To Örebro University

Background: Patients with clinical manifestations suggestive of otitis externa.

Objective: To study the prevalence of mycotic infection in these patients.

Methods: Ear swabs were taken from these patients and both direct examination and culture were done.

Results: The most common pathogenic fungal isolates were Candida species (8.4%) and Aspergillus niger (4.2%). Conclusion: Besides Aspergillus and Candida species isolated in these patients, many bacteria were also seen. Dry climate, wearing head clothes, presence of middle ear infection, wearing hearing aids and swimming were predisposing factors for this infection.

Background: In the diagnostic work up of allergy, determining allergen component-specific immunoglobulin E (IgE) is important for diagnosis, prognosis and choice of treatment.

Objective: The purpose of this study was to evaluate the performance of the immunoblotting assay (Euroline) in detection of IgE antibodies against timothy grass and birch pollen allergen components compared to fluorescent enzyme assay (ImmunoCAP, Phadia 250).

Methods: A total of 128 serum samples from patients allergic to timothy grass and birch pollen were analysed. The levels of IgE antibodies to timothy grass and birch pollen were measured using Euroline DPA-Dx pollen 1 and ImmunoCAP assay. The two methods were then compared on binary (positive vs negative), semi-quantitative (IgE classes) and quantitative (concentration) levels. The two methods were also compared to results from skin prick testing.

Results: The Euroline method showed a positive percentage agreement of 93% and negative percentage agreement of 94% with an overall accuracy of 94% when compared to ImmunoCAP. Kappa analysis showed moderate strength of agreement between the methods in determining IgE classes for 7/11 components tested. All components showed a positive correlation when analysed using Spearman's rank correlation.

Conclusions: Overall, we found that there is good correlation between the Euroline and ImmunoCAP methods in measuring IgE sensitization.

Background:  Patients show varied results to allergen immunotherapy (AIT. The reason for this variability is unclear.

Objective: To describe the relationship between AIT efficacy and demographic characteristics, as well as pre-treatment plasma levels of specific IgE-antibodies to grass and birch pollen.

Methods: A retrospective study was performed based on medical records of 128 patients who received AIT. The patients completed a questionnaire and pre-AIT plasma levels of allergen-specific IgE to grass and birch pollen were measured using EUROLINE DPA-Dx pollen 1 method. Results. Seventy percent of patients classified their allergic symptoms as less severe after AIT. Twenty-seven percent had received AIT targeting only grass pollen, 19% targeting only birch pollen, and 55% targeting both grass and birch. A total of 35 different IgE profiles were found across our study population. On comparison of the demographic characteristics and concentration of allergen-specific IgE-antibodies, no statistically significant differences could be found.

Conclusions: The majority of patients rated their allergic symptoms as less severe after AIT. No clear relationship could be demonstrated between pre-treatment allergen-specific IgE concentration, or demographic characteristics, and effect of AIT. There may be other factors underlying the different responses to AIT.

Covid-19 har funnits i Sverige i drygt ett år nu, och vi lär oss mer och mer om sjukdomen. Covid-19 kan ge många olika symtom, och de som drabbas av sjukdomen blir sjuka i varierande grad. Vissa får bara lukt- och smakbortfall, feber eller huvudvärk, medan andra blir allvarligt sjuka och behöver vårdas länge i respirator. Behandlingen av patienter med covid-19 har utvecklats under året som gått. Att covid-19 kan ge nervpåverkan är nu känt – ett stort antal patienter får neurologiska symtom [1] och det publiceras fortlöpande fler och fler fallbeskrivningar gällande påverkan på en eller flera kranialnerver [2-5]. Den exakta korrelationen mellan de neurologiska komplikationerna och själva infektionen är fortfarande inte helt klargjord. Vi vill här presentera ett patientfall med påverkan på flera kranialnerver i samband med svår covid-19 och intensivvård. 

The coronavirus disease 2019 pandemic has shed light on the post-viral olfactory dysfunction (PVOD). Post-viralolfactory dysfunction is temporary for most people and usually subsides when the common cold symptoms ameliorate. However, in some patients, this condition can persist for several weeks or months. The exact pathological mechanisms of persistent olfactory loss secondary to upper respiratory tract infection (URTI) is unknown, andthere is a lack of effective treatment. An increased understanding of pathology could possibly translate into newtherapeutic regimens. The aim of this systematic review is to synthesize primary data regarding histopathological and neuropathological findings in patients with PVOD secondary to URTI. The Preferred Reporting Items forSystematic Reviews and Meta-Analyses guidelines was followed. Databases PubMed and Web of Science weresearched with keywords and mesh terms to identify relevant articles. The quality of included studies was assessedwith the Newcastle–Ottawa Scale for observational studies. The search yielded a total of 847 articles, after excludingduplicates and articles that were not relevant. A total of 12 studies were selected. The main findings of this reviewwere: olfactory bulb (OB) volume was decreased in patients with PVOD, there was a negative correlation betweenOB volume and duration of olfactory loss, both primary and secondary olfactory cortex changes were found interms of structure and functionality, and the olfactory sensory neurons and nerve bundles were reduced in patientswith PVOD. The mechanisms of PVOD are complex. This review found that viral URTI is attributable to structural andfunctional changes at multiple locations of the olfactory system.

Tonsil tissue not located at known anatomical locations at the entry of aero-digestive system is called ectopic tonsil. This is rarely presented clinically and may lead to diagnostic confusion when encountered. The diagnosis of ectopic tonsil is challenging as it is usually asymptomatic and requires histopathological examination for definitive diagnosis. A case of patient with sublingual ectopic tonsillar tissue is described here. Otolaryngologists colleagues are encouraged to pay attention to this entity and consider it in the differential diagnosis of an unclear lump in the head and neck region especially, in the oral cavity.

Establishing efficient methods for local administration of drugs to the inner ear has great clinical relevance for the management of inner ear disorders. However, the administration route remains a critical issue. The most feasible approach for a non invasive drug delivery to the inner ear is application of medication to the middle ear cavity on the promise that it will diffuse through the thin round window membrane (RWM) separating the inner ear from the middle ear cavity. Gene therapy represents a promising future in otology and offers an exciting therapeutic alternative as it could be used in prevention or management of cochlear disorders. Also for a gene therapy approach, RWM application seems most feasible administration route. Exploring and characterizing the RWM route of administration is thus a fundamentally important area of research for the development of future treatment of inner ear disorders. The objectives of the thesis were to evaluate the efficacy of two drug and gene delivering vehicles to the inner ear, sodium hyaluronate (HYA) and chitosans, which can be applied to the cochlea. Ultimate aim is to establish an efficient drug delivery system and gene transfection for the inner ear. HYA and chitosans loaded with the ototoxic drug neomycin as tracer for drug release have been instilled into the middle ear of the guinea pigs. Effects on RWM and cochlear hair cells were evaluated after a single instillation of HYA (day 7 and 28), chitosans and saline solution (day 7). The hearing organ was analysed for hair cell loss and the thickness and ultrastructural properties of the RWM were analysed by light and transmission electron microscopy. The in vitro transfection efficiency of chitosan was tested by exposing organotypic cultures of the organ of Corti, prepared from postnatal day 2 rats, to chitosan carrying plasmid DNA (pDNA). The in vivo transfection efficiency was tested at one day or seven days after infusing chitosan/pDNA polyplexes with the use of osmotic pumps into the cochlea of adult guinea pigs. Tissue analysis was made by immunohistochemsitry and RT-PCR. HYA and chitosans, especially glycosylated derivative, are safe vehicles that can be used for drug transport into the inner ear through the RWM. Both vehicles successfully released the loaded neomycin, which exerted toxic effects on cochlear hair cells in a degree depending on the concentrations used. The vehicles per se had no noxious effect on the cochlear hair cells but they provoked a comparable effect on the thickness and morphology of the RWM. The thickness of the RWM returned to normal 4 weeks after exposure to HYA. Chitosan as a carrier for inner ear transfection, was associated with inconsistent transfection in vitro and in vivo . The importance of the RWM as a portal for local therapy of inner ear disorders is highlighted in this thesis by focusing on efficiency and effects of the vehicles, applied to the RWM for delivering biologically active agents into the cochlea. The difficulties and variability associated with applying substances to the RWM were explored. The results of this thesis add new knowledge concerning mechanisms of passage of biologically active agents through the RWM and may help us to better understand the role of RWM in the local cochlear therapy and problems of local treatment of inner ear diseases.

Background: Sensorineural hearing loss is a significant problem worldwide and a condition that is not completely cured by currently available therapy. Gene therapy of the inner ear offers an exciting alternative and it has been suggested that this therapeutic modality could be used in treatment aiming at preventing, reversing or managing cochlear disorders. Because of their desired properties as an alternative to the viral vectors, non-viral vectors have been extensively explored for gene delivery. One example is chitosan, a biodegradable cationic polymer.

Objective: To evaluate the in vitro and in vivo transfection efficiency of chitosan as a non-viral gene carrier for gene delivery to cells of the inner ear.

Materials and Methods: Organotypic cultures of the hearing organ, the organ of Corti, were prepared from postnatal day 2 rats, and exposed to chitosan carrying plasmid DNA (pDNA) encoding for green fluorescent protein (GFP) for 24-48 hours. The in vivo transfection efficiency was tested at two time points, at one day or seven days after infusing chitosan/pDNA polyplexes through osmotic pumps into the cochlea of adult guinea pigs (n=41). The tissue was then processed for anti-GFP immunostaining (in vitro and in vivo) and RT-PCR (in vivo).

Results: The in vitro assessment showed prominent GPF transfection after 24-48 hours, while the in vivo GFP transfection in the inner ear was inconsistent and did not show good correlation with the in vitro transfection. Conclusion: It can be concluded that the using chitosan as a carrier for the in vivo transfection, is associated with varying and in consistent degree of transfection.

Objective: This study aimed to elucidate the effect of Staphylococcus aureus and budesonide on the mRNA expression and the biologic role (caspase-1 activation and IL-1β secretion) of NLRP3 inflammasome in primary nasal epithelial cells (NECs) in patients with chronic rhinosinusitis (CRS) and healthy controls.

Material and Methods: Brush biopsies isolated from both patients and healthy controls were denoted respectively for our experiments. These were treated with S. aureus strains (four strains) only and in combination with budesonide (0, 10, 100, 1000 nM). NECs treated with only budesonide (0, 10, 100, 1000 nM) and untreated NECs were used as controls. Expression of NLRP3, caspase-1, IL-1β along with NLRC1/2 was analyzed by qPCR. Caspase-1 activity was measured by fluorogenic substrates Ac-YVAD-AMC. Enzyme-linked immunosorbent (ELISA) assay was performed to measure IL-1β production.

Results: The mRNA levels of NLRC1, NLRC2, caspase-1, and IL-1β significantly increased, while NLRP3 demonstrated a trend toward elevation in the CRS group compared to the healthy controls. Infection with S. aureus increased caspase-1 activity and IL-1β secretion. However, treatment with budesonide decreased mRNA expression of NLRC2 and IL-1β secretion.

Conclusion: Increase in the caspase-1 activity and IL-1β levels, due to possible activation of NLRP3 inflammasomes, upon S. aureus infection, may have an important role in the pathogenesis of CRS.

OBJECTIVE: Establishing methods for topical administration of drugs to the inner ear have great clinical relevance and potential even in a relatively short perspective. To evaluate the efficacy of sodium hyaluronate (HYA) as a vehicle for drugs that could be used for treatment of inner ear disorders.

METHODS: The cochlear hair cell loss and round window membrane (RWM) morphology were investigated after topical application of neomycin and HYA into the middle ear. Sixty-five albino guinea pigs were used and divided into groups depending on the type of the treatment. Neomycin was chosen as tracer for drug release and pharmacodynamic effect. HYA loaded with 3 different concentrations of neomycin was injected to the middle ear cavity of guinea pigs. Phalloidin stained surface preparations of the organ of Corti were used to estimate hair cell loss induced by neomycin. The thickness of the midportion of the RWM was measured and compared with that of controls using light and electron microscopy. All animal procedures were pe rformed in accordance with the ethical standards of Karolinska Institutet.

RESULT: Neomycin induced a considerable hair cell loss in guinea pigs receiving a middle ear injection of HYA loaded with the drug, demonstrating that neomycin was released from the gel and delivered to the inner ear. The resulting hair cell loss showed a clear dose-dependence. Only small differences in hair cell loss were noted between animals receiving neomycin solution and animals exposed to neomycin in HYA suggesting that the vehicle neither facilitated nor hindered drug transport between the middle ear cavity and the inner ear. One week after topical application, the thickness of the RWM had increased and was dependent upon the concentration of neomycin administered to the middle ear. At 4 weeks the thickness of the RWM had returned to normal.

CONCLUSION: HYA is a safe vehicle for drugs aimed to pass into the inner ear through the RWM. Neomycin was released from HYA and transported into the inner ear as evidenced by hair cell loss.

Background: One alternative for controlled drug delivery to the inner ear is application of medication to the middle ear cavity on the promise that it will diffuse through the round window membrane (RWM) into the inner ear.

Objective: to evaluate the efficacy of hyaluronic acid gel (HYA) as a vehicle for drugs that are aimed to treat inner ear disorders.

Methods: in this study the cochlear hair cell loss and RWM morphology were investigated after topical application. Neomycin was chosen as tracer for drug release and the ototoxic effect was evaluated. HYA, (0.5%) loaded with 3 different concentrations of neomycin, was injected to the middle ear cavity of guinea pigs. the ototoxic effect of neomycin in HYA gel was compared to that of neomycin solution applied to the middle ear cavity. Phalloidin stained surface preparation of the organ of Corti was used to estimate hair cell loss induced by neomycin. the thickness of the midportion of the RWM was measured and compared with that of controls using light and electrom microscope.

Result: Neomycin induced a considerable hair cell loss in guinea pigs receiving middle ear injection of HYA loaded with the drug. One week after topical application the thickness of the RWM was dependent upon the concentration of neomycin administered to the middle ear. At 4 weeks the thickness of the RWM had returned to normal.

Conclusion: HYA is a safe vehicle for drugs aimed to pass into the inner ear through the RWM. Neomycin was released from HYA and transported into the inner ear as evidenced by hair cell loss. Key words: Round window membrane (RWM), hyaluronic acid gel, HYA, hair cell loss, thickness.

Background and objectives: Chronic rhinosinusitis (CRS) is an inflammatory disease of the nose and the paranasal sinuses, often associated with an infection by Staphylococcus aureus (S. aureus). Disturbance in the function of ion channels is regarded as an etiological factor for pathogenesis of CRS. The study aims to measure the mRNA expression of the ENaC and CFTR ion channels in nasal epithelial cells (NECs) and to investigate the effect of both the budesonide and S. aureus on these ion channels.

Materials and Method: NECs biopsies obtained from healthy volunteers and patients with CRS. NECs were infected with S. aureus strains and/or budesonide to study the mRNA expression levels of the ENaC and CFTR ion channels.

Results: The mRNA expression level of CFTR was increased while that of ENaC was decreased. S. aureus infection and budesonide treatment induced a significant modulation of ENaC and CFTR ion channels expression.

Conclusion: The CFTR and ENaC ion channel physiology are of importance in the pathogenesis of CRS. Exposure to S. aureus infection and treatment with budesonide modulated the mRNA expression of CFTR and ENaC ion channels. The study is an effort for a better understanding of the pathophysiology of CRS. In order to reach further knowledge, studies on functional properties of the ENaC and CFTR ion channels are necessary.

BACKGROUND: Chronic rhinosinusitis (CRS) is an inflammatory disease of the nose and the paranasal sinuses, often associated with an infection by Staphylococcus aureus (S. aureus). Disturbance in the function of ion channels is regarded as an etiological factor for pathogenesis of CRS.

AIMS: The study aims to measure the mRNA expression of the ENaC and CFTR ion channels in nasal epithelial cells (NECs) and to investigate the effect of both the budesonide and S. aureus on these ion channels.

MATERIALS AND METHOD: NECs biopsies obtained from healthy volunteers and patients with CRS. NECs were infected with S. aureus strains and/or budesonide to study the mRNA expression levels of the ENaC and CFTR ion channels.

RESULTS: The mRNA expression level of CFTR was increased while that of ENaC was decreased. S. aureus infection and budesonide treatment induced a significant modulation of ENaC and CFTR ion channels expression.

CONCLUSION: The CFTR and ENaC ion channel physiology are of importance in the pathogenesis of CRS. Exposure to S. aureus infection and treatment with budesonide modulated the mRNA expression of CFTR and ENaC ion channels.

SIGNIFICANCE: Better understanding of the pathophysiology of CRS.

Development of efficient local delivery systems for the auditory organ has an important role in clinical practice for the management of inner ear disorders using pharmacological means. Chitosan, a biodegradable polymer, is a good drug carrier with bioadhesive properties. The aim of this study was to investigate the feasibility of using chitosan to deliver drugs to the inner ear across the round window membrane (RWM). Three structurally different chitosans loaded with a tracer drug, neomycin, were injected into the middle ear cavity of albino guinea pigs (n=35). After 7 days the effect of chitosans and neomycin was compared among the treatment groups. The hearing organ was analysed for hair cell loss and the RWM evaluated in term of thickness. All tested chitosan formulations successfully released the loaded neomycin which then diffused across the RWM, and exerted ototoxic effect on the cochlear hair cells in a degree depending on the concentrations used. Chitosans per se had no noxious effect on the cochlear hair cells. It is concluded that the chitosans, and especially glycosylated derivative, are safe and effective carriers for inner ear therapy.

Development of efficient local delivery systems for the auditory organ has an important role in clinical practice for the management of inner ear disorders using pharmacological means. Chitosan, a biodegradable polymer, is a good drug carrier with bioadhesive properties. The aim of this study was to investigate the feasibility of using chitosan to deliver drugs to the inner ear across the round window membrane (RWM). Three structurally different chitosans loaded with a tracer drug, neomycin, were injected into the middle ear cavity of albino guinea pigs (n=35). After 7 days the effect of chitosans and neomycin was compared among the treatment groups. The hearing organ was analysed for hair cell loss and the RWM evaluated in term of thickness. All tested chitosan formulations successfully released the loaded neomycin which then diffused across the RWM, and exerted ototoxic effect on the cochlear hair cells in a degree depending on the concentrations used. Chitosans per se had no noxious effect on the cochlear hair cells. It is concluded that the chitosans, and especially glycosylated derivative, are safe and effective carriers for inner ear therapy.

OBJECTIVE: This study comprised a long-term follow-up of a cohort of patients with chronic rhinosinusitis (CRS) regarding clinical features and symptomatology.

METHODS: Data from 42 patients with CRS were available from a previous study. Forty of these patients were alive and were contacted for inclusion after approximately 10 years. Patients completed a questionnaire about disease and symptoms, and underwent a clinical examination.

RESULTS: Thirty-four patients (85%) responded and could be included and evaluated. For the participants in this follow-up study median length of time between initial inclusion (C1) and follow-up (C2) was 11 years (range: 8-15). In some patients the CRS shifted phenotype over time, from CRS with nasal polyposis to CRS without nasal polyposis or vice versa. The median total visual analogue score for combined sinonasal symptoms for all patients was statistically significantly reduced at follow-up. For individual patients, scores for nasal congestion, nasal discharge, facial pressure, and hyposmia were also statistically significantly reduced. The most frequently reported symptom-relieving treatments were nasal steroids and saline rinsing of the nose. Self-reported general quality of life was statistically significantly improved at C2 compared to C1.

CONCLUSION: At long-term follow-up, symptoms were generally reduced and patients reported an improved quality of life. Patients can be given hope for eventual symptom relief. CRS is a chronic condition that seems to harbor the ability to alter its phenotype after several years. Topical corticosteroids and saline rinsing of the nose should be emphasized, since patients consider these treatments to be of high value.