Importance of expression and polymorphisms of Toll-like receptors 2 and 4 in inflammatory diseases of the middle ear and its complications

Uvod: Receptori slični toll-u (eng. Toll like receptors, TLR) imaju bitnu ulogu u aktivaciji početnog imunskog odgovora. Svaki opisani TLR prepoznaje određene mikrobiološke strukture ili molekulske obrasce patogena (eng. pathogen-associated molecular patterns, PAMP) i vezuje specifične ligande. Smatra se da je regulacija ekspresije TLR ima ulogu u patogenezi hroničnog otitisa, i da polimorfizmi u genima za TLR 2 i TLR4 mogu biti asocirani sa podložnošću akutnom otitisu i sekretornom hroničnom otitisu. Ova studija je imala za cilj da utvrdi postojanje ekspresije TLR2 i 4 na izmenjenoj sluznici srednjeg uva kod različitih formi hroničnih otitisa, kao i uticaj polimorfizma Arg753Gln u genu za TLR2 i polimorfizma Thr399Ile u genu za TLR4 i prisustvo bakterija na intenzitet početnog imunskog odgovora. Matrijal i metod istraživanja: Ova studija preseka je sprovedena na Klinici za otorinolaringologiju i maksilofacijalnu hirurgiju Kliničkog centra Srbije; Institutu za patologiju Medicinskog fakulteta Univerziteta u Beogradu i Laboratoriji za Radiobiologiju i molekularnu genetiku Instituta za nuklearne nauke Vinča u Beogradu, Univerziteta u Beogradu, u periodu od januara 2010.do decembra 2013 godine. Svi ispitanici su lečeni na Klinici za otorinolaringologiju i maksilofacijalnu hirurgiju Kliničkog Centra Srbije. Kontrolna grupa za ispitivanje ekspresije gena i proteina na sluznici srednjeg uva obuhvatila je 10 uzoraka makroskopski zdrave sluznice uzetih sa različitih delova srednjeg uva pacijenata lečenih od otoskleroze, kao nezapaljenskog procesa.. Kontrolna grupa za ispitivanje polimorfizama DNK obuhvatila je 100 uzoraka pune krvi pacijenata. Eksperimentalna grupa je obuhvatila uzorke tkiva i pune krvi 85 ispitanika; 45 (15 dece i 30 odraslih) operativno lečenih zbog hroničnog supurativnog otitisa bez holesteatoma; 30 (10 dece i 20 odraslih) zbog hroničnog otitisa sa holesteatomom; 10 bolesnika sa komplikacijama otitisa. Ekspresija proteina TLR 2 i TLR 4 se utvrđivala imunohistohemijskom metodom primenom specifičnih TLR 2 i TLR4 antitela...Introduction: Toll-like receptors (TLRs) have a prominent role in inducing innate immune response. Each of described TLRs recognizes conserved microbial structures or pathogen-associated molecular patterns (PAMP) and has distinct ligands specifities. It has been suggested that regulation of TLRs is involved in the pathogenesis of chronic otitis media, and that TLR2 and TLR 4 polymorphisms were more or less successfully connected with susceptibility to acute otitis and chronic otitis with effusion. In this study, we wanted to establish expression of TLR 2 and 4 on middle ear mucosa in different types of chronic otitis media, and the influence of polymophisms TLR 2 rg753Gln and TLR 4 Thr399Ile and bacterial infection on the intensity of innate immune response. Material and methods: This study was conducted on Clinic for Otorhinolaryngology and Maxillofacial surgery, Clinical Centre of Serbia; Institute of Pathology of Medical Faculty in Belgrade and Laboratory for Radiobiology and Molecular Genetics of Vinca Institute of Nuclear Sciences, in the period from January 2010. to December 2013. All patients and control group subjects were treated on Clinic for otorhinolaryngology and maxillofacial surgery, Clinical Centre of Serbia in Belgrade. Control group for mucosal TLR expression consisted of 10 samles of middle ear mucosa taken from patients with otosclerosis. Control group for DNA polymophism detection consisted of 100 full blood samples in healthy subjects. Experimental group consisted of mucosal tissue samples and full blood samples of 85 patients; 45 treated of chronic otitis without cholesteatoma (15 children and 30 adults), 30 treated of chronic otitis with cholesteatoma (10 children and 20 adults) and 10 patients with complications of chronic otitis media with or without cholesteatoma. Expression of TLR 2 and 4 on middle ear mucosa was determined with immunohistochemical staining..

Uticaj slušnih pomagala na auditivnu percepciju i neposredno verbalno pamćenje kod dece sa bimodalnom stimulacijom

Introduction: The combination of electric stimulation from cochlear implant (CI) with acoustic stimulation from hearing aid (HA), otherwise known as bimodal hearing, may provide several binaural benefits including binaural summation, binaural squelch, reduction of the head shadow effect, and improved localization. Purpose: This study investigated the influence of preoperative rehabilitation and bilateral HA use, bimodal stimulation post-implantation (CI on one ear and HA on the non-implanted ear) and hearing thresholds in the verbal short-term memory. Method: Immediate verbal memory test for Serbian language consisting of four subtests was used for auditory perception testing on 21 pre-lingually deaf children. Results: Duration of bimodal hearing proved to be significant in the terms of auditory perception and verbal short-term memory. Mid- and high-frequency amplified thresholds on the non-implanted ear were correlated with poorer perception and reproduction of monosyllables and nonsense words. Conclusion: Duration of bimodal hearing proved to be significant in the terms of auditory perception, speech reproduction and semantic ability. Patients with a unilateral cochlear implant who have measurable residual hearing in the non-implanted ear should be individually fitted with a hearing aid in that ear, to improve speech perception and maximize binaural sensitivity.Kombinacija električne stimulacije kohlearnog implanta (KI) i akustične stimulacije slušnog pomagala (SP), poznata kao bimodalni sluh, može imati razne binauralne prednosti koje uključuju binauralnu stimulaciju, binauralno sažimanje, redukciju eho efekta i unapređenje lokalizacije izvora zvuka. Cilj: U ovom istraživanju je ispitan uticaj preoperativne rehabilitacije i upotrebe bilateralnog slušnog pomagala, bimodalne stimulacije nakon implantacije (KI na jednom uhu i SP na neimplantiranom uhu) i pragova sluha u implantiranom i neimplantiranom uhu na auditivnu percepciju i verbalno kratkotrajno pamćenje. Metod: Za ispitivanje auditivne percepcije kod dvadeset jednog prelingvalno gluvog deteta korišćen je Test za ispitivanje verbalnog pamćenja za srpski jezik, koji se sastoji od četiri podtesta. Rezultati: Pokazalo se da je trajanje bimodalnog sluha značajno kod auditivne percepcije i verbalnog kratkotrajnog pamćenja. Povećani pragovi srednje i visoke frekvencije na neimplantiranom uhu bili su u korelaciji sa slabijom percepcijom i reprodukcijom jednosložnih i besmislenih reči. Zaključak: Pokazalo se da je trajanje bimodalnog sluha značajno za auditivnu percepciju, reprodukciju govora i semantičku sposobnost. Pacijentima sa unilateralnim kohlearnim implantom, sa merljivim rezidualnim sluhom na neimplantiranom uhu, trebalo bi ugraditi slušno pomagalo u to uho, kako bi se poboljšala percepcija govora i maksimizovala binauralna osetljivost

Toll like receptors 2 and 4 polymorphisms in chronic otitis media

Toll like receptors (TLRs) are the pattern recognition receptors, important for the innate immune system, the first line of defense against bacterial infections which are the main cause of otitis media (OM). Our aim was to investigate the association of TLR4 (Thr399Ile and Asp299Gly) and TLR2 (Arg753Gln) most common polymorphisms with OM prevalence as well as with morphologic changes of middle ear during chronic otitis media. We had 111 controls and 186 patients divided into three groups of COM: nonsuppurative OM, suppurative OM and cholesteatoma. Frequencies of the genotypes of the TLR 2 gene polymorphism were significantly different between men and women having nonsuppurative OM (p=0.03).5th Congress of the Serbian Genetic Society : Book of abstracts; September 28 - October 2, Kladovo, 2014

Receptive speech in early implanted children later diagnosed with autism

Introduction: Incidence of children with autism spectrum disorder (ASD) is rising through the years with estimated 1 in 68 in the US in 2014. This incidence is also rising in the population of congenitally deaf children. Favorable outcome after early cochlear implantation is expected due to plasticity and reorganization capacity of brain in infants and toddlers, but outcomes could be significantly modified in children with diagnosed ASD. Current methods of screening for autism have difficulties in establishing diagnosis in children who have both autism and other developmental delays, especially at such an early age. The aim of the study was to assess the development of auditory perception and speech intelligibility in implanted children with profound congenital hearing loss who were diagnosed with ASD comparing to those who were typically developing. Material and methods: Fourteen children underwent cochlear implantation; four were later diagnosed with ASD and ten were typically developing. All children underwent intensive postoperative speech and hearing therapy. The development of auditory perception and speech intelligibility was assessed using the Categories of Auditory Performance (CAP) and the Speech Intelligibility Rating (SIR) during the 5-years follow-up. Results: In children later diagnosed with ASD, auditory processing developed slowly. Depending on the individual capabilities, by the age of six they could identify environmental sounds or discriminate speech sounds. Speech Intelligibility in children with ASD was at best rated as category 2, with very little or no progress up to the age of six, despite extensive speech and language therapy. Communication skills were strongly affected by a degree of autistic features expression. Conclusion: Preoperative psychological assessment in congenitally deaf infants should be expanded by the use of validated instruments for early detection of autism. The possibility of developing ASD should be kept in mind by all professionals involved in programs for cochlear implantation

Middle ear tuberculosis: Diagnostic criteria

Introduction. Tuberculous otitis is a diagnostic problem due to the difficulty to obtain microbiological, histomorphological and cytological confirmation of the disease. Objective. Our objective was to compare clinical and radiological characteristic and development of otogenic complications in patients with tuberculous otitis and otitis with cholesteatoma as the most destructive form of chronic nonspecific otitis in the purpose of establishing the diagnostic criteria for tuberculous otitis. Methods. Medical records of 12 patients with tuberculous otitis and 163 patients with cholesteatoma treated at the Institute of Otorhinolaryngology and Maxillofacial Surgery in Belgrade during the eight-year period were analyzed. All of the patients underwent otomicroscopic, audiological and radiological examination of the thorax and temporal bone, microbiological examination of the secretion and histomorphological examination of the tissue taken during middle ear surgery. Statistical analysis was done using χ2 test with Yates correction. Results. Otogenic complication as facial palsy and sensorineural hearing loss were more frequent in tuberculous otitis patients, than in cholesteatoma. Also, fistulas of the labyrinth and facial canal bone destruction were also more frequent in tuberculous otitis than in cholesteatoma. A larger extent of temporal bone destruction was noticed on CT scans of the temporal bone in half of the patents with tuberculous otitis. Coexistence with miliary pulmonary tuberculosis was detected in one third of the patients. There were no microbiological or histomorphological confirmations of the disease, except in one case with positive Ziehl-Neelsen staining. Conclusion. Tuberculous otitis media should be considered in patients with serious otogenic complications and with shorter duration of ear discharge, and in association with diagnosed miliary pulmonary tuberculosis and extensive temporal bone destruction. Polymerase chain reaction still is not reliable for diagnosis

Detection and sampling methods for isolation of Candida spp. from oral cavities in diabetics and non-diabetics

The purpose of this study was to detect Candida spp. on the tongue and in the subgingival sites in healthy and type 2 diabetes (T2D) patients with chronic periodontitis (CP), and to compare the accuracy of sampling methods. This study included 131 patients divided into four groups: healthy control (group A), nondiabetics + CP (Group B), diabetics with good metabolic control + CP (group C) and diabetics with poor glycoregulation + CP (Group D). Cotton swab samples from tongue and subgingival samples were obtained from each patient with help of sterile paper points and a sterile curette. Swab cultures were made on Sabouraud dextrose agar. The number of CFUs was counted. The sampling methods for subgingival plaque were compared by Receiving Operator Curve (ROC). The presence of Candida spp. on the tongue was statistically significant among groups (group D vs. others three groups:.2: p lt 0.005 for each group). Positive findings of subgingival Candida spp. did not differ among the groups. There were no significant differences in the quantification of Candida spp., neither on the tongue, nor in the subgingival samples. 17.2% of diabetic patients revealed the presence of Candida spp. in the subgingival samples, with negative finding on tongue. There was a significant difference in the sampling methods for subgingival plaque (p = 0.000). Candida spp. is more prevalent on the tongue of diabetics. The sampling of subgingival plaque by a sterile curette is more accurate than with paper points. Subgingival plaque may represent a reservoir of commensals. It is necessary to standardize the sampling of subgingival plaque

Covid-19 pandemic and hearing loss in children

Pandemija COVID-19 je izazvala ogromne promene u pružanju zdravstvenih usluga na svim nivoima zdravstvene zaštite. Posledično velike su šanse za neblago- vremeno dijagnostikovanje i lečenje pojedinih bolesti. Cilj ovog rada je bio da se, na osnovu dostupne literature, utvrdi kakav je uticaj pandemije COVID-19 na skrining sluha i ranu audiološku procenu, kao i na dostupne načine lečenja oštećenja sluha kod dece. Problemi tokom pandemije su nastali na nivou sprovođenja skrininga sluha, u vidu smanjenog broja dece koja su prošla skirining ili koja su upućena na lečenje dijagnostikovanog oštećenja sluha. Pandemija je izazvala ukidanje ili odlaganje elektivnog operativnog programa, što je doprinelo obustavljanju programa kohlearne implantacije na određene vremenske periode. Problemi sa kojima su se sretali pacijenti i članovi njihovih porodica posle implantacije su bili vezani za same zdravstvene ustanove gde se programi kohlearne implantacije sprovode, za održavanje uređaja i nabavku rezervnih ili zamenskih delova za od- loženo uključivanje ili programiranje uređaja i za propuštanje tretmana re/ habilitacije i za rad kod kuće. Sigurno je da će postojeći zastoji u pružanju na- vedenih zadravstvenih usluga imati dalekosežne posledice za generacije koje je pogodila COVID-19 pandemija. Dalja istraživanja će biti potrebna da se u potpunosti proceni njihov obim i veličina potencijalne štete.The COVID-19 pandemic has caused huge changes in the provision of health services at all levels of health care. As a result, chanses were increased for untimely diagnosis and maltreatment of certain diseases. The aim of this study was to determine, based on the available literature, the impact of the COVID-19 pandemic on hearing screening and early audiological assessment, as well as on available treatments for hearing impairment in children. During the pandemic, reduced number of children underwent screening and were referred for treatment of diagnosed hearing impairments. The pandemic caused the cancellation or postponement of the elective operative program, which contributed to the suspension of the cochlear implantation program for certain periods of time. The problems encountered by patients and their family members after implantation were related to the health facilities where cochlear implantation programs are conducted, to device maintenance or getting replacement parts, delayed programing or switch on appointments and missing Rehabilitation Session. It is certain that the existing delays in the providing these health services will have far-reaching consequences for the generations affected by the COVID-19 pandemic. Further research will be needed to fully assess their extent and magnitude of potential damage

The impact of covid-19 pandemic on hearing loss and tinnitus

Od decembra 2019. godine korona virusna bolest (COVID-19) izazvana novim, do sada nepoznatim, koronavirusom nazvanim SARS-CoV2, ubrzo je postala pandemija koja je promenila svet i ceo zdravstveni sistem. O efektima i dugotrajnim sek- velama COVID-19 infekcije na senzorne organe a posebno na unutrašnje uvo, nema dovoljno podataka jer je primarni cilj lečenja spašavanje života pacijenata sa teškim simptomima. Poznato je da su virusi etiološki faktori različi- tih vrsta oštećenja sluha i pojave tinitusa pa su tako započeta itraživnja negativnog uticaja novog SARS-CoV2 virusa na audio-vestibularne simptome. Mogući patofiziološki mehanizmi oštećenja sluha i tinitusa su: direktna infekcija auditivnog puta i kohlearnog nerva, udaljeni efekti sistemske infekcije sa infekcijom kohlee, vaskularno oštećenje, autoimuni odgovor pro- tiv tkiva unutrašnjeg uva, a moguće je da se neki od ovih mehanizama prepliću. Cilj ovoga rada je bio da se pregledom dostupne literature sagledaju učesta- lost i patofiziološki mehanizmi oštećenja sluha i pojave tinitusa kod pacijenata sa COVID-19 infekcijom.Since December 2019, coronavirus disease (COVID-19) caused by a new hitherto unknown coronavirus called SARS-CoV-2, soon became a pandemic that changed the world and the entire health system. There are insufficient data on the effect and long-term sequelae of COVID-19 infection on sensory organs and especially on the inner ear, because the primary goal of treatment is to save the lives of patients with severe symptoms. It is known that viruses are etiological factors of various types of hearing impairment and the appearance of tinnitus, so the research for the negative impact of the new SARS- CoV2-2 virus on audio-vestibular symptoms has begun. Possible pathophysiological mechanisms of hearing loss and tinnitus are: direct infection on the auditory pathway and cochlear nerve, long term effect of systemic infection with cochlear infection, vascular damage, autoimmune sesponse against inner ear tissue, and some of these meschanisms may intertwine. The aim of this study was to study the frequency and pathophysiological mechanisms of hearing impairment and tinnitus in patients with COVID-19 infection by reviewing the available literature

Rresence of different Candida species at denture wearers with type 2 diabetes and clinically healthy oral mucosa: Pilot study

Background/Aim: The aim of this study was to examine prevalence of different Candida spp. at diabetics and nondiabetics wearing dentures without clinical signs of Denture Stomatitis (DS) and to study if some local and systematic factors are confounders for harboring Candida at these subjects. Material and Methods: Total of 60 subjects wearing partial or complete upper acrylic denture having at least half of palatal mucosa covered by denture were selected and stratified into three experimental groups: systematically health subjects; patients with diagnosed Type 2 Diabetes (T2D) and good glycoregulation; and T2D subjects with poorly regulated blood sugar level. Cotton swab samples were obtained from each patient from hard palate mucosa and denture surface. Swab cultures were made on Sabouraud dextrose agar and ChromAgar Media for distinciton of various Candida spp. Density growth was also measured. Results: Frequency of Candida spp. findings were similar between groups. At healthy subjects, only C.albicans was detected. At diabetics, C.albicans was the most common isolated species, followed by C.glabrata and C.tropicalis. Negative finding of yeasts on palatal mucosa, but positive on denture surface were detected at all groups, with the highest frequency (33.4%) at diabetics with poor glycoregulation. Denture surface was heavier colonized than hard palate mucosa. Duration of diabetes in years were only independent predictors for harboring Candida spp. at denture surface (Exp B=1.186, CI=1.047-1.344, p=0.007). Conclusions: Prosthesis of denture wearers without DS may serve as reservoir of Candida spp. Presence of more pathogenic and resistant non-albicans species are related to diabetics, even without clinical signs of DS

Korelacija između koncentracije receptora 2 faktora nekroze tumora u serumu i destrukcije parodoncijuma kod bolesnika sa dijabetes melitusom tip 2 - studija preseka

Introduction: The role of tumor necrosis factor-α (TNFα) is well documented in pathogenesis of chronic periodontitis (CP) and type 2 diabetes (T2D). Considering short half-life of TNFα, tumor necrosis factor receptor-2 (TNFR2) is used as prosperous surrogate marker of TNFα activity. Objective The aim was to detect TNFR2 serum concentration and correlate it with periodontal destruction in patients with diagnosed T2D and nondiabetics. Methods The study included 85 patients divided into three groups: T2D + CP (group T2D, n = 34); nondiabetics + CP (Group PD, n = 27); and healthy controls (group HC, n = 24). T2D was diagnosed according to WHO criteria (2013) and periodontitis was diagnosed using International Workshop for a Classification of Periodontal Diseases and Conditions criteria (1999). TNFR2 level was measured by enzyme-linked immunosorbent assay (ELISA). Results There was no difference in TNFR2 level among the groups (Kruskal-Wallis, p = 0.482). Significant correlation (Pearson's correlation coefficient) was observed between clinical attachment loss (CAL) and TNFR2 concentration in PD group (rp = -0.460, p = 0.016). In T2D group, correlations were observed between TNFR2 concentration and CaL (rp = 0.363, p = 0.005) and periodontal inflamed surface area (PISA) (rp = 0.345, p = 0.046) and periodontalepithelial surface area (PESA) (rp = 0.578, p = 0.000). Conclusion Higher concentration of TNFR2 was associated with higher CAL, PESA, and PISA scores in T2D group. Contrary to that, nondiabetics with higher values of CAL exhibited lower concentration of TNFR2, presenting potential protective effect on periodontal destruction. These results imply that diabetes may alter TNFR2 secretion originated from periodontium.Uvod: Uloga faktora nekroze tumora-alfa (TNFα) dokazana je u patogenezi hronične parodontopatije (HP) i dijabetesa melitusa tipa 2 (DM tip 2). S obzirom na to da je poluživot TNFα veoma kratak, receptor 2 faktora nekroze tumora (TNFR2) koristi se kao marker aktivnosti TNFα. Cilj rada Cilj ovog rada je određivanje koncentracije TNFR2 u serumu i koreliranje sa parametrima destrukcije parodoncijuma kod zdravih i ispitanika sa dijagnostikovanim DM tip 2. Metode rada U studiju je uključeno 85 pacijenata podeljenih u tri grupe: DM tip 2 + HP (DM grupa, n = 34), zdravi ispitanici + HP (PD grupa, n = 27) i zdrave kontrole (ZK grupa, n = 24). Dijagnoza DM tip 2 postavljena je na osnovu kriterijuma SZO (2013), dok je dijagnoza HP postavljena na osnovu kriterijuma Internacionalne radionice za klasifikaciju stanja i oboljenja parodoncijuma (1999). Koncentracija TNFR2 merena je ELISA metodom. Rezultati Koncentracija serumskog TNFR2 nije se razlikovala među grupama (Kraskal-Volis, p = 0,482). Postoji značajna korelacija (Pirson) između nivoa pripojnog epitela (NPE) i koncentracije TNFR2 u PD grupi (rp = -0,460, p = 0,016). U DM tip 2 grupi, statistički značajna korelacija uočena je između koncentracije TNFR2 i NPE (rp = 0,363, p = 0,005), kao i parametara uticaja inflamacije iz parodoncijuma na sistemsko zdravlje - PISA (rp = 0,345, p = 0,046) i PESA (rp = 0,578, p = 0,000). Zaključak Kod pacijenata sa dijabetesom veće koncentracije TNFR2 odgovaraju većim vrednostima NPE, PESA i PISA. Nasuprot tome, kod sistemski zdravih ispitanika sa HP veće vrednosti NPE su povezane sa manjim koncentracijama TNFR2, što bi moglo govoriti o potencijalnoj zaštitnoj ulozi ovog citokina na destrukciju parodoncijuma. Rezultati govore da dijabetes može uticati na sekreciju TNFR2 iz parodoncijuma