Other Publications (14)
- Mediators of Inflammation
- Zygote (Cambridge, England)
- Journal of Neurotrauma
- PloS One
- World Journal of Gastroenterology
- Experimental Neurology
- Journal of Huntington's Disease
- Biochimica Et Biophysica Acta
- The Journal of Comparative Neurology
- Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]
- Neuro-degenerative Diseases
- Journal of Neuroimmunology
- Molecular Therapy. Methods & Clinical Development
- Surgical Endoscopy
Articles by Stefan Juhas in JoVE
Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice Takahiro Tadokoro1, Atsushi Miyanohara1, Michael Navarro1, Kota Kamizato1, Stefan Juhas2, Jana Juhasova2, Silvia Marsala1, Oleksandr Platoshyn1, Erik Curtis3, Brandon Gabel3, Joseph Ciacci3, Nada Lukacova4, Katarina Bimbova4, Martin Marsala1,4 1Neuroregeneration Laboratory, Department of Anesthesiology, University of California, San Diego, 2Institute of Animal Physiology and Genetics, Czech Academy of Sciences, 3Department of Neurosurgery, University of California, San Diego, 4Institute of Neurobiology, Slovak Academy of Sciences The goal of the present study was to develop and validate the potency and safety of spinal adeno-associated virus 9 (AAV9)-mediated gene delivery by using a novel subpial gene delivery technique in adult mice.
Other articles by Stefan Juhas on PubMed
Effects of a Combination of Thyme and Oregano Essential Oils on TNBS-induced Colitis in Mice Mediators of Inflammation. 2007 | Pubmed ID: 18288268 We examined the anti-inflammatory effects of the combination of thyme and oregano essential oil dietary administered at three concentrations (0.4% thyme and 0.2% oregano oils; 0.2% thyme and 0.1% oregano oils; 0.1% thyme and 0.05% oregano oils) on mice with TNBS-induced colitis. Treatment of colitic animals with the essential oils decreased the mRNA levels of pro-inflammatory cytokines IL-1beta, IL-6, GM-CSF, and TNFalpha, especially after application of the medium dose. The medium dose of the essential oils significantly lowered the amount of IL-1beta and IL-6 proteins too. Moreover, administration of the medium dose decreased the mortality rate, accelerated the body weight gain recovery, and reduced the macroscopic damage of the colonic tissue. Our results indicate that combined treatment with appropriate concentrations of thyme and oregano essential oils can reduce the production of proinflammatory cytokines, and thereby attenuate TNBS-induced colitis in mice.
Apoptotic Processes and DNA Cytosine Methylation in Mouse Embryos Arrested at the 2-cell Stage Zygote (Cambridge, England). Aug, 2009 | Pubmed ID: 19397842 The present study evaluates the role of apoptotic cell death and DNA methylation reprogramming in early developmental failures occurring in embryos at the 2-cell stage. Mouse 2-cell embryos were cultured in vitro and treated with chemicals that cause developmental arrest and apoptosis (alpha-amanitin, actinomycin D, TNF-alpha). After 24 h, 48 h and 72 h culture, embryos were analysed using cell-death assays (annexin V staining, TUNEL labelling and immunodetection of active caspase-3) and genome methylation assay (immunodetection of 5-methylcytosine). The ability of embryos at the 2-cell stage to undergo apoptotic processes was very low. In arrested embryos, the presence of all evaluated features of apoptosis was recorded only after 72 h culture and their incidence was sporadical. Interestingly, the most frequently observed apoptotic sign was nuclear condensation and the timing of its appearance preceded even the phosphatidylserine flip. Both normally developing and arrested embryos displayed reduction in DNA cytosine methylation. In arrested embryos, this process was independent of cellular cleavage, was more pronounced and finished in almost complete demethylation of the embryonic genome. The timing of the demethylation overlapped with the onset of major apoptotic events. Although observed apoptotic cells showed either demethylated or methylated DNA cytosine in their nuclei, at blastocyst stage the demethylated status appeared more frequently in them.
Chronic Spinal Compression Model in Minipigs: a Systematic Behavioral, Qualitative, and Quantitative Neuropathological Study Journal of Neurotrauma. Feb, 2012 | Pubmed ID: 22029501 The goal of the present study was to develop a porcine spinal cord injury (SCI) model, and to describe the neurological outcome and characterize the corresponding quantitative and qualitative histological changes at 4-9 months after injury. Adult Gottingen-Minnesota minipigs were anesthetized and placed in a spine immobilization frame. The exposed T12 spinal segment was compressed in a dorso-ventral direction using a 5-mm-diameter circular bar with a progressively increasing peak force (1.5, 2.0, or 2.5 kg) at a velocity of 3 cm/sec. During recovery, motor and sensory function were periodically monitored. After survival, the animals were perfusion fixed and the extent of local SCI was analyzed by (1) post-mortem MRI analysis of dissected spinal cords, (2) qualitative and quantitative analysis of axonal survival at the epicenter of injury, and (3) defining the presence of local inflammatory changes, astrocytosis, and schwannosis. Following 2.5-kg spinal cord compression the animals demonstrated a near complete loss of motor and sensory function with no recovery over the next 4-9 months. Those that underwent spinal cord compression with 2 kg force developed an incomplete injury with progressive partial neurological recovery characterized by a restricted ability to stand and walk. Animals injured with a spinal compression force of 1.5 kg showed near normal ambulation 10 days after injury. In fully paralyzed animals (2.5 kg), MRI analysis demonstrated a loss of spinal white matter integrity and extensive septal cavitations. A significant correlation between the magnitude of loss of small and medium-sized myelinated axons in the ventral funiculus and neurological deficits was identified. These data, demonstrating stable neurological deficits in severely injured animals, similarities of spinal pathology to humans, and relatively good post-injury tolerance of this strain of minipigs to spinal trauma, suggest that this model can successfully be used to study therapeutic interventions targeting both acute and chronic stages of SCI.
Combinational Spinal GAD65 Gene Delivery and Systemic GABA-mimetic Treatment for Modulation of Spasticity PloS One. 2012 | Pubmed ID: 22291989 Loss of GABA-mediated pre-synaptic inhibition after spinal injury plays a key role in the progressive increase in spinal reflexes and the appearance of spasticity. Clinical studies show that the use of baclofen (GABA(B) receptor agonist), while effective in modulating spasticity is associated with major side effects such as general sedation and progressive tolerance development. The goal of the present study was to assess if a combined therapy composed of spinal segment-specific upregulation of GAD65 (glutamate decarboxylase) gene once combined with systemic treatment with tiagabine (GABA uptake inhibitor) will lead to an antispasticity effect and whether such an effect will only be present in GAD65 gene over-expressing spinal segments.
Natural Orifice Transluminal Endoscopic Surgery Vs Laparoscopic Ovariectomy: Complications and Inflammatory Response World Journal of Gastroenterology. Jul, 2012 | Pubmed ID: 22826620 To compare natural orifice transluminal endoscopic surgery (NOTES) vs standard laparoscopic ovariectomy in mini pigs with respect to technical aspects, complications and parameters of systemic inflammatory response.
Effective Long-term Immunosuppression in Rats by Subcutaneously Implanted Sustained-release Tacrolimus Pellet: Effect on Spinally Grafted Human Neural Precursor Survival Experimental Neurology. Oct, 2013 | Pubmed ID: 23748136 Achievement of effective, safe and long-term immunosuppression represents one of the challenges in experimental allogeneic and xenogeneic cell and organ transplantation. The goal of the present study was to develop a reliable, long-term immunosuppression protocol in Sprague-Dawley (SD) rats by: 1) comparing the pharmacokinetics of four different subcutaneously delivered/implanted tacrolimus (TAC) formulations, including: i) caster oil/saline solution, ii) unilamellar or multilamellar liposomes, iii) biodegradable microspheres, and iv) biodegradable 3-month lasting pellets; and 2) defining the survival and immune response in animals receiving spinal injections of human neural precursors at 6 weeks to 3 months after cell grafting. In animals implanted with TAC pellets (3.4 mg/kg/day), a stable 3-month lasting plasma concentration of TAC averaging 19.1 ± 4.9 ng/ml was measured. Analysis of grafted cell survival in SOD+ or spinal trauma-injured SD rats immunosuppressed with 3-month lasting TAC pellets (3.4-5.1 mg/kg/day) showed the consistent presence of implanted human neurons with minimal or no local T-cell infiltration. These data demonstrate that the use of TAC pellets can represent an effective, long-lasting immunosuppressive drug delivery system that is safe, simple to implement and is associated with a long-term human neural precursor survival after grafting into the spinal cord of SOD+ or spinal trauma-injured SD rats.
A Transgenic Minipig Model of Huntington's Disease Journal of Huntington's Disease. 2013 | Pubmed ID: 25063429 Some promising treatments for Huntington's disease (HD) may require pre-clinical testing in large animals. Minipig is a suitable species because of its large gyrencephalic brain and long lifespan.
Small Molecule Inhibitors of Protein Interaction with Glycosaminoglycans (SMIGs), a Novel Class of Bioactive Agents with Anti-inflammatory Properties Biochimica Et Biophysica Acta. Jan, 2014 | Pubmed ID: 24060749 Small molecule inhibitors of biologically important protein-glycosaminoglycan (GAG) interactions have yet to be identified.
Pig Models of Neurodegenerative Disorders: Utilization in Cell Replacement-based Preclinical Safety and Efficacy Studies The Journal of Comparative Neurology. Aug, 2014 | Pubmed ID: 24610493 An important component for successful translation of cell replacement-based therapies into clinical practice is the utilization of large animal models to conduct efficacy and/or safety cell dosing studies. Over the past few decades, several large animal models (dog, cat, nonhuman primate) were developed and employed in cell replacement studies; however, none of these models appears to provide a readily available platform to conduct effective and large-scale preclinical studies. In recent years, numerous pig models of neurodegenerative disorders were developed using both a transgenic approach as well as invasive surgical techniques. The pig model (naïve noninjured animals) was recently used successfully to define the safety and optimal dosing of human spinal stem cells after grafting into the central nervous system (CNS) in immunosuppressed animals. The data from these studies were used in the design of a human clinical protocol used in amyotrophic lateral sclerosis (ALS) patients in a Phase I clinical trial. In addition, a highly inbred (complete major histocompatibility complex [MHC] match) strain of miniature pigs is available which permits the design of comparable MHC combinations between the donor cells and the graft recipient as used in human patients. Jointly, these studies show that the pig model can represent an effective large animal model to be used in preclinical cell replacement modeling. This review summarizes the available pig models of neurodegenerative disorders and the use of some of these models in cell replacement studies. The challenges and potential future directions in more effective use of the pig neurodegenerative models are also discussed.
Mechanism of Action and Efficacy of RX-111, a Thieno[2,3-c]pyridine Derivative and Small Molecule Inhibitor of Protein Interaction with Glycosaminoglycans (SMIGs), in Delayed-type Hypersensitivity, TNBS-induced Colitis and Experimental Autoimmune Encephalomyelitis Inflammation Research : Official Journal of the European Histamine Research Society ... [et Al.]. Apr, 2016 | Pubmed ID: 26794621 Elucidate the mechanism of action of the small molecule inhibitor of protein binding to glycosaminoglycans, RX-111 and assay its anti-inflammatory activity in animal models of inflammatory disease.
Mutated Huntingtin Causes Testicular Pathology in Transgenic Minipig Boars Neuro-degenerative Diseases. 2016 | Pubmed ID: 26959244 Huntington's disease is induced by CAG expansion in a single gene coding the huntingtin protein. The mutated huntingtin (mtHtt) primarily causes degeneration of neurons in the brain, but it also affects peripheral tissues, including testes.
Revelation of the IFNα, IL-10, IL-8 and IL-1β As Promising Biomarkers Reflecting Immuno-pathological Mechanisms in Porcine Huntington's Disease Model Journal of Neuroimmunology. Apr, 2016 | Pubmed ID: 27049565 Studies on Huntington's disease (HD) demonstrated altered immune response in HD gene carriers. Using multiplexing immunoassay, we simultaneously investigated seven cytokines in secretomes of microglia and blood monocytes, cerebrospinal fluid (CSF) and serum collected from transgenic HD minipigs at pre-symptomatic disease stage. Decline in IFNα and IL-10 was observed in CSF and secretome of microglia whilst elevated IL-8 and IL-1β levels were secreted by microglia. Additionally, IL-8 was increased in serum. The proportion of mutant huntingtin in microglia may have causative impact on cytokine production. IFNα, IL-10, IL-8 and IL-1β represent promising biomarkers reflecting immuno-pathological mechanisms in porcine HD model.
Potent Spinal Parenchymal AAV9-mediated Gene Delivery by Subpial Injection in Adult Rats and Pigs Molecular Therapy. Methods & Clinical Development. 2016 | Pubmed ID: 27462649 Effective in vivo use of adeno-associated virus (AAV)-based vectors to achieve gene-specific silencing or upregulation in the central nervous system has been limited by the inability to provide more than limited deep parenchymal expression in adult animals using delivery routes with the most clinical relevance (intravenous or intrathecal). Here, we demonstrate that the spinal pia membrane represents the primary barrier limiting effective AAV9 penetration into the spinal parenchyma after intrathecal AAV9 delivery. We develop a novel subpial AAV9 delivery technique and AAV9-dextran formulation. We use these in adult rats and pigs to show (i) potent spinal parenchymal transgene expression in white and gray matter including neurons, glial and endothelial cells after single bolus subpial AAV9 delivery; (ii) delivery to almost all apparent descending motor axons throughout the length of the spinal cord after cervical or thoracic subpial AAV9 injection; (iii) potent retrograde transgene expression in brain motor centers (motor cortex and brain stem); and (iv) the relative safety of this approach by defining normal neurological function for up to 6 months after AAV9 delivery. Thus, subpial delivery of AAV9 enables gene-based therapies with a wide range of potential experimental and clinical utilizations in adult animals and human patients.
A New Experimental Model of Calculous Cholecystitis Suitable for the Evaluation and Training of Minimally Invasive Approaches to Cholecystectomy Surgical Endoscopy. Feb, 2017 | Pubmed ID: 27495340 Novel, less invasive approaches such as single-incision laparoscopic cholecystectomy or natural orifice transluminal endoscopic surgery require preclinical evaluation and training. Therefore, there is a need for an experimental model closely mimicking the clinical situation. The aim of our study was to create an experimental model of calculous cholecystitis in a large laboratory animal and test its feasibility for the evaluation of different techniques of cholecystectomy.