicine ,Science - Abstract
MicroRNAs are well characterized in their role in silencing gene expression by targeting 3´-UTR of mRNAs in cytoplasm. However, recent studies have shown that miRNAs have a role in the regulation of genes in the nucleus, where they are abundantly located. We show here that in mouse endothelial cell line (C166), nuclear microRNA miR-466c participates in the regulation of vascular endothelial growth factor a (Vegfa) gene expression in hypoxia. Upregulation of Vegfa expression in response to hypoxia was significantly compromised after removal of miR-466c with CRISPR-Cas9 genomic deletion. We identified a promoter-associated long non-coding RNA on mouse Vegfa promoter and show that miR-466c directly binds to this transcript to modulate Vegfa expression. Collectively, these observations suggest that miR-466c regulates Vegfa gene transcription in the nucleus by targeting the promoter, and expands on our understanding of the role of miRNAs well beyond their canonical role.
- Published
- 2022
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41. Urinary Epidermal Growth Factor as a Marker of Disease Progression in Children With Nephrotic Syndrome
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Debbie S. Gipson, Howard Trachtman, Anne Waldo, Keisha L. Gibson, Sean Eddy, Katherine M. Dell, Tarak Srivastava, Kevin V. Lemley, Larry A. Greenbaum, Sangeeta Hingorani, Kevin E. Meyers, Frederick J. Kaskel, Kimberly J. Reidy, Christine B. Sethna, Cheryl L. Tran, Chia-shi Wang, Katherine R. Tuttle, Gia Oh, Alicia M. Neu, Elizabeth Brown, Jen-Jar Lin, Jennifer Lai Yee, Therese M. Roth, Jonathan P. Troost, Brenda W. Gillespie, Matthew G. Sampson, Matthias Kretzler, and Wenjun Ju
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Diseases of the genitourinary system. Urology ,RC870-923 - Abstract
Introduction: Childhood-onset nephrotic syndrome has a variable clinical course. Improved predictive markers of long-term outcomes in children with nephrotic syndrome are needed. This study tests the association between baseline urinary epidermal growth factor (uEGF) excretion and longitudinal kidney function in children with nephrotic syndrome. Methods: The study evaluated 191 participants younger than 18 years enrolled in the Nephrotic Syndrome Study Network, including 118 with their first clinically indicated kidney biopsy (68 minimal change disease; 50 focal segmental glomerulosclerosis) and 73 with incident nephrotic syndrome without a biopsy. uEGF was measured at baseline for all participants and normalized by the urine creatinine (Cr) concentration. Renal epidermal growth factor (EGF) mRNA was measured in the tubular compartment microdissected from kidney biopsy cores from a subset of patients. Linear mixed models were used to test if baseline uEGF/Cr and EGF mRNA expression were associated with change in estimated glomerular filtration rate (eGFR) over time. Results: Higher uEGF/Cr at baseline was associated with slower eGFR decline during follow-up (median follow-up = 30 months). Halving of uEGF/Cr was associated with a decrease in eGFR slope of 2.0 ml/min per 1.73 m2 per year (P < 0.001) adjusted for age, race, diagnosis, baseline eGFR and proteinuria, and APOL1 genotype. In the biopsied subgroup, uEGF/Cr was correlated with EGF mRNA expression (r = 0.74; P < 0.001), but uEGF/Cr was retained over mRNA expression as the stronger predictor of eGFR slope after multivariable adjustment (decrease in eGFR slope of 1.7 ml/min per 1.73 m2 per year per log2 decrease in uEGF/Cr; P < 0.001). Conclusion: uEGF/Cr may be a useful noninvasive biomarker that can assist in predicting the long-term course of kidney function in children with incident nephrotic syndrome. Keywords: disease progression, epidermal growth factor, focal segmental glomerulosclerosis, nephrotic syndrome, pediatrics
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- 2020
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42. Development of a Facile Approach for Generating Chemically Modified CRISPR/Cas9 RNA
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Tristan Scott, Citradewi Soemardy, and Kevin V. Morris
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Therapeutics. Pharmacology ,RM1-950 - Abstract
The RNA-guided, modified type II prokaryotic CRISPR with CRISPR-associated proteins (CRISPR/Cas9) system represents a simple gene-editing platform with applications in biotechnology and also potentially as a therapeutic modality. The system requires a small guide RNA (sgRNA) and a catalytic Cas9 protein to induce non-homologous end joining (NHEJ) at break sites, resulting in the formation of inactivating mutations, or through homology-directed repair (HDR) can engineer in specific sequence changes. Although CRISPR/Cas9 is a powerful technology, the effects can be limited as a result of nuclease-mediated degradation of the RNA components. Significant research has focused on the solid-phase synthesis of CRISPR RNA components with chemically modified bases, but this approach is technically challenging and expensive. Development of a simple, generic approach to generate chemically modified CRISPR RNAs may broaden applications that require nuclease-resistant CRISPR components. We report here the development of a novel, functional U-replaced trans-activating RNA (tracrRNA) that can be in vitro transcribed with chemically stabilizing 2′-fluoro (2′F)-pyrimidines. These data represent a unique and facile approach to generating chemically stabilized CRISPR RNA.
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- 2020
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43. Out of sight but not out of mind: Size fractionation of plastics bioaccumulated by field deployed oysters
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Francisca Ribeiro, Elvis D. Okoffo, Jake W. O’Brien, Stacey O’Brien, Jonathan M. Harris, Saer Samanipour, Sarit Kaserzon, Jochen F. Mueller, Tamara Galloway, and Kevin V. Thomas
- Subjects
FT-IR ,Py-GC/MS ,Bivalves ,Micro-nanoplastics ,Size fractionation ,Hazardous substances and their disposal ,TD1020-1066 - Abstract
Microplastics contamination has been widely reported in filter feeders yet the < 1 μm size fraction has been largely ignored. In attempt to characterize this sub 1 μm size fraction and better understand the size distribution of microplastics contamination in filter feeders, field deployed oysters were characterised using a combination of size fractionation combined with pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS) as well as Fourier Transform-Infrared Spectroscopy (μFT-IR). Sequential filtration followed by Py-GC/MS identified the 1–22 μm fraction to contain the highest total plastic mass concentration (Ʃ31 mg/g), followed by the 22 μm fraction (Ʃ0.1 mg/g). μFT-IR identified 0.2 particles/g tissue but was limited to particles >150 μm in size. Our results clearly show that an important size fraction of microplastics is being overlooked in almost all studies published to date that rely on FTIR for polymer identification.
- Published
- 2021
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44. AMPK mediates regulation of glomerular volume and podocyte survival
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Khadija Banu, Qisheng Lin, John M. Basgen, Marina Planoutene, Chengguo Wei, Anand C. Reghuvaran, Xuefei Tian, Hongmei Shi, Felipe Garzon, Aitor Garzia, Nicholas Chun, Arun Cumpelik, Andrew D. Santeusanio, Weijia Zhang, Bhaskar Das, Fadi Salem, Li Li, Shuta Ishibe, Lloyd G. Cantley, Lewis Kaufman, Kevin V. Lemley, Zhaohui Ni, John Cijiang He, Barbara Murphy, and Madhav C. Menon
- Subjects
Cell biology ,Nephrology ,Medicine - Abstract
Herein, we report that Shroom3 knockdown, via Fyn inhibition, induced albuminuria with foot process effacement (FPE) without focal segmental glomerulosclerosis (FSGS) or podocytopenia. Interestingly, knockdown mice had reduced podocyte volumes. Human minimal change disease (MCD), where podocyte Fyn inactivation was reported, also showed lower glomerular volumes than FSGS. We hypothesized that lower glomerular volume prevented the progression to podocytopenia. To test this hypothesis, we utilized unilateral and 5/6th nephrectomy models in Shroom3-KD mice. Knockdown mice exhibited less glomerular and podocyte hypertrophy after nephrectomy. FYN-knockdown podocytes had similar reductions in podocyte volume, implying that Fyn was downstream of Shroom3. Using SHROOM3 or FYN knockdown, we confirmed reduced podocyte protein content, along with significantly increased phosphorylated AMPK, a negative regulator of anabolism. AMPK activation resulted from increased cytoplasmic redistribution of LKB1 in podocytes. Inhibition of AMPK abolished the reduction in glomerular volume and induced podocytopenia in mice with FPE, suggesting a protective role for AMPK activation. In agreement with this, treatment of glomerular injury models with AMPK activators restricted glomerular volume, podocytopenia, and progression to FSGS. Glomerular transcriptomes from MCD biopsies also showed significant enrichment of Fyn inactivation and Ampk activation versus FSGS glomeruli. In summary, we demonstrated the important role of AMPK in glomerular volume regulation and podocyte survival. Our data suggest that AMPK activation adaptively regulates glomerular volume to prevent podocytopenia in the context of podocyte injury.
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- 2021
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45. An Introduction to Stereology with Applications to the Glomerulus
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Kevin V. Lemley
- Subjects
Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Published
- 2021
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46. Cocultivation of Anaerobic Fungi with Rumen Bacteria Establishes an Antagonistic Relationship
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Candice L. Swift, Katherine B. Louie, Benjamin P. Bowen, Casey A. Hooker, Kevin V. Solomon, Vasanth Singan, Chris Daum, Christa P. Pennacchio, Kerrie Barry, Vaithiyalingam Shutthanandan, James E. Evans, Igor V. Grigoriev, Trent R. Northen, and Michelle A. O’Malley
- Subjects
RNA-seq ,transcriptomics ,cocultivation ,secondary metabolism ,fungi ,anaerobe ,Microbiology ,QR1-502 - Abstract
ABSTRACT Anaerobic gut fungi (Neocallimastigomycetes) live in the digestive tract of large herbivores, where they are vastly outnumbered by bacteria. It has been suggested that anaerobic fungi challenge growth of bacteria owing to the wealth of biosynthetic genes in fungal genomes, although this relationship has not been experimentally tested. Here, we cocultivated the rumen bacteria Fibrobacter succinogenes strain UWB7 with the anaerobic gut fungi Anaeromyces robustus or Caecomyces churrovis on a range of carbon substrates and quantified the bacterial and fungal transcriptomic response. Synthetic cocultures were established for at least 24 h, as verified by active fungal and bacterial transcription. A. robustus upregulated components of its secondary metabolism in the presence of Fibrobacter succinogenes strain UWB7, including six nonribosomal peptide synthetases, one polyketide synthase-like enzyme, and five polyketide synthesis O-type methyltransferases. Both A. robustus and C. churrovis cocultures upregulated S-adenosyl-l-methionine (SAM)-dependent methyltransferases, histone methyltransferases, and an acetyltransferase. Fungal histone 3 lysine 27 trimethylation marks were more abundant in coculture, and heterochromatin protein-1 was downregulated. Together, these findings suggest that fungal chromatin remodeling occurs when bacteria are present. F. succinogenes strain UWB7 upregulated four genes in coculture encoding drug efflux pumps, which likely protect the cell against toxins. Furthermore, untargeted nonpolar metabolomics data revealed at least one novel fungal metabolite enriched in coculture, which may be a defense compound. Taken together, these data suggest that A. robustus and C. churrovis produce antimicrobials when exposed to rumen bacteria and, more broadly, that anaerobic gut fungi are a source of novel antibiotics. IMPORTANCE Anaerobic fungi are outnumbered by bacteria by 4 orders of magnitude in the herbivore rumen. Despite their numerical disadvantage, they are resilient members of the rumen microbiome. Previous studies mining the genomes of anaerobic fungi identified genes encoding enzymes to produce natural products, which are small molecules that are often antimicrobials. In this work, we cocultured the anaerobic fungus Anaeromyces robustus or Caecomyes churrovis with rumen bacteria Fibrobacter succinogenes strain UWB7 and sequenced fungal and bacterial active genes via transcriptome sequencing (RNA-seq). Consistent with production of a fungal defense compound, bacteria upregulated genes encoding drug efflux pumps, which often export toxic molecules, and fungi upregulated genes encoding biosynthetic enzymes of natural products. Furthermore, tandem mass spectrometry detected an unknown fungal metabolite enriched in the coculture. Together, these findings point to an antagonistic relationship between anaerobic fungi and rumen bacteria resulting in the production of a fungal compound with potential antimicrobial activity.
- Published
- 2021
- Full Text
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47. Opportunities and Challenges of Predictive Approaches for Harnessing the Potential of Genetic Resources
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Johannes W. R. Martini, Terence L. Molnar, José Crossa, Sarah J. Hearne, and Kevin V. Pixley
- Subjects
genetic resources ,germplasm bank collections ,predictive breeding ,genomic selection ,pre-breeding ,Plant culture ,SB1-1110 - Published
- 2021
- Full Text
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48. Essential Oils and Neuropathic Pain
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Imane Ridouh and Kevin V. Hackshaw
- Subjects
neuropathic pain ,essential oils ,lavender ,bergamot ,billy goat weed ,nutmeg ,Botany ,QK1-989 - Abstract
Neuropathic pain is one of the most prominent chronic pain syndromes, affecting almost 10% of the United States population. While there are a variety of established pharmacologic and non-pharmacologic treatment options, including tricyclic antidepressants (TCAs), serotonin-noradrenaline reuptake inhibitors, anticonvulsants, trigger point injections, and spinal cord stimulators, many patients continue to have chronic pain or suboptimal symptom control. This has led to an increased interest in alternative solutions for neuropathic pain such as nutritional supplements and essential oils. In this review, we explore the literature on the most commonly cited essential oils, including lavender, bergamot, rosemary, nutmeg, Billy goat weed, and eucalyptus. However, the literature is limited and largely comprised of preclinical animal models and a few experimental studies, some of which were poorly designed and did not clearly isolate the effects of the essential oil treatment. Additionally, no standardized method of dosing or route of administration has been established. Further randomized control studies isolating the active components of various essential oils are needed to provide conclusive evidence on the use of essential oils for neuropathic pain. In this review, we explore the basis behind some of the essential oils of interest to patients with neuropathic pain seen in rheumatology clinics.
- Published
- 2022
- Full Text
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49. Control of LDL Uptake in Human Cells by Targeting the LDLR Regulatory Long Non-coding RNA BM450697
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Roslyn M. Ray, Anders Højgaard Hansen, Sofie Slott, Maria Taskova, Kira Astakhova, and Kevin V. Morris
- Subjects
Therapeutics. Pharmacology ,RM1-950 - Abstract
Hypercholesterolemia is a condition that is characterized by very high levels of cholesterol in the blood and is a major correlating factor with heart disease. Indeed, high levels of the low-density lipoprotein (LDL) have been causally linked to the development of atherosclerotic cardiovascular disease (ASCVD). A method to specifically reduce cholesterol in the blood in a long-term, stable manner could prove therapeutically relevant. Cholesterol is removed from the blood by the LDL receptor (LDLR) in the liver. Others and we have discovered that a long non-coding RNA (lncRNA; BM450697) functions as an endogenous epigenetic regulator of LDLR and that the repression of this lncRNA by the action of small interfering RNAs (siRNAs) results in the activation of LDLR. We found here, through the interrogation of two siRNAs that can target this lncRNA, both in a transcriptional and post-transcriptional manner, that BM450697 functions as a local scaffold for modulating LDLR transcription. Moreover, we found that conjugation of α-N-acetylgalactosamine (GalNAc) with two lncRNA-directed siRNAs allows for direct liver cell targeting of this lncRNA and functional enhanced uptake of cholesterol. Collectively, these data suggest that targeting the BM450697 lncRNA regulator of LDLR may result in a more specific, long-term, targeted approach to regulating cholesterol in the blood. Keywords: LDL, LDLR, lncRNA, cholesterol, transcriptional gene silencing, Galnac, siRNA, hypercholesterolemia
- Published
- 2019
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50. Trends in nicotine consumption between 2010 and 2017 in an Australian city using the wastewater-based epidemiology approach
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Rachel S. Mackie, Benjamin J. Tscharke, Jake W. O'Brien, Phil M. Choi, Coral E. Gartner, Kevin V. Thomas, and Jochen F. Mueller
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Environmental sciences ,GE1-350 - Abstract
Monitoring smoking prevalence is key to assessing responses to tobacco control measures, and evaluating associated health and economic costs. Estimates of tobacco consumed in Australia are based on various data sources – tax excise clearances, sales, and self-report surveys. There are limitations with each of these data sources which makes triangulation of cigarette use estimates by multiple methods important. Wastewater-based epidemiology, the systematic sampling and analysis of wastewater, is now a routine method to measure and monitor human exposure to a range of chemicals. This study provides a high frequency long-term temporal assessment of exposure to nicotine, the main addictive component of tobacco, using this approach. 291 archived wastewater samples collected from a regional city catchment from 2010 to 2017 were analysed for human-specific nicotine metabolites (cotinine and trans-3′-hydroxycotinine), to estimate per capita nicotine use. Temporal trends in nicotine use determined by wastewater-based epidemiology were compared with national sales and survey data. Wastewater analysis showed a 25% reduction in the mean number of cigarette equivalents consumed from 2010 to 2017, representing a 3% annual decline. These findings are in good agreement with estimates based on surveys and sales data, indicating annual declines of 5% and 4%, respectively. Findings of this study demonstrate WBE to be a relatively cost-effective and objective approach to reporting long-term data on nicotine consumption. When combined with alternative data sources, and valuable sociodemographic information of surveys, wastewater-based epidemiology helps to refine our estimates and understanding of the total impacts of smoking. Keywords: Tobacco consumption, Nicotine, Wastewater-based epidemiology, Temporal monitoring
- Published
- 2019
- Full Text
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