0000000001055546
AUTHOR
Leona Gilbert
showing 32 related works from this author
Sequence variation and regulatory variation in acetylcholinesterase genes contribute to insecticide resistance in different populations of Leptinotar…
2021
Abstract Although insect herbivores are known to evolve resistance to insecticides through multiple genetic mechanisms, resistance in individual species has been assumed to follow the same mechanism. While both mutations in the target site insensitivity and increased amplification are known to contribute to insecticide resistance, little is known about the degree to which geographic populations of the same species differ at the target site in a response to insecticides. We tested structural (e.g., mutation profiles) and regulatory (e.g., the gene expression of Ldace1 and Ldace2, AChE activity) differences between two populations (Vermont, USA and Belchow, Poland) of the Colorado potato beet…
Advanced time-stamped total data acquisition control front-end for MeV ion beam microscopy and proton beam writing
2013
Many ion-matter interactions exhibit [email protected] time dependences such as, fluorophore emission quenching and ion beam induced charge (IBIC). Conventional event-mode MeV ion microbeam data acquisition systems discard the time information. Here we describe a fast time-stamping data acquisition front-end based on the concurrent processing capabilities of a Field Programmable Gate Array (FPGA). The system is intended for MeV ion microscopy and MeV ion beam lithography. The speed of the system (>240,000 events s^-^1 for four analogue to digital converters (ADC)) is limited by the ADC throughput and data handling speed of the host computer.
Effective killing of Borrelia burgdorferi in vitro with novel herbal compounds
2018
Introduction: The tick-borne disease Lyme Borreliosis is caused by Borrelia bacteria. The disease can persist even after treatment with antibiotics, which is why other methods of treatment are needed. Herbal compounds and phytochemicals have been recently examined in relation to eradicating Borrelia bacteria in vitro. Objective: The possible antimicrobial effect of two novel compounds, Biocidin Liquid and LSF Broad-Spectrum Liposomal formulas, was examined in the hopes of discovering an alternative method for eradication of Borrelia bacteria. Methods: The minimum inhibitory concentrations (MICs) and minimum bacterial deaths (MBDs), as well as, time-kill effect of each compound were utilize…
Parvovirus B19V Nonstructural Protein NS1 Induces Double-Stranded Deoxyribonucleic Acid Autoantibodies and End-Organ Damage in Nonautoimmune Mice
2018
Abstract Background Viral infection is implicated in development of autoimmunity. Parvovirus B19 (B19V) nonstructural protein, NS1, a helicase, covalently modifies self double-stranded deoxyribonucleic acid (dsDNA) and induces apoptosis. This study tested whether resulting apoptotic bodies (ApoBods) containing virally modified dsDNA could induce autoimmunity in an animal model. Methods BALB/c mice were inoculated with (1) pristane-induced, (2) B19V NS1-induced, or (3) staurosporine-induced ApoBods. Serum was tested for dsDNA autoantibodies by Crithidia luciliae staining and enzyme-linked immunosorbent assay. Brain, heart, liver, and kidney pathology was examined. Deposition of self-antigens…
Molecular and structural characterization of fluorescent human parvovirus B19 virus-like particles
2005
Although sharing a T = 1 icosahedral symmetry with other members of the Parvoviridae family, it has been suggested that the fivefold channel of the human parvovirus B19 VP2 capsids is closed at its outside end. To investigate the possibility of placing a relatively large protein moiety at this site of B19, fluorescent virus-like particles (fVLPs) of B19 were developed. The enhanced green fluorescent protein (EGFP) was inserted at the N-terminus of the structural protein VP2 and assembly of fVLPs from this fusion protein was obtained. Electron microscopy revealed that these fluorescent protein complexes were very similar in size when compared to wild-type B19 virus. Further, fluorescence cor…
Assembly of fluorescent chimeric virus-like particles of canine parvovirus in insect cells
2004
Canine parvovirus (CPV) is a small non-enveloped ssDNA virus composed of the viral proteins VP1, VP2, and VP3 with a T=1 icosahedral symmetry. VP2 is nested in VP1 and the two proteins are produced by differential splicing of a primary transcript of the right ORF of the viral genome. The VP2 protein can be further proteolytically cleaved to form VP3. Previous studies have shown that VP1 and VP3 are unnecessary for capsid formation and consequently, that VP2 alone is sufficient for assembly. We have hypothesized that insertion of the enhanced green fluorescent protein (EGFP) at the N-terminus of VP2 could be carried out without altering assembly. To investigate the possibility to develop flu…
Mechanisms of cell death in canine parvovirus-infected cells provide intuitive insights to developing nanotools for medicine
2010
Jonna Nykky, Jenni E Tuusa, Sanna Kirjavainen, Matti Vuento, Leona GilbertNanoscience Center and Department of Biological and Environmental Science, University of Jyväskylä, FinlandAbstract: Viruses have great potential as nanotools in medicine for gene transfer, targeted gene delivery, and oncolytic cancer virotherapy. Here we have studied cell death mechanisms of canine parvovirus (CPV) to increase the knowledge on the CPV life cycle in order to facilitate the development of better parvovirus vectors. Morphological studies of CPV-infected Norden laboratory feline kidney (NLFK) cells and canine fibroma cells (A72) displayed characteristic apoptotic events. Apoptosis was f…
Baculovirus entry into human hepatoma cells.
2005
ABSTRACT Autographa californica multiple nucleopolyhedrovirus (AcMNPV), a prototype member of the Baculoviridae family, has gained increasing interest as a potential vector candidate for mammalian gene delivery applications. AcMNPV is known to enter both dividing and nondividing mammalian cell lines in vitro, but the mode and kinetics of entry as well as the intracellular transport of the virus in mammalian cells is poorly understood. The general objective of this study was to characterize the entry steps of AcMNPV- and green fluorescent protein-displaying recombinant baculoviruses in human hepatoma cells. The viruses were found to bind and transduce the cell line efficiently, and electron …
Expression and subcellular targeting of canine parvovirus capsid proteins in baculovirus-transduced NLFK cells
2004
AbstractA mammalian baculovirus delivery system was developed to study targeting in Norden Laboratories feline kidney (NLFK) cells of the capsid proteins of canine parvovirus (CPV), VP1 and VP2, or corresponding counterparts fused to EGFP. VP1 and VP2, when expressed alone, both had equal nuclear and cytoplasmic distribution. However, assembled form of VP2 had a predominantly cytoplasmic localization. When VP1 and VP2 were simultaneously present in cells, their nuclear localization increased. Thus, confocal immunofluorescence analysis of cells transduced with the different baculovirus constructs or combinations thereof in the absence or presence of infecting CPV revealed that the VP1 protei…
Monitoring human parvovirus B19 virus-like particles and antibody complexes in solution by fluorescence correlation spectroscopy
2004
AbstractFluorescence correlation spectroscopy (FCS) was used in monitoring human parvovirus B19 virus-like particle (VLP) antibody complexes from acute phase and pastimmunity serum samples. The Oregon Green 488-labeled VLPs gave an average diffusion coefficient of 1.7x10exp-7 cm(2)s(-1) with an apparent hydrodynamic radius of 14 nm. After incubation of the fluorescent VLPs with an acute phase serum sample, the mobility information obtained from the fluorescence intensity fluctuation by autocorrelation analysis showed an average diffusion coefficient of 1.5x10exp-8 cm(2)s(-1), corresponding to an average radius of 157 nm. In contrast, incubation of the fluorescent VLPs with a pastimmunity se…
High speed microfluidic prototyping by programmable proximity aperture MeV ion beam lithography
2013
Abstract Microfluidics refers to the science and technology for controlling and manipulating fluids that flow along microchannels. For the development of complex prototypes, many microfluidic test structures are required first. Normally, these devices are fabricated via photolithography. This technique requires a photomask for transferring a pattern to photoresists by exposing with UV light. However, this method can be slow when a new structure is required to change. This is because a series of photomasks are needed, which is time consuming and costly. Here, we present a programmable proximity aperture lithography (PPAL) technique for the development of microfluidic prototype in poly(methyl…
Morphological and biochemical features of Borrelia burgdorferi pleomorphic forms
2015
The spirochaete bacterium Borrelia burgdorferi sensu lato is the causative agent of Lyme disease, the most common tick-borne infection in the northern hemisphere. There is a long-standing debate regarding the role of pleomorphic forms in Lyme disease pathogenesis, while very little is known about the characteristics of these morphological variants. Here, we present a comprehensive analysis of B. burgdorferi pleomorphic formation in different culturing conditions at physiological temperature. Interestingly, human serum induced the bacterium to change its morphology to round bodies (RBs). In addition, biofilm-like colonies in suspension were found to be part of B. burgdorferi’s normal in vitr…
Pleomorphic forms of Borrelia burgdorferi induce distinct immune responses.
2016
Borrelia burgdorferi is the causative agent of tick-borne Lyme disease. As a response to environmental stress B. burgdorferi can change its morphology to a round body form. The role of B. burgdorferi pleomorphic forms in Lyme disease pathogenesis has long been debated and unclear. Here, we demonstrated that round bodies were processed differently in differentiated macrophages, consequently inducing distinct immune responses compared to spirochetes in vitro. Colocalization analysis indicated that the F-actin participates in internalization of both forms. However, round bodies end up less in macrophage lysosomes than spirochetes suggesting that there are differences in processing of these for…
Parvovirus B19 nonstructural protein-induced damage of cellular DNA and resultant apoptosis.
2010
Parvovirus B19 is a widespread virus with diverse clinical presentations. The viral nonstructural protein, NS1, binds to and cleaves the viral genome, and induces apoptosis when transfected into nonpermissive cells, such as hepatocytes. We hypothesized that the cytotoxicity of NS1 in such cells results from chromosomal DNA damage caused by the DNA-nicking and DNA-attaching activities of NS1. Upon testing this hypothesis, we found that NS1 covalently binds to cellular DNA and is modified by PARP, an enzyme involved in repairing single-stranded DNA nicks. We furthermore discovered that the DNA nick repair pathway initiated by poly(ADPribose)polymerase and the DNA repair pathways initiated by …
Disassembly of structurally modified viral nanoparticles: characterization by fluorescence correlation spectroscopy.
2005
Abstract Analysis of the breakdown products of engineered viral particles can give useful information on the particle structure. We used various methods to breakdown both a recombinant enveloped virus and virus-like particles (VLPs) from two non-enveloped viruses and analysed the resulting subunits by fluorescence correlation spectroscopy (FCS). Analysis of the enveloped baculovirus, Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV), displaying the green fluorescent protein (GFP) fused to its envelope protein gp64 was performed in the presence and absence of 5 mM SDS and 25 mM DTT. Without treatment, the viral particle showed a diffusion time of 3.3 ms. In the presence of SDS…
Development of the Jyväskylä microbeam facility
2012
Abstract A new microbeam facility is being constructed at the 1.7 MV Pelletron Accelerator in Jyvaskyla. The facility is designed for easy upgrading and incorporates a number of innovative features. Initially, it is based on a Heidelberg doublet with a design capability of a 3 × 5 μm beamspot at PIXE intensities and later upgraded to nanobeam performance. A thermal-expansion compensated rigid frame mounted on a mechanically isolated floor section is used to support the ion optical components. A compact-post focusing electrostatic deflector is used for high linearity beam scanning. This together with a novel time-stamped data collection (TDC) allows dynamic effects in IBIC, fluorescence blea…
Method and apparatus using selected superparamagnetic labels for rapid quantification of immunochromatographic tests
2009
Mika PA Laitinen1, Jari Salmela2, Leona Gilbert1, Risto Kaivola1, Topi Tikkala2, Christian Oker-Blom1, Jukka Pekola3, Matti Vuento11Department of Biological and Environmental Science; 2Department of Physics, University of Jyväskylä, Jyväskylä, Finland; 3Low Temperature Laboratory, Helsinki University of Technology, Helsinki, FinlandAbstract: A rapid method and instrumentation for quantification of immunochromatographic tests (ICT) are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG) present in urine. The test format was a sandwich assay using two distinct monoclonal antib…
Method and apparatus using selected superparamagnetic labels for rapid quantification of immunochromatographic tests
2009
Mika PA Laitinen1, Jari Salmela2, Leona Gilbert1, Risto Kaivola1, Topi Tikkala2, Christian Oker-Blom1, Jukka Pekola3, Matti Vuento11Department of Biological and Environmental Science; 2Department of Physics, University of Jyväskylä, Jyväskylä, Finland; 3Low Temperature Laboratory, Helsinki University of Technology, Helsinki, FinlandAbstract: A rapid method and instrumentation for quantification of immunochromatographic tests (ICT) are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG) present in urine. The test format was a sandwich assay using two distinct monoclonal antib…
No diagnostic utility of antibody patterns against Klebsiella pneumoniae capsular serotypes in patients with axial spondyloarthritis vs. patients wit…
2017
OBJECTIVES: To investigate whether antibody response patterns against Klebsiella pneumoniae capsular serotypes can discriminate patients with axial spondyloarthritis (axSpA) from patients with non-specific low back pain (LBP).METHOD: Immunoglobulin (Ig)G and IgA antibodies against K. pneumoniae capsular serotypes K2, K26, K36, and K50 were measured, and antibody seropositivity compared between groups and analysed for patient correlation in five different groups: (a) 96 patients fulfilling the Assessment of SpondyloArthritis International Society (ASAS) classification criteria for axSpA; (b) 38 patients with either a positive magnetic resonance imaging (MRI) scan as defined by ASAS or a posi…
Human parvovirus B19 induced apoptotic bodies contain altered self-antigens that are phagocytosed by antigen presenting cells.
2013
Human parvovirus B19 (B19V) from the erythrovirus genus is known to be a pathogenic virus in humans. Prevalence of B19V infection has been reported worldwide in all seasons, with a high incidence in the spring. B19V is responsible for erythema infectiosum (fifth disease) commonly seen in children. Its other clinical presentations include arthralgia, arthritis, transient aplastic crisis, chronic anemia, congenital anemia, and hydrops fetalis. In addition, B19V infection has been reported to trigger autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis. However, the mechanisms of B19V participation in autoimmunity are not fully understood. B19V induced chronic dise…
Role of mitochondria in parvovirus pathology.
2014
Proper functioning of the mitochondria is crucial for the survival of the cell. Viruses are able to interfere with mitochondrial functions as they infect the host cell. Parvoviruses are known to induce apoptosis in infected cells, but the role of the mitochondria in parvovirus induced cytopathy is only partially known. Here we demonstrate with confocal and electron microscopy that canine parvovirus (CPV) associated with the mitochondrial outer membrane from the onset of infection. During viral entry a transient depolarization of the mitochondrial transmembrane potential and increase in ROS level was detected. Subsequently, mitochondrial homeostasis was normalized shortly, as detected by rep…
Retraction Note: Evaluating polymicrobial immune responses in patients suffering from tick-borne diseases
2022
There is insufficient evidence to support screening of various tick-borne diseases (TBD) related microbes alongside Borrelia in patients suffering from TBD. To evaluate the involvement of multiple microbial immune responses in patients experiencing TBD we utilized enzyme-linked immunosorbent assay. Four hundred and thirty-two human serum samples organized into seven categories followed Centers for Disease Control and Prevention two-tier Lyme disease (LD) diagnosis guidelines and Infectious Disease Society of America guidelines for post-treatment Lyme disease syndrome. All patient categories were tested for their immunoglobulin M (IgM) and G (IgG) responses against 20 microbes associated wit…
The Putative Metal Coordination Motif in the Endonuclease Domain of Human Parvovirus B19 NS1 Is Critical for NS1 Induced S Phase Arrest and DNA Damage
2011
The non-structural proteins (NS) of the parvovirus family are highly conserved multi-functional molecules that have been extensively characterized and shown to be integral to viral replication. Along with NTP-dependent helicase activity, these proteins carry within their sequences domains that allow them to bind DNA and act as nucleases in order to resolve the concatameric intermediates developed during viral replication. The parvovirus B19 NS1 protein contains sequence domains highly similar to those previously implicated in the above-described functions of NS proteins from adeno-associated virus (AAV), minute virus of mice (MVM) and other non-human parvoviruses. Previous studies have show…
Preparation of graphene nanocomposites from aqueous silver nitrate using graphene oxide’s peroxidase-like and carbocatalytic properties
2020
AbstractThe present study evaluates the role of graphene oxide’s (GO’s) peroxidase-like and inherent/carbocatalytic properties in oxidising silver nitrate (AgNO3) to create graphene nanocomposites with silver nanoparticles (GO/Ag nanocomposite). Activation of peroxidase-like catalytic function of GO required hydrogen peroxide (H2O2) and ammonia (NH3) in pH 4.0 disodium hydrogen phosphate (Na2HPO4). Carbocatalytic abilities of GO were triggered in pH 4.0 deionised distilled water (ddH2O). Transmission electron microscope (TEM), scanning electron microscope (SEM), cyclic voltammetry (CV) and UV-Vis spectroscopy aided in qualitatively and quantitatively assessing GO/Ag nanocomposites. TEM and …
FRI0194 Is There an Association Between Spondyloarthritis and Antibodies Towards Borrelia, Ehrlichia and Chlamydia Species?
2015
Background Several studies suggested that certain microorganisms might contribute to initiation and perpetuation of spondyloarthritis (SpA). Objectives To investigate IgG and IgM antibodies towards Borrelia burgdorferi (Bb), Borrelia garinii (Bg), Borrelia afzelii (Ba), Ehrlichia spp. (Ehr), Chlamydia trachomatis (Ct), and Chlamydia pneumoniae (Cp) in SpA patients, low back pain patients, and healthy subjects and to elucidate whether previous infections could play a role in the onset of SpA. Methods Data collection was based on persons aged 18-40 years referred with low back pain for ≥3 months. They were examined with MRI of the spine and sacroiliac joints, CRP, HLA-B27, and clinical SpA fe…
Transport of Water and Particles in Microfluidics Devices Lithographically Fabricated Using Proton Beam Writing (PBW)
2009
Proton beam writing (PBW) is a MeV ion beam lithography technique that has gained interest in many biological applications such as fabricating microfluidic devices for Lab-On-a-Chip (LOC) applications where capillary forces are important for fluid flow. PBW has a unique capability of being able to direct-write patterns in thick (1-30µm) polymer resist layers with straight vertical sidewalls. It can be used to prepare master stamps and moulds for mass production in polymeric materials. A recent development, where the direct writing of an entire pattern element is carried out in parallel makes PBW especially well suited for Bio-MEMS LOC applications. In this study we have examined the flow dy…
Parvovirus B19 Genotype Specific Amino Acid Substitution in NS1 Reduces the Protein's Cytotoxicity in Culture
2010
A clinical association between idiopathic liver disease and parvovirus B19 infection has been observed. Fulminant liver failure, not associated with other liver-tropic viruses, has been attributed to B19 in numerous reports, suggesting a possible role for B19 components in the extensive hepatocyte cytotoxicity observed in this condition. A recent report by Abe and colleagues (Int J Med Sci. 2007;4:105-9) demonstrated a link between persistent parvovirus B19 genotype I and III infection and fulminant liver failure. The genetic analysis of isolates obtained from these patients demonstrated a conservation of key amino acids in the nonstructural protein 1 (NS1) of the disease-associated genotyp…
Borrelia burgdorferi Outer Membrane Vesicles Contain Antigenic Proteins, but Do Not Induce Cell Death in Human Cells
2022
Like many bacterial species, Borrelia burgdorferi, the pleomorphic bacterium that causes Lyme borreliosis, produces outer membrane vesicles (OMVs). Borrelial OMVs (BbOMVs) have been identified as containing virulence factors, such as outer surface proteins (Osps) A, B, and C, as well as DNA. However, the pathogenicity of BbOMVs in disease development is still unclear. In this study, we characterized purified BbOMVs by analyzing their size and immunolabeling for known antigenic markers: OspA, OspC, p39, and peptidoglycan. In addition, BbOMVs were cocultured with human non-immune cells for cytotoxicity analysis. The results demonstrated that, on average, the vesicles were small, ranging betwe…
Distinctive Evasion Mechanisms to Allow Persistence of Borrelia burgdorferi in Different Human Cell Lines
2021
Lyme borreliosis is a multisystemic disease caused by the pleomorphic bacteria of the Borrelia burgdorferi sensu lato complex. The exact mechanisms for the infection to progress into a prolonged sequelae of the disease are currently unknown, although immune evasion and persistence of the bacteria in the host are thought to be major contributors. The current study investigated B. burgdorferi infection processes in two human cell lines, both non-immune and non-phagocytic, to further understand the mechanisms of infection of this bacterium. By utilizing light, confocal, helium ion, and transmission electron microscopy, borrelial infection of chondrosarcoma (SW1353) and dermal fibroblast (BJ) c…
Development of biotechnological tools for studying infectious pathways of canine and human parvoviruses
2005
Leona Gilbertin väitöskirjatyö antaa uutta tietoa parvovirusten pinnanmuokkauksesta ja hyödyllisyydestä virologian, geeniterapian ja biolääketieteen sovelluksissa. Parvovirukset ovat pieniä, yksisäikeistä DNA:ta sisältäviä eläinviruksia, jotka kykenevät infektoimaan isäntänsä soluja. Ihmisillä ne aiheuttavat ihottumaa sekä nivelkipuja. Tutkimuksessa tuotettiin bakuloviruksen avulla hyönteissoluissa sekä koiran että ihmisen parvoviruksen rakenneproteiineista fluoresloivia fuusioproteiineja. Selvisi, että parvoviruksen pinta sopii hyvin muokkaamiseen ja vieraiden molekyylien ilmentämiseen. Lisäksi havaittiin, että bakuloviruksia voidaan käyttää geeninsiirtovälineenä mm. parvovirusproteiinien …
Development of a microfluidic design for an automatic lab-on-chip operation
2016
Simple and easy to use are the keys for developing lab-on-chip technology. Here, a new microfluidic circuit has been designed for an automatic lab-on-chip operation (ALOCO) device. This chip used capillary forces for controlled and precise manipulation of liquids, which were loaded in sequence from different flowing directions towards the analysis area. Using the ALOCO design, a non-expert user is able to operate the chip by pipetting liquids into suitable inlet reservoirs. To test this design, microfluidic devices were fabricated using the programmable proximity aperture lithography technique. The operation of the ALOCO chip was characterized from the flow of red-, blue- and un-dyed deioni…
Supplementary data to: Sequence and Regulatory Variation in Acetylcholinesterase Genes Contribute to Insecticide Resistance in Different populations …
2021
This is data used in Aigi Margus, Saija Piiroinen, Philipp Lehmann, et al. Sequence and Regulatory Variation in Acetylcholinesterase Genes Contribute to Insecticide Resistance in Different populations of Leptinotarsa decemlineata. Authorea. September 26, 2021. Data contains bioassay data, enzyme activity data, gene expression data and gene sequence data used to research insect herbivores resistance to insecticides. See the attached data description file for more information.