P < 0.001). Linear regression analyses of pure-tone averages at different frequencies, before and after surgery, demonstrated that patients benefitted from a postoperative hearing gain at low and middle frequencies, but their hearing often deteriorated at frequencies of 8000 Hz. As for the impact of the type of tympanoplasty on hearing outcomes, type III-interposition markedly increased hearing gain. The overall rate of postoperative adverse events was 8.8%. We conclude that reconstruction of the ear canal and mastoid via mastoid exclusion/obliteration using bone chips/paté can be considered as an alternative procedure following retrograde mastoidectomy. It gives excellent surgical results and has fewer postoperative adverse events.The aim of this study was to retrospectively review the long-term hearing results and the impact of mastoid exclusion/obliteration in patients with cholesteatoma (102 ears) who underwent retrograde tympanomastoidectomy and in whom bone chips/paté were applied as the sole materials during the procedure. In 79 ears, this was combined with ossiculoplasty in a single-stage procedure. In >71% of ears, the results of audiometric testing were monitored for more than 2 years. The results suggested there was a significant gain in hearing following surgery, with respect to the postoperative change in both air-conduction thresholds and air-bone gaps ( For a long time, the primary objective in the surgical management of cholesteatoma has been the eradication of disease, as well as ensuring a dry and safe ear. Hearing preservation or restoration is also worth considering but is often treated as a secondary goal. Controversies in surgical management of cholesteatoma include the choice in surgical approach, that is, canal wall down (CWD) or canal wall up (CWU), and the need for staged operations , 2.Although cholesteatoma can develop in up to 10% of people with chronic suppurative otitis media, its actual pathogenesis is still unknown . In pastIn 1995, the National Health Insurance System was introduced in Taiwan, which markedly influenced people's behavior in seeking medical advice. As a consequence, more patients with cholesteatoma are presenting with earlier stages of the disease and fewer symptoms of hearing destruction. Therefore, as the clinical characteristics of extensive cholesteatoma have changed, the demand for hearing preservation has increased. This makes hearing preservation and restoration as important as disease eradication when considering surgical removal of cholesteatoma.CWD mastoidectomy allows for better visualization, greater assurance of cholesteatoma eradication, and a lower recidivism rate than CWU, but at the expense of the need for life-long mastoid care. On the other hand, CWU mastoidectomy is not associated with mastoid bowl problems and allows for good hearing outcomes. However, it is associated with a higher recidivism rate than CWD owing to poor visualization, and an increased likelihood of the postoperative formation of a retraction pocket. Retrograde tympanomastoidectomy combines the virtues of both CWD and CWU procedures by the partial or entire removal of the posterior bony canal wall, depending on disease extent , 6. MostEarly attempts to carry out mastoid obliteration mostly used local flaps , 8. SincThis retrospective study aims to evaluate the impact of mastoid exclusion/obliteration in retrograde tympanomastoidectomy for patients with cholesteatoma, using bone chips/paté as the sole materials, and with an emphasis on its long-term postoperative audiometric outcomes.This was a retrospective chart review study, evaluating the surgical outcome and pure-tone audiograms of patients with cholesteatoma, before and after surgical treatment. The research was approved by the Institutional Review Board of Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan (Protocol number 2-103-05-139). Study subjects included patients with cholesteatoma who were treated at the Department of Otolaryngology-Head and Neck Surgery, Tri-Service General Hospital, Taipei, Taiwan, from January 2004 to October 2013. A total of 99 patients (102 ears) were enrolled in this study, all of whom underwent retrograde tympanomastoidectomy; in 79 ears, this was combined with ossiculoplasty in a single-stage procedure .The medical records of patients with cholesteatoma who underwent retrograde tympanomastoidectomy by the senior author (CHW), in conjunction with mastoid exclusion/obliteration surgery using bone chips/pate, were included in the study. Exclusion criteria were as follows: (1) diagnosis other than cholesteatoma, (2) the follow-up hearing test was conducted less than one year after surgery, and (3) there was a greater than 50% destruction of the EAC, which was reconstructed with a transposition of the muscle flaps or a CWD procedure. We performed this technique only in cases involving less than 50% destruction of the EAC following retrograde mastoidectomy, due to the feasibility of EAC reconstruction using bone chips/paté as the sole material in these instances. In the face of extensive cholesteatoma resulting in greater destruction of the EAC or CWD procedure, a meatally based musculoperiosteal flap was modified for mastoid obliteration in conjunction with the use of bone chips/paté, but such cases were excluded from this analysis.We retrospectively reviewed each medical record to obtain the patient demographic, the surgical procedure they underwent, the materials used for ossicular chain reconstruction, the length of follow-up, associated complications, and hearing outcomes.All patients underwent preoperative and postoperative pure tone audiometry. It was suggested that patients should complete hearing tests every 6 months after surgery for the first 2 years, and at yearly intervals thereafter. As mentioned in the exclusion criteria, we excluded individuals with audiometric data covering less than 1 year of postoperative follow-up. In addition, postoperative hearing outcomes were determined using the audiogram records at the latest follow-up. A four-frequency pure-tone average (PTA) was calculated from the average pure-tone audiometry at 500, 1000, 2000, and 4000 Hz. The audiometry results were reported according to the Committee on Hearing and Equilibrium guidelines, except for thresholds at 3000 Hz, which were substituted with thresholds at 4000 Hz . Pure-toThe degree of hearing loss based on AC-PTA is defined by four levels , measureThe procedure usually involves a postauricular incision and general anesthesia. In the early stages of the mastoidectomy, healthy bone chips and bone paté were harvested with chisels and drills, respectively, from an uninvolved area of the mastoid cortex .The retrograde mastoidectomy was initiated by drilling the ear canal and following the extent of the cholesteatoma until noninvolved cavity with healthy mucosa was reached. This approach led to the exposure of cavities of different sizes among individuals following the mastoidectomy . In caseThe ear canal wall was reconstructed with several pieces of curved bony plates placed on the preserved canal wall and tegmen tympani. The isolated mastoid cavity was filled with bone chips . The bonBy contrast, for cases of mastoid obliteration, the mastoid cavity was filled with bone chips and bone paté in combination with the covering of a superiorly based temporalis muscle fascia after mastoid air cells and the mucosa lining had been eradicated .Drain set placement is not necessary after this operation. To clear up the infection, intravenously administered 1000/200 mg of amoxicillin/clavulanic acid three times a day, plus 80 mg of gentamycin two times a day are usually given in the first 3–5 days after the operation. This was followed by 500 mg of oral ciprofloxacin twice daily for 1 to 2 weeks.Patients whose ossicular chain had either been affected by cholesteatoma or had been removed during surgery were subject to an ossiculoplasty in a single-stage tympanomastoidectomy (79 ears). Types of tympanoplasty performed in this study included types I, III, and IV, denoting different statuses of the middle ear and ossicular defects following the removal of the cholesteatoma. Repositioning of sculptured bone grafts or prostheses was utilized in both type III and type IV tympanoplasties to reconstruct the sound conduction mechanism in the tympanic cavity, in accordance with the classification proposed by the Japan Otological Society in 2010 . These tt-test. Mean ± standard deviation (SD), frequency, and percentages were used to describe the characteristics of the study subjects. All statistical analyses were carried out using the SPSS for Windows . P values below the conventional level of statistical significance (P < 0.05) were considered statistically significant.For descriptive analyses, we used a chi-square analysis and McNemar's test for categorical variables and a one-way analysis of variance (ANOVA) for continuous traits (with Least Significant Difference (LSD) test for post hoc comparison). Differences between the measurements of preoperative and postoperative pure-tone audiometry were tested using the paired Patients' ages ranged between 6 to 76 years. Significantly more patients were surgically treated for cholesteatoma in the age range of 21 to 40 years (37.7%) and 41 to 60 years (44.1%) than the other age groups . We did In this study, we presented two types of reconstruction for EAC and mastoid cavity, using mastoid exclusion in 63 ears (61.8%) and mastoid obliteration in 39 ears (38.2%). In mastoid exclusion, bone chips alone or combined with paté were used to completely isolate the mastoid cavity from the middle ear, but the mastoid cavity was not totally obliterated as is done in the mastoid obliteration technique. In other words, we isolated the mastoid cavity but did not totally block its air from communicating with the tympanic cavity and the mastoid cells. We placed bone chips and bony plate fragments in an interlocking position around the aditus ad antrum region, which not only rebuilds the posterior bony external canal wall but also provides greater strength in order to resist further drum retraction.Tympanoplasty type I was performed in 23 of 102 ears (22.5%), type III in 60 of 102 ears (58.9%), and type IV in 19 of 102 ears (18.7%). The mean follow-up period was 30.2 months, with a range of 18 to 72 months. In more than 71% of ears, the audiometric tests were monitored after more than 2 years . PrimaryP < 0.001). The mean postoperative ABG was 22.25 dB, as compared with 29.22 dB preoperatively . The results of these hearing assessments indicate a significant improvement in hearing gain following surgery.To evaluate the overall hearing outcomes after surgery, we analyzed the change in hearing thresholds obtained from AC and ABGs. As shown in To further investigate the postoperative changes in hearing thresholds at each octave frequency, an equation was derived through linear regression analysis of PTA before and after surgery. As demonstrated in P = 0.04).In addition to investigating postoperative hearing gain , we maden = 8), only 2 cases (25.0%) improved to severe or moderate hearing loss and the majority remained in profound hearing loss postoperatively.From a total of 30 cases of mild preoperative hearing loss, 22 of these (73.3%) retained the same degree of hearing loss postoperatively, and only 8 cases (26.7%) declined to moderate hearing loss. From 49 cases of moderate preoperative hearing loss, 19 cases (38.8%) improved to mild hearing loss, 29 cases (59.2%) remained in moderate hearing loss, and 1 case (2.0%) declined to severe hearing loss postoperatively. From 15 cases of severe preoperative hearing loss, 6 cases (40.0%) improved to moderate hearing loss, 6 cases (40.0%) remained in severe hearing loss, and 3 cases (20.0%) declined to profound hearing loss. Of those with profound hearing loss  and a borderline significant trend was found at 250 (P = 0.13), 500 (P = 0.11), and 1000 Hz (P = 0.08), respectively. Upon correlating the postoperative ABG gains in the different types of tympanoplasty, a distinct trend toward significance was shown at 2000 Hz (P = 0.07), and a borderline significant trend was shown at 1000 (P = 0.13). There was no statistically significant difference at 500 and 4000 Hz.To determine the differences in results between the various types of tympanoplasty, postoperative hearing gain in AC and ABG at each octave frequency is given in To assess for significant differences in hearing gain between types of tympanoplasty, the post hoc test indicated that type III-i was better than type I or type IV-c in AC gain across frequencies between 250 and 2000 Hz. Similarly, in ABG gain, the post hoc test revealed that type III-i was better than type IV-i at 1000 Hz, and type III-i was better than type I or type IV-c at 2000 Hz.t-test. The results demonstrated statistically significant differences (P < 0.001) in AC gain in type III-i from 250 to 2000 Hz, and from 500 to 2000 Hz for ABG gain. As shown in P = 0.01 and P = 0.03) and type I (P = 0.04) but not in type IV-i. Taken together, we may conclude that patients benefit from tympanoplasty surgery by exhibiting different gains at various frequencies. The gain increased markedly in type III-i, followed by type III-c, type I, and type IV-c.Analysis of hearing gain for individual ears at different frequencies, for each type of tympanoplasty, was conducted by paired P = 0.213) . In total, five cases (4.9%) had wound infection, three cases (2.9%) had recurrence, and one case (1%) had residual disease . There w= 0.213) . In the Although the idea of temporarily removing the posterior canal wall to allow for better exposure for cholesteatoma extirpation and subsequently reconstructing the canal wall is not new, there is no known perfect solution. Several modifications to the technique are still under development in an attempt to make such an approach technically feasible in terms of desired outcomes , 19–22.In addition to mastoid obliteration, we offered mastoid exclusion for the purpose of reconstructing the EAC and preventing the development of a retraction pocket. This involved partially, but not totally, obliterating the mastoid cavity. For this technique, the mucosal residues do not need to be removed, as for mastoid obliteration. It is worth emphasizing that no more than 50% of the length of the posterior canal wall was partially removed and should be preserved as much as possible during the retrograde mastoidectomy. Otherwise, the reconstruction becomes more difficult for the reasons pointed out by Dornhoffer  and HinoRegardless of whether matter mastoid exclusion or obliteration is performed, both of those procedures help reconstruct the EAC defect with minimal harvest of nearby bone chips, making the procedure easy. We believe that an ideal procedure should be simple and quick. This is the distinguishing feature of the unique technique proposed in this study.For mastoid exclusion, we placed bone chips and bony plates in an interlocking position at the exposed aditus ad antrum and EAC defect regions to effectively close off the middle ear from the attic and mastoid. Unlike similar mastoid exclusion procedures reported in previous literatures , 22, 25,In contrast, a sclerotic mastoid cavity usually implies the coexistence of a dysfunction of the Eustachian tube and middle ear inflammation. This contributes to the suppression of mastoid air cell development from early childhood . In thisIn this study, both AC and ABG values improved significantly after surgery , indicatIn comparing the effect of the types of tympanoplasty on postoperative hearing gain, type III-i was found to be superior to other types . This hiIn our report, the maximum air gain was at 1000 Hz, which then decreased as the frequencies increased. There was a limited hearing gain above 4000 Hz and an adverse impact at 8000 Hz. It is also noteworthy that ossiculoplasty had the greatest impact on hearing gain at low and mid frequencies (from 250 to 2000 Hz), but only a limited impact on frequencies above 4000 Hz, as shown in Many authors performed staged operation as a second-look procedure and staged ossiculoplasty , 22, 24.With regard to the adverse events of surgery, no significant differences could be demonstrated between the two methods. In research on recurrent and residual cholesteatoma, many authors have determined that epitympanoplasty or epitympanic obliteration is crucial in preventing retraction pocket formation and lowering the residual rate , 24, 25.Although infection became the leading cause of complications and a total of five cases were associated with postoperative wound infections, our techniques are relatively safe compared with those in previous studies , 39–41. The limitations of our study include that the hearing assessment data were obtained from different durations of follow-up, although in more than 71% of ears the audiometric tests were monitored for more than 2 years. Moreover, a second-look procedure was lacking; such a procedure has been highly recommended in CWD for a high incidence of residual cholesteatoma. This raises the question of whether the retrograde mastoidectomy technique necessitates a planned second-stage operation or whether it could be substituted for observation. In fact, in our series, recurrence was never observed in the obliterated/excluded mastoid cavity, and no cases required mastoid obliteration/exclusion to be taken down. Furthermore, in six cases, a two-stage ossiculoplasty was performed, and no residual cholesteatoma was found (data not shown). Since the recurrence rates are related to the length of follow-up , 44, theRetrograde tympanomastoidectomy in conjunction with canal wall reconstruction, using mastoid exclusion/obliteration, provides good hearing outcomes with low complication and recidivism rates. Postoperative hearing gains following successful procedures were largely observed in low and mid frequencies, rather than in frequencies above 4000 Hz. In retrograde tympanomastoidectomy, a mastoid exclusion technique can be substituted for mastoid obliteration in patients with a relatively well-pneumatized mastoid cavity.


CAPS is a rare monogenic autoinflammatory syndrome consisting of a spectrum of three conditions: Familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), and the most severe form, NOMID/CINCA. Progressive hearing loss is a characteristic of severe CAPS . PreviouTo characterize the correlation between different NLRP3 mutations and hearing loss in severe CAPS patients and to evaluate whether long-term anakinra treatment stabilizes the progression of hearing loss.A prospective open-label study with long-term extension including 43 patients with severe CAPS was conducted at the National Institutes of Health ,2. The pAltogether 22 patients provided pre and post treatment audiogram data. NLRP3 mutation was identified in 18 (82%) patients, 7 (32%) presented with D303N and 11 (50%) with other locations . In a multivariate analysis, the baseline 4F-PTA  correlated with both age (p=0.032) and location of NLRP3 mutation (p=0.049) so that older patients and patients with mutations outside of D303N presented with more hearing loss. Following the initiation of the anakinra treatment, no significant changes were seen in mean 4F-PTA at any time point up to 5 years in either patients with D303N or patients with other NLRP3 mutations in the MMRM analysis adjusting for age; thus, hearing remained stable. Comparable findings were seen in the worst ear based on air conduction and for best/worst ear based on bone conduction assessments.Mutation-specific risk for hearing loss which is independent of age was seen based on the baseline audiogram data. Anakinra treatment for up to 5 years stabilized the progression of hearing loss regardless of the mutation.NCT00069329M. Leinonen Consultant for: Swedish Orphan Biovitrum, B. Hallén Employee of: Swedish Orphan Biovitrum, H. Olivecrona Employee of: Swedish Orphan Biovitrum.


Age-related hearing impairment (ARHI) is a complex, multifactorial disorder that is attributable to confounding intrinsic and extrinsic factors. The degree of impairment shows substantial variation between individuals, as is also observed in the senescence of other functions. This individual variation would seem to refute the stereotypical view that hearing deterioration with age is inevitable and may indicate that there is ample scope for preventive intervention. Genetic predisposition could account for a sizable proportion of interindividual variation. Over the past decade or so, tremendous progress has been made through research into the genetics of various forms of hearing impairment, including ARHI and our knowledge of the complex mechanisms of auditory function has increased substantially. Here, we give an overview of recent investigations aimed at identifying the genetic risk factors involved in ARHI and of what we currently know about its pathophysiology. This review is divided into the following sections: (i) genes causing monogenic hearing impairment with phenotypic similarities to ARHI; (ii) genes involved in oxidative stress, biologic stress responses, and mitochondrial dysfunction; and (iii) candidate genes for senescence, other geriatric diseases, and neurodegeneration. Progress and prospects in genetic research are discussed. Age-related hearing impairment (ARHI), also known as presbycusis, is the most common sensory impairment among the elderly. Typical features of ARHI are gradual progression later in life and bilaterally symmetrical sensorineural hearing loss which involves the higher frequencies. However, ARHI shows extensive variation, as is also observed in the senescence of other functions. The progression and the severity of hearing impairment vary considerably between individuals. ARHI is a complex, multifactorial trait that is attributable to confounding intrinsic and extrinsic factors. Genetic predisposition could account for a sizable proportion of interindividual variation. Since we published our review on the molecular genetic epidemiology of ARHI in 2011 [genes causing monogenic hearing impairment with phenotypic similarities to ARHI,genes involved in oxidative stress, biologic stress responses, and mitochondrial dysfunction,candidate genes for senescence, other geriatric diseases, and neurodegeneration.This review is divided into the following three areas, although there is likely some overlap between the genes described in each:Recent topic-based genetic contributions, which have been followed by a flurry of additional reports, have been preferentially selected.These genes, which can cause hearing impairment such as pronounced progressive sensorineural hearing loss in the high frequencies, are excellent candidate ARHI susceptibility genes . Many in GRHL2 (grainyhead like 2) gene, which is also known as TFCP2L3 (transcription factor cellular promoter 2-like 3) or BOM , and is a widely expressed transcription factor in human epithelial tissues [ GRHL2 spans approximately 177 kb on chromosome 8q22.3 (NCBI 37/hg19), contains 16 exons, and is translated into a 625-amino-acid protein. It was first associated with the DFNA28 locus through mapping studies involving a five-generation North American family affected with mild to moderate postlingual progressive bilateral sensorineural hearing loss [ GRHL2 (c.1609_1610insC) that results from a truncation in exon 14 is responsible for DFNA28-associated autosomal dominant hearing loss.The DFNA28 (OMIM: 608641) locus is occupied by the tissues . GRHL2 sing loss . A frame GRHL2 (c.1258-1G>A) were reported in a family affected with nonsyndromic hearing loss [ GRHL2 is an ARHI susceptibility gene from the results of an association study performed with 2418 samples from individuals with ARHI at nine centers in seven European countries [ GRHL2, in a region of approximately 16 kb. Particularly, the most significant SNP, rs10955255, was replicated in two of nine independent sample sets; and the odds ratios (ORs) of all nine sample sets had similar implications. However, when Lin et al. tried to replicate these results in a Han Taiwanese population, they found no positive association between the GRHL2 SNP locus (rs10955255: A/G) in intron 1 (coordinate: 102605581) and ARHI [Very recently, a second DFNA28-causing mutation and the first splice-site mutation in GRHL2 c.58-1G>A wing loss . GRHL2 ping loss . Impaireing loss , 12. vanountries . After sThe hearing loss in the family with c.1609-1610insC can be categorized as mild to moderate across all frequencies in the initial stages but progressing toward severe hearing loss of the high frequencies in the fifth decade. The age at onset is variable; the youngest patient was diagnosed in his first decade. DFNA28 hearing loss does not then entirely match the typical features of ARHI. The functional properties of GRHL2 within the inner ear will also critically depend on the presence of its target genes, given that GRHL2 is a transcription factor .KCNQ4 (potassium voltage-gated channel member 4) encodes a voltage-gated potassium channel [+ probably through voltage-gated ion channels. Mutations in these channels have been thought to implicate candidate genes for individuals with slowly progressive hearing loss. channel , which p channel . Signal  KCNQ4 cause an autosomal-dominant type of nonsyndromic hearing loss, DFNA2. It has been reported that DFNA2-associated hearing loss is typically late in onset, is involved in high-frequency hearing loss, and progresses over time [ KCNQ4 gene has 14 exons encoding a protein of 695 amino acids, the mutations seem to cluster around exons 4–7, with missense mutations being the most common [ KCNQ4 mutations, regardless of the particular mutation [Mutations inver time . Althougt common . The promutation . KCNQ4 for association with ARHI by treating ARHI as a quantitative trait in two independent Caucasian populations. For both populations, several SNPs in a region spanning 13 kb in the middle of the KCNQ4 gene were significantly associated with ARHI [van Eyken et al. examined SNPs spread acrossith ARHI . DFNA5. They have concluded that no strong association exists between DFNA5 and ARHI, because no significant linkage was detected between ARHI and microsatellite markers from the DFNA5 region and no significant differences were found in genotypes between individuals with good hearing and those with hearing impairment [DFNA5 was originally discovered in a Dutch family with autosomal dominant nonsyndromic hearing impairment . The audpairment .Different mutations have been found among Korean, Chinese, and Dutch families with DFNA5-associated hearing loss, all exerting a highly specific gain-of-function effect. In these families exon 8 is skipped during splicing, causing a frameshift mutation and premature termination of the protein , 21. Lat DFNA5 as one of several mutations in apoptosis genes that cause monogenic deafness [Apoptosis is a key contributor to the development of presbycusis or ARHI. In a review article focusing on the programmed death of the hair cell, Op de Beeck et al. discusseddeafness . They su COCH (cochlin or coagulation factor C homology) gene, encoding the protein product, cochlin, are responsible for DFNA9-associated hearing loss. COCH mutations have been found in individuals living on four continents and the possibility that they play an important role in presbycusis and disorders of balance has been considered [ COCH gene have been reported, with individuals presenting in the 95th percentile of the air conduction threshold levels for presbycusis for a given age and sex according to ISO 7029 [ COCH-variant T352S (rs 1045644) and ARHI, no significant effect of the SNP on quantitative trait values for ARHI was found [The autosomal dominant nonsyndromic hearing disorder DFNA9 is characterized by late onset bilateral progressive high-frequency sensorineural hearing impairment with associated variable vestibular dysfunction and starts to manifest between 20 and 50 years of age . Its feansidered . PhenotyISO 7029 , 26. In  GIPC3  gene on chromosome 19p13.3.Autosomal recessive deafness 15 (DFNB15), also known as DFNB72 or DFNB95, is caused by homozygous mutations in the ahl5) and audiogenic seizure susceptibility (jams1) in BLSW mice and they screened panels of families of Dutch and Indian origin presenting with autosomal recessive nonsyndromic hearing impairment to examine whether mutations in GIPC3 underlie human sensorineural hearing loss [ ahl5/jams1 and DFNB15/DFNB95 loci and revealed a function for Gipc3 in peripheral auditory signal transmission in mice and humans. They proposed that GIPC3 and its paralogs are excellent candidate genes for ARHI.Charizopoulou et al. identified a sequence polymorphism in the PDZ domain of Gipc3 as the cause of progressive sensorineural hearing loss  or glutamate metabotropic receptor 7 (GRM7) excitotoxicity has been suggested as a risk factor for ARHI –29. CharMYO6 encodes myosin VI, an unconventional actin-based motor protein, expressed in the hair cells of the inner ear that is necessary for maintaining their normal stereociliar structure [ MYO6 has been reported to be responsible for hearing loss associated with loci DFNA22 and DFNB37 [tructure , 32. MYOd DFNB37 , 34. MYO6 with those of three different DFNA22 families published previously [ MYO6 may contribute to presbycusis.Oonk et al. compared the detailed audiometric characteristics of a large Dutch family with autosomal dominant hearing loss caused by a mutation ineviously . HearingOxidative stress plays a major role in the overall aging process. Reactive oxygen species (ROS), such as superoxide anion, hydrogen peroxide, and hydroxyl radicals, are generated as byproducts of mitochondrial oxidative phosphorylation. Overproduction of free radicals and ROS results in damage to mitochondrial DNA (mtDNA). The mitochondrial “vicious cycle” theory of aging proposes that accumulation of mtDNA mutations may lead to enhanced mitochondrial ROS production and subsequent increased oxidative stress during aging .Free radicals, ROS, and associated mitochondrial dysfunction are causes of age-related neurosensory hearing loss. Complex enzymatic and nonenzymatic systems act mutually to counteract ROS-mediated oxidative damage. Oxidative stress in the inner ear, secondary to the decrease in defense mechanisms caused by certain polymorphisms related to a battery of human antioxidant systems, could make individuals more susceptible to ARHI. A number of researchers have focused on several genes and loci involved in oxidative stress and have identified mitochondrial DNA mutations as candidate genes for the development of ARHI .Two classes of antioxidant enzymes are active in the cochlea: enzymes involved in glutathione (GSH) metabolism  and enzymes involved in the breakdown of superoxide anions and hydrogen peroxide  –40. NAT1*4 and NAT2*4 designated as the reference alleles for NAT1 (N-acetyltransferase 1) and NAT2 (N-acetyltransferase 2), respectively [ NAT1 and NAT2 alleles at the website http://nat.mbg.duth.gr. Variant NAT2 alleles or haplotypes possessing combinations of SNPs are segregated into clusters possessing a signature SNP either alone or in combination with others. Common human NAT2 alleles (haplotypes) discussed in the previous studies regarding ARHI are listed .Ünal et al. investigated associations between ARHI and theNAT2*14A . They st GSTM1 , GSTP1 , and GSTT1 , are polymorphic. The GSTT1 polymorphism is caused by deletion of a substantial part of the gene, causing a reduction of GSTT1 activity in all tissues, whereas in individuals homozygous for the GSTM1 deletion the enzyme is absent [The human glutathione S-transferase (GST) family is composed of at least 8 classes  with multiple subfamilies per class , 46. GSTs absent . GSTT1 were more likely to have a “high-frequency steeply sloping” audiogram than participants with the wild-type genotype, suggesting that the basal turn of the cochlea is susceptible to GSTT1-regulated oxidative stress.Angeli et al. investigated the association among audiometric patterns and polymorphisms of the antioxidant enzymes GSTT1, GSTM1, and NAT2 in a hospital-based cohort study of adults with ARHI . ParticiGSTM1, GSTT1, NAT2*5A, NAT2*6A, and NAT2*7A) with a large set of 2,111 independent samples from a general European population and a Finnish population [ NAT2*6A in the European population but no association between GSTM1 or GSTT1. However, in the Finnish population they found significant associations between ARHI and both genes.van Eyken et al. investigated an association between ARHI and genes related to oxidative stress, including polymorphisms in single genes  was performed. The frequency of the NAT2*6A mutant genotype (heterozygous and null genotype combined) was more frequent among patients with presbycusis (60%) than in controls . The GSTT1 null genotype was present in 34% of controls and in 60% of white patients with ARHI . The GSTM1 null genotype was more frequent in patients with ARHI.Bared et al. investigated the association of different polymorphisms oft allele . GenotypWallace and Fan have divided the mtDNA variants clinically relevant to disease susceptibility into three classes: recent deleterious mutations resulting in maternally transmitted disease, ancient adaptive variants that predispose individuals to disease in different environments, and the age-related accumulation of somatic mtDNA mutations that erode function and provide the aging clock . These mmtDNA mutations can exist as point mutations, nucleotide insertions or deletions, and large deletions. mtDNA deletions were reported to accumulate with age in a variety of tissues . Analysehttp://www.mitomap.org/) show that there are 20 recognized mtDNA haplogroups within the European community, 18 in the Asian community and 10 in the African community. Some of these haplotype groups have been associated with ARHI. Manwaring et al. found that haplogroups U and K associated independently with a higher prevalence of ARHI [mtDNA polymorphisms typically reflect different ethnic backgrounds. Specific mtDNA polymorphisms have now been classified into a number of mitochondrial haplogroups. Estimated worldwide haplotype frequencies (Mitomap:  of ARHI . However of ARHI . APOE (Apolipoprotein E) gene encodes apolipoprotein E (ApoE), a multifunctional protein of the lipid and lipoprotein transport system involved mainly in metabolism of dietary lipids. The three major isoforms, human ApoE2, -E3, and -E4, are encoded by three common alleles of APOE called the ε2, ε3, and ε4 alleles. These alleles represent haplotypes of two SNPs in the coding region of APOE. The ε2 allele encodes a cysteine (Cys) at amino acid position 112 and at position 158. Allele ε3 encodes a Cys at 112 and an arginine (Arg) at 158, while the ε4 allele encodes an Arg at 112 and at 158 [ APOE gene were strongly associated with late-onset Alzheimer disease. The ε4 allele is the ancestral and high-risk form, the ε3 allele is the most common in humans and the neutral allele, and the ε2 allele is associated with decreased risk for Alzheimer disease.Thed at 158 . These p APOEε4 allele was less prevalent in a study population of 89 individuals  with nonsyndromic adult-onset sensorineural hearing loss, although they originally assumed that the ε4 allele might predispose individuals to sensorineural hearing loss and ischemic injury in the cochlea by causing atherosclerotic vascular disease [ APOEε4 allele was associated with ARHI overall frequency was demonstrated in the Leiden 85-plus study, a population-based study involving 85-year-olds [ APOEε4 allele was associated with a 2.0-fold increased risk for hearing impairment .O'Grady et al. found that, compared with the general population, the disease . Recentlear-olds . The APO MTHFR (methylenetetrahydrofolate reductase) C677T polymorphism (rs1801133) is one of the most extensively investigated functional polymorphisms. It results in an alanine-to-valine substitution and a 30% decrease in MTHFR enzyme activity in heterozygotes and a 60% decrease in homozygotes [Folate metabolism is essential for cellular functioning. Polymorphisms in genes encoding critical enzymes in folate metabolism play important and interrelated roles in the pathophysiology of numerous disorders such as cancer and cardiocerebrovascular disease. Theozygotes . MTHFR T allele has been shown to be associated with an increased risk for sudden sensorineural hearing loss in several different studies [ methionine synthase (MTR) A2756G and MTHFR C677T gene polymorphisms on the risk for hearing impairment in community-dwelling Japanese adults aged 40–84 years who participated in the National Institute for Longevity Sciences, Longitudinal Study of Aging (NILS-LSA), an ongoing population-based biennial study. Cumulative data  were analyzed with generalized estimating equations. The MTHFR 677T allele was significantly associated with a reduced risk for hearing impairment only when the participants were wild-type homozygotes for MTR A2756G . In addition, a subgroup analysis demonstrated that the favorable effect of the MTHFR 677T allele on the risk of developing hearing impairment was independent of serum folate and plasma total homocysteine levels. We speculated that the MTHFR 677T allele has a potentially advantageous effect of preventing imbalances in the nucleotide pool during DNA synthesis, ensuring that DNA replication occurs with high fidelity.The studies –60. Inte studies , 62. We Endothelin-1, which is known to be an extremely potent vasoconstrictor peptide, is a prominent endothelial mediator. It is widely distributed throughout the mammalian body in cardiovascular and noncardiovascular tissues, including the nervous system, and is also expressed in the spiral modiolar artery, endolymphatic sac, vestibule, stria vascularis, and spiral ganglion cells , 64. EDN1 (endothelin-1) gene and hearing impairment in the aforementioned NILS-LSA participants aged 40 years to 79 years at baseline [We have reported an association between the Lys198Asn (G/T) polymorphism (rs5370) in thebaseline . Mutant Uncoupling proteins (UCPs) are a family of mitochondrial proton transporters and may be regulators of thermal control and energy metabolism. UCPs are thought to be activated by superoxide and considered to decrease generation of mitochondrial free radicals. UCP1 (uncoupling protein 1) and UCP2 (uncoupling protein 2) and hearing impairment in the aforementioned NILS-LSA [ UCP1 A-3826G and UCP2 Ala55Val polymorphisms on hearing impairment. The UCP1 A3826G polymorphism did not exhibit any significant association with ARHI; however, the UCP2 Ala55Val polymorphism did show a significant association with hearing impairment under the dominant, recessive, and additive models (P < 0.05).We reported a relationship between gene polymorphisms inNILS-LSA . Data weGRM7  is thought to be central to maintaining glutamate synaptic transmission and homeostasis in the mammalian cochlea at the synapses between hair cells and the dendrites of afferent auditory nerve fibers . GRM7 locus and ARHI [ GRM7 mRNA expression was also present in the neurons of the spiral ganglion in the mouse cochlea.Friedman et al. identified and subsequently validated a significant risk association between theand ARHI . In a st GRM7 on ARHI [ GRM7 haplotypes (haplotype blocks 6 and 7) and SNP genotypes and various measures of auditory perception, such as the free-field hearing-in-noise test (HINT) and suprathreshold gap detection tasks. They showed that GRM7 alleles were associated primarily with peripheral measures of hearing loss and particularly with speech detection in older adults.Newman et al. have delved further into the effect of on ARHI . They pe GRM7 SNP rs11928865 (TT genotype) occurs more frequently in ARHI patients with certain audiogram-shape subtypes of ARHI, namely, sloping-shaped and 2–4 kHz abrupt loss-shaped phenotype patterns.Luo et al. conducted a case-control candidate gene association study in an elderly male Han Chinese population composed of 982 men with ARHI and 324 controls with normal hearing . The res GRM8 , and the candidate genes located in positive GWAS regions belonged to several different gene families, which overlap only insensibly with those already identified as causing hearing impairments.New candidate loci were reported from a GWAS for hearing quantitative traits among participants recruited from several isolated villages within the international consortium called G-EAR . Eight pEstrogen has recently been identified as a novel modulator of hearing function , 69. The ESRRG  gene was associated with hearing status in the London ARHL cohort, the minor allele being associated with poorer hearing but only in women. This association was replicated in the Carlantino cohort and the combined cohort of isolated populations from Italy and Silk Road countries, both in the direction of the allelic effect and in the female-specific association [ Esrrg knock-out mice revealed worse impairment in female mice (−/−) than in male mice (−/−). These results indicate that ESRRG plays a role in maintenance of hearing in both humans and mice.Nolan et al. conducted an investigation involving a total of 6134 individuals in three independent cohorts: (i) the 1958 British birth cohort; (ii) a London ARHL case-control cohort; and (iii) a cohort from isolated populations of Italy and Silk Road countries to evaluate whether variants in estrogen signaling genes may be risk factors for adult-onset hearing loss. The rs2818964 SNP in theociation . In lineThe degree of ARHI varies substantially among individuals, as is also observed in senescence of other functions. It is not always true that deterioration of hearing with age is inevitable. During the last 10 years or so, tremendous progress has been made through human genetic research into hearing impairment and our understanding of the complex mechanisms of hearing has increased greatly. It might be that certain environmental risk factors are potentially detrimental in a limited number of individuals only, depending on their genetic background. It is hopeful that successful preventive intervention customized according to knowledge of genetic susceptibility will help to preserve good hearing even in the oldest old.


Weight gain may be associated with an imbalance between energy intake and energy expenditure. The resting metabolic rate (RMR) is the main component of total energy expenditure, and is related mainly to lean mass (LM), as well as to other factors such as fat mass (FM), age, sex and genetic factors. A RMR lower than expected may be a risk factor for weight gain. RMR is estimated by equations that use patient's weight, sex, age and height to calculate energy needs. Several studies have shown that these equations have a poor agreement with RMR measured by indirect calorimetry (IC) in obese patients once their excess fat-free mass (FFM) is usually not taken into account.To evaluate the accuracy of five equations in predicting RMR in obese subjects. Results were compared with measured RMR (mRMR) determined by IC.Cross-sectional study was conducted in obese Southern Brazilian volunteers recruited from community. Body mass index (BMI) was calculated by dividing weight (in kilograms) by squared height (in meters). Body composition was evaluated by dual-energy X-ray. RMR was measured by IC (Weir equation) and estimated (eRMR) by Mifflin–St. Jeor, Owen, Harris-Benedict, Ireton-Jones and Horie-Waitzberg & Gonzalez (H & WG) equations Figure . The latSixty individuals  aged 46±13 yrs. range, 21-83 yrs.) were evaluated. Overall, mRMR was 1941±642 kcal/day. mRMR increased along with BMI (Figure 1-83 yrs.In this sample of obese subjects, the available RMR estimate equations that do not take into account the FFM have poor accuracy when compared with mRMR.


The aim of the study was to examine 1) the incidence of disability in Activities of Daily Living (ADL), in persons 78 years and older 2) explore whether being physical active earlier is a significant predictor of being disability free at follow-up and 3) describe the amount of informal and formal care in relation to ADL-disability.Data were used from a longitudinal community-based study in Nordanstig (SNAC-N), a part of the Swedish National Study on Aging and Care (SNAC). To study objectives 1) and 2) all ADL-independent participants at baseline (N = 307) were included; for objective 3) all participants 78 years and older were included (N = 316). Data were collected at baseline and at 3- and 6-year follow-ups. ADL-disability was defined as a need for assistance in one or more activities. Informal and formal care were measured using the Resource utilization in Dementia (RUD)-instrument.The incidence rates for men were similar in the age groups 78-81and 84 years and older, 42.3 vs. 42.5/1000 person-years. For women the incidence rate for ADL-disability increased significantly from the age group 78–81 to the age group 84 years and older, 20.8 vs.118.3/1000 person-years. In the age group 78–81 years, being physically active earlier (aOR 6.2) and during the past 12 month (aOR 2.9) were both significant preventive factors for ADL-disability. Both informal and formal care increased with ADL-disability and the amount of informal care was greater than formal care. The incidence rate for ADL-disability increases with age for women and being physically active is a protective factor for ADL-disability.The incidence rate for ADL-disability increases with age for women, and being physical active is a protective factor for ADL-disability. Human life expectancy is increasing worldwide , 2. As wMost studies are cross sectional and focus on the prevalence of ADL-disability –11. HoweCare of older persons due to disability involves a complex interaction between formal and informal care resources. Care is also highly dependent on the socioeconomic context, on both the micro- (families) and macro level (how care is financed and organized in a society/country). Besides economic strength on both the micro- and macro- level, traditions, politics and culture also contribute to the complex situation.The aims of this paper are first to examine the incidence of impaired physical functioning defined as ADL- disability, in relation to gender; second to explore whether being physically active earlier in life and/or during the past 12 month are significant predictors of being disability free at follow-up. Due to the relationship between ADL-disability and resource use and costs, a third aim is to describe the amounts of informal and formal care in relation to levels of ADL-disability.This study was based on data from the Swedish National Study on Aging and Care in Nordanstig (SNAC-N). SNAC-N is a longitudinal individual population-based ongoing study being conducted in the municipality of Nordanstig in Sweden, which is a rural area in the northern Sweden and had, at the time for the baseline collection, approximately 10 000 inhabitants. This coastal district has no city or central areas–instead there are several small villages covering an area of 1 380 square kilometers. SNAC-N is one of four geographical areas in focus in a larger national study promoted by the Swedish Ministry of Health and Social Affair .In the present study, we used data from participants who were independent in ADL and 78, 81, 84, 87, 90, 93, 96, and 99+ years of age at baseline (N = 307) to study incidence rates. For our analysis of informal and formal care, we used data from all participants who were 78 years and older at baseline and living at home (N = 316).The study was approved by the Ethics Committee of the Karolinska Institutet and the Regional Ethical Review Board in Stockholm.st follow-up three years later and the 2nd follow-up six years later. Data were gathered through interviews and clinical examinations using standardized protocols administered by a trained registered nurse and a licensed practical nurse. A physical examination was also carried out by a physician. Information from a proxy interview was used when the participant was unable to answer or was diagnosed with dementia.Baseline data were collected from March 2001 to March 2003 and data for the 1Sociodemographic factors were age, gender and education. Mortality data were gathered regularly from the Swedish National Death Certificate Register.Basic ADL was measured by interviewing and observing the participants and using the hierarchical scale KATZ index of ADL to assess dependency in basic activities . DisabilAt baseline, study participants were asked whether they had engaged in regular light exercise  1) earlier in life and 2) during the past 12 months. Response alternatives were: every day, several times/ week, 2–3 times/month, less or never.Cognition was measured using the Mini-Mental State Examination (MMSE), a commonly used instrument scoring between 0–30 points, where 30 points represents no impairment .Parts of the Resource Utilization in Dementia (RUD) instrument was used to calculate the amount of formal and informal care at baseline and follow-ups in the participant´s residence . In the Age- and gender- specific incidence rates were calculated at both the 3-year and 6-year follow-up, using as the numerator all cases with a diagnosis of ADL-disability (needing assistance in one or more or two or more ADL-activities), and as the denominator, the examined population. Age- and gender- specific incidence rates were calculated as the number of new cases divided by the person-years at risk. The 95% confidence intervals (CI) were based on the Poisson distributions. Person-years for non-disabled subjects were calculated as the time between baseline examination and the follow-up examinations. For the ADL-disabled subjects, half this time was assumed, due to the uncertainty of disability onset. Furthermore, a subject who developed any type of disability was considered to no longer be at risk. The incidence rate was calculated in two separate analysis, needing assistance in one or more (ADL1+), or two or more (ADL2+) ADL-activities. Logistic regression analyses were used to estimate the association between physical activity and ADL-disability adjusted for gender and cognition. The results are presented as adjusted odds ratios (aOR), and 95% CIs. Mean values for the number of hours per month of informal and formal care that the persons received were calculated using univariate analyses of variance (One-Way ANOVA).The IBM SPSS Statistics version 20.0 for Windows  was used for modeling analyses and statistical testsnd follow-up after six years, 135 (43.4%) were re-examined, of those 135 participants 18 were not examined at the 1st follow-up (refused or missing). Between baseline and the 2nd follow-up 40.0% of the participants had died, more men than women (44.7% vs. 36.6%). The study population was divided into two age groups, one in the age of 78 and 81 year at baseline (n = 155) and the second 84 years and older at baseline (n = 152). At the 2nd follow-up, 60.0% of the men and 47.6% of the women had died (non-significant)  for men and 83.0 (SD 4.5) for women. At the 2ificant) .st and 2nd follow-up. In the age group 84 years and older, more women (57.6%) than men (23.8%) had become ADL-disabled at 2nd follow-up in terms of needing assistance with one or more activities (p<0.05). Both at 1st and 2nd follow-up and needing assistance with one or more activities (ADL 1+), or two or more (ADL 2+), women had become more ADL-disabled in the age group 84 years and older than women in the age group 78–81 (p<0.05). There were no significant differences in ADL-disability between men in the two age groups (After three years 10.6% had become ADL-disabled (needing assistance with one or more activities) and after six years 28.1%. There was a non-significant tendency for more men than women in the age group 78–81 to become ADL-disabled, at both the 1e groups .The overall incidence rate for ADL-disability was higher for women, 59.3/1000 person-years, than for men 42.4/1000 person-years. The incidence rate for disability increased with age especially for women . The incOf the 155 persons in the age group 78–81 years, 126 answered the question about physical activity, and in the age group of 84 years and older, 84 of 152 answered the question. Of those who answered the question, 2% in the age group 78–81 and 3,8% in the age group 84 years and older reported never having been physically active earlier in life, and 7.9% and 10.7% respectively, reported never having been physically active during the past 12 months. In the age group 78–81 years, there was a significant association between being physically active during the past 12 month prior to baseline examination, aOR 2.9 (1.3–6.2) and earlier in life, aOR 6.2 (1.3–29.3) and being non ADL-disabled at the follow-ups. No association was found between physical activity and ADL-disability in persons 84 years and older, see In the study population of both ADL-disabled and non-ADL-disabled persons, the mean hours per month was for the informal care 15.7 and for the formal care 4.6 hours/month. Among the persons still living at home at the follow-ups, the amount of both informal and formal care increased with the number of ADL activities the persons were required help with, and the differences in amount of care between the ADL groups were significant (p<0.05). When the formal and informal care is divided into assistance in IADL and ADL, the analysis showed that the informal care mainly consisted of assistance with IADL and the formal care of assistance with both IADL and ADL. Between baseline and the two follow-ups, 34 persons had moved to residential care. Their ADL-dependency varied between 0–5. Thirty persons were living alone at home before they moved and only four persons were living together with their wife and were dependent in 4 or 5 ADL-activities .The aims of the present study were to examine the incidence of impaired physical functioning defined as ADL disability, to explore whether being physical active earlier is a significant predictor of being disability free at follow-up and to describe the amount of informal and formal care received in relation to ADL-disability. The main findings from the study can be summarized as follows:The incidence rates for men were almost identical in the age group 78–81 and the age group 84 years and older 42.3 vs. 42.5/1000 person-years. For women the incidence rate for ADL-disability increased significantly from the age group 78–81 to the age group 84 years and older, both for ADL1+ (20.8 vs. 118.3/1000 person-year) and ADL 2+ (8.5 vs. 81.5/1000 person-year).In the age group 78–81 years, being physically active earlier (aOR 6.2) and during the past 12 month (aOR 2.9) were both significant preventive factors for ADL-disability.The amount of both informal and formal care increased with ADL-disability, and the amount of informal care was greater than that of formal careWe found that, of the non-disabled population 78 years and older at baseline, 10.6% had become AD-disabled after three years and 28.1% after six years. This is almost the same result as in the study from the Netherlands .The inciThe overall incidence rate for ADL-disability was higher for women, 59.3/1000 person-years, than for men 42.4/1000 person-years. Higher incidence rate in women than in men was found in the study from Brazil also. The incidence rate was lower in the Brazilian study due to the younger study population, 60 years and older . DividinIn the age group 78–81 years, physical activity earlier in life and/or during the past 12 months prior to baseline examination was shown to be a protective factor for ADL-disability at the follow-ups. In the present study, physical activity referred to activities such as biking, walking in the forest, golfing and other not particularly intensive physical activities. Other studies have also shown that physical activity can reduce disability in old age , 33 and The amount of care increased with the number of ADL-activities the person received help with. For persons who were independent in ADL and needed help with IADL-activities, help was mainly informal. When persons were more disabled, the pattern was different. Help with IADL was mostly informal and help with ADL was mostly formal. When persons were disabled in two or more ADL-activities, the amount of formal care was greater than the amount of informal care, mainly due to help with ADL. In the present study, nine persons moved to residential care between the follow-ups. Those persons received help with 0–5 ADL-activities, but all were living alone at home before moving. Of those in the study who were still living at home and were receiving help with two or more ADL-activities, all were living with a spouse. The present study shows that the informal care provided by co-habiting persons strongly contributes to allowing older people to stay at home longer before moving to residential care. Informal care is also important for society as a whole, because it replaces formal care and without it, older persons would probably have to move to residential care earlier. The amount of informal and formal care varies between countries. A study examining community care in 11 different countries in Europe found that formal care was small in Italy, while in the UK it was more than double the European average across all levels of dependency . AnotherThe limitation of the present study may be the size of the study population. Another limitation can be that we have not taken into account whether those who died during follow-up had developed disability as we lack the data. This might have led to an underestimation of disability-incidence. The reason why there were so few people receiving assistance in two or more ADL activities, was probably because the majority of people with disability in two or more ADL activities had moved to residential care. The strengths of the present study is its longitudinal individual population-based design, which is preferable to cross-sectional designs. The data collection team members were almost identical at baseline and the follow-ups, thus minimizing reliability problems and that the study use validated instruments e.g. MMSE, Katz index of ADL, RUD.The incidence rate for ADL-disability increases with age for women, and being physical active is a protective factor for ADL-disability.


Muckle-Wells-syndrome (MWS) is an autoinflammatory disease characterized by systemic and organ-specific inflammation due to excessive interleukin (IL)-1 release. Inner ear inflammation results in irreversible sensorineural hearing loss, if untreated. Early recognition and therapy may prevent deafness. The aims of the study were to characterize the spectrum of hearing loss, optimize the otologic assessment for early disease and determine responsiveness to anti-IL-1-therapy regarding hearing.A single center prospective cohort study of children and adults with MWS was performed. Standardized clinical, laboratory and otologic assessments including standard pure tone audiometry, additional high tone thresholds, vestibular organ testing, tinnitus evaluation and functional disability classes were determined serially. Pure-tone-average models were developed and evaluated. Risk factors for hearing loss and the impact of anti-IL-1 treatment were determined.p = 0.008). Treatment with IL-1 blockade resulted in improved or stable hearing in 91 % of patients.A total of 23 patients with genetically confirmed MWS were included, of whom 63 % were females; 52 % were children. At baseline all patients had active MWS; 91 % reported clinically impaired hearing with 74 % having an abnormal standard assessment (0.5–4 kHz). In contrast, high frequency pure tone averages (HF-PTA) were abnormal in all symptomatic patients including those with early hearing loss (sensitivity 100 %). Females were at highest risk for hearing loss even after adjustment for age  contains supplementary material, which is available to authorized users. NLRP3-gene (CIAS1) on chromosome 1q44 encoding the protein NLRP3 (cryopyrin) in MWS [Muckle-Wells syndrome (MWS) is an autosomal dominant autoinflammatory disease in the clinical spectrum of cryopyrin-associated periodic syndrome (CAPS). CAPS comprise the mildest form, familial cold autoinflammatory syndrome (FCAS), the intermediate MWS and the most severe phenotype chronic infantile neurological cutaneous and articular syndrome (CINCA) or neonatal-onset multisystem inflammatory disease (NOMID) . First d) in MWS . Subsequ) in MWS . In CAPS) in MWS , causing) in MWS . Devasta) in MWS .Sensorineural hearing loss in MWS often rapidly progresses from mild high-tone deficits to complete deafness , 11. EarEarly detection of imminent hearing loss is crucial, yet challenging. Traditionally, pediatric and adult screening audiograms determine individual hearing thresholds at the frequencies 0.5, 1.0, 2, and 3 or 4 kHz reflecting those frequencies most relevant for speech discrimination. Hearing thresholds at each frequency are determined, and averaged in a single value, the so-called 4 pure tone average . This commonly used approach has significant limitations for the early detection of hearing loss in MWS, since the frequencies affected earliest are above the test range and therefore not included in the evaluation. However, early detection of imminent hearing loss and immediate initiation of targeted therapy may prevent progression to deafness in children and adults with MWS. Thus, a tailored assessment tool for detection of early hearing loss in MWS is urgently needed.Therefore, the aims of the study were 1) to characterize the distinct pattern of hearing loss at diagnosis of MWS, 2) to modify the established standard 4PTA assessment tool to the hearing loss characteristics of MWS patients and assess its sensitivity in detecting hearing loss and 3) to determine risk factors associated with hearing loss in MWS and the effects of IL-1-inhibition.NRLP3 mutation. The clinical diagnosis was based on the presence of MWS typical features of fever, non-purulent conjunctivitis, urticaria-like rash, sensorineural hearing loss, arthralgia/arthritis, fatigue coupled with raised inflammatory markers. Mutations were determined as previously described [NLRP3 gene. The study was approved by the local ethics committee .A single-center cohort study of consecutive patients diagnosed with MWS between 4/2004 and 12/2007 was performed. Pediatric and adult patients had to have a clinical diagnosis of MWS and genetic confirmation of a escribed . ExclusiNLRP3 gene mutation. Clinical characteristics were documented as previously reported [All MWS patients were diagnosed and followed according to a standardized protocol by an interdisciplinary tertiary care team. Demographic information included age at diagnosis, gender, ethnicity and type of reported . Review reported . MWS-DASAll patients had serial comprehensive otolaryngologic assessments. Pure-tone audiometry was serially performed for each ear separately using conventional, play or behavioral audiograms. All adult patients were additionally tested using speech audiometry, caloric vestibular stimulation and tinnitus handicap questioning. The audiograms of members of the same family were compared in order to capture the family-specific risk of progressive hearing loss. The audiologic examination included air and bone conduction threshold for pure tone frequencies of 0.125, 0.25, 0.5, 1, 2, 4, 6 and 8 kHz and middle ear functional testing by impedance audiometry. Hearing threshold is defined as the dB value at which sound is perceived at a given frequency. In order to account for normal age-related hearing loss, measured age-adjusted hearing thresholds were calculated . Values were considered abnormal if they differed by ≥ 10 dB from normative hearing threshold levels . Values were considered abnormal if they differed by ≥ 10 dB from normative hearing threshold levels  Fig. . WHO graPatients were treated with anakinra  at dose of 100 mg/day for  ≥40 kg body weight or 1–2 mg/kg/day in patients <40 kg body weight. In children with persistent disease activity, the anakinra dose was stepwise increased to a maximum of 8 mg/kg/day. Anakinra was self-administered by subcutaneous injection once daily.Patients received canakinumab subcutaneously 150 mg or 2 mg/kg for body weight <40 kg every 8 weeks. In case of residual symptoms, patients were maintained on a more intense dosing regimen (increase dose up to 600 mg s.c. or 8 mg/kg s.c and/or increase of dosing frequency).Hearing loss was defined as increased threshold of ≥10 dB for any of the age-adjusted pure tone average for the frequencies 0.5, 1, 2 and 4 kHz (4-PTA) and the two high frequencies 6 and 8 kHz (HF-PTA).1) Improved or stable hearing was defined as a) decrease in hearing threshold by ≥ 20 dB in one frequency or by ≥10 dB in two, or more consecutive frequencies for improved hearing or b) absence of worsening of hearing at last follow-up. 2) Worsening of hearing was defined as increase in hearing threshold by ≥20 dB in one frequency, or by ≥10 dB in two or more consecutive frequencies, in accordance with definitions used in previous studies .p <0.05 as statistically significant. Statistical analyses were performed using SAS statistical software for Windows Version 9.3 .Descriptive statistics were performed; results were reported as frequencies with percentages, means with standard deviations, or medians with ranges. Univariate parametric analyses utilized Fisher’s exact and chi-square analyses for categorical data and students T-test for continuous variables. Non-parametric tests were performed where appropriate. Paired univariable analyses were conducted to determine treatment responses. Repeated measures linear regression analysis was performed for comparative and predictive modeling of hearing thresholds to account for paired ears within patients. All tests considered NLRP3 mutations were found in this cohort: 61 % had the c.937G>A (p.Glu313Lys) formerly known as E311K mutation, 22 % the c.1049C>T (p.Thr350Met), formerly known as T348M and 17 % the c.598G>A , formerly known as V198M mutation  in nine and c.1049C>T (p.Thr350Met) and c.598G>A  in two respectively.All patients had active disease at diagnosis reflected in a median MWS-DAS of 8. Most commonly seen clinical features were arthralgia (96 %), eye involvement 91 %) and headache (83 %). Clinical hearing loss was reported by 91 % of patients. Other features included oral ulcers (78 %), rash (65 %) and myalgia (52 %). At baseline, the mean SAA level was 68.7 mg/dl, mean CRP level 2.2 mg/dl (Table  % and hep <0.001). All patients had normal tympanic membrane mobility, normal middle ear pressure by impedance audiometry and normal caloric vestibular stimulation. Seven adult patients reported tinnitus.At baseline, 21/23 MWS patients (91 %) had sensorineural hearing loss by audiogram, correspondingly 42/46 hearing assessments (single ears) were abnormal. All 11 adults had abnormal hearing assessments , while only 10/12 children  had evidence of sensorineural hearing loss. Hearing thresholds at baseline varied significantly across ages and frequencies  MWS hearing assessments demonstrated an abnormal 4PTA, including 22/22 adult patient hearing assessments (100 %) and 12/24 (50 %) children ears. The median baseline 4PTA was 28 (range 0–72); at last follow-up, the median 4PTA was 26 (range 0–56) . An abnormal HF-PTA was found in 22/22 adult assessments (100 %) and in 20/24 (83 %) in children. The median baseline HF-PTA was 45 (range 0–114); at last follow-up, the median HF-PTA was 43 (range 0–114) , two patients, both adults, showed slight impairment (WHO grade I), eight – four children and four adults – had moderate (grade II), four had severe hearing impairment (WHO grade III), all were adults, and one adult had profound hearing impairment (WHO grade IV).At last follow-up, 9 patients – all children – presented with no hearing impairment (WHO grade 0), one adult showed slight (WHO grade I), nine patients, six adults and three children, had moderate (grade II), three patients, all adults (WHO grade III), and one adult still had profound hearing impairment (WHO grade IV). So one child reversed hearing loss during treatment.p = 0.008), earlier age at diagnosis (p <0.001), and affection of high compared to low frequencies (p = <0.001) were found to be significantly associated with hearing loss in a repeated measures linear regression model to account for paired ears within patients. Young age was primarily associated with high frequency hearing loss, while older patients had evidence of abnormal hearing thresholds across all frequencies. Female gender was an independent risk factor for hearing loss in MWS.Female gender  patient ears had improved or stable hearing assessments as defined above when treated with anti-IL1 therapy. Improvement was documented in 11 ears, six of which were treated with canakinumab and five with anakinra. Stable hearing was documented for 33 ears, 20 of which were treated with canakinumab and 13 with anakinra. Only two ears worsened significantly; these include one of an 18-year-old male and one of a 37-year-old female, both treated with anakinra. The contralateral ears for the two patients remained stable . Therefore, optimized testing for early hearing loss in MWS has to include high frequency hearing thresholds of 6 and 8 kHz (HF-PTA) in order to significantly increase the sensitivity of the hearing assessment and to allow for the recognition of early MWS associated hearing loss due to inflammatory injury. This is particularly important in children, since early on in life MWS solely affects high frequencies. In fact, the study determined that earlier age at diagnosis was significantly associated with the development of hearing loss. Rapid onset of therapy may control the inflammation thus greatly reducing the risk of inner ear damage and deafness in MWS . The stuOnset and progression of hearing loss in MWS has a distinct pattern Fig. . The infNLRP3 mutations [Prevention of deafness as a result of inner ear inflammation in children and adults is a key task when treating patients with MWS. Anti-IL-1 therapy was effective in stabilizing progression of hearing loss in this cohort, similarly to what was reported by others caring for patients with CAPS , 32–34. utations . Overallutations . Reasonsutations . InteresNLRP3 gene mutation types regarding impact on hearing [There are several imitations to the study. The number of patients tested is small, only 23 children and adults were included. However, MWS is a very rare condition. A comprehensive, standardized assessment and therapy protocol was applied prospectively to all patients allowing for the capture of detailed clinical and audiological data at baseline and following therapy. The age spectrum was wide with relatively few patients per age group threatening the generalizability of the results. However, the clinical spectrum of CAPS is also wide and varies further between  hearing . The stuDiagnostic evaluation and monitoring of hearing abilities in patients with MWS mandates a comprehensive otolaryngologic approach. The addition of HF-PTA to the standard 4PTA pure tone threshold testing allows for early recognition of imminent hearing loss and should prompt the rapid start of anti-IL-1 therapy for prevention of damage in particular in the high risk MWS populations.


In vivo, the effects of anti-T20 on a lethal anaphylaxis model using PGN-challenged OVA allergic mice, including the rectal temperature and mortality, and serum levels of TNF-α, IL-6, and LTC4 were assayed. The results showed that anti-T20 specifically bound to TLR2 and significantly inhibited PGN, LTA, and Pam3CSK4-driven TNF-α and IL-6 production by RAW264.7 cells. Also, anti-T20 protected OVA allergic mice from PGN-induced lethal anaphylaxis, and the serum levels of TNF-α, IL-6, and LTC4 of anti-T20 treated PGN-challenged OVA allergic mice were decreased as compared to isotype control of anti-T20 treated mice. In summary, this study produced a new antibody against the specific extracellular domain of TLR2 which has protective effect on TLR2 agonists-driven inflammatory and allergic response.Specific blocking strategies of TLR2-mediated inflammatory signaling and hypersensitivity reactions may offer novel therapeutic strategies to prevent a variety of diseases. In this study, we investigated the blocking effects of a new anti-TLR2 antibody anti-T20 against a 20 mer peptide T20 located in the extracellular specific domain of mouse TLR2. In addition, the effects of the anti-T20 Toll-like receptors (TLR) are known to be major actors of the innate response, most of them principally recognize pathogen-associated molecular pattern molecules (PAMPs), and are able to transduce signals through different pathways. These signals are the first step towards cell pathway activation and towards a more specific response. One of the important TLRs, TLR2, is widely distributed on the surface of monocyte-macrophages, dendritic cells, mast cells, basophils, and other cells –4 and caSimilar properties of cell-mediated signal transduction activity and analogous inflammatory cytokine release following ligands interaction with TLR2 have been seen. Agonists of TLR2 can generate both antimicrobial and antitumorigenic effects with subsequent benefits in terms of protective host immunity, while they also can induce hyperinflammatory and hypersensitivity reactions, which have the potential to promote detrimental effects on human health , 9. Acco in vitro and on OVA-specific allergic mice from PGN-induced lethal anaphylaxis, which provides a novel strategy to prevent TLR2 agonist-mediated inflammation and promotion of allergic immune responses.In this study, we produced a new anti-TLR2 antibody and investigated its blocking effects on agonists-stimulated mouse macrophage cell line RAW264.7 ad libitum. The animal experiments were approved by the Ethics Committee for Experimental Animals at Guangdong Medical College and were performed according to relevant national and international guidelines. In all experiments, efforts were made to minimize suffering, and any animals appearing moribund during the course of experimentation were immediately anesthetized with pentobarbital sodium . RAW264.7 cells were obtained from the Tissue Type Culture Collection of the Chinese Academy of Sciences .New Zealand white rabbits and BALB/c mice were obtained from the Experimental Animal Center of Southern Medical University  and the Experimental Animal Center of Guangdong Medical College . The animals were maintained at 25°C in a 12 h equal light : dark cycle with 50% humidity and were fed with commercial feed and sterile waterα, and IL-6 ELSIA kits were all obtained from Wuhan Boster Biological Technology, Ltd. . Leukotriene C4 (LTC4) EIA kits were obtained from Cayman Chemical Company .Ovalbumin (OVA), bovine serum albumin (BSA), and Freund's adjuvant were obtained from Sigma-Aldrich Chemical Co. . Blue-carrier protein (BC), carbodiimide (EDC), the BCA kit, and anti-mouse IgE-HRP were obtained from Pierce Biotechnology . Sepharose 4B was obtained from Amersham Pharmacia Biotech , and commercialized mouse TLR2 antibody (T2.5) was obtained from BioLegend . PGN, LTA, and Pam3CSK4 were obtained from InvivoGen . Irrelevant rabbit control antibody, anti-mouse IgG, IgE-HRP, FITC-goat anti-mouse and FITC-goat anti-rabbit, TNF-http://imed.med.ucm.es/Tools/antigenic.pl) was used to predict epitopes in this sequence.A 20 mer peptide (named as T20) encoding the amino acid sequence (DSQS LKSI RDIH HLTL HLSE) on the basis of the predicted B cell dominant epitope of mouse TLR-2 was synthesized by Hybio Pharmaceutical Company  . PredictIn the presence of EDC, T20 was cross-linked with BC, BSA, and rabbit IgG, respectively, to obtain the cross-linked T20-BC, T20-BSA, and the rabbit IgG-T20 (rIgG-T20) by conventional methods. T20-BC acted as an immunogen, and two New Zealand white rabbits were immunized six times with this immunogen, at a dose of 1 mg each time by conventional immunizing methods.6 RAW264.7 cells were treated with anti-T20 (1 μg/test), or anti-T20 (1 μg/test) plus T20 (1 μg/test), or T2.5 (1 μg/test) at room temperature for 30 min. Irrelevant rabbit antibody was used as a control and washed three times with PBST containing 1% BSA, at 350 g for 5 min by centrifugation, following which FITC-goat anti-mouse antibody or FITC-goat anti-rabbit antibody was added, and incubated in the dark at room temperature for 30 min, washed twice, and then resuspended in 200 μL PBST containing 1% BSA and detected by flow cytometry. The expression of TLR2 protein in RAW264.7 cells was detected by anti-T20 (1 μg/mL), or anti-T20 (1 μg/mL) plus T20 (1 μg/mL), or T2.5 (1 μg/mL) using western blot routinely with β-actin as internal reference. The BCA kit was used for quantitative protein measurements of the collected samples.Anti-T20 was purification with a cross-linked product Sepharose 4B-rIgG-T20 column. RAW264.7 cells were cultured and collected by trypsin digestion and then detected by flow cytometry and western blot. 1 × 106 RAW264.7cells/well were seeded into six-well plates, in IMDM medium containing 5% bovine serum, at 37°C in 5% CO2 incubator, and grouped by PGN (1 μg/mL), PGN (1 μg/mL) plus anti-T20 antibody (1 μg/mL), PGN (1 μg/mL) plus anti-T20 antibody (5 μg/mL), and PGN (1 μg/mL) plus anti-T20 antibody (25 μg/mL), or LTA (0.1 μg/mL), LTA (0.1 μg/mL) plus anti-T20 antibody (0.1 μg/mL), LTA (0.1 μg/mL) plus anti-T20 antibody (0.5 μg/mL), and LTA (0.1 μg/mL) plus anti-T20 antibody (2.5 μg/mL); Pam3CSK4 (0.1 μg/mL), Pam3CSK4 (0.1 μg/mL) plus anti-T20 antibody (0.1 μg/mL), Pam3CSK4 (0.1 μg/mL) plus anti-T20 antibody (0.5 μg/mL), and Pam3CSK4 (0.1 μg/mL) plus anti-T20 antibody (2.5 μg/mL) following culture, and at 6 h and 12 h time-points, supernatants were collected, respectively, from each of the groups and frozen at −40°C until detected by commercial ELISA kits.Approximately 10μg) emulsified with AL(OH)3 on days 0, 10, and 20. The anti-OVA IgG and IgE antibody levels were detected on day 21. The dose of OVA was intravenously administered at 30 mg/Kg.BALB/c mice of six-week-old BALB/c mice were multipoint immunized in the abdominal subcutaneous areas with OVA (100 μg/mouse) experimental group; Group 3, the OVA (30 mg/Kg) plus PGN (100 μg/mouse) plus anti-T20 antibody (100 μg/mouse) group, which was the intervention experimental group; Group 4, the OVA (30 mg/Kg) plus anti-T20 antibody (100 μg/mouse) group; and Group 5, the isotype negative control group of OVA (30 mg/Kg) plus PGN (100 μg/mouse) plus rabbit irrelevant antibody (100 μg/mouse), and also normal mice as control group. Changes in rectal temperature and mortality were determined, respectively.Then, the OVA allergic mouse model was established and divided into the following groups: Group 1, the OVA (30 mg/Kg) challenged group; Group 2, the OVA (30 mg/Kg) plus PGN , and anti-T20 IgG was approximately a titer of 1 : 100. Anti-T20  was purified with Sepharose 4B-rIgG-T20 affinity column. Flow cytometry showed that rabbit anti-T20 (r-anti-T20) could bind to RAW264.7 cells and that this combination could be blocked by T20 peptide. Fluorescence microscopy also showed that anti-T20 (r-anti-T20) could bind to RAW264.7 cells .OVA-specific IgG titers were approximately 1 : 500,000 and IgE titers were approximately 1 : 400 detected by ELISA method using purified OVA as the coating antigen  were significantly increased at 30 min and 60 min after OVA challenge as compared with OVA allergic mice (OVA model), and anti-T20 treatment (OVA model plus PGN plus r-anti-T20) markedly attenuated this increase, but not isotype control of anti-T20 (OVA model plus PGN plus rIgG)  . Also, sus rIgG) .TLR2-mediated inflammatory signaling and hypersensitivity reactions may be blocked by at least two ways: first, the intracellular domain is prone to gene mutation or deletion or by blocking the intracellular signal transduction pathway. However, although this does not affect the extracellular segment recognition , 12 and Currently, the study of TLR2-mediated identification of its agonists still cannot show the exact role of the different domains of the TLR2 extracellular domain in the ligand recognition process. In addition to the polyclonal antibody of the TLR2 extracellular domain targeted against the 26 peptide (179L-204I), other commercial and laboratory prepared anti-TLR2 monoclonal antibody preparations and polyclonal antibody against the TLR2 extracellular domain remain mostly unclear. In this study, we used a protein epitope prediction system and synthetic peptide technology that helped to predict B cell dominant epitope of mouse TLR-2, which can clearly show the target domain and avoid using the full-length and extracellular domain of mouse TLR-2 as an immunogen. in vitro and in vivo.For TLR2 agonists, we used three kinds of them (i.e., Pam3CSK4 , LTA 1717, 18, aα and IL-6 in PGN-challenged OVA allergic mice were significantly increased as compared with only OVA allergic mice, but anti-T20 treatment markedly attenuated their increase. The results also showed that anti-T20 only reduces PGN/OVA mediated anaphylaxis but has no effect on only OVA induced anaphylaxis. Also, serum levels of LTC4 had similar tendencies. We noted that the role of the TLR2 signaling pathway in anaphylaxis and its presence are still controversial, and this might be related with the usage of the type and dose of the specific TLR2 agonist or antagonist, the target cell type, and the type of inflammatory factors, as well as the use of different allergic animal model [ S. aureus-derived PGN causes mast cells to release TNF-α, IL-4, IL-5, and IL-6, degranulation and to open Ca2+ channels by binding to TLR2 on the surface of mast cells [In view of gradually rising incidences of allergic diseases that are often associated with an infection, which can exacerbate allergic reaction , here, wal model –29. McCust cells . Moreovest cells . Meanwhist cells –34. FurtIn summary, this study produced a new antibody against the specific extracellular domain of TLR2 which has protective effect on TLR2 agonists-driven inflammatory and allergic response.


Octodon degus) is a diurnal rodent with dichromatic color vision. Its retinal structure is similar to that in humans in many respects, therefore, it is well suited to study retinal aging. Histological, cell type-specific and ultrastructural alterations were examined in 6-, 12- and 36-months old degus. The characteristic layers of the retina were present at all ages, but slightly loosened tissue structure could be observed in 36-month-old animals both at light and electron microscopic levels. Elevated Glial fibrillary acidic protein (GFAP) expression was observed in Müller glial cells in aging retinas. The number of rod bipolar cells and the ganglion cells was reduced in the aging specimens, while that of cone bipolar cells remained unchanged. Other age-related differences were detected at ultrastructural level: alteration of the retinal pigment epithelium and degenerated photoreceptor cells were evident. Ribbon synapses were sparse and often differed in morphology from those in the young animals. These results support our hypothesis that (i) the rod pathway seems to be more sensitive than the cone pathway to age-related cell loss; (ii) structural changes in the basement membrane of pigment epithelial cells can be one of the early signs of degenerative processes; (iii) the loss of synaptic proteins especially from those of the ribbon synapses are characteristic; and (iv) the degu retina may be a suitable model for studying retinal aging.The retina is sensitive to age-dependent degeneration. To find suitable animal models to understand and map this process has particular importance. The degu ( The vertebrate retina, like other parts of the central nervous system, is subjected to degenerative changes caused by aging. The retina is also the site of diseases for which age is a major risk factor, including macular degeneration and glaucoma . This latter species presents several advantages for studying different pathological conditions. The degu is a diurnal, highly visual South American hystricomorph rodent native to Chile, which in old age expresses cognitive deficits, anxiety . Since degu retina is more similar to the human retina than to the retina of other rodents, this description will provide a strong foundation for future studies where experimental manipulations and/or neuroprotective agents can be studied.n = 8), 12 (n = 8), and 36-months of age (n = 12) were used. This latter group is considered as aging (but not old) group. Degus were housed individually in opaque glass cages (40 × 25 × 25 cm) at the animal facilities of the University of Murcia. Throughout the study, the experimental room was maintained under controlled temperature (21 ± 1°C) and 12 h light/dark cycle (lights on at 7:00 a.m. and off at 19:00 p.m.). The floors of the cages were covered with wood shavings that were changed once a week. Food and water were provided ad libitum by placing 120 g food pellets  per day and water bottles on a grid located on the top of the tank. The water in the tank was changed daily. All experiments were performed in accordance with relevant regulatory standards, experimental guidelines and procedures complied with the European Community Council Directive (2010/63/UE) and the ethical committee of the University of Murcia.A total of 28 female degus (body weight 180–270 g) of 6 , administered with a continuous flow vaporizer , and then sacrificed by decapitation. Both eyes were immediately removed and distinctly disposed according to histological or electron microscopic procedure to be performed.n = 2–5 measurements from one tissue block). The following parameters were measured: (i) cross-section of the retina from the outer limiting membrane (OLM) to the inner limiting membrane (ILM); (ii) the width of individual retinal layers. Statistical comparisons were made using one-way ANOVA test followed by Tukey-B posthoc analysis. Data were presented as mean ± SEM (GraphPadPrism5.0).For histology eyes were fixed in 4% paraformaldehyde  dissolved in 0.1M phosphate buffer . The eyecups were dissected and embedded in epoxy resin  as we previously described . Sections were cut at 70 nm in Reichert Ultracut S and counterstained with Reynold’s lead citrate. Samples were examined and photographed in a JEOL 1200EX electron microscope.Electron microscopy was performed on eyes fixed with 4% PFA supplemented with 1% glutaraldehyde dissolved in 0.1M PB. After washing in PB, tissue samples were treated with 1% OsOEyes were dissected immediately after sacrifice in ice-cold phosphate buffer with saline (PBS) and fixed in 4% PFA at room temperature. Tissues were then washed in PBS and cryoprotected in 20% sucrose at 4°C. For cryostat sectioning, retinas were embedded in tissue-freezing medium , cut in a cryostat  at 10 μm radially. Sections were mounted on subbed slides. Primary antibodies and peanut agglutinin-conjugated with FITC (PNA) were used overnight at room temperature .The number of RGCs , outer nuclear layer (ONL), outer plexiform layer (OPL), inner nuclear layer (INL), inner plexiform layer (IPL) and GCL . The number of RGCs (Brn3a-positive cells)/100 μm retina length was significantly decreased in the 36-month-old degu retina -positivity was selectively localized to endfeet of Müller cells Figures . Müller Basement membrane proliferation of the RPE in the areas of age-related retinal peripheral degeneration was observed in the 36-month-old degus retinas. The thickening of Bruch’s membrane and fibrosis of the choriocapillary were evident. The somas of the RPE cells were pressed toward this thinner basement membrane in the 36-month-old retinas, suggesting an initial age-related alteration of the RPE. Early peripheral changes of the RPE included increased basal infoldings, phagolysosomes and lipofuscin deposits, as well as atrophy and whirling extensions of the basement membrane into the cytoplasm Figure . The relWe observed that the cones composed a nest between the rods in all age groups, in accordance with the cone-dominant retinal structure of the degu  were more electron dense and cup-shaped spherules, while cone terminals were bowl-shaped, less dense pedicles Figures . The onlPhotoreceptors transmit their signals at ribbon synapses in the OPL, the first synaptic region in the retina, whereas bipolar cells make their ribbon synaptic contacts in the IPL. A large number of regularly aligned synaptic vesicles were tethered to the ribbon. The rod photoreceptor ribbon synapses had horseshoe-shaped structure in the OPL, however, the cone photoreceptor (OPL) and bipolar cell ribbons (IPL) had a dot-like appearance at light microscopic level. We analyzed the structure of the OPL with retina-specific ribbon synapse markers presynaptic CtBP2 and Bassoon labeling. The rod ribbon synaptic profiles were marked with horseshoe-shaped ribbons by CtBP2 and continuous distribution of punctate staining by Bassoon protein Figures . Near thThe pan-bipolar cell marker Chx10 reveals the organization of bipolar cells in the INL of the retina . The most prominent difference was at the age of 36 months with a significant reduction in these parameters  rod/cone ratio is similar to the human ratio; (ii) the diurnal behavior is charactheristic to this species and its activity pattern resembles that of the human; (iii) its lifespan is considerably longer than those of the other experimental models, therefore the age-related changes can be better monitored and compared to those described in the human retina; and (iv) other, age-related diseases  often appear spontaneously in this species  depends on the underlying structure of neural networks. Any loss of the pre- and/or postsynaptic profiles of the retinal neurons causes changes in their morphology and function. As a consequence, these retained neurons may be capable to establish new synaptic contacts , so the retina may undergo marked remodeling. In the aging degu retina, the synaptic rearrangements/alterations in the OPL and potentially concomitant synaptic alterations in the IPL efficacy would reduce transmission, while the loss of function of the RPE cells would alter the homeostasis of the retinas. The neuronal elements of the vertical pathway such as rod bipolar and ganglion cells were seen affected already at 36-month of age. We think that these structural changes become more obvious, will reach other cells and will more seriously affect the synaptic complexes with advancing age.The proven similarities between the degus and the human retina in their structure and aging processes offer a possibility to develop potential treatments and therapies for retinal age-related alterations and diseases.KS researched data and wrote, edited and reviewed the manuscript; CE, EF-V, ET, VI and GS Jr. researched data; RG and MTH researched data, reviewed and edited the manuscript; DR and AT reviewed the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.