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This thesis summarizes the research conducted in the field of core-multishell nanocarriers for the application in dermal drug delivery. While the CMS nanocarrier as a platform for drug delivery has been established by several publications, the goal of my thesis was to specialize the CMS nanocarrier as a bioresponsive nanocarrier for applications in the skin. The skin is a yet poorly characterized biological environment regarding its chemistry, especially in the state of disease. However, the penetration and successful delivery of the fluorescent model drug nile red into deeper skin layers, compared to a standard nile red cream, has been stated in a previous study. The increased penetration was the starting point for the general hypothesis on the CMS nanocarrier being a potential drug delivery platform. However, the fate of the nanocarrier inside the skin was identified as a potential toxicity issue. Hence, the first project of this PhD work aimed at establishing a biodegradable CMS nanocarrier. Based on a diacid and a triol, a hyperbranched polyester as the core molecule of a CMS nanocarrier was synthesized and used as a macroiniiator for the ring-opening polymerization of Epsilon-caprolactone. Surrounded by a water-soluble mPEG outer shell, this nanocarrier showed a complete enzymatic degradation in seven days, while surprisingly, hydrolysis of the ester-based CMS nanocarrier did not take place under physiologically relevant conditions. The low molecular weight of the core molecule and the low branching of the resulting CMS nanocarrier are hypothesized to be the reason for the poor drug loading capacity of this biodegradable nanocarrier. Aiming at the redox-barrier of skin, we hypothesized the natural gradient of reductants and oxidants in the skin to be a suitable trigger for stimulus-initiated release of drugs at sites of inflammation. A complementary set of a reduction-sensitive and an oxidationsensitive CMS nanocarrier has been synthesized. Based on a hyperbranched polyglycerol as a core building block, and a water-soluble mPEG outer shell, the alkyl inner shell contained either a disulfide moiety, or a thioether moiety. The reduction-sensitive disulfide-containing rsCMS and its oxidation-sensitive thioether-containing osCMS counterpart were tested for the triggered release of the encapsulated dye nile red in vitro. A physiologically relevant concentration of 10 mM GSH was not able to trigger a fast release of NR, possibly due to the limited reduction capacity of GSH in the absence of redox-mediating enzymes. However, the reductant TCEP led to 90 % release in 24 h. The incubation of osCMS with 1 % H2O2 triggered a sustained release of 80 % NR over the same period. Skin penetration experiments of NR@rsCMS on ex vivo human skin supported our hypothesis on the natural GSH gradient in the skin. Also, the synthesis of an EPR-labelled dexamethasone was established, and the PCA-labelled version will be used in future studies on the distribution of ROS in skin as the trigger for stimulated release of drugs from osCMS.
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Ocular infections in animals can be result in blindness. The infections are most caused by bacteria particular microbiota from the patient’s conjunctiva, eyelid, and nose. When damage to ocular tissue occurs, opportunistic bacteria may invade ocular tissues causing infection that may be difficult to treat, especially if the bacteria are antibiotic resistant like MRSA. Therefore, the usage of antibiotics has to be carefully monitored to prevent antibiotic resistance and a global reduction in a use of antibiotics a practical aim. Povidone-Iodine (PI) is generally known for being effective against bacteria, fungi, viruses, and protozoa. Moreover, an ideal solution as it is inexpensive, and no irritation is caused when diluted below 1%. One study indicated that Neosporin eye drops and PI administration before ophthalmic surgery caused similar decreases in the number of colonies and species cultured. Therefore, PI can be offered as an alternative to antibiotics. Forty eyes: (24 dogs and 16 cats) were divided into four groups of ten according to the concentrations of PI 0.1%, 0.2%, 1% and 5%. Seventy-four eyes (48 dogs and 8 cats) were included for assessment of the antimicrobial activity of 0.2% PI. Conjunctival swabs were collected from 44 horses (46 eyes) to determine the occurrence of MRSA on the ocular surface of the horse. In the first study, ocular swabs were collected before aseptic preparation (t0), after aseptic preparation (t1) and after surgery (t2). Tissue reaction was monitored at t1. Specimens were analyzed for cultivable aerobic and anaerobic bacteria and fungi. Positive culture swabs were used to measure the effectiveness of PI. In the second study, the conjunctival swabs that were collected before ophthalmic surgery were immediately processed with testing of colonies suspected to be S. aureus was carried out using Vitek2 system (bioMérieux, Lyon) and the presence of MRSA was verified by PCR. Genetic relatedness of all MRSA isolates was investigated by pulsed-field gel electrophoresis (PFGE) and spa-typing. The result of the first study indicated that 0.2% and 1% PI solutions could be used to eliminate bacterial contamination of ocular surface tissue without tissue reaction. However, 0.2% PI proven to be more effective than more concentrated solutions (p= 0.02). Additionally, 0.2% PI concentration significantly reduced Bacillus sp. and coagulase negative Staphylococcus sp. at t1 and t2. MRSA was not positive in our first study. That is why the effectiveness of PI on eliminating MRSA on the ocular surface of animals still undetermined. The results of the second study indicated that the isolation rates of MRSA on the ocular surface of horses were high (13%) 6/46 samples. Molecular typing revealed that all detected MRSA were the spa-type t011. This type is commonly associated with the European livestockassociated MRSA lineage of the clonal complex CC 398 that recently emerged in horses in Germany. Additionally, PFGE revealed that the six MRSA had a very closely related relationship (>90%). According to the history of the patients and the result of the microbiological examination, it can be suggested that this phenomenon was a nosocomial infection. MRSA reported in our study exhibited further resistance towards antibiotics frequently used for topical eye treatment, e.g. fluoroquinolones, tetracycline's, kanamycin and gentamicin. The most effective concentration of PI was proven to be 0.2%. It demonstrated excellent antibacterial activity against Bacillus sp. and coagulase-negative Staphylococcus sp. This study demonstrated a high prevalence of MRSA on the ocular surfaces of equine ophthalmic patients supporting the need for constant bacterial surveillance. More studies are needed on the source of MRSA in equine ophthalmic patients including an environmental factor, standard operation procedures (SOPs), manipulations and diagnostic approaches. The evaluation of the bactericidal activity of antiseptic agents against MRSA on the ocular surface of animals at either higher concentrations of PI or alternative protocols and active components to reduce ocular surface contamination are still needed.,Augeninfektionen bei Tieren können zur Erblindung führen. Die Infektionen können durch Bakterien, insbesondere die Mikrobiota der Bindehaut, des Augenlides und der Nase des Patienten verursacht werden. Bei Beschädigung des Augengewebes, können die opportunistischen Bakterien in das Auge eindringen und zu einer Infektion führen, welche schwierig zu behandeln ist, insbesondere dann, wenn die Bakterien antibiotikaresistent sind, wie MRSA. Deshalb muss der Einsatz eines Antibiotikums sorgfältig überwacht werden, um eine Antibiotikaresistenz zu verhindern. Den Einsatz von Antibiotika zu reduzieren, ist ein globales Ziel. Povidone-Iodine (PI) ist bekannt dafür, dass es gegen Bakterien, Pilze, Viren und Protozoen wirksam ist. Darüber hinaus ist es günstig und verursacht keine Irritation, wenn es unter 1% verdünnt wird. In einer Studie konnte gezeigt werden, dass Neosporin-Augentropfen und die Verabreichung von PI vor Augenoperationen eine ähnliche Reduktion an kultivierten Bakterienkolonien und -spezies verursachten. Deshalb kann PI als Alternative zu Antibiotika genutzt werden. Vierzig Augen: (24 Hunde und 16 Katzen) wurden in je vier Zehnergruppen entsprechend der Konzentrationen von PI 0,1%, 0,2%, 1% und 5% eingeteilt. Vierundsiebzig Augen (48 Hunde und 8 Katzen) wurden zur Beurteilung der antimikrobiellen Aktivität von 0,2% PI beurteilt. Von 44 Pferden (46 Augen) wurden Konjunktivalabstriche gesammelt, um das Auftreten von MRSA auf der Augenoberfläche des Pferdes zu bestimmen. In der PI Studie wurden Augenabstriche vor der aseptischen vorbereitung (t0), nach der aseptischen vorbereitung (t1) und nach der Operation (t2) entnommen. Die Gewebsreaktion wurde zum Zeitpunkt t1 überwacht. Die Proben wurden im Hinblick auf kultivierbare aerobe und anaerobe Bakterien und Pilze analysiert. Die Prävalenz von positiven Kulturabstrichen wurde verwendet, um die Wirksamkeit von PI zu messen. In der MRSA Studie wurden die Bindehautabstriche gesammelt, unmittelbar bevor ophthalmochirurgische Eingriffe erfolgten, und die Identifizierung und Empfindlichkeitsprüfung von S. aureus verdächtigen Kolonien, wurde mit Vitek2 (bioMérieux, Lyon) durchgeführt und das Vorhandensein von MRSA durch PCR verifiziert. Die genetische Verwandtschaft aller MRSA-Isolate wurde mittels Pulsfeld-Gelelektrophorese (PFGE) und Spa-Typisierung untersucht. Das Ergebnis der PI Studie an Kleintiere zeigte, dass Lösungen mit 0,2% und 1% PI verwendet werden könnten, um eine bakterielle Kontamination von Augenoberflächengewebe ohne Gewebereaktion zu eliminieren. 0,2% PI sind hierbei wirksamer als konzentriertere Lösungen (p = 0,02). Zusätzlich reduzierte eine Konzentration von 0,2% PI Bacillus sp. und Koagulase-negative Staphylococcus sp. Zu den Zeitpunkten t1 und t2. In diesen Teil der Studie würde kein MRSA nachgewiesen. Aus diesem Grund wird die Wirksamkeit von PI zur Eliminierung von MRSA auf der Augenoberfläche von Tieren nicht beurteilt werden. Die Ergebnisse der MRSA Studie zeigten, dass die Isolierungsraten mit 6/46 Proben hoch waren (13%). Die molekulare Typisierung ergab, dass alle detektierten MRSA vom Typ t011 sind. Dieser Typ wird üblicherweise mit der europäischen, mit Tieren assoziierten MRSALinie des klonalen Komplexes CC 398 in Verbindung gebracht, die kürzlich auch bei Pferden in Deutschland aufgetreten ist. Darüber hinaus konnte mit der PFGE nachgewiesen werden, dass die sechs MRSA-Keime sehr nah miteinander verwandt waren (> 90%). Nach der Anamnese der Patienten und dem Ergebnis der mikrobiologischen Untersuchung, lässt sich vermuten, dass dieses Phänomen durch eine nosokomiale Infektion begründet war. Die in unserer Studie kultivierten MRSA zeigten weitere Resistenzen gegenüber Antibiotika, die häufig zur topischen Augenbehandlung verwendet werden, wie z.B. Fluorchinolone, Tetracycline, Kanamycin und Gentamicin. Die effektivste Konzentration von PI wurde mit 0,2% nachgewiesen. Es zeigte eine ausgezeichnete antibakterielle Aktivität gegen Bacillus sp. und Koagulase-negative Staphylococcus sp. beim Kleintier. Diese Studie zeigte eine hohe Prävalenz von MRSA auf den Augenoberflächen von equinen Patienten, die die Notwendigkeit einer konstanten bakteriellen Überwachung unterstützen. Weitere Studien zur Quelle von MRSA bei equinen ophthalmologischen Patienten, einschließlich Umweltaspekten, standardisierten Vorgehenssverfahren (SOPs) und Manipulationen sowie diagnostische Ansätze sind erforderlich. Es ist notwendig, die bakterizide Aktivität von Antiseptika gegen MRSA auf der Augenoberfläche des Tieres entweder mit höheren Konzentrationen von PI oder alternativen Protokollen und aktiven Komponenten zur Verringerung der Kontamination der Augenoberfläche zu beurteilen.,
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Sensory circuits are typically established during early development, yet how circuit specificity and function are maintained during organismal growth has not been elucidated. To gain insight we quantitatively investigated synaptic growth and connectivity in the Drosophila nociceptive network during larval development. We show that connectivity between primary nociceptors and their downstream neurons scales with animal size. We further identified the conserved Ste20-like kinase Tao as a negative regulator of synaptic growth required for maintenance of circuit specificity and connectivity. Loss of Tao kinase resulted in exuberant postsynaptic specializations and aberrant connectivity during larval growth. Using functional imaging and behavioral analysis we show that loss of Tao-induced ectopic synapses with inappropriate partner neurons are functional and alter behavioral responses in a connection-specific manner. Our data show that fine-tuning of synaptic growth by Tao kinase is required for maintaining specificity and behavioral output of the neuronal network during animal growth.
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An overt pro-inflammatory immune response is a key factor contributing to lethal pneumococcal infection in an influenza pre-infected host and represents a potential target for therapeutic intervention. However, there is a paucity of knowledge about the level of contribution of individual cytokines. Based on the predictions of our previous mathematical modeling approach, the potential benefit of IFN-γ- and/or IL-6-specific antibody-mediated cytokine neutralization was explored in C57BL/6 mice infected with the influenza A/PR/8/34 strain, which were subsequently infected with the Streptococcus pneumoniae strain TIGR4 on day 7 post influenza. While single IL-6 neutralization had no effect on respiratory bacterial clearance, single IFN-γ neutralization enhanced local bacterial clearance in the lungs. Concomitant neutralization of IFN-γ and IL-6 significantly reduced the degree of pneumonia as well as bacteremia compared to the control group, indicating a positive effect for the host during secondary bacterial infection. The results of our model-driven experimental study reveal that the predicted therapeutic value of IFN-γ and IL-6 neutralization in secondary pneumococcal infection following influenza infection is tightly dependent on the experimental protocol while at the same time paving the way toward the development of effective immune therapies.
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It is essential for scholars to reflect on their research practices and critically assess scientific rigor. In the current article, we aim to critically review the state of qualitative research in organization studies by focusing on trends in sample sizes. Organizational scholars presenting qualitative, interview-based manuscripts tend to face the ongoing challenge of how many interviews are enough. The research reported in this article, covering 11 years and investigating 855 interview-based studies, provides empirical evidence that, across the years, the number of interviews seems to be rather high. The total sample included studies with more than 100 interviews (8% of the sample), more than 50 interviews (34%) and studies with more than 30 interviews (62%). Furthermore, when studies start to increase in sample size, they often do so at the expense of homogeneity across respondents. We conclude by giving some possible explanations for why we are facing such a situation today.,Forum Qualitative Sozialforschung / Forum: Qualitative Social Research, Vol 20, No 3 (2019): Qualitative Content Analysis I,
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Introduction Despite the fact that ileostomy reversal is considered a comparatively small surgical procedure there are some postoperative complications following this procedure, which may lead to a prolonged hospital stay. The presented bi-centered prospective randomized clinical trial DRASTAR investigated the use of subcutaneous suction drains in patients undergoing elective ileostomy reversal with regard to a potential reduction of length of hospital stay and postoperative complications. Materials and methods All patients included in the trial were randomized into two parallel study groups. One group underwent ileostomy reversal with insertion of a subcutaneous suction drain, the other group underwent ileostomy reversal (ISR) without insertion of a drain. There was a prospective postoperative follow-up-period of three months with a documentation of all complications and the individual patient satisfaction. The statistical analysis of the collected data was done using the software IBM SPSS Statistics 21 ® (IBM, Armonk, New York, USA). Length of hospital stay in days was used as primary outcome measure. Results A total of 118 patients could be included in the trial. 59 patients were randomly appointed to receive ileostomy reversal with insertion of a subcutaneous suction drain, 59 patients were intended for ileostomy reversal without insertion of a drain. After the follow-up-period of three months, 50 patients with subcutaneous suction drain and 53 patients without drain could be analyzed. Median length of hospital stay among patients with drain was eight days versus nine days in the group without subcutaneous suction drain (p=0.17). A postoperative infection of the wound, the necessity of re-operation and anemia could be identified as statistically significant risk factors for a prolonged hospital stay. There were no significant differences between both study groups in terms of wound infection rate and number of required re-interventions. 14 % of patients with insertion of a subcutaneous suction drain developed wound infections versus 17% of patients without insertion of a drain (p=0.68). Six patients in the group with drain needed revision in general anesthesia versus eight patients without subcutaneous suction drain (p=0.13). Conclusion Omission of subcutaneous suction drains in ileostomy reversal is not inferior to the insertion of a subcutaneous suction drain in terms of length of hospital stay and postoperative complication rat,Einleitung Postoperative Komplikationen nach Ileostomarückverlagerung sind trotz des vergleichsweise geringen Umfanges dieses Eingriffes häufig und führen zu einer verlängerten Krankenhausliegedauer. Die hier vorgelegte prospektive Bi-Center-Studie DRASTAR untersuchte den Einsatz subkutaner Redondrainagen bei der elektiven, konventionellen Ileostomarückverlagerung hinsichtlich einer potentiellen Reduktion postoperativer Komplikationen und der hiermit assoziierten Krankenhausliegedauer. Methodik Es handelt sich um eine prospektiv-randomisierte Studie mit zwei parallelen Studienarmen. Eine Patientengruppe erhielt im Rahmen ihrer Ileostomarückverlagerung eine subkutane Redondrainage, die andere Gruppe nicht. Es erfolgte eine Nachbeobachtung der Patienten unter Dokumentation aufgetretener Komplikationen und der individuellen Patientenzufriedenheit bis drei Monate postoperativ. Die Auswertung der gesammelten Daten mit dem primären Endpunkt „Krankenhausliegedauer in Tagen“ erfolgte mit der Software IBM SPSS Statistics 21 ® (IBM, Armonk, New York, USA). Ergebnisse Es konnten primär 118 Patienten in die Studie eingeschlossen werden. 59 Patienten wurden dem Studienarm mit Einlage einer Redondrainage zugeordnet, 59 Patienten dem Studienarm ohne Redondrainage. Nach drei Monaten Follow-up konnten 50 Patienten mit und 53 Patienten ohne Redondrainage in die Analyse aufgenommen werden. Die mediane Krankenhausliegedauer im Studienarm mit Redondrainage betrug acht Tage versus neun Tage im Studienarm ohne Redondrainage (p=0,17). Das Auftreten von Wundinfektionen, die Notwendigkeit von Revisionseingriffen sowie das Vorliegen einer Anämie konnten als statistisch signifikante Risikofaktoren für die Verlängerung der Krankenhausliegedauer identifiziert werden. Die Wundinfektionsrate lag bei 14% in der Gruppe mit Redondrainage versus 17% in der Gruppe ohne Redondrainage (p=0,68). Hinsichtlich der Zahl an notwendigen Revisionseingriffen zeigte sich mit sechs revidierten Patienten in der Gruppe mit Redondrainage versus acht revidierten Patienten in der Gruppe ohne Redondrainage ebenfalls kein statistisch signifikanter Unterschied zwischen beiden Studienarmen (p=0,13). Schlussfolgerung Die Studie konnte zeigen, dass der Verzicht auf die Einlage einer subkutanen Redondrainage bei der Ileostomarückverlagerung der Redon-Drainageneinlage hinsichtlich postoperativer Krankenhausliegedauer und postoperativer Komplikationsrate nicht unterlegen ist.,
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Der für diese Arbeit gewählte Titel, »Der Wert im Inneren der künstlerischen Produktivität«, verweist auf die Analyse der Kunst unter dem Aspekt der kapitalistischen Wertproduktion. Hierüber versteht sich die vorliegende Arbeit als ein Beitrag zur Debatte über Kunst und Ästhetik vor dem Hintergrund der marxistischen Gesellschaftslehre und Philosophie. Die Fragestellung lautet, was heißt eine Kunstphilosophie, die auf den Instrumentenkoffer des reifen Marx zurückgreift. Anders gesagt, was heißt, die gewöhnlichen Kategorien der Kunsttheorie durch die Kategorien der Kritik der politischen Ökonomie zu durchdringen. Die vorliegende Arbeit besteht in erster Linie aus dieser Durchdringung, ohne damit eine Reduktion der Ästhetik auf das bloß Ökonomische zu beabsichtigen. Aus dieser kritischen Intervention ergeben sich drei Thesen: eine historischeThese, die unter der Behauptung der relativen Unabhängigkeit oder Autonomie der Kunst als Gefüge oder Ebene ihr Verhältnis zur gesamtgesellschaftlichen Struktur untersucht. Die ökonomische These besagt, dass die neoklassische Wirtschaftslehre gesellschaftlich blind ist in Bezug auf die Klassenverhältnisse im Kunstbetrieb oder hinsichtlich des Reproduktionsprozesses der Kunstwelt. Letztendlich legt die auf die Wertsubstanz bezogene These Nachdruck auf das Eigentum an den Produkten der eigenen Arbeit als Voraussetzung des Kauf-Verkauf-Verhältnisses. Dies macht die eigentümliche Wertform der Kunstproduktion im Kapitalismus aus. Im letzten Teil der Arbeit wird die Umkehrung der Prämissen der vorhergehenden Kapitel vollzogen und das Kunstwerk als Gebrauchswert für Andere postuliert.Dabei wird die Dimension des Werkes als Praxis hervorgehoben. Diese Dimension erlaubt ein Hantieren mit dem Werk, das eine ikonoklastische Handlung mit dem Werk über die Herrschaft der Warenform und die Werkherrschaft des privaten Eigentums hinaus vollkommen plausibel macht.
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In this study, we report our initial results on in situ biosynthesis of S-allyl-l-homocysteine (Sahc) by simple metabolic conversion of allyl mercaptan in Escherichia coli, which served as the host organism endowed with a direct sulfhydration pathway. The intracellular synthesis we describe in this study is coupled with the direct incorporation of Sahc into proteins in response to methionine codons. Together with O-acetyl-homoserine, allyl mercaptan was added to the growth medium, followed by uptake and intracellular reaction to give Sahc. Our protocol efficiently combined the in vivo synthesis of Sahc via metabolic engineering with reprogrammed translation, without the need for a major change in the protein biosynthesis machinery. Although the system needs further optimisation to achieve greater intracellular Sahc production for complete protein labelling, we demonstrated its functional versatility for photo-induced thiol-ene coupling and the recently developed phosphonamidate conjugation reaction. Importantly, deprotection of Sahc leads to homocysteine-containing proteins—a potentially useful approach for the selective labelling of thiols with high relevance in various medical settings.
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“Diversity in the Cultures of Physics” was an Erasmus+ funded Strategic Partnership launching several key actions aimed at improving the gender balance in physics and its subfields. The Strategic Partnership consisted of six universities in four countries: Freie Universität Berlin in Germany, Universitat Autònoma de Barcelona and Universitat de Barcelona in Spain, the University of Manchester and the University of Sheffield in the United Kingdom and Uppsala Universitet in Sweden. One of the central activities of the transnational network was an annual international summer school series for female physicists transitioning from Undergraduate/Master programmes to a PhD study. This document presents the curriculum that has been developed for those summer schools and that has been used and evaluated in each round of the summer school series. Thus it is based on the insights and evaluation of all summer schools implemented. The curriculum consists of four core pillars of modules: 1) research stays at physics departments; 2) visits to physics research institutions; 3) empowerment and gender equality policy; 4) gender studies and physics. A transversal pillar is composed of sessions concerning group-building processes and group dynamics. For each pillar, a manifold variety of modules that pursue the learning objective of the respective pillar have been designed, planned and carried out. The curriculum describes aims and functions of the pillars and provides an overview of formats and designs of the modules that have been part of each pillar. For some prominent modules that have been carried out in almost every summer school and might also be practicable in future summer schools, more details are provided. It is pointed out which instructional recommendations are to be considered, what challenges are to be expected and how long the module typically lasts.
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Objective: To test the hypothesis that olfactory (OF) and gustatory function (GF) is disturbed in patients with autoimmune encephalitides (AE). Methods: The orthonasal OF was tested in 32 patients with AE and 32 age- and sex-matched healthy controls (HC) with the standardized Threshold Discrimination Identification (TDI) score. This validated olfactory testing method yields individual scores for olfactory threshold (T), odor discrimination (D), and identification (I), along with a composite TDI score. The GF was determined by the Taste Strip Test (TST). Results: Overall, 24/32 (75%) of patients with AE, but none of 32 HC (p < 0.001) had olfactory dysfunction in TDI testing. The results of the threshold, discrimination and identification subtests were significantly reduced in patients with AE compared to HC (all p < 0.001). Assessed by TST, 5/19 (26.3%) of patients with AE, but none of 19 HC presented a significant limitation in GF (p < 0.001). The TDI score was correlated with the subjective estimation of the olfactory capacity on a visual analog scale (VAS; r(s) = 0.475, p = 0.008). Neither age, sex, modified Rankin Scale nor disease duration were associated with the composite TDI score. Conclusions: This is the first study investigating OF and GF in AE patients. According to unblinded assessment, patients with AE have a reduced olfactory and gustatory capacity compared to HC, suggesting that olfactory and gustatory dysfunction are hitherto unrecognized symptoms in AE. Further studies with larger number of AE patients would be of interest to verify our results.
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