UFGI publication round-up week 5/14
Circumferential-scanning endoscopic optical coherence tomography probe based on a circular array of six 2-axis MEMS mirrors.
Author information
- 1
- Department of Electronic Engineering, Tsinghua University, Beijing, 100084, China.
- 2
- Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518000, China.
- 3
- University of Florida, Gainesville, FL, 26110-613, USA.
- 4
- School of Physics and Electronic Engineering, Zhengzhou University of Light Industry, Zhengzhou, 450000, China.
- 5
- Wuxi WiO Technologies Co. Ltd., Wuxi, 214000, China.
- 6
- Department of General Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515 China.
- 7
- Foshan Optomedic Technologies Co., Ltd. Foshan, 280000, China.
- 8
- Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, 215000, China.
- 9
- lhuo@mail.tsinghua.edu.cn.
- 10
- hkx@ufl.edu.
Abstract
We present a novel circumferential-scan endoscopic optical coherence tomography (OCT) probe by using a circular array of six electrothermal microelectromechanical (MEMS) mirrors and six C-lenses. The MEMS mirrors have a 0.5 mm × 0.5 mm mirror plate and a chip size of 1.5 mm × 1.3 mm. Each MEMS mirror can scan up to 45° at a voltage of less than 12 V. Six of those mirrors have been successfully packaged to a probe head; full circumferential scans have been demonstrated. Furthermore, each scan unit is composed of a MEMS mirror and a C-lens and the six scan units can be designed with different focal lengths to adapt for lesions with uneven surfaces. Configured with a swept source OCT system, this MEMS array-based circumferential scanning probe has been applied to image a swine’s small intestine wrapped on a 20 mm-diameter glass tube. The OCT imaging result shows that this new MEMS endoscopic OCT has promising applications in large tubular organs.
A study on the safety and efficacy of reveglucosidase alfa in patients with late-onset Pompe disease.
Author information
- 1
- University of Florida, School of Medicine, 1600 SW Archer Road, Gainesville, FL, 32607, USA. bbyrne@ufl.edu.
- 2
- Department of Pediatrics, University of Florida, P.O. Box 100296, Gainesville, FL, 32610, USA. bbyrne@ufl.edu.
- 3
- University Hospital Birmingham, Mindelsohn Way, Edgbaston, Birmingham, B15 2GW, UK.
- 4
- University of California San Diego Health System, 4168 Front Street, San Diego, CA, 92103, USA.
- 5
- Kansas University Medical Center, 3901 Rainbow Blvd/MSN 2012, Kansas City, KS, 66160, USA.
- 6
- Royal Free London NHS Foundation & University College London Department of Hematology, Pond St, London, NW3 2QG, UK.
- 7
- SA Pathology, Frome Rd, Adelaide, SA, 5000, Australia.
- 8
- University Hospital of Nice, Pasteur Hospital, Nice, France.
- 9
- Paris-Est Neuromuscular Center, INSERM U974, UPMC, Hôpital Pitié-Salpêtrière, 47-83 boulevard de l’Hôpital, 75013, Paris, France.
- 10
- Johannes Gutenberg University, Langenbeckstr. 1, 55131, Mainz, Germany.
- 11
- Salford Royal NHS Foundation Trust, M6 8HD, Salford, UK.
- 12
- BioMarin Pharmaceutical, 105 Digital Drive, Novato, CA, 94949, USA.
Abstract
BACKGROUND:
Late-onset Pompe disease is a rare genetic neuromuscular disorder caused by lysosomal acid alpha-glucosidase (GAA) deficiency that ultimately results in mobility loss and respiratory failure. Current enzyme replacement therapy with recombinant human (rh)GAA has demonstrated efficacy in subjects with late-onset Pompe disease. However, long-term effects of rhGAA on pulmonary function have not been observed, likely related to inefficient delivery of rhGAA to skeletal muscle lysosomes and associated deficits in the central nervous system. To address this limitation, reveglucosidase alfa, a novel insulin-like growth factor 2 (IGF2)-tagged GAA analogue with improved lysosomal uptake, was developed. This study evaluated the pharmacokinetics, safety, and exploratory efficacy of reveglucosidase alfa in 22 subjects with late-onset Pompe disease who were previously untreated with rhGAA.
RESULTS:
Reveglucosidase alfa plasma concentrations increased linearly with dose, and the elimination half-life was <1.2 h. Eighteen of 22 subjects completed 72 weeks of treatment. The most common adverse events were hypoglycemia (63%), dizziness, fall, headache, and nausea (55% for each). Serious adverse events included hypersensitivity (n = 1), symptomatic hypoglycemia (n = 2), presyncope (n = 1), and acute cardiac failure (n = 1). In the dose-escalation study, all treated subjects tested positive for anti-reveglucosidase alfa, anti-rhGAA, anti-IGF1, and anti-IGF2 antibodies at least once. Subjects receiving 20 mg/kg of reveglucosidase alfa demonstrated increases in predicted maximum inspiratory pressure (13.9%), predicted maximum expiratory pressure (8.0%), forced vital capacity (-0.4%), maximum voluntary ventilation (7.4 L/min), and mean absolute walking distance (22.3 m on the 6-min walk test) at 72 weeks.
CONCLUSIONS:
Additional studies are needed to further assess the safety and efficacy of this approach. Improvements in respiratory muscle strength, lung function, and walking endurance in subjects with LOPD may make up for the risk of hypersensitivity reactions and hypoglycemia. Reveglucosidase alfa may provide a new treatment option for patients with late-onset Pompe disease.
Administration of low dose triclosan to pregnant ewes results in placental uptake and reduced estradiol sulfotransferase activity in fetal liver and placenta.
Author information
- 1
- Department of Medicinal Chemistry, University of Florida, Gainesville, Florida, 32610, USA.
- 2
- Department of Medicinal Chemistry, University of Florida, Gainesville, Florida, 32610, USA. Electronic address: mojames@ufl.edu.
- 3
- Department of Physiology and Functional Genomics, University of Florida, Gainesville, Florida, 32610, USA.
Abstract
Sulfonation is a major pathway of estrogen biotransformation with a role in regulating estrogen homeostasis in humans and sheep. Previous in vitro studies found that triclosan is an especially potent competitive inhibitor of ovine placental estrogen sulfotransferase, with Kic of <0.1 nM. As the placenta is the main organ responsible for estrogen synthesis in pregnancy in both women and sheep, and the liver is another site of estrogen biotransformation, this study examined the effects of triclosan exposure of pregnant ewes on placental and hepatic sulfotransferase activity. Triclosan, 0.1 mg/kg/day, or saline vehicle was administered to late gestation fetal sheep for two days either by direct infusion into the fetal circulation or infusion into the maternal blood. On the third day, fetal liver and placenta were harvested and analyzed for triclosan and for cytosolic estradiol sulfotransferase activity. Placenta contained higher concentrations of triclosan than liver in each individual sheep in both treatment groups. There was a negative correlation between triclosan tissue concentration (pmol/g tissue) and cytosolic sulfotransferase activity (pmol/min/mg protein) towards estradiol. These findings demonstrated that in the sheep exposed to very low concentrations of triclosan, this substance is taken up into placenta and reduces estrogen sulfonation.
Non-accidental Trauma Work-up: Unusual Retinal Finding Leads to a Rare Diagnosis.
Author information
- 1
- From the *Shands Children’s Hospital, Department of Pediatrics, †College of Medicine, ‡Division of Genetics and Metabolism, §Raymond C. Philips Unit, Division of Genetics and Metabolism, Department of Pediatrics, and ∥Congenital Heart Center, Department of Pediatrics, University of Florida, Gainesville, FL.
Abstract
Lipoprotein lipase (LPL) deficiency is an autosomal recessive condition due to absent or decreased activity of LPL enzyme. The LPL deficiency is a rare condition that is mainly diagnosed in children, but there is no standard screening method at this time. In our report, we describe a 6-day-old male infant who was found to have hypertriglyceridemia after lipemia retinalis was diagnosed from a fundoscopic examination for nonaccidental trauma work-up. After dietary modification was done, his triglyceride levels decreased significantly, and there were no complications. When diagnosed later in life, recurrent pancreatitis can be a significant complication.
The Post-injury Inflammatory State and the Bone Marrow Response to Anemia.
Author information
- 1
- University of Florida, 3463, Surgery, Gainesville, Florida, United States.
- 2
- Uninversity of Florida, Surgery, Gainesville, Florida, United States.
- 3
- University of Florida, Gainesville, Florida, United States.
- 4
- University of Florida, Surgery, Gainesville, Florida, United States.
- 5
- University of Florida, Surgery, Gainesville, Florida, United States ; Alicia.mohr@surgery.ufl.edu.
Abstract
RATIONALE:
The pathophysiology of persistent injury-associated anemia is incompletely understood, and human data are sparse.
OBJECTIVE:
To translate pre-clinical findings by characterizing injury-associated anemia among critically ill trauma patients. This study compared elective hip repair patients with an acute stress response to severely injured trauma patients with a prolonged stress response and evaluated the bone marrow response to anemia.
METHODS:
A translational prospective observational cohort study comparing severely injured, blunt trauma patients who had operative fixation of a hip or femur fracture (n=17) with elective hip repair patients (n=22) and healthy donors (n=8).
MEASUREMENTS AND MAIN RESULTS:
During admission, trauma patients had 625 mL operative blood loss, five units of red blood cell transfusions, and hemoglobin decreased from 10.5 to 9.3 g/dL. Compared with hip repair, trauma patients had higher plasma norepinephrine (21.9 vs. 8.9 ng/mL*) and hepcidin concentrations (56.3 vs. 12.2 ng/mL*). Bone marrow erythropoietin and erythropoietin receptor expression were increased among hip repairs (23% increase* and 14% increase*, respectively), but not trauma patients (3% increase and 5% increase, respectively) compared with healthy controls. Trauma patients had lower bone marrow transferrin receptor expression than hip repair (57% decrease*). Erythroid progenitor growth was decreased in trauma patients (39.0 colonies/plate*) compared with hip repair (57.0 colonies/plate) and healthy controls (66.5 colonies/plate).
CONCLUSIONS:
Elective hip repair was associated with a mild neuroendocrine activation and an appropriate bone marrow response to anemia. Severe blunt trauma was associated with exaggerated neuroendocrine activation, erythropoietin dysfunction, iron dysregulation, erythroid progenitor growth suppression and persistent injury-associated anemia. Clinical trial registration available at www.clinicaltrials.gov, ID NCT02577731.
Toxicity of functionalized fullerene and fullerene synthesis chemicals.
Author information
- 1
- University of Florida, Department of Environmental Engineering Sciences, Black Hall, Gainesville, FL 32611, USA. Electronic address: indeglia@gmail.com.
- 2
- University of Florida, Particle Engineering Research Center, FL, USA.
- 3
- University of Florida, Center for Environmental and Human Toxicology, FL, USA.
- 4
- University of Florida, Department of Environmental Engineering Sciences, Black Hall, Gainesville, FL 32611, USA.
Abstract
Fullerene is one of the most studied carbon-based nanoparticles due to its unique structure and potential for diverse applications. This study focuses on toxicological effects of two fullerene nanomaterials, contributing to ecological as well as human risk assessment strategies. The biological responses from two basic fullerene materials, aqueous-nanoC60 and alkaline-synthesized fullerenol, were examined using four model organisms. Bioassays were conducted on bacteria (Pseudomonas aeruginosa and Staphylococcus aureus) to determine population impacts and to assess mechanisms of cellular effects for both Gram-negative and Gram-positive species. LC50 of aqu-nC60 stirred for 28 days for P. aeruginosa was estimated to be 1336 mg/L; however, toxicity of the same aqu-nC60 preparation for S. aureus was insignificant. Freshwater green algae Raphidocelus subcapitata and invertebrate Ceriodaphnia dubia were exposed to 28-day stirred aqu-nC60 with no significant toxicological impact. Aqu-nC60 stirred for 14 days bore no toxicity within two orders of magnitude greater than the highest concentration administered. LC50 for organisms exposed to alkaline-synthesized fullerenol prepared in the laboratory was 2409 mg/L for P. aeruginosa with no determinable toxicity to S. aureus, and 1462 mg/L and 45.2 mg/L for R. subcapitata and C. dubia, respectively. Toxicity thresholds for commercially-prepared fullerenol were lower for all species, an impact attributed to the presence of impurities. Mechanistic analysis of membrane damage on bacteria by laboratory-prepared fullerenol indicated necrotic and apoptotic responses with and without photoactivation. Toxicological responses from fullerenol synthesis by-products were only determinable for C. dubia with effects attributable to impurities.
Keeping time without a spine: what can the insect clock teach us about seasonal adaptation?
Author information
- 1
- Departments of Entomology and Evolution, Ecology and Organismal Biology, Ohio State University, Columbus, OH 43210, USA denlinger.1@osu.edu.
- 2
- Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611, USA.
- 3
- Department of Biology, Texas A&M University, College Station, TX, 77843, USA.
- 4
- Institute of Ecology and Evolution, University of Oregon, Eugene, OR, 97403, USA.
Abstract
Seasonal change in daylength (photoperiod) is widely used by insects to regulate temporal patterns of development and behaviour, including the timing of diapause (dormancy) and migration. Flexibility of the photoperiodic response is critical for rapid shifts to new hosts, survival in the face of global climate change and to reproductive isolation. At the same time, the daily circadian clock is also essential for development, diapause and multiple behaviours, including correct flight orientation during long-distance migration. Although studied for decades, how these two critical biological timing mechanisms are integrated is poorly understood, in part because the core circadian clock genes are all transcription factors or regulators that are able to exert multiple effects throughout the genome. In this chapter, we discuss clocks in the wild from the perspective of diverse insect groups across eco-geographic contexts from the Antarctic to the tropical regions of Earth. Application of the expanding tool box of molecular techniques will lead us to distinguish universal from unique mechanisms underlying the evolution of circadian and photoperiodic timing, and their interaction across taxonomic and ecological contexts represented by insects.This article is part of the themed issue ‘Wild clocks: integrating chronobiology and ecology to understand timekeeping in free-living animals’.
Multicentre study of maternal and neonatal outcomes in individuals with Prader-Willi syndrome.
Author information
- 1
- Division of Genetics and Metabolism, Department of Pediatrics, University of California, Irvine, California, USA.
- 2
- Division of Neonatology, Department of Pediatrics, University of California, Irvine, California, USA.
- 3
- Pediatrics, Children’s Hospital of Orange County, Orange, California, USA.
- 4
- Division of Genetics and Metabolism, Department of Pediatrics, Loma Linda University Medical School, Loma Linda, California, USA.
- 5
- Department of Pediatrics, College of Medicine, University of Florida, Gainesville, Florida, USA.
- 6
- Vanderbilt Kennedy Center for Research on Human Development, Vanderbilt University, Nashville, Tennessee, USA.
- 7
- Health Informatics Institute, University of South Florida, Tampa, Florida, USA.
- 8
- Departments of Psychiatry, Behavioral Sciences, and Pediatrics, Kansas University Medical Center, Kansas City, Kansas, USA.
- 9
- Center of Epigenetics, College of Medicine, University of Florida, Gainesville, Florida, USA.
- #
- Contributed equally
Abstract
INTRODUCTION:
Prader-Willi syndrome (PWS) is a complex genetic disorder associated with three different genetic subtypes: deletion of the paternal copy of 15q11-q13, maternal UPD for chromosome 15 and imprinting defect. Patients are typically diagnosed because of neonatal hypotonia, dysmorphism and feeding difficulties; however, data on the prenatal features of PWS are limited.
OBJECTIVE:
The aim of the study was to identify and compare frequencies of prenatal and neonatal clinical features of PWS among the three genetic subtypes.
METHODS:
Data from 355 patients with PWS from the Rare Diseases Clinical Research Network PWS registry were used to analyse multiple maternal and neonatal factors collected during an 8-year multisite study.
RESULTS:
Among our cohort of 355 patients with PWS (61% deletion, 36% UPD and 3% imprinting defect) 54% were born by caesarean section, 26% were born prematurely and 34% with a low birth weight (frequencies 32%, 9.6% and 8.1%, respectively, in the general population). Fetal movements were reported as decreased in 72%. All babies were hypotonic, and 99% had feeding difficulties. Low Apgar scores (<7) were noted in 17.7% and 5.6% of patients, respectively, compared with 1% and 1.4%, respectively, in the general population. Maternal age and pre-pregnancy weight were significantly higher in the UPD group (p=0.01 and <0.001, respectively).
CONCLUSION:
We found a higher rate of perinatal complications in PWS syndrome compared with the general population. No significant differences in the genetic subtypes were noted except for a higher maternal age and pre-pregnancy weight in the UPD subgroup.
A Defective mRNA Cleavage and Polyadenylation Complex Facilitates Expansions of Transcribed (GAA)n Repeats Associated with Friedreich’s Ataxia.
Author information
- 1
- Department of Biology, Tufts University, Medford, MA 02421, USA.
- 2
- Department of Developmental, Molecular and Chemical Biology, Tufts University School of Medicine, Boston, MA 02111, USA.
- 3
- Department of Biology, Tufts University, Medford, MA 02421, USA; Laboratory of Amyloid Biology, St. Petersburg State University, St. Petersburg 199034, Russia.
- 4
- Department of Biology, Tufts University, Medford, MA 02421, USA; The Broad Institute of MIT and Harvard, Cambridge, MA 02139, USA.
- 5
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA 02139, USA; Center for Neurogenetics, University of Florida, Gainesville, FL 32610, USA.
- 6
- The David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA 02139, USA.
- 7
- Department of Biology, Tufts University, Medford, MA 02421, USA. Electronic address: sergei.mirkin@tufts.edu.
Abstract
Expansions of microsatellite repeats are responsible for numerous hereditary diseases in humans, including myotonic dystrophy and Friedreich’s ataxia. Whereas the length of an expandable repeat is the main factor determining disease inheritance, recent data point to genomic trans modifiers that can impact the likelihood of expansions and disease progression. Detection of these modifiers may lead to understanding and treating repeat expansion diseases. Here, we describe a method for the rapid, genome-wide identification of trans modifiers for repeat expansion in a yeast experimental system. Using this method, we found that missense mutations in the endoribonuclease subunit (Ysh1) of the mRNA cleavage and polyadenylation complex dramatically increase the rate of (GAA)n repeat expansions but only when they are actively transcribed. These expansions correlate with slower transcription elongation caused by the ysh1 mutation. These results reveal an interplay between RNA processing and repeat-mediated genome instability, confirming the validity of our approach.
Critical Role of COI1-Dependent Jasmonate Pathway in AAL toxin induced PCD in Tomato Revealed by Comparative Proteomics.
Author information
- 1
- Key Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Ministry of Agriculture, Department of Horticulture, Zhejiang University, Hangzhou 310058, China.
- 2
- Proteomics and Mass Spectrometry, Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, FL 32610, USA.
- 3
- Department of Biology, Genetics Institute, University of Florida, Gainesville, FL 32610, USA.
- 4
- Thermo Fisher Scientific, West Palm Beach, Florida 33407, USA.
- 5
- Zhejiang Provincial Key Laboratory of Horticultural Plant Integrative Biology, Department of Horticulture, Zhejiang University, Hangzhou 310058, China.
Abstract
Alternaria alternata f.sp. Lycopersici (AAL) toxin induces programmed cell death (PCD) in susceptible tomato (Solanum lycopersicum) leaves. Jasmonate (JA) promotes AAL toxin induced PCD in a COI1 (coronatine insensitive 1, JA receptor)-dependent manner by enhancement of reactive oxygen species (ROS) production. To further elucidate the underlying mechanisms of this process, we performed a comparative proteomic analysis using tomato jasmonic acid insensitive1 ( jai1), the receptor mutant of JA, and its wild type (WT) after AAL toxin treatment with or without JA treatment. A total of 10367 proteins were identified in tomato leaves using isobaric tags for relative and absolute quantitation (iTRAQ) quantitative proteomics approach. 2670 proteins were determined to be differentially expressed in response to AAL toxin and JA. Comparison between AAL toxin treated jai1 and its WT revealed the COI1-dependent JA pathway regulated proteins, including pathways related to redox response, ceramide synthesis, JA, ethylene (ET), salicylic acid (SA) and abscisic acid (ABA) signaling. Autophagy, PCD and DNA damage related proteins were also identified. Our data suggest that COI1-dependent JA pathway enhances AAL toxin induced PCD through regulating the redox status of the leaves, other phytohormone pathways and/or important PCD components.
Culex quinquefasciatus (Diptera: Culicidae) From Florida Transmitted Zika Virus.
Author information
- 1
- Florida Medical Entomology Laboratory, Department of Entomology and Nematology, IFAS, University of Florida, Gainesville, FL, United States.
- 2
- Infectious Diseases & Immunology, College of Veterinary Medicine, University of Florida, Gainesville, FL, United States.
Abstract
We report a laboratory colony of Culex quinquefasciatus mosquitoes were experimentally able to salivate Zika virus (ZIKV, Flaviviridae; Flavivirus) at 16 days post infection (dpi). ZIKV RNA was detected in bodies and in saliva deposited on filter paper cards with subsequent studies demonstrating the presence of live ZIKV in saliva.
Structure-guided evolution of antigenically distinct adeno-associated virus variants for immune evasion.
Author information
- 1
- Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
- 2
- Department of Genetics, The University of North Carolina at Chapel Hill, Chapel Hill, NC; 27599.
- 3
- Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32611.
- 4
- Center for Structural Biology, The McKnight Brain Institute, University of Florida, Gainesville, FL 32611.
- 5
- Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599; aravind@med.unc.edu.
- 6
- Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599.
Abstract
Preexisting neutralizing antibodies (NAbs) against adeno-associated viruses (AAVs) pose a major, unresolved challenge that restricts patient enrollment in gene therapy clinical trials using recombinant AAV vectors. Structural studies suggest that despite a high degree of sequence variability, antibody recognition sites or antigenic hotspots on AAVs and other related parvoviruses might be evolutionarily conserved. To test this hypothesis, we developed a structure-guided evolution approach that does not require selective pressure exerted by NAbs. This strategy yielded highly divergent antigenic footprints that do not exist in natural AAV isolates. Specifically, synthetic variants obtained by evolving murine antigenic epitopes on an AAV serotype 1 capsid template can evade NAbs without compromising titer, transduction efficiency, or tissue tropism. One lead AAV variant generated by combining multiple evolved antigenic sites effectively evades polyclonal anti-AAV1 neutralizing sera from immunized mice and rhesus macaques. Furthermore, this variant displays robust immune evasion in nonhuman primate and human serum samples at dilution factors as high as 1:5, currently mandated by several clinical trials. Our results provide evidence that antibody recognition of AAV capsids is conserved across species. This approach can be applied to any AAV strain to evade NAbs in prospective patients for human gene therapy.
A Rationally Engineered Capsid Variant of AAV9 for Systemic CNS-Directed and Peripheral Tissue-Detargeted Gene Delivery in Neonates.
Author information
- 1
- Horae Gene Therapy Center, University of Massachusetts Medical School, Worcester, MA 01605, USA.
- 2
- Li Weibo Institute for Rare Diseases Research, University of Massachusetts Medical School, Worcester, MA 01605, USA.
- 3
- Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA 01605, USA.
- 4
- Department of Biochemistry and Molecular Biology, University of Florida, Gainesville, FL 32610, USA.
- 5
- Viral Vector Core, University of Massachusetts Medical School, Worcester, MA 01605, USA.
- 6
- New Iberia Research Center, University of Louisiana at Lafayette, New Iberia, LA 70560, USA.
- 7
- Department of Pediatrics, University of Massachusetts Medical School, Worcester, MA 01605, USA.
- 8
- West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Abstract
Adeno-associated virus (AAV) has provided the gene therapy field with the most powerful in vivogene delivery vector to realize safe, efficacious, and sustainable therapeutic gene expression. Because many clinically relevant properties of AAV-based vectors are governed by the capsid, much research effort has been devoted to the development of AAV capsids for desired features. Here, we combine AAV capsid discovery from nature and rational engineering to report an AAV9 capsid variant, designated as AAV9.HR, which retains AAV9’s capability to traverse the blood-brain barrier and transduce neurons. This variant shows reduced transduction in peripheral tissues when delivered through intravascular (IV) injection into neonatal mice. Therefore, when IV AAV delivery is used to treat CNS diseases, AAV9.HR has the advantage of mitigating potential off-target effects in peripheral tissues compared to AAV9. We also demonstrate that AAV9.HR is suitable for peripheral tissue-detargeted CNS-directed gene therapy in a mouse model of a fatal pediatric leukodystrophy. In light of recent success with profiling diversified natural AAV capsid repertoires and the understanding of AAV capsid sequence-structure-function relationship, such a combinatory approach to AAV capsid development is expected to further improve vector targeting and expand the vector toolbox for therapeutic gene delivery.
Atomic structure of a rationally engineered gene delivery vector, AAV2.5.
Author information
- 1
- Department of Biochemistry and Molecular Biology, College of Medicine, McKnight Brain Institute, University of Florida, Gainesville, Florida 32610, United states.
- 2
- Biological Science Imaging Resource, Department of Biological Sciences, The Florida State University, 89 Chieftan Way, Rm 119, Tallahassee Fl 32306, USA.
- 3
- Department of Molecular Genetics and Microbiology and Powell Gene Therapy Center,College of Medicine, University of Florida, Gainesville, Florida 32610, United states.
- 4
- Department of Pharmacology, Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
- 5
- Department of Biochemistry and Molecular Biology, College of Medicine, McKnight Brain Institute, University of Florida, Gainesville, Florida 32610, United states. Electronic address: mckenna@ufl.edu.
Abstract
AAV2.5 represents the first structure-guided in-silico designed Adeno-associated virus (AAV) gene delivery vector. This engineered vector combined the receptor attachment properties of AAV serotype 2 (AAV2) with the muscle tropic properties of AAV1, and exhibited an antibody escape phenotype because of a modified antigenic epitope. To confirm the design, the structure of the vector was determined to a resolution of 2.78 Å using cryo-electron microscopy and image reconstruction. The structure of the major viral protein (VP), VP3, was ordered from residue 219 to 736, as reported for other AAV structures, and the five AAV2.5 residues exchanged from AAV2 to AAV1, Q263A, T265 (insertion), N706A, V709A, and T717N, were readily interpretable. Significantly, the surface loops containing these residues adopt the AAV1 conformation indicating the importance of amino acid residues in dictating VP structure.
Environmental and health effects of the herbicide glyphosate.
Author information
- 1
- Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611, USA; Department of Plant Pathology, IFAS, University of Florida, Gainesville, FL 32610, USA. Electronic address: ahcvanbruggen@ufl.edu.
- 2
- Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611, USA; Department of Life and Environment Science, Hangzhou Normal University, Zhejiang 310036, China.
- 3
- Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611, USA; Department of Plant Pathology, IFAS, University of Florida, Gainesville, FL 32610, USA.
- 4
- Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611, USA.
- 5
- Faculty of Organic Agricultural Sciences, Ecological Plant Protection, University of Kassel, 37213 Witzenhausen, Germany.
- 6
- Emerging Pathogens Institute, University of Florida, Gainesville, FL 32611, USA; Department of Medicine, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
Abstract
The herbicide glyphosate, N-(phosphonomethyl) glycine, has been used extensively in the past 40years, under the assumption that side effects were minimal. However, in recent years, concerns have increased worldwide about the potential wide ranging direct and indirect health effects of the large scale use of glyphosate. In 2015, the World Health Organization reclassified glyphosate as probably carcinogenic to humans. A detailed overview is given of the scientific literature on the movement and residues of glyphosate and its breakdown product aminomethyl phosphonic acid (AMPA) in soil and water, their toxicity to macro- and microorganisms, their effects on microbial compositions and potential indirect effects on plant, animal and human health. Although the acute toxic effects of glyphosate and AMPA on mammals are low, there are animal data raising the possibility of health effects associated with chronic, ultra-low doses related to accumulation of these compounds in the environment. Intensive glyphosate use has led to the selection of glyphosate-resistant weeds and microorganisms. Shifts in microbial compositions due to selective pressure by glyphosate may have contributed to the proliferation of plant and animal pathogens. Research on a link between glyphosate and antibiotic resistance is still scarce but we hypothesize that the selection pressure for glyphosate-resistance in bacteria could lead to shifts in microbiome composition and increases in antibiotic resistance to clinically important antimicrobial agents. We recommend interdisciplinary research on the associations between low level chronic glyphosate exposure, distortions in microbial communities, expansion of antibiotic resistance and the emergence of animal, human and plant diseases. Independent research is needed to revisit the tolerance thresholds for glyphosate residues in water, food and animal feed taking all possible health risks into account.
The Plant Phenology Ontology: A New Informatics Resource for Large-Scale Integration of Plant Phenology Data.
Author information
- 1
- Florida Museum of Natural History, University of Florida, Gainesville, FL, United States.
- 2
- Berkeley Natural History Museums, University of California, Berkeley, Berkeley, CA, United States.
- 3
- USA National Phenology Network, The University of Arizona, Tucson, AZ, United States.
- 4
- Unit for Field-based Forest Research, Swedish University of Agricultural Sciences, Lammhult, Sweden.
- 5
- CyVerse, The University of Arizona, Tucson, AZ, United States.
Abstract
Plant phenology – the timing of plant life-cycle events, such as flowering or leafing out – plays a fundamental role in the functioning of terrestrial ecosystems, including human agricultural systems. Because plant phenology is often linked with climatic variables, there is widespread interest in developing a deeper understanding of global plant phenology patterns and trends. Although phenology data from around the world are currently available, truly global analyses of plant phenology have so far been difficult because the organizations producing large-scale phenology data are using non-standardized terminologies and metrics during data collection and data processing. To address this problem, we have developed the Plant Phenology Ontology (PPO). The PPO provides the standardized vocabulary and semantic framework that is needed for large-scale integration of heterogeneous plant phenology data. Here, we describe the PPO, and we also report preliminary results of using the PPO and a new data processing pipeline to build a large dataset of phenology information from North America and Europe.
Editorial overview: Parasites/parasitoids/biological control: Bee health in the modern age: a major concern.
Author information
- 1
- Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C, Denmark.
- 2
- Department of Entomology and Nematology, University of Florida, Gainesville, FL 32611, USA.
Employing hypothesis testing and data from multiple genomic compartments to resolve recalcitrant backbone nodes in Goodenia s.l. (Goodeniaceae).
Author information
- 1
- Department of Organismal Biology & Ecology, Colorado College, Colorado Springs, CO 80903, USA; Department of Biology, Rhodes College, Memphis, TN 38112, USA. Electronic address: rjabaily@coloradocollege.edu.
- 2
- Western Australian Herbarium, Department of Biodiversity, Conservation and Attractions, Kensington, WA 6151, Australia. Electronic address: kelly.shepherd@dbca.wa.gov.au.
- 3
- Department of Biology, Rhodes College, Memphis, TN 38112, USA. Electronic address: prycemichener@gmail.com.
- 4
- Department of Biology, Rhodes College, Memphis, TN 38112, USA. Electronic address: buscj-19@rhodes.edu.
- 5
- Department of Biology, University of Florida, Gainesville, FL 32607, USA; Department of Natural Resources and Environmental Management, University of Hawaii- Mānoa, Honolulu, HI 96822, USA. Electronic address: rodrigo25@ufl.edu.
- 6
- Department of Biological Sciences, California State University, Stanislaus, One University Circle, Turlock, CA 95382, USA. Electronic address: agardner1@csustan.edu.
- 7
- Department of Biology, University of Florida, Gainesville, FL 32607, USA; Genetics Institute, University of Florida, Gainesville, FL 32607, USA. Electronic address: emilysessa@ufl.edu.
Abstract
Goodeniaceae is a primarily Australian flowering plant family with a complex taxonomy and evolutionary history. Previous phylogenetic analyses have successfully resolved the backbone topology of the largest clade in the family, Goodenia s.l., but have failed to clarify relationships within the species-rich and enigmatic Goodenia clade C, a prerequisite for taxonomic revision of the group. We used genome skimming to retrieve sequences for chloroplast, mitochondrial, and nuclear markers for 24 taxa representing Goodenia s.l., with a particular focus on Goodenia clade C. We performed extensive hypothesis tests to explore incongruence in clade C and evaluate statistical support for clades within this group, using datasets from all three genomic compartments. The mitochondrial dataset is comparable to the chloroplast dataset in providing resolution within Goodenia clade C, though backbone support values within this clade remain low. The hypothesis tests provided an additional, complementary means of evaluating support for clades. We propose that the major subclades of Goodenia clade C (C1-C3 + Verreauxia) are the result of a rapid radiation, and each represents a distinct lineage.
CTCF boundary remodels chromatin domain and drives aberrant HOX gene transcription in acute myeloid leukemia.
Author information
- 1
- Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville, FL, United States.
- 2
- Department of Hematology and Oncology,, The Affiliated Zhongda Hospital, Southeast University Medical School, Nanjing, China.
- 3
- Department of Hematology, The First Affiliate Hospital of Xiamen University, Xiamen, China.
- 4
- Department of Genetics, Southern Medical University, Guangzhou, China.
- 5
- Department of Anatomy and Cell biology, University of Florida College of Medicine, Gainesville, FL, United States.
- 6
- Department of Biochemistry and Molecular Biology, UM Sylvester Comprehensive Cancer Center, University of Miami Miller School of Medicine, Miami, FL, United States.
- 7
- Department of Physiological Sciences, University of Florida, Gainesville, FL, United States.
- 8
- Division of Hematology/Oncology, Department of Medicine, University of Florida College of Medicine, Gainsville, FL, United States.
- 9
- Division of Hematology/Oncology, Department of Medicine, University of Florida College of Medicine, Gainesville, FL, United States.
- 10
- Center for Autoimmune Genomics and Etiology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, United States.
- 11
- UF Health Cancer Center, University of Florida College of Medicine, Gainesville, FL, United States.
- 12
- Macau Institute for Applied Research in Medicine and Health, State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Techn, Taipa, Macao sumingh@ufl.edu.
Abstract
HOX gene dysregulation is a common feature of acute myeloid leukemia (AML). The molecular mechanisms underlying aberrant HOX gene expression and associated AML pathogenesis remain unclear. The nuclear protein CCCTC-binding factor (CTCF), when bound to insulator sequences, constrains temporal HOX gene expression patterns within confined chromatin domains for normal development. Here, we employed targeted pooled CRISPR-Cas9 knockout library screening to interrogate the function of CTCF boundaries in the HOX gene loci. We discovered that the CTCF binding site located between HOXA7 and HOXA9 genes (CBS7/9) is critical for establishing and maintaining aberrant HOXA9-HOXA13 gene expression in AML. Disruption of the CBS7/9 boundary resulted in spreading of repressive H3K27me3 into the posterior active HOXA chromatin domain that subsequently impaired enhancer/promoter chromatin accessibility and disrupted ectopic long-range interactions among the posterior HOXA genes. Consistent with the role of the CBS7/9 boundary in HOXA locus chromatin organization, attenuation of the CBS7/9 boundary function reduced posterior HOXA gene expression and altered myeloid specific transcriptome profiles important for pathogenesis of myeloid malignancies. Furthermore, heterozygous deletion of the CBS7/9 chromatin boundary in the HOXA locus reduced human leukemic blast burden and enhanced survival of both transplanted AML cell xenograft and patient derived xenograft (PDX) mouse models. Thus, the CTCF boundary not only constrains the normal gene expression program, but also plays a role in maintaining the oncogenic transcription program for leukemic transformation. The CTCF boundaries may serve as novel therapeutic targets for the treatment of myeloid malignancies.
Strength in Numbers: Opportunities for Enhancing the Development of Effective Treatments for Type 1 Diabetes-The TrialNet Experience.
Author information
- 1
- Clinical Research Center, Diabetes Research Program, Benaroya Research Institute at Virginia Mason, Seattle, WA.
- 2
- Diabetes Center and Pediatric Epidemiology Center, Morsani College of Medicine, University of South Florida, Tampa, FL.
- 3
- Barbara Davis Center for Diabetes, University of Colorado School of Medicine, Aurora, CO.
- 4
- Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL.
- 5
- Departments of Immunobiology and Internal Medicine, Yale University, New Haven, CT.
- 6
- Department of Pediatrics, College of Medicine, University of Florida, Gainesville, FL atkinson@ufl.edu.
- 7
- Department of Pathology, College of Medicine, University of Florida, Gainesville, FL.
Abstract
The early to mid-1980s were an inflection point in the history of type 1 diabetes research. Two landmark events occurred: the initiation of immune-based interventions seeking to prevent type 1 diabetes and the presentation of an innovative model describing the disorder’s natural history. Both formed the basis for hundreds of subsequent studies designed to achieve a dramatic therapeutic goal-a means to prevent and/or reverse type 1 diabetes. However, the need to screen large numbers of individuals and prospectively monitor them using immunologic and metabolic tests for extended periods of time suggested such efforts would require a large collaborative network. Hence, the National Institutes of Health formed the landmark Diabetes Prevention Trial-Type 1 (DPT-1) in the mid-1990s, an effort that led to Type 1 Diabetes TrialNet. TrialNet studies have helped identify novel biomarkers; delineate type 1 diabetes progression, resulting in identification of highly predictable stages defined by the accumulation of autoantibodies (stage 1), dysglycemia (stage 2), and disease meeting clinical criteria for diagnosis (stage 3); and oversee numerous clinical trials aimed at preventing disease progression. Such efforts pave the way for stage-specific intervention trials with improved hope that a means to effectively disrupt the disorder’s development will be identified.
A Pilot Study of Inhaled CO Therapy in Neonatal Hypoxia-Ischemia: Carboxyhemoglobin Concentrations and Brain Volumes.
Author information
- 1
- Department of Pediatrics, University of California, San Francisco, San Francisco, CA, United States.
- 2
- Department of Anesthesiology, Center for Translational Research in Neurodegenerative, McKnight Brain Institutive, University of Florida, Gainesville, FL, United States.
- 3
- Department of Pediatrics, University of Florida, Gainesville, FL, United States.
- 4
- Department of Agricultural and Biological Egineering, University of Florida, Gainesville, FL, United States.
- 5
- Department of Neurology, Center for Translational Research in Neurodegenerative, McKnight Brain Institutive, University of Florida, Gainesville, FL, United States.
- 6
- Department of Psychiatry, Center for Translational Research in Neurodegenerative, McKnight Brain Institutive, University of Florida, Gainesville, FL, United States.
- 7
- Department of Neuroscience, Center for Translational Research in Neurodegenerative, McKnight Brain Institutive, University of Florida, Gainesville, FL, United States.
Abstract
Objective: The objective of this pilot study was to start evaluating the efficacy and the safety (i.e., carboxyhemoglobin concentration of carbon monoxide (CO)) as a putative neuroprotective therapy in neonates. Study Design: Neonatal C57BL/6 mice were exposed to CO at a concentration of either 200 or 250 ppm for a period of 1 h. The pups were then sacrificed at 0, 10, 20, 60, 120, 180, and 240 min after exposure to either concentration of CO, and blood was collected for analysis of carboxyhemoglobin. Following the safety study, 7-day-old pups underwent a unilateral carotid ligation. After recovery, the pups were exposed to a humidified gas mixture of 8% oxygen and 92% nitrogen for 20 min in a hypoxia chamber. One hour after the hypoxia exposure, the pups were randomized to one of two groups: air (HI+A) or carbon monoxide (HI+CO). An inhaled dose of 250 ppm of CO was administered to the pups for 1 h per day for a period of 3 days. At 7 days post-injury, the pups were sacrificed and the brains analyzed for cortical and hippocampal volumes. Results: CO exposure at 200 and 250 ppm produced a peak carboxyhemoglobin concentration of 21.52 ± 1.18% and 27.55 ± 3.58%, respectively. The carboxyhemoglobin concentrations decreased rapidly, reaching control concentrations by 60 min post exposure. At 14 days of age (7 days post injury), the HI+CO (treated with 1 h per day of 250 ppm of CO for 3 days post injury) had significant preservation of the ratio of ipsilateral to contralateral cortex (median 1.07, 25% 0.97, 75% 1.23, n = 10) compared the HI+A group (p < 0.05). Conclusion: CO exposure of 250 ppm did not reach carboxyhemoglobin concentrations which would induce acute neurologic abnormalities and was effective in preserving cortical volumes following hypoxic-ischemic injury.
Neonatal sepsis in rural India: timing, microbiology and antibiotic resistance in a population-based prospective study in the community setting.
Author information
- 1
- Department of Epidemiology and Pediatrics, Center for Global Health and Development, College of Public Health, University of Nebraska Medical Center, Omaha, NE, USA.
- 2
- Department of Environmental, Agricultural, &Occupational Health, Center for Global Health and Development, College of Public Health, University of Nebraska Medical Center, Omaha, NE, USA.
- 3
- Division of Biostatistics &Epidemiology, RTI International, Research Triangle Park, NC, USA.
- 4
- Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India.
- 5
- Division of Social Policy, Health, &Economics Research, RTI International, Research Triangle Park, NC, USA.
- 6
- Department of Pediatrics, SCB Medical College, Cuttack, Odisha, India.
- 7
- Social Pediatrics and Minority Health, Asian Institute of Public Health, Bhubaneswar, Odisha, India.
- 8
- Neonatal Unit, Department of Pediatrics, Capital Hospital, Bhubaneswar, Odisha, India.
- 9
- Center for Translational Research, Asian Institute of Public Health, Bhubaneswar, Odisha, India.
- 10
- Disease Surveillance Laboratories, Asian Institute of Public Health, Bhubaneswar, Odisha, India.
- 11
- Department of Microbiology, Indira Gandhi Medical College &Research Institute, Puducherry, India.
- 12
- Department of Pathology, Immunology and Laboratory Medicine, Emerging Pathogens Institute, University of Florida, Gainesville, FL, USA.
- 13
- Department of Medicine, College of Medicine, Emerging Pathogens Institute, University of Florida, Gainesville FL, USA.
- 14
- Department of Pediatrics, Division of Neonatology, Sparrow Hospital – Neonatology, Lansing, MI, USA.
Abstract
OBJECTIVE:
To examine the timing and microbiology of neonatal sepsis in a population-based surveillance in the Indian community setting.
STUDY DESIGN:
All live born infants in 223 villages of Odisha state were followed at home for 60 days. Suspect sepsis cases were referred to study hospitals for further evaluation including blood culture.
RESULTS:
Of 12 622 births, 842 were admitted with suspected sepsis of whom 95% were 4 to 60 days old. Culture-confirmed incidence of sepsis was 6.7/1000 births with 51% Gram negatives (Klebsiella predominating) and 26% Gram positives (mostly Staphylococcus aureus). A very high level of resistance to penicillin and ampicillin, moderate resistance to cephalosporins and extremely low resistance to Gentamicin and Amikacin was observed.
CONCLUSION:
The bacterial burden of sepsis in the Indian community is not high. Judicious choice of empiric antibiotics, antibiotic stewardship and alternate modalities should be considered for the management or prevention of neonatal sepsis in India.
Preface: Translational Gene Therapy Coming of Age!
Author information
- 1
- College of Medicine, University of Florida Gainesville, FL, United States.
Non-toxigenic environmental Vibrio cholerae O1 strain from Haiti provides evidence of pre-pandemic cholera in Hispaniola.
Author information
- 1
- Emerging Pathogens Institute, University of Florida, Gainesville, USA.
- 2
- Department of Environmental and Global Health, College of Public Health and Health Profession, University of Florida, Gainesville, Florida, USA.
- 3
- Department of Pathology, Immunology and Laboratory Medicine, University of Florida, Gainesville, USA.
- 4
- Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy.
- 5
- Department of Public Health and Infectious Diseases, Sapienza University of Rome, Rome, Italy.
- 6
- University Hospital Campus Bio-Medico, Italy.
- 7
- Interdisciplinary Center for Biotechnology Research, University of Florida, Gainesville, Florida, USA.
- 8
- Department of Medicine, College of Medicine, University of Florida, Gainesville, Florida, USA.
Abstract
Vibrio cholerae is ubiquitous in aquatic environments, with environmental toxigenic V. cholerae O1 strains serving as a source for recurrent cholera epidemics and pandemic disease. However, a number of questions remain about long-term survival and evolution of V. cholerae strains within these aquatic environmental reservoirs. Through monitoring of the Haitian aquatic environment following the 2010 cholera epidemic, we isolated two novel non-toxigenic (ctxA/B-negative) Vibrio cholerae O1. These two isolates underwent whole-genome sequencing and were investigated through comparative genomics and Bayesian coalescent analysis. These isolates cluster in the evolutionary tree with strains responsible for clinical cholera, possessing genomic components of 6th and 7th pandemic lineages, and diverge from “modern” cholera strains around 1548 C.E. [95% HPD: 1532-1555]. Vibrio Pathogenicity Island (VPI)-1 was present; however, SXT/R391-family ICE and VPI-2 were absent. Rugose phenotype conversion and vibriophage resistance evidenced adaption for persistence in aquatic environments. The identification of V. cholerae O1 strains in the Haitian environment, which predate the first reported cholera pandemic in 1817, broadens our understanding of the history of pandemics. It also raises the possibility that these and similar environmental strains could acquire virulence genes from the 2010 Haitian epidemic clone, including the cholera toxin producing CTXϕ.
Islet Microvasculature Alterations With Loss of Beta-cells in Patients With Type 1 Diabetes.
Author information
- 1
- Department of Pathology, Immunology and Laboratory Medicine, University of Florida, Gainesville, Florida.
- 2
- Department of Pediatrics, College of Medicine, University of Florida, Gainesville, Florida.
Abstract
Islet microvasculature provides key architectural and functional roles, yet the morphological features of islets from patients with type 1 diabetes are poorly defined. We examined islet and exocrine microvasculature networks by multiplex immunofluorescence imaging of pancreases from organ donors with and without type 1 diabetes ( n=17 and n=16, respectively) and determined vessel diameter, density, and area. We also analyzed these variables in insulin-positive and insulin-negative islets of 7 type 1 diabetes donors. Control islet vessel diameter was significantly larger (7.6 ± 1.1 μm) compared with vessels in diabetic islets (6.2 ± 0.8 μm; p<0.001). Control islet vessel density (number/islet) was significantly lower (5.3 ± 0.6) versus diabetic islets (9.3 ± 0.2; p<0.001). Exocrine vessel variables were not significantly different between groups. Islets with residual beta-cells were comparable to control islets for both vessel diameter and density and were significantly different from insulin-negative islets within diabetic donors ( p<0.05). Islet smooth muscle actin area had a significant positive correlation with age in both groups ( p<0.05), which could negatively impact islet transplantation efficiency from older donors. These data underscore the critical relationship of islet beta-cells and islet vessel morphology in type 1 diabetes. These studies provide new knowledge of the islet microvasculature in diabetes and aging.