Dissertations / Theses: 'Intradermal'

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Liu, John (John Hsiao-Yung). "Intradermal needle-free powdered drug injection." Thesis, Massachusetts Institute of Technology, 2012. http://hdl.handle.net/1721.1/74993.

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Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2012. Cataloged from PDF version of thesis. Includes bibliographical references (p. 65-67). This thesis presents a new method for needle-free powdered drug injection. The design, construction, and testing of a bench-top helium-powered device capable of delivering powder to controllable depths within the dermis is presented. This device uses a jet of gas undergoing choked flow to entrain powder and subsequently penetrates through the skin for delivery of the powder. Different nozzle designs and orifice geometries are also explored. In vitro injection of polymer beads (1-5 [mu]im in diameter) into porcine tissue demonstrate the device's capability of drug delivery to depths of 260 to 5000 [mu]m. The jet parameters of nozzle orifice diameter and applied pressure are shown to affect injection depth, shape, and success rate. The presented device has the potential to be implemented with stabilized formulations of vaccines to address the cold chain problem-the cost and risk of transporting temperature sensitive vaccines to developing countries. by John Liu. S.M.

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Vosseler, Michael [Verfasser], and Roland [Akademischer Betreuer] Zengerle. "Transdermal chronopharmaceutical drug delivery: microneedles, intradermal infusion experiments and a delivery device = Transdermale chronopharmazeutische Medikamentengabe: Mikronadeln, intradermale Infusionsversuche und ein Verabreichungssystem." Freiburg : Universität, 2014. http://d-nb.info/1123479208/34.

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Ranamukhaarachchi, Sahan. "Skin mechanics, intradermal delivery and biosensing with hollow metallic microneedles." Thesis, University of British Columbia, 2017. http://hdl.handle.net/2429/61168.

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Microneedles (MNs) have gained significant attention over the past decade in drug delivery and biosensing due to their minimally-invasive and less painful nature of use compared to intramuscular/subcutaneous injections, and significant biological benefits. Several fundamental processes enabling MN functionality have not been completely understood, including mechanical interaction between MNs and skin for targeted depth penetration; and precise quantification of fluid delivery in the skin. This thesis presents novel materials, and methodologies for evaluating MN interactions with skin, and investigates the performance of hollow MNs in both intradermal fluid drug delivery and biosensing. A micromechanical comparison between human skin and porcine skin was performed using to determine their mechanical behavior affecting MN insertions. Stratum corneum (SC) of human skin was significantly stiffer (117 ± 42 MPa) than porcine skin (81 ± 32 MPa), requiring higher force of MN insertion to rupture the SC in human skin (107 ± 17 mN) than porcine skin (96 ± 23 mN). An artificial mechanical skin model was developed layer-by-layer to simulate tough human skin (MN insertion force 162 ± 11 mN) and to study the dynamics of MN insertion. Key factors that affected MN insertions into skin, including velocity of impact and total energy delivered to the skin, were identified. ID fluid delivery by hollow MNs was assessed using a novel method involving the low-activity radiotracer technetium-99m pertechnetate (⁹⁹mTcO₄₋). Its delivery allowed accurate quantification of fluid delivered into the skin, back-flowed to the skin surface, and total fluid ejected from the syringes via ID devices with sub-nanoliter resolution. Hollow MNs performed more accurate ID injections than conventional needles (93% vs. 69-87% of fluid per 0.1 mL injection volume). A MN-optofluidic biosensing platform capable of eliminating blood sampling was developed with MNs that can access dermal interstitial fluid that contains numerous drugs at concentrations comparable to blood. The MN lumen was functionalized to collect, trap and detect drugs in 0.6 nL of sample. The optofluidic components provided specific high-sensitivity absorbance measurements for drug binding using enzyme-linked assays. Streptavidin-horseradish peroxidase (LoD = 60.2 nM) and vancomycin (LoD = 84 nM) binding validated this point of care system. Applied Science, Faculty of Electrical and Computer Engineering, Department of Graduate

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Ivory, Matthew Owen. "Characterising skin immune cells to inform development of intradermal vaccines and therapeutics." Thesis, Cardiff University, 2016. http://orca.cf.ac.uk/96969/.

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Epidermal Langerhans cells (LCs) and multiple subsets of dermal dendritic cells (dDCs) make skin a valuable route for vaccination, offering the potential for antigen-sparing immunisation. The interconnected immunological functions of dDC subsets and LCs are not fully understood however. This Thesis therefore aimed to explore the interactions of skin immune cells with viral pathogens and vaccines to inform the development of future therapeutics and intradermal vaccines. LCs and dDCs were isolated from ex vivo human skin tissue using a walkout protocol which allowed the enrichment of the migratory cells from the tissue. LCs and dDCs were infected with a lentiviral vector encoding GFP, allowing study of post-entry HIV viral restriction. The study uncovered the existence of a SAMHD-1-independent antiviral factor unique to LCs. LCs and dDCs from ex vivo skin were used to examine the cross-presentation of an inactivated influenza virus-derived matrix peptide to CD8+ T-cells. Two CD11c+ subsets of dDCs were found to potently cross present the antigen. Delivery of VLPs, which lack genetic material, markedly reduced cross-presentation, suggesting that viral genetic material is vital for dDCs to activate cross-presentation pathways. Future work is required to determine if this is true of other influenza peptides or pathogens. Vaccine delivery studies performed using murine and human models found that dDCs were responsible for the greatest uptake of ovalbumin peptide antigen and LCs did not migrate out of the epidermis in the first 4 hours after inactivated influenza virus vaccine delivery respectively. Collectively, this work highlighted the importance of dDCs in antigen uptake and cross-presentation to prime cytotoxic T-cell responses. Innovative delivery methods such as microneedles offer a means of accessing the dermal compartment in a pain-free manner, though further work is required to determine the optimal combination of vaccine formulation and delivery method to harness the immunostimulatory abilities of dDCs.

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Ferguson, Peter Charles. "Intradermal injection of autologus dermal fibroblasts improves wound healing in irradiated skin." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape11/PQDD_0005/MQ40803.pdf.

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Donate, Amy Lynn. "Development of a Non-Invasive Electrode for Intradermal Electrically Mediated DNA Vaccination." Scholar Commons, 2011. http://scholarcommons.usf.edu/etd/3077.

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Current progress in the development of vaccines has decreased the incidence of fatal and non-fatal infections and increased longevity. However, new technologies need to be developed to combat an emerging generation of infectious diseases. DNA vaccination has been demonstrated to have great potential for use against a wide variety of diseases. Alone, this vaccine technology does not generate a significant immune response for vaccination, but combined with delivery by electroporation (EP), can enhance plasmid expression and immunity against the expressed antigen. Most EP systems, while effective, can be invasive and painful making them less desirable for use in vaccination. Our lab recently developed a non-invasive electrode known as the multi-electrode array (MEA), which lies flat on the surface of the skin without penetrating the tissue. This study evaluated the use of the MEA for the development of DNA vaccines. We assessed the appropriate delivery conditions for gene expression and the development of humoral immunity. We used both B. anthracis and HBV as infectious models for our experiments. Our results indicated that the MEA can enhance gene expression in a mouse model with minimal to no tissue damage. Optimal delivery conditions, based on generation of antibodies, were determined to be 125-175V/cm and 150ms with 200ug and a prime boost protocol administered on Day 0 and 14. Under these conditions, end-point titers of 20,000-25,000 were generated. Neutralizing antibodies were noted in 40-60% of animals. Additionally, we utilized a guinea pig model to assess the translation potential of this electrode. The plasmid encoding HBsAg, pHBsAg, was delivered intradermally with the MEA to guinea pig skin. The results show increased protein expression resulting from plasmid delivery using the MEA as compared to injection alone. Within 48 hours of treatment, there was an influx of cellular infiltrate in the experimental groups. Humoral responses were also increased significantly in both duration and intensity as compared to the injection only groups. Results from both experimental models demonstrate that protective levels of humoral immunity can be generated and that this electrode should translate well to the clinic.

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Anamur, Cihad [Verfasser], and Gerhard [Akademischer Betreuer] Winter. "Novel formulation approaches for ballistic intradermal vaccination / Cihad Anamur. Betreuer: Gerhard Winter." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2015. http://d-nb.info/1079140247/34.

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Abrams, Stephanie B. "Evaluation of Veterinary Allergen Extract Content and Resultant Canine Intradermal Threshold Concentrations." The Ohio State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=osu1524066093298819.

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Omolu, A. M. N. "Evaluation of a novel intradermal delivery system for the treatment of pressure ulcers." Thesis, University College London (University of London), 2018. http://discovery.ucl.ac.uk/1561632/.

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Background: Pressure ulcers, a form of chronic wound, represent a significant health and resource burden in elderly and immobilised patient populations. Pressure ulcers have been shown to exhibit high matrix metalloproteinase (MMP) activity which prevents normal wound healing and doxycycline, as an MMP inhibitor, offers a potential novel treatment. Intradermal delivery of doxycycline could help treat the earliest stages of wound development and prevent further wound progression. The skin, however, has a natural barrier function which prevents the diffusion of large exogenous molecules. Microneedle rollers offer a minimally invasive technique to transiently permeabilise the skin, creating microscopic pores that act as conduits for doxycycline diffusion. / Methods: The research described in this thesis focusses on the repurposing of existing microneedle rollers for the intradermal delivery of doxycycline as a pressure ulcer treatment. Firstly, the effect of microneedle length and application method on micropore formation and rate of drug permeation was investigated using the recently-launched artificial membrane Strat-MTM and compared with excised biological tissue. Next, the biological effects of doxycycline and its transmembrane delivery were modelled in a dermal tissue equivalent (DTE) model, formed from collagen and dermal fibroblasts, by assessing changes DTE contractile behaviour and matrix metalloproteinase (MMP) activity. This simplified in vitro model was further developed to better emulate the pressure ulcer microenvironment by introducing: (i) mechanical loading using a bespoke metal weight, (ii) glucose deprivation through the use of glucose-free media, and (iii) inflammatory cells, specifically macrophages, through co-culture. Investigations progressed to a preliminary in vivo surgical wound model using compression by magnets to determine the biological effects of pressure in a whole organism. This pre-existing model was established in our lab for future investigations of microneedle-mediated doxycycline delivery. Finally, the repurposing of the existing non-invasive imaging technology optical coherence tomography (OCT) for pressure ulcer evaluation was explored in an observational clinical audit. / Results: The results demonstrate that microneedle rollers significantly enhance the transmembrane delivery of doxycycline with significant effect on tissue-equivalent contraction and MMP activity. Results from the pressure ulcer models corroborate previous findings that the pressure ulcer microenvironment augments MMP activity. Lastly, OCT is shown to detect subsurface biomarkers of skin in the earliest stages of pressure ulcer development, most suitable for treatment with doxycycline.

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Kennedy, Joakim. "Pharmaceutical engineering of microneedle-mediated intradermal nanoparticle delivery device : potential for lymphatic targeting." Thesis, Queen's University Belfast, 2016. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.706993.

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Microneedles (MNs) are minimally invasive drug delivery devices that can be used for transdermal drug delivery (TDD). They have the potential to enhance drug delivery for therapeutic agents which are currently difficult to deliver, costly, or cause a wide range of side effects. The therapeutic agent must normally overcome the highly hydrophobic barrier of the skin’s outermost layer, the stratum corneum (s.c.), in order to be delivered transdermally. MNs have a height of 50 m to 1500 m and overcome the barrier by penetrating the s.c. and creating aqueous pores. They can be fabricated from different materials, such as silicon, metal, glass, sugars, and polymers. Dissolving polymeric MNs are used to penetrate the s.c. and release the drug incorporated in the polymeric matrix into the epidermis when they dissolve by taking up skin interstitial fluid (ISF). This means that when nanoparticles (NPs) are delivered using dissolving MNs, the amount delivered is dependent upon the amount localized in the needle portion of the array. The present study developed and characterized dissolving MNs using a novel two-step process which improves the loading distribution of the therapeutic agent in the needle portion of the array. The two-step process fabricates the needles and the baseplate individually, allowing both to be fabricated from different for- mulations and optimized separately. The MNs were loaded with a model protein, ovalbumin (OVA), and poly(lactic-co-glycolide acid) (PLGA) NPs. When the loading distribution was measured for the NPs, it was found that about 80% of the NPs were localized in the needles. This is almost double the localization compared to arrays using the standard fabrication method. The MNs loaded with NPs were used in an in vivo mouse model. The NPs were successfully delivered into the skin of the mice and were detected in the draining lymph nodes. This showed that the developed TDD device can be used for targeted delivery of the lymphatic system. This study was overall successful in engineering a MN-mediated intradermal NP delivery device that can be used to target the lymphatic system.

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Torrisi, Barbara Maria. "Liquid loaded microneedles for the intradermal delivery of botulinum toxin for Primary Focal Hyperhidrosis." Thesis, Cardiff University, 2012. http://orca.cf.ac.uk/39693/.

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Primary focal hyperhidrosis (PFHH) is a medical condition characterised by overactivity of the eccrine sweat glands, primarily occurring on palmar, plantar and axillary regions. PFHH can have a significant adverse impact on a patient’s quality of life. Multiple intradermal injections of a commercial formulation of botulinum toxin A (BTX A) (Botox®) is the most effective non-surgical treatment currently licensed in the UK for cases of severe PFHH. Although effective, intradermal BTX A injections are associated with considerable pain and discomfort for the patient and are time-consuming for the administering clinician. This study aims to evaluate the potential of using pocketed microneedle devices for minimally invasive intradermal delivery of BTX A, as a liquid formulation, into human skin. Pocketed microneedles, metallic 700 μm-long needles containing a cavity within the needle shaft, were selected as an appropriate and relatively untested intradermal delivery device. Pocketed microneedle devices (PMDs) were liquid loaded by immersion into a ‘Botox® like’ formulation that mimicked the composition of the commercial Botox® formulation, with the exception of BTX A, which was replaced by the model macromolecular protein β-galactosidase (~465 kDa). A water-soluble dye was also included to enable visualisation. Microneedles were assessed for loading uniformity by light microscopy and the formulation residency time was evaluated by monitoring evaporation using a digital camera. The microneedle loading capacity was determined using an established quantitative assay for β-galactosidase. Studies using excised human breast skin, maintained in organ culture, examined delivery of the model β-galactosidase from liquid loaded PMDs and the time-dependent diffusion of the protein within the dermal tissue. A more clinically representative model of BTX A, formaldehyde inactivated BTX A, i.e., botulinum toxoid, was used to determine the deposition pattern of the therapeutic within the skin. Following skin delivery the toxoid was detected by immnohistochemical staining and fluorescence imaging, following its conjugation to an appropriate fluorophore. Immersion of the PMD into a ‘Botox® like’ formulation resulted in successful uptake and retention of the model protein solution. Quantitative studies indicated that nanogram quantities (~100 ng/microneedle array) of the β-galactosidase model can be loaded and retained on individual microneedles, in a liquid state. These results suggest that the loading capacity of the microneedle device is appropriate for therapeutic botulinum toxin formulations, although loading uniformity will need to be addressed. Histological analysis revealed effective delivery of the model β-galactosidase from a PMD to the epidermal and the dermal layers of the skin. Rapid and extensive diffusion of the protein within the deeper dermis was also demonstrated. Further, immunohistochemical and fluorescence studies indicated effective PMD loading and successful delivery of botulinum toxoid to the dermis of human skin. These data suggest that it should be possible for BTX A to access its therapeutic target (the eccrine sweat glands) following delivery via PMDs. This study has demonstrated for the first time that pocketed microneedles represent a viable, minimally invasive alternative, for the intradermal delivery of botulinum toxin A (Botox®). Future pre-clinical and clinical studies are now required to test and optimize a microneedle-based delivery system that is most suited to clinical practice.

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Wong, David Michael. "The Intradermal Skin Test in the Horse: Value as a Diagnostic Modality in Equine Allergies." Thesis, Virginia Tech, 2003. http://hdl.handle.net/10919/9916.

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Recent studies have provided conflicting results in regards to equine intradermal skin testing and its use in defining causative antigens in IgE mediated diseases such as equine recurrent airway obstruction (RAO). This study was divided into two experiments. In the first experiment of this study, the hypothesis tested was normal horses would have minimal variability in the wheals formed by intradermal injection of positive control stimulants. This was evaluated by examining the repeatability of skin test wheals created by 5 concentrations of histamine, compound 48/80, and phaseolus vulgaris (PHA) within a normal horse and between 12 normal horses at 0.5 hours, 4 hours, and 24 hours post injection. Minimal variability was detected within individual horses and between 12 horses for histamine and compound 48/80 at 0.5 hours and for PHA at 4 hours. This information suggests that the intradermal injection of positive control substances is a repeatable test in normal horses. In the second experiment of this study, the hypothesis tested was normal horses react differently to intradermal injection of positive control stimulants (histamine, compound 48/80, PHA) and/or an environmental antigen (Aspergillus) in comparison to horses affected with RAO. This was evaluated by identifying differences in wheal responses between normal horses and RAO affected horses. Concentration response curves were created in normal and RAO affected horses to the aforementioned stimulants at 0.5 hours, 4 hours, and 24 hours post injection. No statistically significant differences were noted in concentration response between normal and ROA affected horses when compound 48/80 and PHA were evaluated. RAO affected horses demonstrated a greater slope at the 0.5 hour time when compared to normal horses suggesting that RAO affected horses are hypersensitive to intradermal injection of histamine. Injection of Aspergillus mix at 4000 protein nitrogen units/ml caused an intradermal wheal reaction at the 24-hour time in 4/5 RAO horses. This reaction was not noted in normal horses. This information suggests that there may be a positive relationship between causative antigens (i.e. Aspergillus) that may induce clinical RAO and positive intradermal skin test results. An additional aspect that was evaluated in both experiments involved histologic examination of skin biopsies taken from wheals created by intadermal injection of histamine, compound 48/80, PHA, and Aspergillus at various times post injection. In the first experiment, intradermal injection of histamine caused severe dermal edema and margination of neutrophils and eosinophils at 0.5 hours. Compound 48/80 demonstrated mild to modest dermal edema at 0.5 hours while PHA demonstrated severe dermal edema, hemorrhage, and lymphactic ectasia at 4 and 24 hours. PHA also demonstrated a neutophilic inflammation at 4 hours that progressed to a mixed lymphohistiocytic and neutrophilic inflammation at 24 hours. In the second experiment, no edema and modest to moderate neutrophilic inflammation was noted in normal horses after intradermal injection of Aspergillus at 24 hours. In contrast, RAO affected horses demonstrated mild to modest edema and a mild to moderate mixed inflammatory response (lympho-histocytic, neutrophilic, eosinophilic) after intradermal injection of Aspergillus at 24 hours suggesting a delayed type response. Master of Science

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Ferraro, Bernadette. "Intradermal Delivery of Plasmids Encoding Angiogenic Growth Factors by Electroporation Promotes Wound Healing and Neovascularization." [Tampa, Fla] : University of South Florida, 2009. http://purl.fcla.edu/usf/dc/et/SFE0002823.

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Engelke-Mücksch, Laura [Verfasser], and Julia [Akademischer Betreuer] Engert. "Cutaneous vaccination - investigation of different approaches of intradermal vaccine delivery / Laura Engelke-Mücksch ; Betreuer: Julia Engert." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2018. http://d-nb.info/1168590825/34.

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Donadei, Agnese. "Skin receptors agonists as novel adjuvants for Meningococcal glycoconjugate vaccines: the impact of antigen-agonist conjugation and evaluation of intradermal vaccine delivery as an alternative to the intramuscular route." Doctoral thesis, Università degli studi di Padova, 2016. http://hdl.handle.net/11577/3424824.

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Vaccines are complex multi-component products, in which an interdependent relationship among constituents exists: the concomitant inclusion of antigens, adjuvants, and excipients is essential to guarantee the efficacy of the final product. The selection of adjuvants and formulation criteria is a fundamental aspect, having implications towards efficacy, safety and stability of vaccines. In this field, the work was aimed at the design, formulation and characterization of novel vaccines suitable for intradermal delivery and able to induce earlier onset of immune response as well as more potent and longer lasting immune responses in comparison to conventional vaccines. With this purpose two different model antigens were used to evaluate the intradermal route as immunological target for vaccination: MenC-CRM_197 glycoconjugate and CRM_197 protein (detoxified diphtheria toxin). Moreover different adjuvants were tested to potentiate and modulate the immune responses towards a desired quality profile. The impact of adjuvant-antigen conjugation on immune response against the antigen was investigated in comparison to the adjuvant/antigen co-delivery. For the first time, the intradermal delivery was evaluated as route for the administration of a MenC-CRM_197 glycoconjugate vaccine. LT mutant (LTK63), MPL (TLR4a), Toll like receptor 7 agonist (TLR7a), αGalCer analog, Dectin-1 receptor agonist (β-(1-3)glucan hexasaccharide) and an oil in water (o/w) emulsion have been chosen to target the skin-resident antigen-presenting cells and to enhance the immune response against the antigen. The in vivo evaluation of such novel formulations provided encouraging results with respect to quantity, quality and functionality of the induced immune responses. Notably, the intradermal delivery of MenC-CRM_197 glycoconjugate showed superiority in term of immunogenicity and serum bactericidal titers compared to the intramuscular administration, highlighting the power of the intradermal route for glycoconjugate vaccine delivery. Moreover, the addition of LTK63, TLR4a, TLR7a, αGalCer analog adjuvants allowed the reduction of the number of doses administered and especially LTK63, TLR4a, TLR7a adjuvants were able to modulate the quality of the immune response towards a more beneficial, for the model antigen used, Th1 response. No adjuvant effect was observed when formulations were combined with (β-(1-3) glucan hexasaccharide. When MenC-CRM_197+TLR4a formulation was intradermally delivered in combination with the o/w emulsion adjuvant no additional improvements in immunogenicity were observed in comparison to the single adjuvant effect. The immune system activation was also investigated using a protein model antigen 〖(CRM〗_197) intradermally; however no improvements in term of immunogenity were observed in comparison to the intramuscular administration. This finding has specified the interest in selecting a glycoconjugate antigen to exploit the advantages of the intradermal route in terms of immunogenicity. To further investigate the role of adjuvant and to ensure codelivery of adjuvant/antigen, the conjugations of TLR7a to the glycoconjugate model antigen have been investigated. Two different approaches were developed: the conjugation of TLR7a to MenC polysaccharide and to CRM_197 carrier protein. When the conjugation of TLR7a to CRM_197 carrier protein was tested in vivo, results clearly showed that conjugation of the TLR7a enhances the anti-carbohydrate response. This effect was less pronounced than after co-administration of a commercial glycoconjugate with a standard dose of TLR7a adsorbed on AlumOH. Conjugation of the small immunopotentiator molecule was particularly suited for vaccination via intradermal delivery, where insoluble salts of aluminum cannot be used because of their reactogenicity. These findings have opened the path to the rational design of improved adjuvanted glycoconjugate vaccines for intradermal routes. Regarding the conjugation of TLR7a to MenC polysaccharide tested in vivo, the obtained results have shown a negative impact of the conjugation to MenC polysaccharide receptor recognition and consequently a reduction of the anti-MenC carbohydrate response. In order to investigate the potential use of a short β-glucan chain as adjuvant for intradermal vaccination toward the CRM_197 protein model antigen, in comparison to the previously presented glycoconjugate model antigen, the results obtained by the in vitro and in vivo evaluation of the β-glucan conjugation to the CRM_197 protein were reported. The results have demonstrated that receptor activation was significantly impacted by the presentation of the glucan conjugated to the protein antigen. Considering that glycoconjugate vaccines are some of the safest and most effective vaccines for reducing, even eradicating, infectious diseases, the conjugation of well-defined synthetic glucans could represent a useful strategy for developing new glycoconjugate vaccines for intradermal delivery, with inherent adjuvant properties. The combination of antigens and adjuvants needs to be rationally defined to develop stable formulations and to give the safest and most efficient response with respect to the considered pathogen and route of administration. In the present work, formulation science has been proposed as a key element of novel vaccine development suitable for intradermal delivery. I vaccini sono prodotti complessi costituiti da più componenti, nei quali esiste un rapporto di interdipendenza tra i costituenti: la concomitante presenza di antigeni, adiuvanti, ed eccipienti è essenziale per garantire l'efficacia del prodotto finale. La selezione degli adiuvanti e i criteri di formulazione sono un aspetto fondamentale, avendo implicazioni su efficacia, sicurezza e stabilità dei vaccini. In questo ambito, il lavoro è stato finalizzato alla progettazione, formulazione e caratterizzazione di nuovi vaccini atti alla consegna per via intradermica e capaci di indurre uno sviluppo precoce, più intenso e più duraturo della risposta immunitaria rispetto ai vaccini convenzionali. Con questo scopo due diversi antigeni sono stati utilizzati come modello per valutare la via di somministrazione intradermica per la vaccinazione: il glicoconiugato MenC-CRM_197 e la proteina CRM_197 (tossina difterica detossificata). Inoltre diversi adiuvanti sono stati usati per potenziare e modulare le risposte immunitarie verso un profilo di qualità desiderata. In aggiunta è stato valutato l'impatto che la coniugazione dell’adiuvante all’antigene ha sulla risposta immunitaria contro l'antigene in confronto alla semplice miscelazione. La via di immunizzazione intradermica è stata valutata, per la prima volta, per la somministrazione del glicoconiugato MenC-CRM_197. Per dirigere l’antigene verso le cellule APC presenti nella pelle e migliorare la risposta immunitaria verso l’antigene, sono stati testati diversi adiuvanti, quali la tossina enterica detossificata di Escherichia coli (LTK63), un agonista del recettore Toll Like 4 (TLR4a) il monofosfolipide A (MPL), un agonista del recettore Toll like 7 (TLR7a), un analogo dell’αGalCer, un agonista del recettore Dectin-1 (β- (1-3) hexasaccharide glucano) e l’emulsione olio/acqua (o/w). La valutazione in vivo di tali nuove formulazioni ha fornito alcuni risultati incoraggianti riguardanti la quantità e la qualità delle risposte immunitarie indotte. In particolare, i risultati immunologici ottenuti hanno dimostrato una netta superiorità della via di immunizzazione intradermica rispetto a quella intramuscolare quando è utilizzato il glicoconiugato MenC-CRM_197. Inoltre, l'aggiunta degli adiuvanti LTK63, TLR4a, TLR7a, l’analogo dell’αGalCer ha consentito la riduzione del numero di dosi somministrate e soprattutto le formulazioni adiuvantate con LTK63, TLR4a e TLR7a sono state in grado di modulare la qualità della risposta immunitaria verso una più vantaggiosa risposta Th1. Nessun effetto adiuvante è stato osservato utilizzando il glucano esasaccarico come adiuvante miscelato all’antigene. Il glicoconigato MenC-CRM_197 adiuvantato con il TLR4a è stato consegnato per via intradermica anche in combinazione con l’emulsione o/w, ma non è stato osservato alcun effetto adiuvante aggiuntivo della miscela in confronto all’effetto dei singoli immunopotenziatori. In aggiunta è stata valutata l’immunogenicità della proteina CRM_197 a seguito della consegna intradermica ma non è stato osservato alcun miglioramento in termini di immunogenicità in confronto alla somministrazione intramuscolare. Questo risultato ha messo in evidenza l'importanza dell’adeguata scelta dell’antigene per sfruttare i vantaggi della via di immunizzazione intradermica: un antigene glicoconiugato è preferibile ad un antigene proteico. Per meglio definire l’effetto adiuvante e garantire la concomitante somministrazione di antigene e adiuvante, sono stati svilippati e studiati due approcci di coniugazione del TLR7a all'antigene modello MenC-CRM_197: la coniugazione del TLR7a prima al polisaccaride MenC e poi alla proteina CRM_197. I risultati ottenuti dai tests in vivo del coniugato TLR7a alla proteina CRM_197 hanno dimostrato che la coniugazione del TLR7a migliora la risposta anti-MenC. L’effetto adiuvante del TLR7a coniugato si è rivelato meno pronunciato rispetto alla somministrazione di una dose standard di TLR7a adsorbito su AlumOH. Bisogna però sottolineare che il sale di alluminio non può essere consegnato per via intradermica, dato la sua reattogenicità, perciò ne risulta che la coniugazione del TLR7a alla proteina del glicoconiugato MenC-CRM_197 può essere molto utile per la vaccinazione intradermica. I risultati ottenuti costituiscono la base per una razionale progettazione di vaccini glicoconiugati adiuvantati da somministrare per via intradermica. Per quanto riguarda l’approccio di coniugazione del TLR7a al MenC, i risultati ottenuti in vivo hanno portato alla luce un impatto negativo della coniugazione sul riconoscimento recettoriale del polisaccaride e di conseguenza una riduzione della risposta anti-MenC. Il glucano esasaccaridico è stato utilizzato per studiare il suo effetto adiuvante quando coniugato ad un antigene proteico a seguito di una somministrazione intradermica. I risultati ottenuti dalla sperimentazione in vitro ed in vivo dell’esasaccaride coniugato alla proteina CRM_197 hanno dimostrato che l'attivazione del recettore Dectin-1 è significativamente influenzata dalla presenza del glucano. Considerando che i vaccini glicoconiugati sono alcuni dei vaccini più sicuri e più efficaci per ridurre e sradicare le malattie infettive, la coniugazione di glucani sintetici potrebbe rappresentare una strategia utile per lo sviluppo di nuovi vaccini glicoconiugati adatti alla somministrazione intradermica, con intrinseche proprietà adiuvanti. La combinazione appropriata di antigeni e adiuvanti deve guidare la formulazione di vaccini stabili che forniscano una risposta più sicura ed efficace contro il patogeno desiderato mediante la via di somministrazione scelta. Nel presente lavoro, la scienza di formulazione è stata proposta come un elemento chiave dello sviluppo di nuovi vaccini adatti alla consegna intradermica.

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Korkmaz, Emrullah. "Biodissolvable Microneedle Arrays for Effective Transdermal and Intradermal Delivery of Vaccines and Therapeutics: Manufacturing and Cutaneous Applications." Research Showcase @ CMU, 2016. http://repository.cmu.edu/dissertations/761.

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Transdermal delivery of biologically active molecules offers many attractive advantages over prevailing oral and parenteral drug delivery approaches toward addressing the low bioavailability of oral drugs and the inconvenience of hypodermic needle injections. However, transdermal delivery is hampered by the inability of the vast majority of drugs to penetrate through skin due to the physical barrier imposed by the skin’s outer layer, stratum corneum (SC), which allows only very small molecules (< 500 Da) to pass through. This places significant limits on the types of vaccines and therapeutics that can be directly delivered through skin. To mitigate these limitations while retaining the advantages of transdermal delivery, microneedle arrays (MNAs) have been developed to mechanically penetrate the SC for enabling the delivery of bioactive micro- and macromolecules through skin. In addition to transdermal applications of MNA technology that deliver drugs systemically through the skin’s microvasculature and lymphatic flow, MNAs have been used for intradermal applications to deliver drugs locally to the targeted skin microenvironments. Specifically, dissolvable MNAs that incorporate drug (and dissolve when inserted into skin) provide an effective and minimally-invasive means to deliver vaccines and therapeutics to and through skin. Despite recent advancements in MNA technology, there still remain considerable challenges in design and manufacturing, which hinders rapid and low cost fabrication of dissolvable MNAs with clinically-relevant materials and geometries, thereby limiting optimal skin-targeting immunization and treatment strategies. In this Ph.D. research, to address the issues with the current MNA fabrication technologies, which impose strict limitations on manufacturable MNA designs, a novel micro- manufacturing technique based on mechanical diamond micromilling and micromolding is proposed. The overarching objective of this Ph.D. thesis research is to design and evaluate a new and comprehensive manufacturing strategy for successful creation of dissolvable MNA-based trans/intradermal delivery platforms that enable efficient, precise, and reproducible delivery of bioactive molecules to and through skin toward effective vacci- nation strategies and skin-targeted therapies. The specific objectives include: (1) to devise a novel micromilling/elastomer molding/spin-casting based fabrication approach for accurate and reproducible manufacturing of dissolvable MNAs with diverse and unique geometries, and from a myriad of biodissolvable and biodegradable materials, (2) to investigate the intradermal delivery characteristics of the created dissolvable MNAs through ex vivo and in vivo studies in mouse and human skin, and (3) to evaluate the use of the fabricated dissolvable MNAs for effective intradermal delivery of a number of vaccines (e.g., antigens and adjuvants), therapeutics (e.g., anti-cytokine biologics), and genetic materials (e.g., recombinant DNA and RNA) relevant to a broad range of novel cutaneous applications, including enhanced intradermal immunization strategies and skin-targeted therapies.

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Gagnaire, Aurelie. "Rôle de la voie COX-2 au cours de l'infection par Brucella." Thesis, Aix-Marseille, 2016. http://www.theses.fr/2016AIXM4054.

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Brucella est une bactérie intracellulaire facultative à Gram négatif responsable d’une zoonose, la brucellose. Pour persister dans l’organisme, Brucella agit comme un pathogène furtif en modulant la réponse immunitaire de l’hôte. La cyclooxygénase 2 (COX-2) est l’enzyme responsable de la synthèse des prostanoïdes, des médiateurs lipidiques dérivés de l’acide arachidonique (AA) présentant des propriétés immunorégulatrices. Cette thèse est centrée sur l’étude de cette voie métabolique au cours de l’infection par Brucella in vitro dans des cellules dendritiques (DC) humaines et murines ainsi qu’in vivo chez la souris en comparant différentes routes d’infection. Nous avons mis en évidence la capacité de l’infection à stimuler in vitro la production d’AA ainsi que l’expression de Ptgs2. In vivo, la comparaison des différentes routes d’inoculation a montré que l’infection intradermale induit une signature génique inflammatoire caractérisée par l’expression de Ptgs2 et d’Ifng. L’utilisation de NS-398, un inhibiteur spécifique de COX-2 stimule la clairance bactérienne dans les ganglions cervicaux (CLN) drainant le site d’infection. Ces résultats ouvrent ainsi la voie à de nouvelles stratégies thérapeutiques dans le traitement de la brucellose. La seconde partie de la thèse traite de l’implication des infections bactériennes dans l’initiation des processus oncogéniques. Nous y présentons une revue répertoriant l’ensemble des mécanismes pouvant contribuer à l’initiation oncogénique ainsi qu’un projet que nous développons au laboratoire portant sur l’initiation d’un lymphome folliculaire à la suite d’une stimulation antigénique chronique suite à l’infection par B. abortus Brucella is a facultative intracellular Gram-negative bacterium, responsible for a zoonosis called brucellosis. To persist into the host, Brucella acts as a stealthy pathogen by modulating the host immune response. The cyclooxygenase 2 (COX-2) is the enzyme responsible for the synthesis of prostanoids, a family of lipid mediators derived from the arachidonic acid (AA) and presenting immunomodulatory properties. Here, we have studied the impact of this pathway during Brucella infection in vitro in human and murine dendritic cells (DCs) as well as in vivo by comparing different infection routes. We have highlighted the ability of the infection to stimulate the AA synthesis and Ptgs2 expression. In vivo, by comparing different inoculation routes we showed that intradermal infection induces a specific inflammatory gene signature characterized by an important expression of Ptgs2 and Ifng. The use of NS-398, a specific inhibitor of COX-2 stimulates the bacterial clearance in the cervical lymph nodes (CLN) draining the site of infection. These results might open the way to new therapeutic strategies in the treatment of brucellosis. In a second part of the thesis, we discuss the involvement of bacterial infections in initiating oncogenic processes. Here, we present a review listing all the mechanisms that contribute to the oncogenic initiation and a project that we are developing in the laboratory dealing with the initiation of follicular lymphoma following chronic antigenic stimulation during B. abortus infection

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Roberts, Holly Ann. "Equine Intradermal Test Threshold Concentrations for House Dust Mite and Storage Mite Allergens and Identification of Stable Fauna." The Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1396694230.

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Roxhed, Niclas. "A Fully Integrated Microneedle-based Transdermal Drug Delivery System." Doctoral thesis, Stockholm : Kungliga Tekniska högskolan, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-4484.

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Thiebaud, Robert S. "The Role of Nitric Oxide, Acetylcholine, and Vasoactive Intestinal Peptide on Skin Blood Flow During In-Vivo Electrical Field Stimulation." BYU ScholarsArchive, 2010. https://scholarsarchive.byu.edu/etd/2451.

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The purpose of this study was to characterize a novel technique to study neurogenic control of cutaneous vasodilation. We monitored skin blood flow (SkBF) during in-vivo electrical stimulation (e-stim) intended to activate cutaneous nerves and used intradermal microdialysis to deliver receptor antagonists to characterize their contribution to cutaneous vasodilation. We examined the role of acetylcholine receptors (RACh), nitric oxide (NO), and vasoactive intestinal peptide receptors (RVIP) on the cutaneous vasodilation induced by e-stim in the absence of the sympathetic adrenergic nervous system. Six men and three women participated in the study. Three intradermal microdialysis probes were placed in the skin of the dorsal side of their forearm. The adrenergic nervous system was eliminated by delivery of a cocktail of phentolamine (0.01 mg/ml), propranolol (1 mM), and BIBP-3226 (10 µM). At one skin site atropine (0.1 mg/ml) was delivered to block RACh. At a second site we blocked nitric oxide synthase (NOS, 10 mM L-NAME) and RACh. Finally at the third site, we blocked RACh, NOS, and RVIP (0.47 mg/ml VIP10-28). The SkBF response to 1 minute stages of graded increases in frequency (0.2, 1, 2, 4, 8, and 32 Hz) at a current of 1.0 ± 0.1 mA was used to generate a stimulus-response curve before and after drug delivery. At skin site 1 RACh blockade decreased the area under curve (AUC) by 4% from 614 ± 279 to 591 ± 331 (p>0.05). Nitric oxide synthase and RACh blockade reduced the vasodilator response to e-stim by 23% from 1036 ± 457 to 801 ± 448 AUC (p>0.05). Combined NOS, RACh, and RVIP blockade reduced the vasodilator response by 48% from 802 ± 412 to 418 ± 268 AUC (p=0.07). RACh blockade had no effect on the vasodilator response to e-stim. However, in these preliminary studies both NOS and RVIP blockade provided some attenuation of the cutaneous vasodilator response associated with e-stim. Additional studies will focus on these two neurotransmitters as this novel method is refined. Advantages of e-stim include activating the sympathetic nervous system without activating local and humoral factors and studying neurotransmitters in an in-vivo setting during rest, thermal stress, or exercise.

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Beffort, Lisa. "Evaluation der Verträglichkeit und Wirksamkeit der nadellos, intradermal applizierten Vakzine Porcilis® M Hyo ID ONCE gegen Mycoplasma hyopneumoniae unter Feldbedingungen." Diss., Ludwig-Maximilians-Universität München, 2015. http://nbn-resolving.de/urn:nbn:de:bvb:19-180119.

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In der vorliegenden Studie wurde die Sicherheit und Wirksamkeit einer nadellosen, intradermalen Vakzination von Ferkeln im Alter von 21 Tagen mit dem Inaktivatimpfstoff Porcilis® M Hyo ID ONCE unter Feldbedingungen evaluiert. Hierfür wurde ein geschlossener Betrieb in Nordostdeutschland mit einer nachgewiesenen M. hyopneumoniae Infektion ausgewählt. 420 klinisch gesunde Saugferkel wurden randomisiert in zwei Vakzine- und eine Kontrollgruppe eingeteilt. Die Tiere der M Hyo ID ONCE-Gruppe erhielten einmalig, nadellos mit dem IDAL Gerät, 0,2 ml Porcilis® M Hyo ID ONCE intradermal. Die Tiere der zweiten Vakzinegruppe wurden intramuskulär mittels Kanüle mit 2 ml M+PAC® geimpft. Die Kontrolltiere erhielten Diluvac® Forte. Zur Überprüfung der Wirksamkeit wurden die Gewichte der Tiere erhoben und der prozentuale Anteil des makroskopisch veränderten Lungengewebes jedes Lungenlappens am Schlachthof beurteilt. Die Evaluierung der Verträglichkeit des Impfstoffes zeigte, dass die nadellose, intradermale Vakzination signifikant größere, härtere und gerötetere u./ o. wärmere lokale Hautreaktionen hervorrief, als die alleinige intradermale Injektion des Adjuvans Diluvac® Forte. Nach der Applikation von Porcilis® M Hyo ID ONCE wurden signifikant größere und härtere lokale Hautreaktionen beobachtet als nach der Injektion von M+PAC®. Insgesamt waren die beobachteten Hautreaktionen jedoch von geringem Durchmesser bis maximal 1,5 cm und transienter Natur. Die klinischen Untersuchungen, die Temperaturverlaufskontrollen und die Mortalität ergaben keine signifikanten Unterschiede zwischen der Applikation von Porcilis® M Hyo ID ONCE und der Injektion des Placebos. Die Gewichtsdaten betreffend waren die mittleren täglichen Zunahmen der vakzinierten Tiere sowohl in der Mastphase, als auch über die gesamte Studienzeit signifikant um 37 g beziehungsweise 28 g höher. Die intradermal vakzinierten Tiere hatten gegenüber der Kontrollgruppe eine um 1,6 Tage verkürzte Mastperiode. Die Mastleistungsdaten unterschieden sich nicht signifikant zwischen den beiden Vakzinegruppen. Bezüglich der EP-typischen Lungenläsionen, reduzierte die Impfung mit Porcilis® M Hyo ID ONCE signifikant den prozentualen Anteil des makroskopisch veränderten Lungengewebes im Vergleich zur ungeimpften Kontrollgruppe. Folglich hatten signifikant mehr Tiere der M Hyo ID ONCE-Gruppe (31,3 %) makroskopisch unveränderte Lungen, als Tiere der Placebogruppe (7,7 %). Zusätzlich konnte nach der intradermalen Applikation von Porcilis® M Hyo ID ONCE eine signifikante Reduktion der Prävalenz und des Grades EP-typischer Lungenläsionen gegenüber der intramuskulären Vakzination mit M+PAC® nachgewiesen werden. Abschließend kann die nadellose, intradermale Applikation von Porcilis® M Hyo ID ONCE bei 21 Tage alten Ferkeln als sicher und wirksam beurteilt werden.

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Schenkel, Marc. "Intradermal skin testing in cats using fluorescein reveals a high association with abdominal skin lesions as a possible hallmark for atopic cats /." [S.l.] : [s.n.], 2001. http://www.ub.unibe.ch/content/bibliotheken_sammlungen/sondersammlungen/dissen_bestellformular/index_ger.html.

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23

Ganier, Clarisse. "Potentiel thérapeutique des cellules stromales mésenchymateuses dans l'épidermolyse bulleuse dystrophique récessive Intradermal injection of bone marrow-MSCs corrects recessive dystrophic epidermolysis bullosa in a xenograft model Intradermal injection of human umbilical cord-MSCs shows less efficacy than bone marrow-MSCs to correct recessive dystrophic epidermolysis bullosa in a xenograft model." Thesis, Sorbonne Paris Cité, 2018. https://wo.app.u-paris.fr/cgi-bin/WebObjects/TheseWeb.woa/wa/show?t=2117&f=15515.

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L'épidermolyse bulleuse dystrophique récessive (EBDR) est une maladie génétique cutanée due à des mutations de perte de fonction du gène COL7A1 codant pour le collagène VII. Le collagène VII forme les fibres d'ancrage, structures essentielles pour l'adhésion de l'épiderme au derme sous-jacent. Les patients EBDR développent dès la naissance, des décollements bulleux de la peau et des muqueuses responsables de plaies chroniques et de graves complications locales et systémiques. La survenue de carcinomes épidermoïdes cutanés agressifs reste la première cause de décès. Il n'existe pas de traitement à ce jour. Les cellules stromales mésenchymateuses (CSM) sont des cellules multipotentes, isolées à partir de tissus adultes (moelle osseuse, tissu adipeux) ou de tissus périnataux (cordon ombilical). Des travaux antérieurs ont montré que les injections locales et systémiques de CSM issues de la moelle osseuse (CSM-MO) allogéniques ont un potentiel pour réduire l'inflammation cutanée et améliorer la cicatrisation des plaies chez les patients EBDR. Ces améliorations cliniques sont cependant transitoires et les mécanismes d'action des CSM-MO dans l'EBDR ainsi que leur durée de vie après injection sont mal connus. Les CSM-MO pourraient agir via leurs propriétés immunomodulatrices, anti-fibrotiques, pro-angiogéniques, par un effet paracrine permettant l'expression de collagène VII endogène et/ou la sécrétion de collagène VII par les CSM-MO injectées. L'objectif de cette thèse a été d'étudier le potentiel thérapeutique des CSM dans des modèles précliniques de l'EBDR. Nous avons tout d'abord montré que les CSM-MO expriment une quantité d'ARNm de COL7A1 et de collagène VII comparable aux fibroblastes dermiques sains en culture. Nous avons ensuite évalué la capacité des CSM-MO humaines à survivre et produire du collagène VII à la jonction dermo-épidermique (JDE) à long terme après une injection locale dans des peaux équivalentes humaines EBDR greffées sur souris nude. L'injection intradermique (ID) de CSM-MO in vivo a permis de restaurer l'expression du collagène VII ainsi que la formation de fibres d'ancrage à la JDE jusqu'à 6 mois après l'injection. Les CSM-MO sont retrouvées dans la peau équivalente jusqu'à 4 mois après l'injection. Ces résultats montrent qu'une injection ID unique de CSM-MO in vivo permet de rétablir une production prolongée de collagène VII synthétisé par les cellules injectées et d'améliorer l'adhésion dermo-épidermique de la peau équivalente EBDR. Nous avons ensuite comparé l'efficacité des CSM issues de la gelée de Wharton de cordon ombilicaux (CSM-CO) humains aux CSM-MO suivant la même méthodologie que précédemment. Les CSM-CO expriment en culture une quantité d'ARNm de COL7A1 et de collagène VII supérieures aux CSM-MO et fibroblastes dermiques sains. Une injection unique ID de CSM-CO dans la peau EBDR équivalente greffée permet de rétablir une faible expression de collagène VII jusqu'à 4 mois après l'injection. Les CSM-CO sont détectées dans la peau équivalente jusqu'à 2 mois après l'injection. Ces données montrent que les CSM-CO ont une capacité moindre à restaurer l'expression du collagène VII à la JDE comparativement aux CSM-MO injectées dans le même modèle de xénogreffes EBDR. Ces résultats ouvrent la perspective d'une thérapie génique ex vivo utilisant des CSM-MO murines Col7a1-/-. Les souris Col7a1-/- reproduisent les lésions cutanées et muqueuses observées chez les patients EBDR. L'espérance de vie de ces animaux est très réduite. Les CSM-MO murines Col7a1-/- transduites en culture à l'aide d'un vecteur rétroviral SIN exprimant COL7A1 produisent en moyenne 30 fois plus de collagène VII que les CSM-MO murines WT. Des expériences in vivo sont nécessaires pour déterminer si l'injection de CSM-MO génétiquement corrigées ont le potentiel de traiter des lésions cutanées et muqueuses, pour définir la dose optimale et la durée de l'effet chez l'animal. Ceci constituerait une étape importante vers la clinique Recessive dystrophic epidermolysis bullosa (RDEB) is a severe skin disease caused by loss-of-function mutations in COL7A1 encoding type VII collagen. Type VII collagen forms anchoring fibrils which are essential structures for dermal-epidermal adherence. Patients with RDEB suffer since birth from skin and mucosal blistering and develop severe complications. The development of aggressive squamous cell carcinomas is the first cause of demise of these young patients. To date, there is no treatment. Mesenchymal stromal cells (MSC) are multipotent cells, isolated from adult tissue (bone marrow, adipose tissue) or perinatal tissue (umbilical cord). Previous works have shown that local and systemic injections of allogeneic bone marrow-derived MSC (BM-MSC) have a potential to reduce skin inflammation and to improve wound healing in RDEB patients. However, clinical improvement was transient and the mechanisms of action of BM-MSC in RDEB and also their survival after injection are still poorly understood. BM-MSC could act through immunomodulation, anti-fibrotic and angiogenic proprieties, paracrine effects leading to type VII collagen production in the host tissues and/or type VII collagen secretion by injected BM-MSC. The aim of our work was to study the therapeutic potential of MSC for RDEB in preclinical models. We first showed that BM-MSC produce COL7A1 mRNA and type VII collagen levels comparable to healthy dermal fibroblasts in culture. We then assessed the long-term capacity of human BM-MSC to survive, produce and deposit type VII collagen at the dermal-epidermal junction (DEJ) after local injection in human RDEB skin equivalents transplanted onto nude mice. In vivo intradermal (ID) injection of a single dose of human BM-MSC led to the production and deposition of human type VII collagen at the DEJ and allowed anchoring fibrils formation for at least six months post-injection. Injected human BM-MSC were found in the skin at least four months post-injection. These data show that intradermally injected human BM-MSC have the potential to improve dermal-epidermal adhesion of RDEB skin equivalents through sustained deposit of type VII collagen molecules and subsequent anchoring fibrils formation. We then compared the efficacy of human Umbilical Cord Wharton's Jelly-MSC (UC-MSC) with BM-MSC using the same methodology as previously described. UC-MSC showed in vitro a significantly higher amount of COL7A1 mRNA and type VII collagen compared to BM-MSC and healthy dermal fibroblasts in culture. ID injection of a single dose of UC-MSC in vivo led to the production and deposition of low levels of human type VII collagen at the DEJ for four months post-injection. Injected human UC-MSC were found in the skin two months post-injection. These data disclosed a lower efficacy of UC-MSC to restore collagen VII at the DEJ compared to BM-MSC injected in the same xenograft RDEB model. These data open the perspective of using gene-corrected BM-MSC from a Col7a1-/- RDEB murine model to restore normal dermal-epidermal adhesion. Col7a1-/- mice reproduce cutaneous and mucosal lesions observed in RDEB patients. The life expectancy of these animals is very short. We could show that transduction of Col7a1-/- murine BM-MSC in culture using a COL7A1-expressing SIN retroviral vector led to type VII collagen expression levels which were 30-fold higher on average than in BM-MSC from WT mice. In vivo data are required to determine whether the injection of gene-corrected BM-MSC has the potential to treat skin and mucosal lesions in RDEB mice and to define the optimal dose and duration of the effect in vivo. Restoration of type VII collagen expression and anchoring fibrils formation in Col7a1-/- mice would represent an important step towards clinical translation

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Tsairidou, Smaragda. "Genetics of disease resistance : application to bovine tuberculosis." Thesis, University of Edinburgh, 2016. http://hdl.handle.net/1842/25397.

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Bovine Tuberculosis (bTB) is a disease of significant economic importance, being one of the most persistent animal health problems in the UK and the Republic of Ireland and increasingly constituting a public health concern especially for the developing world. Limitations of the currently available diagnostic and control methods, along with our incomplete understanding of bTB transmission, prevent successful eradication. This Thesis addresses the development of a complementary control strategy which will be based on animal genetics and will allow us to identify animals genetically predisposed to be more resistant to disease. Specifically, the aim of my PhD project is to investigate the genetic architecture of resistance to bTB and demonstrate the feasibility of whole genome prediction for the control of bTB in cattle. Genomic selection for disease resistance in livestock populations will assist with the reduction of the in herd-level incidence and the severity of potential outbreaks. The first objective was to explore the estimation of breeding values for bTB resistance in UK dairy cattle, and test these genomic predictions for situations when disease phenotypes are not available on selection candidates. Through using dense SNP chip data the results of Chapter 2 demonstrate that genomic selection for bTB resistance is feasible (h2 = 0.23(SE = 0.06)) and bTB resistance can be predicted using genetic markers with an estimate of prediction accuracy of r(g, ĝ) = 0.33 in this data. It was shown that genotypes help to predict disease state (AUC ≈ 0.58) and animals lacking bTB phenotypes can be selected based on their genotypes. In Chapter 3, a novel approach is presented to identify loci displaying heterozygote (dis)advantage associated with resistance to M. bovis, hypothesising underlying non-additive genetic variation, and these results are compared with those obtained from standard genome scans. A marker was identified suggesting an association between locus heterozygosity and increased susceptibility to bTB i.e. a heterozygote disadvantage, with the heterozygotes being significantly more in the cases than in the controls (x2 = 11.50, p < 0.001). Secondly, this thesis focused on conducting a meta-analysis on two dairy cattle populations with bTB phenotypes and SNP chip genotypes, identifying genomic regions underlying bTB resistance and testing genomic predictions by means of cross-validation. In Chapter 4, exploration of the genetic architecture of the trait revealed that bTB resistance is a moderately polygenic, complex trait with clusters of causal variants spread across a few major chromosomes collectively controlling the trait. A region was identified on chromosome 6, putatively associated with bTB resistance and this chromosome as a whole was shown to contribute a major proportion (hc 2= 0.051) of the observed variation in this dataset. Genomic prediction for bTB was shown to be feasible even when only distantly related populations are combined (r(g,ĝ)=0.33 (SE = 0.05)), with the chromosomal heritability results suggesting that the accuracy arises from the SNPs capturing linkage disequilibrium between markers and QTL, as well as additive relationships between animals (~80% of estimated genomic h2 is due to relatedness). To extend the analysis, in Chapter 5, high density genotypes were inferred by means of genotype imputation, anticipating that these analyses will allow the identification of genomic regions associated with bTB resistance more closely, and that would increase the prediction accuracy. Genotype imputation was successful, however, using all imputed genotypes added little information. The limiting factor was found to be the number of animals and the trait definitions rather than the density of genotypes. Thirdly, a quantitative genetic analysis of actual Single Intradermal Comparative Cervical Test (SICCT) values collected during bTB herd testing was conducted aiming to investigate if selection for bTB resistance is likely to have an impact on the SICCT diagnostic test. This analysis demonstrated that the SICCT has a negligibly low heritability (h2=0.0104 (SE = 0.0032)) and any effect on the responsiveness to the test is likely to be small. In conclusion, breeding for disease resistance in livestock is feasible and we can predict the risk of bTB in cattle using genomic information. Further, putative QTLs associated with bTB resistance were identified, and exploration of the genetic architecture of bTB resistance revealed a moderately polygenic trait. These results suggest that given that larger datasets with more phenotyped and genotyped animals will be available, we can breed for bTB resistance and implement the genomic selection technology in breeding programmes aiming to improve the disease status and overall health of the livestock population. Using the genomics this can be continued as the epidemic declines.

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Pereira, Hugo Miguel Lino. "Alergia à picada de Himenópteros em cães." Master's thesis, Universidade de Lisboa, Faculdade de Medicina Veterinária, 2016. http://hdl.handle.net/10400.5/11845.

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Dissertação de Mestrado Integrado em Medicina Veterinária Todos os anos, uma quantidade significativa de cães tem manifestações alérgicas após ser vítima de picada por himenópteros, não sendo conhecida a prevalência desta alergia em Portugal. Adicionalmente, em medicina veterinária, existem poucos estudos sobre os métodos de diagnóstico disponíveis para estudar a alergia aos himenópteros, incluindo a sua eficácia e segurança. Os testes intradérmicos (IDs) são, em medicina humana, meios de diagnóstico seguros de elevada sensibilidade, sendo os resultados apoiados por testes in vitro. Na primeira fase do trabalho procurou-se recolher informações sobre a casuística desta alergia em Portugal. A partir de inquéritos a centros veterinários do país, concluiu-se que a maioria dos casos ocorre na primavera-verão, que o agente envolvido na picada geralmente não é identificado, que os adultos jovens estão mais predispostos, sendo comummente picados na face e que as picadas acontecem maioritariamente em animais de exterior, sendo que, não parecem existir raças mais predispostas. Concluiu-se ainda que o angioedema é o sinal clínico de maior ocorrência, que o tratamento mais usado pelos clínicos são os glucocorticóides e que os donos não sabem como prevenir o problema. Neste estudo não se encontrou relação entre o número de casos de cães alérgicos picados e as zonas de maior densidade populacional de Apis melífera (p>0,05). A segunda fase deste trabalho incidiu na realização de testes IDs com três venenos específicos de himenópteros (Apis melífera, Vespula spp. e Polistes spp.) para avaliar a rentabilidade diagnóstica e a segurança. O protocolo utilizado foi baseado nos protocolos sequencial e simultâneo descritos em medicina humana e, tanto quanto sabemos, este protocolo foi usado pela primeira vez em cães. Os resultados apresentados revelam que os IDs podem ser realizados com segurança e que são eficazes no sentido de identificar hipersensibilidade à picada de himenópteros. O estudo revela também que, além do agente suspeito, alguns dos pacientes tendem a apresentar dupla positividade nos IDs, ficando por saber qual a influência das reações cruzadas nestes resultados. Os animais responderam positivamente a concentrações entre 0,01 a 1 μg/ml. O grupo controlo não apresentou qualquer tipo de reatividade a essas concentrações. Estudos futuros já planeados, com recurso a testes serológicos, poderão permitir estudar a existência de reatividade cruzada. Este estudo abre novas portas no diagnóstico de cães alérgicos a venenos de himenópteros, permitindo desenhar novos tratamentos, nomeadamente com recurso à imunoterapia específica. ABSTRACT - HYMENOPTERA STING ALLERGY IN DOGS - Although its prevalence is still not known in Portugal, every year dogs are stung by hymenoptera and show allergic reactions. To our knowledge, there are only a couple of studies regarding hymenoptera allergy and its diagnosis in dogs. In human patients, the use of intradermal tests is both safe and sensitive and results are sometimes supported by in vitro tests. In this work, firstly, we performed surveys in veterinary centres in order to gather as much information as possible about this allergy in Portugal. We have concluded that most hymenoptera stings happen in the spring/summer season; that usually the owners don’t recognise the insect which causes the allergy; that young adult dogs living outside are more often stung and that this occurs mainly on the face. It also seems there is no breed predisposition for this allergy. We also found that angioedema is the most reported clinical sign and that corticosteroids are often used to treat this allergy. Interestingly, owners are not given any instructions on how to prevent the following sting. The regions with higher density of Apis melífera in Portugal did not relate with an increase number of dogs stung (p>0,05). In the second part of this work, intradermal tests using three distinct venoms (Apis melífera, Vespula spp. and Polistes spp.) were performed to evaluate both the diagnostic yield and test safety. As far as we know, the protocol used in this study was adapted from human sequential and simultaneous protocols and it was, for the first time, used in dogs. The results revealed that intradermal testing is both efficient and safe to use in dogs and helpful in the diagnosis of hymenoptera stings in allergic animals. The study also reveals that although dogs did react to the main sting suspect, some of them also tend to be positive to other venoms. It’s yet to be evaluated, in dogs, what kind of role cross-reactivity plays in this type of tests. Dogs have positively reacted to 0,01 a 1 μg/ml of venom concentration intervals. The control group hasn’t shown any reaction to these concentrations. Further investigations in dogs with serological testing will allow studying the presence and nature of cross-reactivity. It is hoped that our research in hymenoptera allergy in dogs will improve the understanding of the disease and allows developments in both diagnosis and management options, particularly the use of specific venom immunotherapy as prevention.

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Beffort, Lisa [Verfasser], and Mathias [Akademischer Betreuer] Ritzmann. "Evaluation der Verträglichkeit und Wirksamkeit der nadellos, intradermal applizierten Vakzine Porcilis M Hyo ID ONCE gegen Mycoplasma hyopneumoniae unter Feldbedingungen / Lisa Beffort. Betreuer: Mathias Ritzmann." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2015. http://d-nb.info/1068767189/34.

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Cunha, Victor do Espirito Santo. "Avalia??o da sensibilidade de c?es com dermatite al?rgica aos extratos alerg?nicos padronizados de ?caros da poeira domiciliar." Universidade Federal Rural do Rio de Janeiro, 2006. https://tede.ufrrj.br/jspui/handle/tede/741.

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Made available in DSpace on 2016-04-28T20:15:24Z (GMT). No. of bitstreams: 1 2006- Victor do Espirito Santo Cunha.pdf: 1132781 bytes, checksum: 5edd69d626ede6488b658d61c07e111b (MD5) Previous issue date: 2006-02-22 Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico The objective of this case-control study was to evaluate whether allergenic extracts from five species of house dust mites standardized for humans may be taken into account in the diagnosis of the canine atopic dermatitis. Extracts of Dermatophagoides pteronyssinus, D. farinae, Blomia tropicalis, Lepidoglyphus destructor and Tyrophagus putrescentiae have been evaluated through intradermal testing on 45 dogs, from which 20 belonged to the control group and 25 suffered from allergic dermatitis. There was a significant difference on the response pattern between the two groups (p<0,05). Only one dog (5%) from the control group has reacted to the intradermal test, whereas from the allergic group, 14 dogs (56%) have presented at least one positive reaction (odds ratio = 24,2). Most of the positive reactions observed in the allergic group were to the extracts of T. putrescentiae or L. destructor, each one inducing reactions on ten dogs (40%). The D. farinae, D. pteronyssinus e B. tropicalis extracts were responsible for positive reactions on 7 (28%), 3 (12%) and 3 (12%) dogs, respectively. The intradermal testing sensitivity and specificity were 56% and 95%, respectively, and the positive predictive value and the negative predictive value were 93% and 63%, respectively. O presente estudo do tipo caso-controle teve como objetivo avaliar se extratos alerg?nicos de cinco esp?cies de ?caros da poeira domiciliar padronizados para humanos podem ser utilizados no diagn?stico da dermatite at?pica canina. Extratos de Dermatophagoides pteronyssinus, D. farinae, Blomia tropicalis, Lepidoglyphus destructor e Tyrophagus putrescentiae foram avaliados atrav?s de testes intrad?rmicos em 45 c?es, dos quais 20 controles e 25 com dermatite al?rgica. Uma diferen?a significativa foi observada no padr?o de respostas entre os dois grupos (p<0,05). Apenas um animal (5%) do grupo controle reagiu ao teste cut?neo, enquanto que no grupo dos al?rgicos 14 c?es (56%) apresentaram pelo menos uma rea??o positiva (odds ratio = 24,2). As maiores freq??ncias de rea??es positivas observadas no grupo dos al?rgicos foram aos extratos de T. putrescentiae ou L. destructor, cada um induzindo rea??es em 10 (40%) c?es. Os extratos de D. farinae, D. pteronyssinus e B. tropicalis foram respons?veis por rea??es positivas em 7 (28%), 3 (12%) e 3 (12%) c?es, respectivamente. A sensibilidade e a especificidade dos testes intrad?rmicos foram de 56% e 95%, respectivamente e, o valor preditivo positivo e valor preditivo negativo de 93% e 63%, respectivamente. .

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Ferreira, Rafael Rodrigues. "Avaliação de diferentes concentrações de histamina e extratos alergênicos em cães sadios submetidos a teste intradérmico." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2013. http://hdl.handle.net/10183/81225.

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Teste intradérmico avalia reação de hipersensibilidade a diversos agentes que possam apresentar poder reacional alérgico e são comumente utilizados para complementar o diagnóstico da dermatite atópica canina (DAC). Ainda não existe consenso sobre as concentrações de histamina e extratos alergênicos a serem utilizadas. Para determinar a concentração ideal de histamina, como controle positivo, e do limiar irritativo de extratos alergênicos em teste intradérmico é necessário que diversas concentrações sejam avaliadas em uma população bem numerosa de cães hígidos. O objetivo desta pesquisa foi avaliar em 160 cães sadios submetidos a teste intradérmico, quais seriam as concentrações de histamina e de extratos alergênicos consideradas ideais. A solução contendo 0,1 mg/mL de histamina foi considerada como parâmetro ideal, provocando reações cutâneas com diâmetro médio, mediana e desvio padrão, de 15,18 mm, 14,97 mm e 2,07 mm, respectivamente. A partir do estabelecimento da concentração de histamina, foram determinadas as concentrações ótimas dos extratos alergênicos, expressas em PNU/mL: 1.000 para Dermatophagoides pteronyssinus, 500 para D. farinae, 125 para Blomia tropicalis e 2.000 para Malassezia pachydermatis. Futuros estudos devem ser conduzidos em cães atópicos para verificação da acurácia dos testes intradérmicos realizados com essas concentrações. Intradermal testing evaluates hipersensitivity reaction to different agents that can present allergic reactivity power. It is commonly used to complement canine atopic dermatitis diagnosis. There is still no consensus about histamine concentrations and allergen extracts to be used. The determination of the histamine ideal concentration as positive control and the irritant threshold of allergen extracts for intradermal testing, requires evaluation of different concentrations on a large population of healthy dogs. The purpose of this research was to evaluate the ideal histamine and allergen extracts concentrations on 160 healthy dogs submitted to intradermal testing. A histamine solution 0,1 mg/mL was considered the ideal parameter. It caused cutaneous reactions with average diameter, median measure and standard deviation of 15.18 mm, 14.97 mm and 2.07 mm, respectively. From the histamine concentration establishment, the optimum allergen extracts concentrations were determined, expressed by PNU/mL: 1.000 for Dermatophagoides pteronyssinus, 500 for D. farinae, 125 for Blomia tropicalis and 2.000 for Malassezia pachydermatis. Future studies have to be conducted on atopic dogs to verify the accuracy of the intradermal testing with these concentrations.

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Bulcão, Rachel Picada. "Avaliação toxicológica in vivo de nanocápsulas poliméricas biodegradáveis." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2013. http://hdl.handle.net/10183/143464.

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Nanopartículas poliméricas biodegradáveis têm recebido atenção como carreadores de fármacos ao longo dos últimos anos. Em muitos casos, a segurança destes nanocarreadores não foi demonstrada e pouco se sabe sobre a relação entre as suas características físico-químicas e suas propriedades toxicocinéticas e toxicodinâmicas. A nanotoxicologia está emergindo como uma especialidade importante da nanotecnologia e/ou toxicologia, e refere-se ao estudo da interação de nanoestruturas com sistemas biológicos. Nos últimos anos, a maioria das pesquisas foi centrada em estudos in vitro, entretanto, os resultados destes estudos necessitam também ser avaliados em experimentos in vivo para o avanço na utilização de nanocarreadores na área biomédica. Com isso, o objetivo deste trabalho foi avaliar a toxicidade de nanocápsulas de núcleo lipídico (LNC), de poli(-caprolactona), após administração intraperitoneal (i.p.) e intradérmica (i.d.) em ratos Wistar. Para a avaliação toxicológica aguda, foi administrada dose única em que se observaram sinais clínicos e fisiológicos, em ambas as vias. Após 14 dias, os animais foram eutanasiados e análises macroscópicas e histopatológicas foram realizadas. Além disso, sangue e urina foram coletados para análises laboratoriais e avaliação de funções teciduais. A avaliação toxicológica subcrônica foi procedida da mesma forma, exceto pela administração de doses repetidas diárias durante 28 dias. As suspensões de nanocápsulas foram preparadas pelo método de precipitação do polímero pré-formado, as quais apresentaram tamanho médio de partícula inferior a 250 nm, índice de polidispersão (IPD) < 1, potencial zeta negativo e pH em torno de 6,7. Os animais tratados pela via i.p. (n=6/grupo) receberam para avaliação da toxicidade aguda: solução salina ou polissorbato 80 (PS80) (12 ml/kg), utilizados como controles e diferentes doses de LNC (18,03, 36,06, e 72,12 × 1012 LNC/kg); no teste de toxicidade subcrônica foram utilizados os mesmos controles porém com doses de 3mL/kg e 6,01, 12,02 ou 18,03 × 1012 LNC/kg. Nos testes de toxicidade aguda, nos animais administrados pela via i.p., foi observada diminuição significativa de peso nos grupos tratados com LNC mesmo após 14 dias da administração (p<0,05). Entretanto no teste subcrônico esta alteração foi transitória, e ocorreu apenas no grupo que recebeu a maior dose até o quinto dia de administração (p<0,05). Houve aumento no peso relativo do baço nos animais que receberam a dose mais alta de LNC (p<0,05) no tratamento agudo. A análise histopatológica em ambos os tratamentos, demonstrou a presença de um granuloma de tipo corpo estranho no fígado e no baço dos animais que receberam a dose mais alta, provavelmente devido ao volume de LNC administrado. Não houve alteração nas análises bioquímicas de dano hepático, renal, dentre outros em todos os grupos tratados. Os dados hematológicos apresentaram uma leve alteração, entretanto foi demonstrada interferência metodológica, evidenciada por testes preliminares in vitro. Além disso, foram avaliados biomarcadores do estresse oxidativo (EO), marcadores inflamatórios e de genotoxicidade. Os resultados dos biomarcadores de oxidação de proteínas e lipídios não foram suficientes para iniciar um processo oxidativo, visto que não houve peroxidação lipídica. Ainda, não houve depleção de antioxidantes, dano ao DNA ou alteração nos marcadores inflamatórios. Nos ratos tratados pela via i.d., foi utilizada solução salina 1,2 ml/kg como grupo controle e uma dose de 7,2 × 1012 LNC/kg de LNC, para um estudo preliminar agudo e solução salina ou PS 80 (0,9ml/kg) e três doses de LNC (1,8, 3,6 ou 5,4 × 1012 LNC/kg) para avaliação da toxicidade subcrônica. No teste de toxicidade aguda, não houve alteração do peso corpóreo, entretanto no teste de toxicidade subcrônica houve uma diminuição reversível do peso no grupo que recebeu PS80 (p<0,05). Os dados histopatológicos não apresentaram alteração. Não houve alteração nos parâmetros bioquímicos, exceto uma leve diminuição da atividade da butirilcolinesterase no grupo que recebeu a dose mais alta (p<0,05). Por outro lado, houve aumento nos leucócitos no grupo que recebeu LNC, no teste de toxicidade aguda e nos grupos que receberam PS 80 e 5,4 × 1012 LNC/kg (p<0,05) após doses repetidas. Em relação à avaliação sanguínea e tecidual dos biomarcadores do EO e dos marcadores inflamatórios, foi observada uma indução nos marcadores de oxidação de proteínas juntamente com uma indução enzimática nos ratos que receberam a dose mais alta, além de uma diminuição dos níveis do IL-10 nos grupos que receberam PS80 e a dose mais alta (p<0.05). Pode-se concluir que nas condições dos experimentos, tanto pela via i.p. quanto pela via i.d., não foram demonstrados danos teciduais, pois os achados laboratoriais foram condizentes com os achados histopatológicos. Além disso, os mecanismos de reparo foram suficientes para contrabalançar eventuais danos oxidativos ou inflamatórios. Assim, o presente trabalho contribui para futuras avaliações toxicológicas de nanocápsulas poliméricas, visto que foram realizadas avaliações agudas e subcrônicas sistemáticas, com marcadores de dano renal precoce e possíveis mecanismos de toxicidade envolvidos após administração por ambas as vias. O aumento na utilização destas nanocápsulas e as lacunas nas informações toxicológicas fazem com que desafios importantes devam ser superados para permitir sua incorporação segura. Com isso, estudos nesta linha podem embasar a avaliação da resposta tóxica e, consequentemente, levar ao estabelecimento de regulamentações para avaliação da toxicidade da maioria das nanopartículas poliméricas biodegradáveis utilizadas como carreadoras de fármacos. Biodegradable polymeric nanoparticles have received attention as drug carriers over the past years. In many cases, the safety of nanocarriers has not been demonstrated and little is known about the relationship of its physicochemical characteristics and their toxicokinetic and toxicodynamic properties. Nanotoxicology is emerging as an important field of nanotechnology and toxicology, and refers to the study of the interaction of nanostructures with biological systems. In recent years, most research has focused on in vitro studies; however, the results of these studies should also be evaluated trough in vivo experiments, in order to advance in biomedical application of nanocarriers. Thus, the objective of this study was to evaluate the toxicity of lipid-core nanocapsules (LNC), prepared with poly(ɛ-caprolactone), after intraperitoneal (i.p.) and intradermal (i.d.) administration in rats. For acute toxicological evaluation, it was administered a single dose, i.p. and i.d., clinical signs and physiological effects were observed. After 14 days, animals were euthanized and macroscopic and histopathological analyses were done. In addition, blood and urine were collected for laboratory analysis and evaluation of tissue functions. Subchronic toxicological evaluation was similar, except for the administration of repeated doses for 28 days. The suspension of nanocapsules were prepared by interfacial deposition of polymer, which had particle size less than 250 nm, polydispersity index (IPD) <1, negative zeta potential and pH around 6.7. Animals were treated via i.p. (N = 6/group), the doses used for acute toxicity test were: saline or polysorbate 80 (PS80) (12 ml/kg) as controls or three different doses of LNC (18.03, 36.06, e 72.12 × 1012 LNC/kg); for subchronic toxicity test, same controls were used but the doses were 3 ml/kg and 6.01, 12.02 ou 18.03 × 1012 LNC/kg administered daily for 28 days. In acute toxicity test, with i.p. administration, groups treated with LNC presented a significant reduction in relative weight even after 14 days of administration (p<0.05); however in the subchronic test, this change was transient, and occurred only in the group receiving the highest dose until the fifth day of administration (p<0.05). There was an increase in relative weight of spleen in animals that received the highest dose of LNC (p<0.05) in acute treatment. Histopathological analysis in both the treatments, showed a granulomatous foreign body reaction in liver and spleen of animals receiving the highest dose, probably because the volume of LNC administered. There were no changes in biochemical parameters of liver or kidney damage, among all treated groups. Hematological data showed a slight change; however it was demonstrated an interference of the methodology, further evidenced by preliminary in vitro tests. Furthermore, we evaluated biomarkers of oxidative stress (OS), inflammatory and genotoxicity markers. The results of the oxidation of proteins and lipids biomarker were not sufficient to initiate an oxidative process, since no lipid peroxidation occurred. Still, no depletion of antioxidants, DNA damage or change in inflammatory markers was observed. In rats treated via i.d., saline was used as control (1.2 ml/kg) and a dose of 7.2 × 1012 LNC/kg of LNC to a preliminary acute study, and saline or PS 80 (0.9ml/kg) used as controls or three doses of LNC (1.8, 3.6 ou 5.4 × 1012 LNC/kg) for subchronic toxicity evaluation. In acute toxicity test, there was no change in relative body weight, however, a decreased was found for the group receiving PS 80 in subchronic test (p <0.05). No histopathological alteration was found. Also, there was no change in biochemical parameters, except a slight decrease of butyrylcholinesterase activity in the group receiving the highest dose (p<0.05). Moreover, in acute toxicity test, it was found an increase in white blood cells in group receiving LNC; these increasing also occurred after repeate dose test, in PS 80 and 5.4 × 1012 LNC/kg of LNC groups (p <0.05). Regarding blood and tissue biomarkers of OS and inflammatory markers, an induction in protein oxidation marker along with antioxidant induction in rats which received the highest dose were observed, also reduced levels of IL-10 in rats that received the higher dose and PS80 (p <0.05). It can be concluded that, under the experimental conditions, for i.p. and i.d. administration, tissue damage was not found, since laboratorial analysis results were consistent with histopathological findings. Furthermore, mechanisms of repair were sufficient to offset oxidative damage or inflammation.Thus, this study contributes to future toxicological evaluations of polymeric nanocapsules, since a systematic acute and subchronic evaluation with early renal damage markers and possible mechanisms of toxicity involved after ip and id routes were performed. The increase in the use of these nanocapsules and the gaps in toxicological information make important to overcome these challenges in order to allow its safe incorporation. Thus, studies in this line are important to evaluate toxic response, and lead to establishing rules for evaluating the toxicity of most biodegradable polymer nanoparticles used as carrier of drugs.

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Lee, Nigel. "Comparison of a single versus a four intradermal sterile water injection technique for the relief of lower back pain for women in labour: A mixed methods study." Thesis, Australian Catholic University, 2013. https://acuresearchbank.acu.edu.au/download/fb8991208095920185a907088b92dba49532b490c2fcc90be0531a679c1ed24c/5032178/LEE_NIGEL_2013.pdf.

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Significant back pain is often experienced by women in labour and may increase the need for pharmacological pain relief, often with associated side effects including excessive sedation and restriction to mobility. Sterile water injections (SWIs) are a simple, safe, effective, non-pharmacological technique for relieving back pain in labour; however, the number of injections required to achieve optimal analgesia is unknown. Furthermore, administration of SWI causes a brief, but intense, pain which may influence the acceptability of the procedure to labouring women. There is limited data from previous trials on how women view SWI, and the benefits of this particular analgesic option, versus the pain associated with administration. No previous studies have examined how midwives regard the prospect of causing pain to labouring women; albeit to relieve pain. The aim of this research was to determine if a single injection of sterile water was clinically similar to four injections in terms of degree of analgesia and to examine the experiences of labouring women and midwives receiving, and administering, SWIs...

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Matias, Daniela Ferreira. "Avaliação de uma nova técnica em alergologia veterinária : testes cutâneos por picada em cães." Master's thesis, Universidade Técnica de Lisboa. Faculdade de Medicina Veterinária, 2013. http://hdl.handle.net/10400.5/5285.

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Dissertação de Mestrado Integrado em Medicina Veterinária A dermatite atópica (DA), uma das doenças alérgicas mais comuns no cão, é uma doença pruriginosa e inflamatória resultante da interacção de factores genéticos, ambientais e imunológicos. Dada a incidência crescente e o impacto na qualidade de vida dos pacientes, o seu diagnóstico deve ser uma prioridade. Este deve ser baseado na história e sinais clínicos dermatológicos, aos quais se seguem a exclusão dos diagnósticos diferenciais, como é o caso da sarna sarcóptica. O tratamento é, sempre que possível, etiológico, além de sintomático. A selecção dos alergénios relevantes para cada paciente deve ser fundamentada em provas de sensibilização. Estas incluem testes cutâneos e/ou serológicos, sendo o teste intradérmico (TID) normalmente eleito. Em alergologia humana, este foi, praticamente, substituído pelo teste cutâneo por picada (TCP) que, para além de mais específico, permite uma execução mais simples, rápida, segura, económica e confortável. O principal objectivo deste estudo é averiguar se estas vantagens também se verificam nos cães. Numa primeira fase, 10 cães não atópicos foram testados pela técnica de picada com 21 extractos alergénicos de uso em alergologia humana (ALK-Abelló®). Concluiu-se que as concentrações utilizadas, não sendo responsáveis por reacções irritantes nestes cães, podem ser aplicadas naqueles com DA. Antes dos testes, os mesmos animais foram, aleatoriamente, divididos em dois grupos: sedados e não sedados. Nestes últimos, durante e após os TCP, foi adaptada a forma abreviada da escala de dor de Glasgow, confirmando- -se que a técnica é pouco dolorosa. Em ambos os grupos, as concentrações séricas de cortisol, antes e após os TCP, ficaram dentro dos valores normais de referência, pelo que a validade da técnica não parece ser comprometida por activação do eixo hipotálamo- -pituitária-adrenal. Numa segunda fase, 13 cães atópicos foram sujeitos a TID com 28 extractos alergénicos (Greer Laboratories®) e, na mesma sedação, a TCP com os extractos da fase anterior. Todos os cães com DA desenvolveram reacções positivas nos TCP. Os TCP parecem ser seguros e de fácil execução mesmo em cães não sedados. Os níveis de cortisol permaneceram dentro dos limites normais, antes e após os TCP, comprovando indirectamente que a técnica é pouco dolorosa. Os extractos da ALK-Abelló® não provocam reacções irritantes no cão. A elevada especificidade desta técnica relativamente aos TID poderá vir a ser um dos seus grandes benefícios. Estudos adicionais deverão ser desenvolvidos de forma a encontrar as concentrações óptimas para a introdução dos TCP em alergologia veterinária, uma vez que as usadas podem ser demasiado baixas. No entanto, os resultados são bastante promissores. ABSTRACT - EVALUATION OF A NEW TECHNIQUE IN VETERINARY ALLERGOLOGY: SKIN PRICK TESTS IN DOGS - Atopic dermatitis, one of the most common allergic conditions in the dog, is a pruritic and inflammatory disease resulting from the interaction of genetic, environmental and immunological factors. Of increasing incidence and with great impact on the patient quality of life, its recognition and treatment are of extreme importance. Diagnosis is based on compatible history and clinical signs and exclusion of other pruritic diseases, such as scabies. Whenever possible, etiological treatment should be used in addition to symptomatic. This implies knowledge of patient individual sensitizations, which can be achieved by skin or serological tests. In veterinary medicine, the intradermal test (ITD) is usually used. In human allergology, it was largely replaced by skin prick test (SPT) because of its higher specificity and pratical reasons such as being easier to perform, safer, quicker and less expensive. Also, patients report less discomfort. In this study we want to perform this technique and see if advantages reported for Man also occur in the dog. Initially (stage one), 10 non-atopic dogs were tested by SPT with 21 allergenic extracts for use in human allergology (ALK-Abelló®). We concluded that the concentrations used are not responsible for irritants false positive reactions in these dogs. Therefore, it can be applied in atopic dogs. Before testing, the same animals were randomly divided into two groups: sedated and non-sedated dogs. Adapted form of the Glasgow composite pain scale was apllied to non-sedated-group during and after the SPT. This technique was shown to be well tolerated. In both groups, serum cortisol levels were within the normal reference values before and after the SPT; thus, the validity of the technique does not appear to be compromised by activation of the hypothalamic-pituitary-adrenal axis. In a second stage, we perform IDT with 28 allergenic extracts (Greer Laboratories®) and SPT with the extracts from the preceding stage in 13 sedated atopic dogs. All dogs with atopic dermatitis had some positive reactions in SPT. SPT appear to be safe and easy to perform even in non-sedated dogs. Cortisol levels remained within normal limits before and after the procedure, indirectly implying no significant pain or discomfort was experienced. ALK-Abelló® human extracts didn’t cause irritant reactions. Higher specificity of SPT comparing to IDT could be in the future one of the great benefits of this technique. Further studies are needed, especially regarding optimal concentrations of the extracts to be used in veterinary, before SPT could be proposed for routine use in veterinary allergology. Results are nevertheless very promising.

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Kelly, Robert Francis. "Epidemiology of bovine tuberculosis and influence of liver fluke co-infection in Cameroon, Central Africa." Thesis, University of Edinburgh, 2017. http://hdl.handle.net/1842/29560.

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Despite Africa accounting for ~20% of the global cattle population, prevalence estimates and related risk factors of bovine tuberculosis (bTB), caused by Mycobacterium bovis, are still poorly quantified in many countries across the continent. Control of bTB in Africa is difficult due to poor monitoring of cattle movements and limited abattoir surveillance. Also M. bovis is zoonotic and risk factors for transmission include living in close contact with cattle and consumption of unpasteurised milk. Cattle keeping is integral to some rural populations in Cameroon and understanding the epidemiology of bTB in cattle populations is important both to bovine and public health. Detection of bTB in cattle is difficult due to variability of immune responses to M. bovis infection. The interferon-γ (IFN-γ) assay maybe useful to estimate bTB prevalence and identify bTB risk factors in Cameroon. However its performance can vary at different stages of bTB pathogenesis and in different cattle populations. Recently Fasciola hepatica co-infections have been reported to suppress IFN-γ responses in M. bovis infected cattle but the potential effect with F. gigantica co-infections on bTB prevalence estimates in Cameroon is unknown. An abattoir study was conducted in Cameroon to assess the performance of the IFN-γ assay. In 2012-13; 2064 slaughtered cattle were sampled from Bamenda abattoir (North West Region; NWR) and Ngaoundere abattoir (Vina Division; VD). Individual animal data was collected from routine meat inspection including identification of bTB and Fasciola pathology. Cattle were also tested for bTB using the IFN-γ assay and an M. bovis antibody ELISA. In the absence of a gold-standard diagnostic, the IFN-γ assay was compared to other diagnostic tests to assess agreement and identify factors that affected performance of the assay. Agreement between IFN-γ assay, TB lesion identification and an M. bovis antibody ELISA was poor-moderate, probably partly related to differences in immune response detected. A presence of Fasciola gigantica also increased the odds of false negative IFN-γ assay results. On further investigation co-infected cattle had increased odds of TB lesions and reduced IFN-γ responses that potentially could lead to ~20% reduction in test sensitivity. In an attempt to take into account the potential impact of F. gigantica, when estimating bTB prevalence, an antibody ELISA was developed to detect the exposure in live cattle. To highlight the awareness of disease in cattle-rearing communities, estimate prevalence and identify risk factors of bTB in cattle populations; two cross-sectional studies were conducted in 2013. A stratified clustered cross-sectional study of pastoral cattle herds, in the NWR and the VD, sampled 1448 pastoral cattle reared by 100 pastoralists. A smaller cross-sectional study sampled 60 dairy cattle from 46 small-holder co-operative dairy farmers. Individual animal data and herd-level data were collected and animals were screened by both the single comparative intradermal skin test (SCITT) and IFN-γ assay. Awareness of zoonotic TB was low yet consumption of raw milk was high in cattle-keeping communities highlighting the need for accurate bTB prevalence estimates. Despite the high awareness of the clinical presentation of bTB, clinical signs identified by pastoral herdsmen were not associated with cattle being bTB positive. The SCITT was used to compare two manufacturers cut offs for the IFN-γ assay, ≥0.05 and ≥0.1, and highlighted that these two diagnostics may detect different populations of bTB positive cattle. Using the IFN-γ assay at ≥0.1, bTB prevalence was highest in dairy cattle (21.67%) and was also present in pastoral cattle in the NWR and VD (11.33% and 6.55% respectively). Importantly, as F. gigantica is endemic in Cameroon and its influence could mean the true prevalence of bTB could be higher. Female pastoral cattle were at lower odds of being IFN-γ assay positive potentially due to immunosuppressive factors had lower odds of disease. Husbandry practices also decreased the odds of being IFN-γ assay positive such as drinking from streams, antelope and contact with herds at grazing. Age increased the odds of pastoral cattle being IFN- assay positive potentially being a confounder to chronicity of bTB and other co-infections may influence IFN-γ responses. Dairy cattle herds had different risk factors for being IFN- positive likely due to differences in husbandry practices. Considering the potential risk to public health of M. bovis this thesis highlights the extent of bTB across two major cattle keeping regions in Cameroon and the public health risk in cattle-rearing communities. Furthermore the relationship between Fasciola co-infection and IFN- responses to M. bovis described has potential implications for bTB diagnosis in cattle populations where the parasite is present across the globe.

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Archibald, Timothy, Stacy Brown, and Alexei Gonzalez-Estrada. "Refrigerated Stability of Diluted Succinylcholine, Pancuronium, and Atracurium." Digital Commons @ East Tennessee State University, 2018. https://dc.etsu.edu/asrf/2018/schedule/88.

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Refrigerated Stability of Diluted Succinylcholine, Pancuronium, and Atracurium. T. Archibald1, S. Brown1, A. Gonzalez-Estrada2 1College of Pharmaceutical Sciences, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN2Quillen College of Medicine, Allergy and Clinical Immunology, East Tennessee State University, Johnson City, TN The purpose of this study is to investigate the stored stability of dilutions of neuromuscular blocking agents (NMBAs), namely succinylcholine, pancuronium, and atracurium, for skin prick/intradermal testing. Concentrations of NMBAs were monitored by liquid chromatography-mass spectrometry (LC-MS/MS) for a period of 14 days. Dilutions of NMBAs were prepared in saline by factors of 10x, 100x, 1,000x, 10,000x, and 100,000x as sensitivity of the assay allowed. Each drug was prepared with an n = 5 for each dilution, using a different newly opened product for each series. Diluted drug products were stored in a laboratory refrigerator until sampling. On sampling days (day 0, 1, 2, 4, 7, and 14), one milliliter aliquots of each dilution were removed, filtered, and analyzed against freshly prepared set of reference dilutions. The results are expressed as beyond use date (BUD), defined as recovery of drug versus the reference (90-110%). Based on the LC-MS/MS data, the BUD for succinylcholine diluted by 10x and 100x is 48 and 24 hours, respectively. The1000x dilution is also stable for 24 hours.Higher dilutions of succinylcholine (10,000x to100,000x) should be used immediately following preparation (within less than 24 hours), as the potency of these dilutions had decreased below 90% at the 24 hr sampling. .Pancuronium diluted by 10x and 100x, had a BUD of 48 hours, and the1,000x dilution was stable for 24 hours.As with the succinylcholine, the 10,000x and 100,000x dilutions expressed potency of <90% at 24 hours. .Atracurium diluted to 10x had a BUD of 96 hours, the100x dilution is stable for 24 hours yet higher dilutions (1,000x to 10,000x) do not persist beyond 24 hours. . The 100,000x dilution of atracurium was unknown, given than the signal intensity was too weak to monitor by our LC-MS/MS method. With increasing dilution factors, the stability of these drugs in saline decreases, associated with an increasing deviation between samples and freshly prepared references. The most stable dilutions for each of the drugs tested were 10x and 100x. Stability of these drugs is likely compromised by hydrolysis of the ester bonds in the drug molecules.

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Weiblen, Carla. "Soroprevalência da pitiose equina no rio grande do sul, diagnóstico e controle da pitiose em modelo experimental." Universidade Federal de Santa Maria, 2015. http://repositorio.ufsm.br/handle/1/10237.

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Pythiosis is a granulomatous disease of humans and animals caused by an aquatic oomycete named Pythium insidiosum. In Brazil, equine species is the most affected and the highest occurrence of pythiosis is observed in Brazilian Pantanal. Nowadays, in Rio Grande do Sul (RS), in South of Brazil, there has seen an increase in the number of equine pythiosis. Nevertheless, there is no information about seroprevalence in Brazil. Early diagnosis is the key to success in controlling this infection. Currently, immunotherapy with proteic antigens has been studied as a promising alternative of treatment. The intradermal (ID) route to apply proteic antigen is an option to be evaluated, for both early diagnosis and pythiosis treatment, by using proteic antigens of P. insidioum. In this study it was investigated the seroprevalence of equine pythiosis in the RS State and also it was evaluated and compared the use of proteic antigens of P. insidiosum by the ID route in the diagnosis and control of pythiosis in an experimental model (Oryctolagus cuniculus). To determine the prevalence of equine pythiosis in RS it was used 1002 equine serum samples evaluated by indirect ELISA test. Of the total samples tested, 11.1% (111/1002) were seropositive for equine pythiosis. For the diagnosis it was applied 0.1mL (1.7mg) of proteic antigen by ID route in rabbits (O.cuniculus) (n = 15) divided into three groups: negative control (n = 5), with experimental pythiosis (n = 5) and animals previously immunized with the proteic antigen (n = 5). In order to verify and compare the use of proteic antigen of P. insidiosum by ID as an option of treatment it was used ten rabbits with pythiosis. Five rabbits with pythiosis were treated with 0.1mL proteic antigen applied through ID and other five rabbits by subcutaneous (SC) route with 2mL (34mg) (n = 5). The ID test was able to detect cutaneous reactions in 24h and 72h in all animals exposed to P. insidiosum, as well as it demonstrated sensitivity and specificity comparable with the pattern of diagnosis of pythiosis (indirect ELISA). The treatment proteic antigen of P. insidiosum, by ID and SC route, did not observed difference (P> 0.05) in the size of the lesions. However, the clinical cure affect as 40% of rabbits in both groups. Thus, protein antigen of P. insidiosum by ID may be used in therapeutic protocols of pythiosis in rabbits, also this route as a protocol of pythiosis has other advantages such as lower protein concentration and volume of antigen to control pythiosis. However, research is needed for evaluation of ID route for diagnosis and control of pythiosis in naturally affected species. A pitiose é uma doença granulomatosa de humanos e animais, causada pelo oomiceto aquático Pythium insidiosum. A maior ocorrência da enfermidade no Brasil é na espécie equina, especialmente no Pantanal Mato-Grossense, porém no Rio Grande do Sul (RS) observa-se um aumento dos casos da doença. Todavia, não há dados de soroprevalência da pitiose no Brasil. O diagnóstico precoce da enfermidade é fundamental para sucesso no controle da pitiose. Atualmente, a imunoterapia com antígenos proteicos vem sendo estudada como uma alternativa promissora de tratamento. A via intradérmica (ID) é uma opção a ser avaliada tanto para um diagnóstico precoce quanto para o tratamento da pitiose empregando antígenos proteicos de P. insidiosum. Neste estudo investigou-se: a soroprevalência da pitiose equina no estado do RS, bem como avaliou-se e comparou-se o uso de antígenos proteicos de P. insidiosum pela via ID no diagnóstico e controle da pitiose em modelo experimental. Para determinar a soroprevalência da pitiose equina no RS foram utilizadas 1002 amostras de soro equino avaliadas pelo método de ELISA indireto. Do total das amostras testadas, 11,07% (111/1002) foram soropositivas para a pitiose equina. Para o diagnóstico aplicou-se 0,1 mL (1,7mg) de antígeno proteico pela via ID, em coelhos (Oryctolagus cuniculus) (n=15) separados em três grupos: controle negativo (n=5), com pitiose experimental (n=5) e animais previamente imunizados com antígeno proteico (n=5). A fim de verificar e comparar a aplicação do antígeno proteico pela via ID como opção de tratamento utilizaram-se dez coelhos com pitiose. Cinco animais foram tratados com 0,1mL de antígeno proteico aplicado via ID e outros cinco coelhos foram tratados pela via subcutânea (SC) com 2mL (34mg) do antígeno. O teste ID foi capaz de detectar reações cutâneas em 24h e 72h nos animais previamente expostos ao agente, demonstrando sensibilidade e especificidade comparáveis ao teste padrão de diagnóstico da pitiose (ELISA indireto). No tratamento com antígeno proteico pelas vias ID e SC, embora não tenha sido observada diferença (P>0,05) no tamanho das lesões, a cura clínica em 40% dos coelhos de ambos os grupos demonstrou que a via ID pode ser utilizada em protocolos terapêuticos da pitiose em modelo experimental, pois também apresenta outras vantagens como menor concentração e volume de antígeno proteico para o controle da pitiose. No entanto, são necessárias pesquisas para o emprego da via ID no diagnóstico e controle da pitiose nas espécies naturalmente acometidas.

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Jegan, Vanessa. "Avaliação do teste intradérmico em equinos com extratos alergênicos de pólens, insetos e três concentrações de histamina." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2017. http://hdl.handle.net/10183/172630.

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O teste alérgico intradérmico (TID) é uma importante ferramenta no diagnóstico dos alérgenos implicados nas reações de hipersensibilidade mediada por IgE. O objetivo deste estudo foi avaliar o TID em eqüinos não alérgicos com extratos alergênicos de Cynodon dactylon, Lolium multiflorum, Paspalum notatum, Culex sp. e Aedes aegipty, e três concentrações de histamina, em dois volumes diferentes, e baseado nos resultados, verificar a acurácia do TID em eqüinos alérgicos. Foram realizados TID em 17 eqüinos não alérgicos com três concentrações de cada extrato (1:2000v/w, 1:4000v/w e 1:8000v/w) e três concentrações de histamina (0,1mg/ml, 0,05mg/ml e 0,025mg/ml), em dois volumes (0,1 e 0,05ml). Baseado nos resultados obtidos, foram realizados TID em seis eqüinos alérgicos utilizando os extratos na concentração de 1:2000v/w, a histamina na concentração de 0,025mg/ml, em um volume de 0,05ml. Nos eqüinos não alérgicos, nenhuma das três concentrações dos extratos testados nos dois volumes mostrou-se irritativa. Concentrações maiores de histamina provocaram halos maiores e levemente mais túrgidos, e quanto maior o volume injetado, maiores as reações Os TID realizados em eqüinos alérgicos mostraram confiabilidade dos resultados por evidenciar hipersensibilidade individual e não provocar reações irritativas. Em conclusão, as diferentes concentrações dos estratos testados em eqüinos não alérgicos não provocaram reações falso positivas (irritantes). O volume de 0,05ml é mais recomendado para realização dos TID pois as aplicações com 0,1ml produziram reações maiores, prejudicando a sensibilidade do teste. A concentração de 0,025mg/ml da solução de histamina provocou a formação de halos menores, permitindo maior acurácia do cut off no TID. Os extratos alergênicos testados em equinos alérgicos não provoraram reações em todos os animais, o que poderia ser considerado uma reação irritativa. Os equinos alérgicos apresentaram reações positivas diferentes, de acordo com a hipersensibilidade individual, demonstrando sensibilização alérgica verdadeira. The intradermal test (IDT) is an important tool in the diagnosis of allergens involved in IgE-mediated hypersensitivity reactions. The aim of this study was to evaluate the IDT in nonallergic horses with allergenic extracts of Cynodon dactylon, Lolium multiflorum, Paspalum notatum, Culex sp. and Aedes aegypti, and three histamine concentrations, in two different volumes, and based on the results, verify the accuracy of IDT in allergic horses. IDT was performed on 17 nonallergic horses with three concentrations of each extract (1:2000v/w, 1:4000v/h and 18000v/w) and three histamine concentrations (0,1mg/ml, 0,05mg/ml and 0,025mg/ml) in two volumes (0,1 and 0,05ml). Based on the results obtained, IDT was performed in six allergic horses using extracts at the concentration of 1:2000v/w, histamine at a concentration of 0.025mg/ml, in a volume of 0,05ml. In nonallergic horses, none of the three concentrations of the extracts tested in the two volumes were irritant. Larger concentrations of histamine provoked larger and slightly more turgid wheals, and the larger the volume injected, the greater the reactions The IDT performed in allergic horses showed reliability of the results because they demonstrated individual hypersensitivity and did not provoke irritative reactions. In conclusion, the different concentrations of extracts tested in nonallergic horses did not provoke false positive (irritant) reactions. The volume of 0,05ml is more recommended for execution of IDT because the applications with 0,1ml produced larger reactions, impairing the sensitivity of the test. The concentration of 0,025mg/ml of the histamine solution caused the formation of smaller wheals, allowing a better accuracy of the cut off in the IDT. The allergenic extracts tested in allergic horses did not provoke reactions in all the animals, what could be considered an irritative reaction. Allergic horses presented different positive reactions, according to individual hypersensitivity, demonstrating true allergic sensitization.

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Kannanayakal, Thomas Joseph. "Inflammatory Cytokines in Jet Propulsion Fuel-8 Induced Irritant Contact Dermatitis in Male Fischer Rats." Wright State University / OhioLINK, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=wright1242168249.

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Tarantola, Arnaud. "Epidemiology as a tool to improve prevention of human rabies : local and global health implications of postexposure prophylaxis data, Institut Pasteur du Cambodge, 2003-2014." Thesis, Sorbonne Paris Cité, 2018. http://www.theses.fr/2018USPCC031/document.

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La rage entraîne plus de 60,000 décès par an dans le Monde, dont 800 au Cambodge, pays fortement endémique pour la rage canine.La mort survient dans près de 100% des cas de rage, maladie évitable dans presque 100% des cas par l’accès à une prophylaxie post-exposition (PPE) antirabique adéquate et en temps utile. L’amélioration de l’accès à une PPE dans les zones rurales des pays endémiques permettra d’épargner des vies humaines à court terme. Cette thèse en épidémiologie a tiré parti des données collectées auprès des patients consultant au centre antirabique et les chiens testés à l’Institut Pasteur du Cambodge (IPC), Phnom Penh. Suite à un bilan épidémiologique de la situation et des obstacles auxquels sont confrontés les patients cherchant à accéder à la PPE adéquate et en temps utile, elle vise à contribuer à améliorer 1/ l’accès géographique et 2/ l’accès financier à une PPE pour les populations rurales du Cambodge. Nous avons développé une stratégie originale d’identification des poches de populations à haut risque d’incomplétude vaccinale après une exposition potentielle à la rage. Ceci devrait permettre d’améliorer l’accès géographique à la PPE et se concrétiser par l’ouverture en Juillet 2018 d’un centre périphérique de prévention de la rage dans l’Ouest du Cambodge. Cette stratégie d’identification de difficultés d’accès aux soins est applicable à d’autres thématiques de santé, sous certaines conditions. Notre rappel des patients et l’analyse des décès par rage parmi les patients n’ayant pas complété de leur propre chef le protocole PPE de 4 sessions intradermales sur 1 mois ne permettent pas de mettre en évidence une différence de mortalité par rage parmi les patients n’ayant reçu que 3 sessions sur 1 semaine, par rapport à au moins 4 sessions/1mois. Le raccourcissement du protocole à 1 semaine permet de réduire les coûts directs et indirects et l’absence de revenus pendant la durée du traitement en capitale. La mise en place de ce protocole doit s’accompagner d’un suivi d’au moins 6 mois des patients après leur prise en charge initiale. L’ensemble de ces travaux a des implications qui dépassent le cadre du Cambodge: Dans ses recommandations d’Avril 2018, l’OMS recommande désormais ce nouveau protocole IPC– le premier protocole PPE antirabique abrégé à 1 semaine Rabies causes more than 60,000 deaths worldwide each year, including 800 in Cambodia, where canine-mediated rabies virus circulates. Death occurs in nearly 100% of rabies cases, a disease which is nearly 100% avoidable by timely and adequate rabies post-exposure prophylaxis (PEP). Improving access to PEP in rural areas of endemic countries will spare human lives in the short term. This epidemiology PhD used the data collected in patients referred to the rabies prevention clinic and tested dogs at Institut Pasteur du Cambodge (IPC), Phnom Penh. After a baseline assessment of access to and obstacles to access timely and adequate PEP in Cambodia, this PhD aims to contribute to improving: 1/ geographical access and 2/ financial access to PEP for rural populations in Cambodia. We developed an original strategy to identify populations with a high risk of PEP noncompletion after a bite by a potentially rabid dog. This should help improve geographical access to PEP following the implementation in July 2018 of a peripheral rabies prevention center in Western Cambodia. This strategy can be applied to identify difficulties in accessing health services relevant to other health issues, under certain conditions. After patient callback and analysis of rabies deaths among those who did and did not complete the 4-sessions/1-month intradermal PEP regimen of their own accord, we were unable to demonstrate a difference in rabies mortality among patients who only received 3 vaccine sessions over the first week compared to those receiving at least 4 sessions/one month. Abridging the protocol to one week would reduce direct and indirect costs and the loss of income during PEP in the Capital. The adoption of this abridged regimen must be associated with a strengthened clinical monitoring system for at least 6 months following patients’ initial PEP.The work presented in this PhD has implications which reach beyond Cambodia: WHO recommends this new IPC regimen – the first approved one-week, abridged rabies PEP regimen – in its April 2018 guidelines

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Pereira, Desydere Trindade. "Estudo da sensibilização de cães com dermatite atópica na região central do Rio Grande do Sul." Universidade Federal de Santa Maria, 2015. http://repositorio.ufsm.br/handle/1/10196.

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Canine atopic dermatitis (CAD) is a common dermatosis, defined as a genetic-based disease, which predisposes to cutaneous inflammation and pruritus, mediated by class IgE immunoglobulins directed against specific antigens in most cases. Clinical diagnosis may be later complemented by skin allergic and/or serological tests (ELISA). The aim of these tests is to identify possible allergens in order to enable the clinicians to select candidate antigens for allergen specific immunotherapy. This work aimed to identify the sensitization profile of 58 dogs with atopic dermatitis diagnosis. All animals were submitted to intradermic test (IDT) and screened for the presence of antibodies against different allergens using a serologic test. House dust mites are described as the most frequent allergens in all continents. However, the positivity to C. dactylon is not commonly described and may be characteristic for the region. With this work it was possible to identify the main allergens involved in the immunologic response of atopic dogs residing in Rio Grande do Sul, pointing to the importance to include C. dactylon in screening tests for allergy. A dermatite atópica canina (DAC) é uma dermatose comum, definida como uma doença de cunho genético que predispõe à inflamação e ao prurido cutâneo, mediada por imunoglobulinas da classe IgE dirigidas contra antígenos específicos na maior parte dos casos. O diagnóstico da DAC é clínico e pode ser posteriormente complementado por testes alérgicos cutâneos e/ou sorológicos. O objetivo desses testes é identificar possíveis alérgenos e, com isso, possibilitar ao clínico a seleção de antígenos candidatos para a imunoterapia alérgeno-específica. No presente trabalho buscou-se identificar o perfil de sensibilização de 58 cães diagnosticados com dermatite atópica. Todos os animais foram submetidos ao teste intradérmico (TID) e à detecção de anticorpos específicos para diferentes alérgenos através de teste sorológico (ELISA). Os ácaros domiciliares são descritos como os alérgenos mais frequentes em todos os continentes. Entretanto, a positividade ao C. dactylon não é usualmente descrita e pode ser característica da região. Com esse trabalho foi possível identificar os principais alérgenos envolvidos na resposta imunológica de cães atópicos residentes no Rio Grande do Sul, ressaltando-se a importância da inclusão do extrato de C. dactylon em testes alérgicos.

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Loth, Andreas [Verfasser], and Heinz [Akademischer Betreuer] Lehr. "Entwicklung von Verfahren und Applikatoren für den intradermalen Wirkstoffeintrag / Andreas Loth. Betreuer: Heinz Lehr." Berlin : Technische Universität Berlin, 2012. http://d-nb.info/1066160260/34.

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Streker, Meike [Verfasser], and Martina [Akademischer Betreuer] Kerscher. "Randomisierter Halbseitenvergleich eines hyaluronsäurehaltigen Gels nach intradermaler Injektion mittels neuartigen Injektorsystems an Gesicht, Hand und Dekolleté / Meike Streker. Betreuer: Martina Kerscher." Hamburg : Staats- und Universitätsbibliothek Hamburg, 2011. http://d-nb.info/1020458577/34.

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Göller, Manuel Erhard [Verfasser], Nicole [Akademischer Betreuer] Kemper, Nicole [Gutachter] Kemper, and Isabel [Gutachter] Hennig-Pauka. "Bewertung des Einsatzes der intradermalen Applikation von Impfstoffen bei Saugferkeln / Manuel Erhard Göller ; Gutachter: Nicole Kemper, Isabel Hennig-Pauka ; Betreuer: Nicole Kemper." Hannover : Stiftung Tierärztliche Hochschule Hannover, 2021. http://d-nb.info/1237684951/34.

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Naderer, Anna Magdalena [Verfasser]. "Untersuchung zur Bewertung der Verträglichkeit und Immunantwort einer intradermalen Applikation von Porcilis® PRRS an unterschiedlichen Applikationsstellen bei Jungsauen unter Feldbedingungen / Anna Magdalena Naderer." München : Verlag Dr. Hut, 2015. http://d-nb.info/107976898X/34.

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Santos, Camila Mathias dos. "Avaliação das propriedades imunomoduladoras de toxinas termo-lábeis do tipo II produzidas por Escherichia coli enterotoxigênica (ETEC) administradas por via transcutânea." Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/42/42132/tde-23102009-113942/.

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Toxinas termo-lábeis expressas por Escherichia coli enterotoxigênica (LT-I, LT-IIa e LT-IIb) são adjuvantes sistêmicos e de mucosa. Essas proteínas apresentam reduzida identidade (<14%) em suas subunidades B e ligação a receptores distintos, o que pode resultar em propriedades biológicas diferenciais. O objetivo do trabalho foi avaliar a resposta imune induzida pela toxina LT-IIa e seu pentâmero B (LT-IIaB) administradas pelas vias transcutânea e intradérmica. Inicialmente as proteínas foram expressas em E. coli, purificadas e avaliadas quanto à funcionalidade in vitro. Numa segunda etapa, as propriedades imunomoduladoras de LT-IIa e LT-IIaB, imunogenicidade e atividade adjuvante, foram avaliadas em modelo murino. A resposta imune (humoral e celular) induzida contra ovalbumina (OVA), aplicada como antígeno, foi determinada. Os resultados demonstram que as atividades adjuvantes induzidas por LT-IIa e LT-IIaB variam de acordo com a via de inoculação e que as holotoxinas LT-I e LT-IIa induzem graus de inflamação e ativação de resposta imune distintos em camundongos. Heat-labile toxins expressed by enterotoxigenic Escherichia coli (LT-I, LT-IIa and LT-IIb) are potent systemic and mucosal adjuvants. These proteins have low identity (<14%) in their B subunits and bind to different receptors, which may result in differential biological properties. The objective of this work was to evaluate the immune response induced by LT-IIa toxin and its pentameric B subunit (LT-IIaB) delivered by transcutaneous and intradermic routes. Initially the proteins of interest were expressed in recombinant E. coli strains, purified and tested for functionality in vitro. In a second moment, the immunomodulatory properties of LT-IIa and LT-IIaB, immunogenicity and adjuvant activity, were evaluated in mouse model. The humoral and cellular immune responses induced against ovalbumin (OVA) used as antigen were determined. The results show that the adjuvant activity of LT-IIa and its B pentamer depends on the route of inoculation and that LT-I and LT-IIa differ both on induction of inflammation and activation of immune responses in mice.

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Lahm, Katja. "Investigation on intradermal delivery of drug powder by needle-free injection /." 2005. http://www.gbv.de/dms/bs/toc/501260501.pdf.

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Luo, Yong Cun, and 羅永村. "Studies on non-specific reaction of intradermal tuberculin test in dairy cows." Thesis, 1995. http://ndltd.ncl.edu.tw/handle/69216192950845500786.

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Hsu, Wen-Lin, and 許文麟. "Intradermal Glucose-Responsive Insulin Delivery by pH-Sensitive Vesicles incorporated Microneedle Patches." Thesis, 2019. http://ndltd.ncl.edu.tw/handle/774v63.

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碩士 國立中山大學 醫學科技研究所 107 Diabetes mellitus has become a serious non-communicable disease in the World. As of 2014, Diabetes mellitus affects about 387 million people and the number is estimated to increase to over then 500 million by 2035. Typically, subcutaneous insulin injection is a key of glycemic control but it is also inconvenient and painful. These weakness lie bad blood glucose monitoring and disseminating diabetes complications. To improve the weakness, inhaled and oral insulin delivery has been introduced in the early years. However, they cannot be an effective insulin delivery system in clinical diabetes care now due to their side effects, high cost as well as low bioavailability. Therefore, we designed a new intradermal drug delivery system, which combines microneedle (MN) patch and nanoparticles, as a novel long-term glycemic control strategy. Microneedles, a pain-free and minimally invasive drug delivery system made from biocompatible and biodegradable polymer (i.e., polyvinyl alcohol or polyvinylpyrrolidone), is portable to improve the poor patient compliance of glycemic control for diabetes homecare. Furthermore, we also designed a glucose-sensitive nanoparticles to achieve the long-term glycemic control and avoid the hypoglycemia risk. This smart insulin patch offers a clinical opportunity for closed-loop delivery of insulin in a long-term glycemic control, pain-free, and safe manner.

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Kung, Chien-Chia, and 龔建嘉. "Environmental Mycobacterium Flora in Dairy Farms and the Effect on Intradermal Tuberculin Test Results." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/50796501276269704010.

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碩士 國立臺灣大學 獸醫學研究所 99 Among the more than 100 known species of the genus Mycobacterium, majority of them are described as “environmental mycobacteria” (EM), present as saprophytes in water or soil worldwide. Several species of EM are known as opportunistic pathogens and may lead to false positive diagnosis of bovine tuberculosis by intradermal tuberculin test (ITT). In Taiwan, ITT is a standard diagnostic tool for tuberculosis eradication program in dairy herds and cattle showing positive reaction are culled immediately. However, from April of 2009 to August of 2010, approximately 32％ (37/115) of the cattle culled did not exhibit typical tubercle lesions grossly or histopathologically, nor M. bovis or M. tuberculosis was detected by polymerase chain reaction (PCR). In the present study, we used PCR to detect the heat shock protein 65 (hsp65) gene fragments of EM present in various environmental samples to evaluate the distribution of EM flora in dairy farms displaying ITT-positive result, as well as in farms with ITT-negative result as the control group. A total of 16 dairy farms were included and divided into three groups. Group A included farms which had ITT-positive history and M. bovis could be isolated from the retropharyngeal lymph node; within the 105 environmental samples of group A, the positive rate of hsp65 PCR was 50.58%. Group B were farms which had ITT-positive history but no M. bovis could be isolated from the retropharyngeal lymph nodes, indicating that these farms might be ITT false-positive; within the 194 environmental samples of group B, the positive rate of hsp65 PCR was 78.35%. Group C were farms in which dairy cattle displayed ITT-negative history; within the 88 environmental samples of group C, the positive rate of HSP65 PCR was 60.23%. The results showed that the positive rate of hsp65 PCR from Group B was clearly higher than those of group A and C (p&lt;0.05), the cattle in these farms might have a higher chance to contact EM resulting in non-specific sensitization. To further investigate where in the farms that the cattle had higher risk of exposing to EM, each farm was divided into several different regions and it was found that the sport fields in these farms had much higher positive rates of hsp65 PCR than in other regions. The hsp65 PCR products were sequenced and subjected to phylogenetic analysis. It was found that fourteen mycobacterial flora could only be found in all of the six farms of group B, suggesting that the EM in group B was really different from group A and group C. The results of phylogenetic analysis of the sequences of the hsp65 PCR products from environmental samples and from the milk, tissue, and blood of cattle from these farms of group B strongly supported that the EM flora should contribute to the ITT-false positivity. Such false-positive reactors have caused severe, unnecessary economic losses not only to the producers but also to the government. Therefore, knowing the EM flora present in cattle farms and whether theses EM flora may have influence on the ITT results is rather important.

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Barcaro-Machado, Barbara H. "Design and Assessment of a Laser-Assisted Intradermal Medication: A Prospective Randomised Case-control Clinical Trial." Thesis, 2020. https://arro.anglia.ac.uk/id/eprint/706803/1/Barcaro-Machado_2020.pdf.

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Human skin is a potential route for drug administration. Although some inherent cutaneous pathways assist in transporting drugs into the skin, so effective is the stratum corneum that most of the drugs applied do not exhibit the necessary lipophilicity or are too large to permeate this barrier in a significant concentration. Research aimed at optimising device-assisted transcutaneous medication is a novel paradigm and essential to expanding the drugs suitable for application via the skin. The importance of this research is emphasised by the gap in the literature concerning the use of fractional ablative lasers as physical penetration enhancers to bypass the barrier to drug penetration offered by the stratum corneum, allowing for a significant increase in dermal bioavailability of many pharmaceutical agents. The literature has suggested that drugs can diffuse into the microchannels produced by the lasers and penetrate directly to the dermis but usually do not investigate comparative studies on clinical outcomes. Convincing data has been restricted to the oncology field, and larger, controlled trials are necessary to evaluate the impact on the skin surface and the quantitative and qualitative effectiveness of the method. Despite the promising results, the lack of standard methodology regarding laser protocols and for analysing the results reinforces the need for comprehensive research to establish laser-assisted medication as a standard therapeutic modality. This is the first study that has validated and employed three-dimensional stereophotogrammetry as a tool to provide accurate quantification of the skin’s microtopography before and after laser-assisted medication. The stereophotogrammetry system provided accurate numerical data corresponding to the influence of laser-assisted medication on the surface of wrinkles and scars. The thesis original contribution to knowledge included the design of a reproducible, active, laser-assisted, intradermal delivery of medication. An in vivo, prospective, randomised, double-blind, comparative clinical trial was designed. Consenting patients presenting wrinkles and scars were randomised and subjected to one session of laser skin resurfacing with a 2,940 nm Er:YAG (Erbium dopped Yttrium Aluminium Garnet) fractional ablative laser followed by immediate topical application of the medication. The laser parameters were standardised for all patients. This is the first robust study that has compared the effect of different substances on the skin surface after laser-resurfacing and on different conditions of skin integrity, scars and wrinkles. A control group received vitamin C immediately after laser skin resurfacing and another study group received vitamin C plus a cosmeceutical containing growth factors. Most patients treated presented an improvement in the skin condition. However, the groups receiving the cosmeceutical containing growth factors exhibited statistically significantly better results compared to the control groups receiving vitamin C only. This finding demonstrates the positive effect of laser-assisted medication on skin surface and that the addition of growth factors to formulae enhances the result of the treatment.

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Lin, Heng-Ching, and 林恒慶. "Diagnosis and Phylogenetic Analysis of Mycobacterial Infection and Mycobacterium bovis from Intradermal Tuberulolin Test-Positive Herbivore." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/95596970579206998538.

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碩士 國立嘉義大學 獸醫學系研究所 99 Tuberculosis (TB) is a zoonotic disease affecting mammal worldwide. Herbivore’s TB is determined by Mycobacterium bovis and other nontuberculous mycobacteria (NTM). NTM may interfere with the result of intradermal tuberculin test (ITT), and cause fake-positive. According to law, the diagnosis and euthanasia of TB is using ITT in Taiwan. The aim of this study was to understand the pathogenicity, distribution of pathogens detection, and analyze of diagnosis method of ITT positive animal. We sampled from ITT positive animal (cattle and deer) for mycobacterial isolation, pathological examination, acid-fast stain, molecular diagnosis and antibody ELISA. ITT-positive sample included 145 dairy cattle and 12 deer with, and 166 dairy cattle serum and 94 deer serum in this study. Mycobacterium spp. could be isolated from 47 animal (47 / 144, included cattle 132 and deer 12; 32.64 %), 31 out of 144 (21.53%) were identified as Mycobacterium bovis. Gross lesion (gross lesion and histopathology, acid-fast stain and culture) was 56.69 % (89 / 157) positive rate. Most of gross lesions were retropharynx lymph node and mediastinal lymph nodes. The PCR detection rate was higher at hilar lymph nodes, mediastinal lymph nodes and retropharynx lymph node. Those organs could suggest being the sample to diagnosis TB using. Analyze detection rate of Mb-ELISA, Mb-Rp-ELISA and PCR for ITT positive cattle. The result were 4.76 % (4 / 84), 4.76 % (4 / 84) and 86.9 % (73 / 84), respectively. We found that Mb-ELISA or Mb-Rp-ELISA was high specificity but low sensitivity. To detect the mycobacterial infection humoral immunity response did not produce antibody. On the other hand, the sensitivity and specificity of PCR was better than others, and could be an aid for TB diagnosis when compared to ITT result. The sequence of M. bovis was no different significantly between ITT positive animals. Same genotype may infect cattle among different farms. Conclusion of this study, most of ITT positive animals were infected by M. bovis. Retropharynx lymph node, hilar lymph nodes and mediastinal lymph nodes were most lesions found in cattle. The lesions in contrast of deer were found in retropharynx lymph node and lung. Ante mortem diagnosis for TB was primarily relied on ITT test. Other diagnosis method could be an aid for. The DNA sequences variation of M. bovis isolated from deer and cattle were no different significantly, M. bovis infection shared similar phylogenetic relationship.

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Hsu, Ming-Yi, and 許名宜. "Studies of intradermal tests of house dust and dust mite in canine atopic dermatitis at Taipei area." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/35091847924706999255.

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碩士 國立臺灣大學 獸醫學研究所 93 Host dust and dust mite are important allergens in canine atopic dermatitis. The purpose of this study was to investigate sensitivity to house dust and dust mite in atopic dogs in Taipei city. One hundred atopic dogs were enrolled according to history, clinical signs ,physical and dermatological exams. Intradermal test with house dust and dust mite were performed and investigated. The diagnosis of atopic dermatitis of each dog was based upon the clinical manifestations compatible to criteria, and results of intradermal skin tests. fifty females and fifty males aged from one to forteen years old with twenrt-three breeds . The mean age of this series was 5.7years. Among these One hundred dogs, eighty six dogs (86%) exhibited positive reactions to either house dust or dust mite, or both. Positive reactions to house dust and dust mite were 15% (15/100) and 3% (3/100) respectively. Positive reaction to both allergens was 68% (68/100). No breed, age predilection found in this series. Sex and body weight were statistical related to positive reaction (either house or dust mite), Analyses between sex, age, body weight and positive reaction to house dust, positive reaction to dust mite, and positive reaction either to house dust or dust mite reveals statistically related between body weight and positive reaction. Percentage of cases exhibited positive reaction to both allergens was significantly higher than that of cases reacted to either allergen alone. This suggested that positive reaction to either allergen would be associated to each other. Further investigation would be needed to clarify the cross-reaction between house dust and dust mites. This study suggests the high positive rate of intradermal tests to house dust and dust mite. House dust and dust mite are common and important allergens in atopic dogs at Taipei area.

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