The burden of hearing loss is on the increase, especially in low-income countries such as South Africa. The need for urgent action to prevent ear and hearing problems is a priority, especially as in many cases permanent hearing loss is preventable. In South Africa, as in other developing countries, there is a limited number of hearing health professionals and audiological resources. The lack of hearing health services may impact the general public’s awareness of hearing and hearing health. Limited information is available on the South African public’s knowledge of audiologists and the services they provide, especially in underserved rural communities.The aim of this study was to describe individuals’ awareness of the audiology profession, hearing and hearing loss, and hearing health in a rural area of the Limpopo Province.A cross-sectional survey design was employed for the purpose of this study. Using a random sampling strategy, 297 households in four rural villages were selected and a self-developed questionnaire was administered to one individual (18 years and older) per household. The questionnaire consisted of 23 questions targeting awareness of the audiology profession, as well as knowledge of hearing, hearing loss and hearing health.Only 14% of participants were aware of the audiology profession, indicating that individuals living in rural communities are not aware of the role of audiologists and the services they provide. Doctors and nurses were identified by participants as the individuals who assist them with hearing-related problems. Although most participants (87%) acknowledged that it is very important to undergo a hearing test, only 5% have previously visited an audiologist. Most participants were aware that ear infections and excessive noise exposure can cause hearing loss. The majority also believed that ears must be kept clean at all times and used cotton buds to maintain ear hygiene.There is a general lack of public awareness of audiologists and the services they offer. This study highlighted the need for the National Department of Health in collaboration with professional associations and hearing health professionals to develop and implement effective strategies to increase the South African public’s awareness of the profession and the services they provide. South African universities can also play a significant role in teaching students to develop context-relevant strategies to increase awareness of the profession. Audiologists are health care professionals who assess, diagnose and treat hearing- and balance-related problems  recently highlighted the importance of intensifying action to prevent deafness and hearing loss as worldwide approximately 360 million people live with disabling hearing impairmentInformation on the prevalence and causes of hearing impairment (across the lifespan) is available for the South African population with the bulk of the research conducted in urban areas and/or within the private health care sector  area of the Sekhukhune District in the southern part of the Limpopo Province. The Sekhukhune District is one of the poorest districts in the country and is characterised by poor infrastructure and lack of safe water supply  and the majority  were females. The home language of most participants was isiZulu , followed by Sepedi , whilst the remaining 21% (n = 63) speak a combination of Xitsonga, Tshivenda, isiNdebele and Shona. Most participants  were unemployed. Only 51% (n = 151) of the participants had some form of high school education, and 12% (n = 36) had no formal education.A total of 297 individuals were included in the study. The average age of the participants was 38.5 years . Each research assistant was required to administer the questionnaire on five community members who met the same inclusion criteria as for the main study. The 30 pilot study participants resided in the same communities where the research was conducted. The objectives of the pilot study were to ensure the face and content validity of the questionnaire as well as to ensure that there were no discrepancies with regard to the documentation of participant responses.Ethical clearance was obtained from the Human Research Ethics Committee  of the University of Witwatersrand before the study commenced (Protocol number M121006). Permission to conduct the study was obtained from the relevant tribal authorities (e.g. chiefs). Every third household in each of the villages was visited by the research team. Individuals in each of the selected households were informed of the study and only one individual per household was requested to participate in the study. Participants were fully informed about the nature of the study, and assured of confidentiality and their rights to withdraw from the study at any time without negative consequences. Only participants who gave written consent were included in the study. Participants who required hearing health care were referred to their local primary health care clinic or the audiology department. All completed questionnaires were verified for completeness by the audiologist and data were captured on a password-protected Excel spreadsheet. Raw data were stored in a locked cabinet and will be destroyed after a period of 5 years.The results will be presented as it relates to the: (1) awareness of the audiology profession, (2) hearing and hearing loss and (3) hearing health.n = 255) were not aware of the audiology profession. A large majority indicated that other health professionals can test hearing  of the participants were aware that audiologists are the health professionals responsible for testing hearing (n = 15) and to help with ear problems (n = 22). These participants indicated word-of-mouth  and information from other health workers  as their sources of information.Only 14% (n = 257) agreed that it was ‘greatly important’ to have their hearing tested. Only two participants indicated that it was ‘not important at all’. The majority of participants  agreed that everyone’s hearing can be tested regardless of age. Interestingly, only 5% (n = 16) of participants have previously visited an audiologist for a hearing test.Despite the lack of awareness about the audiology profession, many of the participants  indicated that ear infections can cause hearing loss. Most participants  specified that they ‘hardly ever’ have ear infections, whilst only 5% (n = 15) have ear infections ‘frequently’ and 3% (n = 9) ‘almost always’. Interesting responses were recorded regarding the actions that participants take when they have an ear infection. The majority 38% (n = 114) indicated that they consult with medical professionals at a clinic or hospital, whilst 7% (n = 22) reported that they do nothing. Twelve per cent (n = 37) reported that they insert oil in their ears. The types of oil inserted included sweet oil (n = 12), cooking oil (n = 9), fish oil (n = 6), chicken fat or oil (n = 5), glycerine (n = 4) and castor oil (n = 1). Only one participant reported consulting a traditional healer for treatment of hearing loss or/and ear infection.When asked whether an ear infection can cause hearing loss, the majority of the participants  agreed that excessively loud noise can damage hearing. Participants were required to indicate which type of noise typically found in their village can cause damage , lowering the volume of the music they are listening to  and using some form of ear protection, such as cotton wool and ear plugs .The last question probed how participants thought they can protect their hearing from excessive noise. The results are presented in n = 190) also use it when their ears are itchy.Most of the participants believed that ears must be kept clean at all times and indicated that they use cotton buds, matchsticks and a variety of other items to clean their ears . Of the There is a general lack of awareness of audiology as a profession in the community where the research was conducted. This is despite the availability of audiological services at the two hospitals and one local non-government clinic in the EMLM area. It is postulated that health care workers  based at the two hospitals and various primary health care clinics are not aware of the availability of audiological services in the EMLM. Participants who were aware of the audiology profession indicated that they were made aware of the services offered primarily by other health care workers and also through word-of-mouth. This supports the notion that the primary source of health-related information is through health workers who do health talks in hospitals and clinics  of the participants did not know what to do if a hearing problem is noted. The majority of participants  correctly indicated that they will consult a health care worker  at either the clinic or the hospital. Ten per cent (n = 28) also mentioned treatment options such as being fitted with a hearing aid or undergoing an operation. The consultation sources and suggested treatment options mentioned by participants are appropriate. However, without referral by health professionals to audiologists, the early identification and appropriate management of hearing impairment  may impact the quality of life of individuals with hearing impairment  into the ear. The practice of inserting oil to treat ear infection has also been reported in rural African communities  . However, some participants offered inappropriate suggestions  or did not know how to protect hearing from excessive noise.Ear hygiene practices comprised mostly the use of objects  to clean or scratch ears. This is despite the potential complications associated with the use of these objects in ears. Common complications associated with the use of cotton buds include otitis externa, otomycosis, laceration in the external auditory meatus, accumulation of wax and perforated tympanic membrane  the audiology profession and its role in the identification and management of hearing- and balance-related problems, (2) the availability of audiological services in the area, (3) the impact of excessive noise exposure on hearing and strategies on how to protect hearing and (4) ear hygiene, specifically why the use of cotton buds is not advised.It is, however, imperative that the National Department of Health prepare, implement and monitor a national plan for the prevention and control of major causes of avoidable hearing impairment within the framework of primary health care (Olusanya et al., It is further recommended that similar studies should be conducted in a variety of other contexts to determine the extent of education required to prevent avoidable hearing- and balance-related problems.


This special issue contains a collection of papers highlighting the collaborative research taking place at Macquarie University’s Australian Hearing Hub. Included in this introduction is a brief outline of the challenges in the hearing health and communication space and a brief description of the Australian Hearing Hub and its members, alongside an overview of the studies included in this special issue. It is estimated that 360 million people worldwide have a hearing loss , impactiThe Australian Hearing Hub is an initiative of Macquarie University and the Commonwealth Government’s Education Investment Fund. It brings together leading researchers and health-care organizations to drive research, education, and innovation to improve the lives of people with hearing, communication, or mental health disorders. Its innovative framework of engagement and collaboration enables researchers, educators, and clinicians to identify and address challenges in hearing health and communication over the life span, conduct joint research projects, and implement effective long-term changes in policy and practice. Major partners in the Australian Hearing Hub include Cochlear Limited, whose world headquarters are located adjacent to the Australian Hearing Hub, Australian Hearing, the largest provider of government-funded hearing services, and its world-recognized research division, the National Acoustic Laboratories. Other key partners include the Royal Institute for Deaf and Blind Children and its associated cochlear-implant program, the Sydney Cochlear Implant Centre, The Shepherd Centre and the Cooperative Research Centre, and The HEARing CRC. Allied with Macquarie’s leadership in the Australian Research Council’s Centre for Cognition and its Disorders, a pan-Australian, indeed global, partnership of research centers dedicated to understanding human cognition and communication, the partners work together to improve communication over the life course.The success of the Australian Hearing Hub collaboration lies not only in the generation of published articles but also advancing clinical best practices and the development of new technologies to improve hearing and communication over the life course. This special issue collates a range of studies from partners within the Australian Hearing Hub. It not only demonstrates the truly collaborative nature of the Hub—each has two or more Hub partners working on the project—but also examines hearing health and communication issues with the specific intent of using evidence-based research to improve the lives of people with hearing loss and communication disorders now and in the future. The Australian Hearing Hub regards evidence-based practice as paramount to developing a successful long-term hearing health-care model. One study  investigThe overarching theme of the papers in this special issue is of collaboration—collaboration between universities and government agencies, research partnerships with commercial entities, and engagement with charities and clinical organizations, each with a mission that spans basic science to delivery of hearing health care. This holistic approach to the investigation of hearing and deafness is integral to the increasingly shared vision of the Hub partners as they seek to develop new therapeutic interventions and care pathways. It also sets the agenda for the next stage of the Hub’s development, where joint research frameworks between partners will be employed to leverage significantly more resources for research and innovation than can be achieved separately. Australia’s research-funding ecosystem, its well-resourced health programmes, and the public visibility of iconic Australian “brands” such as Cochlear Limited and Australian Hearing, make this a real possibility. An immediate goal is to ensure hearing becomes Australia’s 10th National Health Priority. From that vantage point, the Australian Hearing Hub can become a model of best practice, pioneering the way in which new therapies and interventions are rapidly assessed for efficacy, and implemented into practice, not just in Australia, but globally.


CIB2 is widely expressed in various human and animal tissues, mainly in skeletal muscle, nervous tissue, inner ear, and retina. The CIB2 protein is responsible for maintaining Ca2+ homeostasis in cells and interacting with integrins—transmembrane receptors essential for cell adhesion, migration, and activation of signaling pathways. Calcium signaling pathway is crucial for signal transduction in the inner ear, and integrins regulate hair cell differentiation and maturation of the stereocilia. To date, mutations detected in CIB2 are causative for nonsyndromic hearing loss (DFNB48) or Usher syndrome type 1 J. Patients harboring biallelic CIB2 mutations suffer from bilateral, early onset, moderate to profound HI. In the paper, we summarize the current status of the research on CIB2.Hearing plays a crucial role in human development. Receiving and processing sounds are essential for the advancement of the speech ability during the early childhood and for a proper functioning in the society. Hearing loss is one of the most frequent disabilities that affect human senses. It can be caused by genetic or environmental factors or both of them. Calcium- and integrin-binding protein 2 (CIB2) is one of the recently identified genes, involved in HI pathogenesis.  Receiving and processing sounds are essential for a proper development of communication skills during the early childhood. Hearing loss is one of the most frequent disabilities that affect human senses. According to the World Health Organization , over 5 More than a half (50–60 %) of the congenital hearing loss cases are due to genetic factors. To date, approximately 300 genes are considered related to the process of hearing . Most ofCIB2  gene has been added to the extensive list of genes associated with hearing, loss [Recently, ng, loss .CIB2 gene (MIM# 605564) is localized on chromosome 15 (15q25.1) [2+ is a crucial molecule in cellular signaling pathways and also takes part in signal transduction in the inner ear in the organ of Corti. It is one of the factors determining the transmission of sound and balance information through the hair cell apical mechanosensitive transduction (MET) channels to the ribbon synapse at the bottom of the hair cells. MET channels participate in the transport of potassium and calcium ions from the endolymph to the hair cells. Ca2+ is a second messenger that induces conformational changes in effector molecules regulating the sensitivity of the cochlear amplifier, which is a mechanism of increasing the amplitude and frequency selectivity of sound waves. Consequently, the intracellular Ca2+ concentration is crucial for MET channels adaptation, frequency tuning, hair bundle twitching, outer hair cells (OHC), electromotility, and afferent synaptic transmission [The ene MIM# 05564 is 15q25.1) . Ca2+ issmission –10.Fig. Integrins are essential for cell adhesion and activation of intra- and extracellular signaling pathways. They are α/β heterodimeric transmembrane receptors for ligands in the extracellular matrix Fig. b. IntegrCIB gene family started in the 1990s when two independent research groups found a novel protein that was named according to its putative function, i.e., CIB  [2+ and undergo N-myristoylation [CIB2, the human genome contains three other genes  encoding highly homologous proteins, which together form a family of calcium-binding proteins and contain elongation factor-hand (EF-hand) domains [Studies on the protein)  or KIP (protein) , respectprotein) , 15–17. oylation . The latoylation . Except  domains .CIB2 mRNA was transcribed in a wide variety of human tissues, including brain, lungs, skeletal muscles [CIB2 in the human ear or retina. Further studies in mice confirmed that expression of Cib2 included also the inner ear (cochlea) and retina [Nonquantitative reverse transcription PCR analysis of human tissues showed that  muscles , and hum muscles , but so d retina . Particud retina . When exd retina –27. It id retina .Cib2 gene are present in embryos, throughout postnatal development as well as in adult mice [Cib2 expression was observed in forebrain, midbrain, hindbrain, spinal cord, somites, inner ear, vibrissae, gut, and musculature in mouse embryos. In the ear, Cib2 transcripts were detected in the cytoplasm of adult supporting cells, inner hair cells, OHC, cuticular plate , and along the stereocilia. The signal for Cib2 was usually more intense in the shorter row of stereocilia tips than in the longer row. The transcript was first observed at postnatal day 2 in the developing organ of Corti and vestibular organs. Until postnatal day 8, it was limited to supporting cells in the organ of Corti. In the retina, Cib2 was localized in inner and outer segments of photoreceptor cells and in retinal pigment epithelium. A signal of diffused immunoreactivity was also detected in the inner plexiform layers, outer plexiform layers, and the ganglion cell layer [Cib2 mRNA is mainly expressed in skeletal muscles. Within the muscle, Cib2 is expressed in sarcolemma, enriched in the myotendinous junctions and neuromuscular junctions. Lower levels of expression were noticeable in brain and in lungs [Transcripts of the ult mice . Using ill layer . Semiquain lungs .Cib2 transcript was observed in several areas and the highest level of expression was detected in the hippocampus (cornu ammonis area three regions and dentate gyrus). Furthermore, it was also found in sensory, entorhinal, and prefrontal cortex. Intracellular localization of the Cib2 protein in rats is considered to be the Golgi apparatus and in nerve cells the protein localizes mainly to neurites [In the rat brain tissue, the neurites .Cib2 mRNA was detected in many various tissues, mainly in stomach, heart, and ovary [Cib2 in zebrafish is detected throughout the development. Drosophila gene CG9236, encoding a protein which is in 71 % similar and in 59 % identical to the human CIB2, is expressed in several larval and imago tissues, therein the adult eye [In sheep, the expression of nd ovary . Express2+ and Mg2+ ions. They are considered as regulatory motifs that mediate responses to changes in calcium concentration and fulfill a role of intracellular calcium signaling mediators. CIB2 contains three EF-hand domains and through the second and third domains, it is able to bind Ca2+  have been discovered: c.97C>T (p.Arg33*), c.192G>C (p.Glu64Asp), c.196C>T (p.Arg66Trp), c.272T>C (p.Phe91Ser), c.297C>G (p.Cys99Trp), c.368T>C (p.Ile123Thr), and c.556C>T (p.Arg186Trp) [CIB2 mutations can be assigned to isoform B, it indicates that this particular isoform is the most significant one for the process of sound transduction, but further research is necessary to confirm the presumption.To date, seven mutations in the g186Trp) . All of g186Trp) , while cCIB2 gene present a recessive pattern of inheritance. The majority of them lead to hearing loss (DFNB48) and only p.Glu64Asp was identified in a family with Usher syndrome type 1 J. All individuals harboring homozygous or compound heterozygous CIB2 variants suffered from the early onset, bilateral, moderate to profound HI [CIB2 mutations present Pakistani and Turkish origin, further research revealed also Dutch and Caribbean Hispanic CIB2 families. It indicates that individuals originating from other than Pakistani and Turkish populations may carry mutations in the CIB2 gene [To date, all mutations discovered in the found HI , 38, 39.IB2 gene , 38, 39.CIB2 [2+ binding by the second EF-hand domain. On the contrary, the c.368T>C variant probably magnifies the affinity of Ca2+ binding. None of the mutations mentioned above induce significant changes in CIB2 distribution in tissues [The c.192G>C, c.272T>C, and c.297C>G variants probably reduce the interaction of CIB2 with integrins and modify their activation. The c.192G>C mutation changes the protein conformation, thereby it affects binding affinity or kinetics of integrin. All these amino acids substitutions may cause slight changes in subcellular location of the protein which possibly affects the efficiency of calcium sequestration . NeverthCIB2 . The c.2 tissues . The c.5 tissues . Variant tissues , 40.CIB2 gene. The deletion of this region was reported in patients suffering from abnormalities of the ears  hypotonia and developmental delay. Linkage analysis in a Pakistani family with spasticity, severe mental retardation and visual impairment, pointed that the CIB2 gene may be involved in the pathogenesis of these abnormalities [CIB2 expression may also play a protective role in cardiovascular diseases by decreasing the pace of the vascular calcification [Different disorders other than HI or Usher syndrome have also been linked to the 15q24 locus, containing the malities , 41, 42.fication .CIB2 gene, its function still remains unclear and is to be fully discovered. Mutations in CIB2, which so far have been revealed, segregate with both DFNB48 and USH1 J. Thus, CIB2 is a causative gene for both disorders. Nevertheless, further studies are required for a better understanding of the role of the CIB2 gene in human. Clarification of its function and associated molecular mechanisms will be the next step towards better prevention and treatment of hearing loss or cardiovascular diseases in patients, thereby towards improved living standards of people at risk of CIB2-associated diseases.Although some research has already been performed on the


This study evaluated the prevalence of minimal hearing loss (MHL) in South Korea based on the 2010 to 2012 Korea National Health and Nutrition Examination Survey. A total of 16,630 representative individuals (older than 12 years) who completed ear examinations and structured questionnaires were analyzed. Only participants who had normal tympanic membranes were included. MHL was categorized into the following three groups: 1) unilateral sensorineural hearing loss  ≥ 15 dB in the affected ear), 2) bilateral sensorineural hearing loss , and 3) high-frequency sensorineural hearing loss . To evaluate clinical symptoms, subjective hearing status, tinnitus, and quality of life of each MHL group were compared to those of normal-hearing listeners. The use of hearing aids (HAs) was also investigated in the MHL population. The prevalence of normal hearing and MHL were 58.4% and 37.4%, respectively. In univariate analyses, the prevalence of MHL increased with age. It was significantly increased in males. Regarding clinical symptoms, 13.0% and 92.1% of participants with MHL reported difficulties with hearing and annoying tinnitus, respectively. In multivariate analyses, these proportions were significantly higher in the MHL groups than in normal-hearing listeners. Participants with MHL also showed significantly lower Euro Qol-5D index scores than did normal-hearing listeners. Regarding hearing rehabilitation, among minimally hearing impaired participants with subjective hearing loss, only 0.47% of individuals used HAs. Our results reveal that MHL is common in South Korea. It is associated with significant subjective hearing loss, tinnitus, and poor quality of life. Therefore, clinicians need to pay attention to this special group and provide proper counselling and rehabilitative management. The prevalence of hearing impairment is increasing owing to an aging society and growing use of personal listening devices , 2. HearIn an earlier study, Bess et al. 1998) categorized minimal hearing loss (MHL) into three distinct groups  and demonstrated an association between MHL and educational performance and functional status in school-aged children , 8. Alth categoriThus, the objective of this study was to determine the prevalence of MHL in South Korea based on national survey data obtained from the 2010 to 2012 Korea National Health and Nutrition Examination Survey (KNHANES) and assess the quality of life of people with MHL. The definition of MHL used in this study was based on the previous study by Bess et al. (1998).This study used the data from the fifth KNHANES. The KNHANES is a nationwide survey conducted annually by the Korea Centers for Disease Control and Prevention to investigate the health and nutritional status of a representative Korean population. Every year, about 10,000 individuals in 3,840 households are selected from a panel to represent the population through a multistage clustered and stratified random sampling method based on the National Census Data. A total of 576 survey areas were drawn from the population and housing census by considering the proportion of each subgroup. The participation rate of selected households was about 80%. From 2010 to 2012, a total of 23,621 individuals  agreed to participate in the health surveys. They underwent ear, nose, and throat (ENT) examinations. To exclude mixed or conductive hearing loss, individuals with tympanic membrane perforation and cholesteatomatous conditions including retraction pocket, otitis media with effusion, and insertion of a ventilation tube were excluded. Among 19,864 participants who had normal tympanic membranes, 16,630 participants completed both the audiometric measurement and the ENT questionnaire.All participants provided written informed consent before completing the survey. KNHANES followed the tenets of the Declaration of Helsinki for biomedical research. It was approved by the Institutional Review Board of the Korean Centers for Disease Control and Prevention . Written informed consent was also obtained from the next of kin, caretakers, or guardians of minors/children enrolled in this survey. Approval for this research study was obtained from the Institutional Review Board of Samsung Medical Center (IRB No. 2016-02-076).Pure tone threshold was measured in a sound-proof booth using an automatic audiometer . Otolaryngologists who had been trained to operate the audiometer provided instructions to participants and obtained recordings. Audiometry was performed for participants over 12 years of age. Only air conduction thresholds were measured. Supra-auricular headphones were used in the soundproof booth. The otolaryngologist provided basic instructions to participants regarding the automated hearing test. Automated testing was programmed using a modified Hughson-Westlake procedure with a single pure tone for 1–2 seconds. The lowest pure tone level at which the subject’s response rate was 50% was set as the threshold. Participants responded by pushing a button when they heard a tone. Results were automatically recorded. The following frequencies were tested: 0.5, 1, 2, 3, 4, and 6 kHz. An ear examination was conducted with a 4 mm 0°-angled rigid endoscope attached to a Charge-Coupled Device (CCD) camera by trained otolaryngologists. Endoscopic examination was performed to identify tympanic membrane perforation, cholesteatoma (including retraction pocket), and otitis media with effusion (including the presence of a ventilation tube).Bess et al. (1998): unilateral sensorineural hearing loss (USHL), bilateral sensorineural hearing loss (BSHL), and high-frequency sensorineural hearing loss (HFSHL) [Minimal sensorineural hearing loss was categorized into three distinct groups according to t al. 199: unilateTo determine clinical symptoms, participants completed a questionnaire asking about their hearing and whether they had any symptoms of tinnitus. Subjective hearing status was measured by asking the following survey question: “Which sentence best describes your hearing status (while using no HAs)?”. There were four answers for the question: (1) “Don’t feel difficulty at all,” (2) “A little bit difficult,” (3) “Very difficult,” and (4) “Can’t hear at all.” Subjective hearing loss was indicated when the response was (2), (3), or (4). Participants were also asked about their experience with tinnitus. In response to the question “Within the past year, did you ever hear a sound  originating in your ear?”, examiners were instructed to record “yes” if a participant reported that they heard an odd or unusual noise at any time in the past year. Participants who responded positively to this question were then queried concerning the resulting annoyance in their lives using the following questions: “How severe is this noise in daily life?” . Participants were assigned to the group with annoying tinnitus if the severity of tinnitus was annoying or severely annoying. Regarding quality of life, the Euro Qol-5D (EQ-5D) was used to evaluate all participants aged 18 years or older. The EQ-5D is a standard tool used to measure patients’ health status in the following five dimensions: mobility, self-care, usual activities, pain/discomfort, and anxiety/depression , 15. EacP-values were two-sided. Bonferroni’s correction was applied to P-values and the corresponding confidence intervals owing to multiple testing. Statistical significance was considered when an adjusted P-value was less than 0.05.All statistical analyses were performed by taking into account the weights from the complex sampling design according to the guidelines for analysis of KNHANES data obtained by the Korea Centers for Disease Control and Prevention. The survey design created a sample weight assigned to each sample individual through the following three steps so that the total sample would represent the population (on average) for the 3-year period (2010–2012): calculating the base weight of the inverse of the final probability of an individual being selected, adjusting for non-response, and post-stratification adjustment to match previous census population control totals. The weights in the 2010, 2011, and 2012 surveys were combined and the average weight (weight for each year/3) was calculated. Statistical analyses were performed using SAS version 9.4 . The prevalence of MHL was then estimated. The chi-squared test was used to compare the prevalence of MHL among age groups and according to sex. Logistic regression or linear regression was used to compare the prevalence of MHL to normal hearing according to the responses to the questionnaires. Of 16,630 participants, 58.4% had normal hearing, while 37.4% had MHL . BSHL acP < 0.0001) differed among age groups based on chi-squared testing and post-hoc analysis, except amongst those in their forties (40 to 49 years of age) or fifties (50 to 59 years of age). The prevalence of MHL increased with age until the 6th decade of life (60 to 69 years of age) and decreased afterwards . It was higher in males  . However, sex was associated with the prevalence of MHL only for HFSHL. HFSHL was significantly predominant in males . The prevalence of MHL in the HFSHL subgroup was also higher in males, regardless of whether the ear was affected unilaterally  or bilaterally .After excluding participants who had worse than moderate hearing loss (PTA > 40 dB HL either in one ear or both ears), a total of 15,569 participants were analyzed to compare the prevalence of MHL to that of normal hearing using linear and logistic regression analyses. In univariate analyses, the prevalence of MHL increased with age (7–1.534) . The preP < 0.0001, OR: 2.729, 95% CI 2.217–3.360) after adjusting for age and sex. While 13.0% of participants with MHL reported difficulties with their hearing, only 3.1% of participants with normal hearing reported subjective hearing loss. In multivariate analyses, participants with MHL also complained of tinnitus significantly more than did normal hearing participants . Annoying tinnitus was observed significantly more often in participants with MHL compared to those with normal hearing . In the subgroups, participants with mild-to-profound USHL complained of their hearing difficulties the most , followed by participants with mild BSHL . Regarding tinnitus, participants with mild BSHL mostly reported tinnitus , while participants with mild-to-profound USHL mostly reported annoying tinnitus .Comparisons of participants with MHL and normal hearing regarding subjective complaints of hearing loss and tinnitus are summarized in P < 0.0001) lower mean EQ-5D index score in the linear regression analysis after adjusting for age and sex. The average EQ-5D indices in the normal hearing and MHL groups were 0.972 and 0.935, respectively. In the subgroups, the mild BSHL group had the lowest average score on the EQ-5D (0.889), followed by the minimal BSHL group (0.926).A total of 13,730 participants completed the EQ-5D survey. Compared to the normal hearing group, the MHL group had a significantly  in logistic regression analysis after adjusting for age and sex.The use of HAs among those with MHL is shown in Using data from the KNHANES 2010–2012, we found that the weighted prevalence of MHL in the South Korean population aged 12 years or older was 37.4%. When MHL was divided into subgroups , the prevalence of USHL, BSHL, and HFSHL were 14%, 16%, and 7.4%, respectively. These prevalence rates are similar to those found in a previous survey, although there might be some differences in the definitions of hearing loss. The WHO prevalence statistics from 2012 were based on a definition of mild hearing loss as an average threshold at 0.5, 1, 2, and 4 kHz of between 26 and 40 dB HL . The BSHIn the present study, BSHL was the most prevalent category (48.5%), followed by USHL (34.6%) and HFSL 16.8%). Compared to a population-based study on Canadian children aged 0 to 18 years, the prevalence of each subgroup was slightly different. Although BSHL was the most prevalent category in both studies, USHL was more prevalent in adults aged 12 years or older than in children aged 18 years or younger  6.8%. Com.The present study found that the proportion of participants complaining about their hearing with MHL was significantly higher than the proportion of normal hearing participants complaining about their hearing in univariate and multivariate analyses . ParticiDespite suffering from subjective hearing loss, only 0.47% of participants with MHL reported that they currently use HAs . A previIn conclusion, MHL is common in South Korea. It is associated with significant hearing problems, including subjective hearing discomfort, tinnitus, and poor quality of life. Nevertheless, hearing rehabilitation is extremely limited for patients with MHL. Therefore, minimally hearing-impaired patients, especially those with hearing handicaps, might be considered as candidates for auditory rehabilitation, including counselling regarding communication strategies and the option to evaluate the potential benefits of sound amplification.


Importance. Rituximab was not effective in ameliorating the hearing loss in a patient with atypical Cogan's syndrome. Observations. We report the case of a patient who developed acute bilateral uveitis and sensorineural hearing loss. A diagnosis of atypical Cogan's syndrome was made. The patient's hearing loss did not improve despite high dose steroids and azathioprine. Rituximab was administered given a recent report of its efficacy in a patient with refractory disease; however, our patient's hearing loss did not improve. Conclusion. Hearing loss in Cogan's syndrome is difficult to treat. Though rituximab was ineffective in our case, earlier administration in the disease course could be effective for future patients.  Cogan's syndrome is a rare disease characterized by inflammatory eye disease and vestibuloauditory dysfunction. The ocular symptoms are usually amenable to treatment but the vestibuloauditory symptoms can persist despite aggressive therapy. Rituximab has been proposed as a treatment option for persistent hearing loss associated with Cogan's syndrome; however, we report a case in which a patient's sensorineural hearing loss was not responsive to this agent.3/μL). She was treated with IV hydration, ciprofloxacin, and metronidazole, with improvement in her clinical and laboratory parameters. However, on hospital day 2, she developed acute onset bilateral hearing loss, associated with low grade tinnitus. She denied otalgia, otorrhea, vertigo, or ataxia. Concurrently, she endorsed bilateral eye redness, pain with eye movement, and decreased vision.A 43-year-old female was transferred to our institution because of bilateral hearing loss associated with bilateral eye pain and blurry vision. She had been in her usual state of health until five days earlier when she developed fevers, diarrhea and vomiting, dizziness, and diffuse myalgia/arthralgia. She presented to an outside hospital and was found to be tachycardic to 110 beats/minute with acute kidney injury  and thrombocytopenia  and Mycoplasma IgM was positive. MRI/MRA of the brain was unremarkable. CT angiography of the chest showed no vascular or pulmonary abnormalities.On exam, she was afebrile with a normal heart rate and blood pressure in no acute distress. There were no facial rashes and her neurologic exam including cranial nerves was intact. Audiometry demonstrated bilateral sensorineural hearing loss see  and opht Chlamydia and Mycoplasma titers. Her hearing loss stabilized but did not improve after 3 months. Given the suggestion of efficacy of rituximab in sensorineural hearing loss [2 weekly × 4 doses. At 6-month follow-up, however, her hearing had not improved and audiogram was unchanged. She is currently clinically stable and functions well with hearing aids. Given the inflammatory eye disease and bilateral sensorineural hearing loss and lack of signs of other autoimmune diseases, the patient was diagnosed with atypical Cogan's syndrome. She was treated with methylprednisolone 1 gm daily IV × 3 days and subsequently started on prednisone 1 mg/kg daily with azathioprine titrated to 150 mg daily as a steroid sparing agent. She was also treated with one month of doxycycline for her positiveing loss , and recIn 1945, ophthalmologist David Cogan reported a series of four patients with nonsyphilitic interstitial keratitis and audiovestibular dysfunction  and anot Chlamydia in particular has been proposed to be a triggering agent based on its ability to cause ocular infections as well as serologic evidence of recent Chlamydia infections in a high number of patients in previous series of Cogan's syndrome [ Chlamydia IgG, so we elected to treat our patient with antibiotics, based on data from reactive arthritis, which suggests clinical benefit if the underlying inflammatory stimulus is treated [ Mycoplasma IgM antibody titer. While Mycoplasma has not been specifically identified as a precipitating factor in Cogan's syndrome, associations have been reported both with anterior uveitis [The etiology of Cogan's syndrome is unknown. A large percentage of patients seem to have a preceding infectious illness, as in our patient.syndrome . Our pat treated . Interes uveitis  and with uveitis .The pathophysiology of the vestibular symptoms in Cogan's syndrome has been unclear. Some studies have found an association with autoantibodies. In 2002, Lunardi et al.  found IgThough the ocular manifestations of Cogan's syndrome usually respond to topical therapies or systemic corticosteroids, the treatment of vestibuloauditory manifestations can be challenging. Up to half of patients become permanently deaf despite treatment , and pro


To evaluate a set of psychometric properties  of the Parkinson's disease Activities of Daily Living Scale (PADLS) in people with Parkinson's disease (PD). Specific attention was paid to the association between PADLS and PD severity, according to the Hoehn & Yahr (H&Y) staging. The sample included 251 persons with PD (mean age 70 [SD 9] years). Data collection comprised a self-administered postal survey, structured interviews, and clinical assessments at home visits.rs > 0.5) with perceived functional independence, ADL dependency, walking difficulties, and self-rated PD severity than with variables such as PD duration and cognitive function (rs < 0.5). PADLS scores differed across H&Y stages . Those in H&Y stages IV-V had more ADL disability than those in stage III , whereas there were no significant differences between the other stages. Data completeness was 99.6% and the mean PADLS score was 2.1. Floor and ceiling effects were 22% and 2%, respectively. PADLS scores were more strongly associated ( PADLS revealed excellent data completeness, acceptable targeting, and external construct validity. It seems to be well suited as a rough estimate of ADL disability in people with PD. The ability to perform activities of daily living (ADL) is essential for independent living. ADL includes activities such as feeding, dressing, bathing, cooking, cleaning, and shopping. People with Parkinson's disease (PD) often experience ADL limitations already early during the disease course . Poor ADrs 0.74–0.86) [r 0.63–0.77) [rs 0.45–0.61) [r 0.42–0.47) [rs 0.40) [rs 0.18–0.23) [rs 0.16) [r 0.37–0.68; rs 0.52–0.87) [r 0.54–0.59; rs 0.36–0.83) [There are various ways of assessing ADL. The assessment can be based on observations of actual ADL performance, interviews, or self-ratings. Assessments can address difficulties in performing ADL, dependence on assistance from others and from assistive devices, or a combination of both . Several63–0.77) , 6. Lowe52–0.87) . Large v r 0.89) and external construct validity in terms of associations with, for example, motor symptoms , complications of PD therapy (rs 0.56), depressive symptoms (r 0.43), PD duration , and frequency of social activities (r 0.02–0.44). A recent review listed PADLS as a “suggested” rating scale for assessment of ADL in people with PD, and it was argued that more psychometric studies are needed before PADLS can be classified as “recommended” [The Parkinson's disease Activities of Daily Living Scale (PADLS) is a single-item self-reported rating scale targeting ADL in people with PD . Since ioms r 0.4, PD durammended” ) and themmended” ).Thus, this study aimed to evaluate a set of psychometric properties  of PADLS scores in people with PD. Specific attention was paid to the association between PADLS and PD severity according to H&Y.We utilized cross-sectional baseline data of the project “Home and Health in People Ageing with PD.” Details regarding the project design and methods have been published elsewhere . The pron = 10), severe cognitive difficulties (n = 91), or other reasons that made them unable to give informed consent or take part in the majority of the data collection . Fifty-eight persons were excluded since they lived outside Skåne County. The remaining 437 persons were invited to participate, but 22 of those were unreachable and two had a revised diagnosis. Out of the remaining 413 participants, 157 (38%) declined. Another five persons were excluded since they had not responded to the PADLS by themselves or not responded within two months from the home visit (part of the data collection) or due to extensive missing data. This resulted in a final study sample of 251 participants . Further participant characteristics are presented in Participants were recruited from three hospitals in Skåne County, Sweden. A detailed flow chart of the recruitment procedure has been published . A totalThe data collection is comprised of a self-administered postal survey and a subsequent home visit, which included interview-administered questionnaires and questions as well as clinical assessments. The home visits were conducted by two registered occupational therapists who had undergone project-specific training. More details regarding the procedure have been described elsewhere .The self-administered postal survey included the PADLS, a single-item self-reported rating scale that addresses perceived ADL difficulties and dependence on others as well as on assistive devices during various ADL . RespondIn addition, the postal survey included a question on self-rated general health   and the The structured interview during the home visit included a question on PD duration and three study-specific questions targeting social activities. The latter were self-rated by asking about the frequency of visiting/receiving visits from friends/family . Moreover, depressive symptoms were self-rated using the Geriatric Depression Scale  . PerceivMotor symptoms were clinically assessed according to the UPDRS part III , whereas complications of PD therapy were assessed according to the UPDRS part IV  . Cognitip values were used and the level of statistical significance was set to p < 0.05.Statistical analyses were performed in IBM SPSS Statistics, version 24. Analyses included data completeness, targeting, and external construct validity of the PADLS. Two-tailed Data completeness refers to the degree to which a rating scale is completed , 34 and Targeting refers to the scale's ability to mirror the levels of the targeted variable  in the study sample . The mears). The hypotheses were based on clinical reasoning and previous studies regarding associations between ADL and other variables [rs) between PADLS and walking difficulties in daily life, perceived functional independence, self-rated PD severity, ADL dependency, motor symptoms, and complications of PD therapy were anticipated to be >0.5 [External construct validity of a rating scale is supported when scores are more strongly associated with related constructs and more weakly associated with nonrelated constructs . In thisariables –10, 16.  be >0.5 , 10, 16. be >0.5 , 8, 10. U tests were used to explore whether PADLS scores differed between H&Y stages. H&Y stages IV and V were merged due to few participants in H&Y stage V (n = 6).Kruskal-Wallis and Mann–Whitney All but one participant responded to the PADLS, resulting in 99.6% data completeness. The score distribution is presented in p < 0.001). Specifically, those in H&Y stages IV and V had higher PADLS scores than those in H&Y stage III , whereas there were no significant differences between the other H&Y stages (PADLS scores correlated >0.5 with walking difficulties in daily life, perceived functional independence, self-rated PD severity, and ADL dependency. The associations between PADLS scores and other studied variables were weaker . The KruY stages .This study confirmed that the PADLS has satisfactory psychometric properties for use in the PD population. The PADLS revealed excellent data completeness; only one participant left the form blank. This indicates that the scale is easy to understand and perceived as relevant  by peoplExternal construct validity of the PADLS was generally supported by largely expected associations with other variables. However, the associations with motor symptoms and complications of PD therapy were lower than expected. In comparison to the present sample, the participant characteristics in previous studies , 10, 16 Indeed, varying results in previous studies imply a challenge when stating a priori hypotheses, which is essential for the exploration of a scale's external construct validity . We do fAlthough PADLS scores increased with increasing H&Y stages, the differences across these stages were small. The finding that PADLS scores differed only between H&Y stage III versus stages IV and V is not surprising. That is, the definition of H&Y stage III states that “patients are still physically capable of leading independent lives” whereas stages IV-V define a “severely disabling” PD .Data completeness might have been affected by the study design. That is, our data collectors were instructed to screen all self-administered ratings at the subsequent home visit. In case of missing values, the data collectors were instructed to ask the participants to add responses. However, another psychometric study based on the same data collection did report missing values that were close to those collected in another sample not using the same procedure .rs when studying the external construct validity of the PADLS. This is since rs and r tend to attenuate estimated correlations between ordinal data [One could argue that using polychoric and polyserial correlation coefficients would have been a theoretically better choice than nal data , 41, sucnal data , 16, we Our decision to use the ADL Staircase  and not It needs to be emphasized that the PADLS is a single-item rating scale and as such, it only gives a rough, global estimate of a person's ADL disabilities. Still, it should be noted that although the abbreviated response categories are phrased, for example, “mild difficulties in day-to-day activities,” the PADLS captures both perceived difficulties, dependence on others and on assistive devices during ADL performance. Moreover, it includes both personal and instrumental ADL. As such, the PADLS can be useful in clinical practice as well as in research for purposes of providing a crude categorization of levels of ADL disabilities. However, a more comprehensive rating scale is needed if a more thorough ADL assessment is warranted, and especially so for those with less severe ADL disabilities. This is in agreement with recommendations from the developers of the PADLS, who stated that the scale is not suitable for use in isolation but should be considered a complement to existing scales .We found the self-reported, single-item rating scale PADLS to yield scores with excellent data completeness, acceptable targeting, and external construct validity. The PADLS seems to be well suited for providing a rough indicator of ADL disability in people with PD. As psychometric testing is a continuous process, further studies focusing on additional aspects, such as responsiveness, minimal important difference, and response category functioning, are warranted.


Gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) refer to a group of heterogeneous cancers of neuroendocrine cell phenotype that mainly fall into one of two subtypes: gastroenteropancreatic neuroendocrine tumors  or gastroenteropancreatic neuroendocrine carcinomas . Although originally defined as orphan cancers, their steadily increasing incidence highlights the need to better understand their etiology. Accumulating epidemiological and clinical data have shed light on the pathological characteristics of these diseases. However, the relatively low number of patients has hampered conducting large-scale clinical trials and hence the development of novel treatment strategies. To overcome this limitation, tractable disease models that faithfully reflect clinical features of these diseases are needed. In this Review, we summarize the current understanding of the genetics and biology of these diseases based on conventional disease models, such as genetically engineered mouse models (GEMMs) and cell lines, and discuss the phenotypic differences between the models and affected humans. We also highlight the emerging disease models derived from human clinical samples, including patient-derived xenograft models and organoids, which may provide biological and therapeutic insights into GEP-NENs. Summary: Gastroenteropancreatic neuroendocrine neoplasms have been an understudied entity with limited treatment options. We discuss the disease models, including emerging organoids, that may provide biological and therapeutic insight into these diseases. Chronic atrophic gastritis: chronic inflammation of the gastric mucosa usually associated with a persistent Helicobacter pylori infection.Complete response (CR): the state of a patient whose cancer disappears because of the treatment.Gastrointestinal adenocarcinoma: gastrointestinal-derived malignant tumor of differentiated, mucus-secreting glands.Hypergastrinemia: elevated levels of the digestive hormone gastrin, which induces gastric acid secretion (gastrin is released by G cells in the antrum of the stomach).Median survival: the length of time from either the date of diagnosis or the start of treatment for a disease to when half of the patients are still alive.Neuroendocrine marker: an immunoreactive marker indicating the neuroendocrine differentiation of a tissue. Chromogranin A, synaptophysin and CD56 are used as neuroendocrine markers for GEP-NENs. Chromogranin A is a neuroendocrine secretory protein, synaptophysin is a synaptic vesicle glycoprotein present in neuroendocrine cells and CD56 is a neural cell adhesion molecule.Refractory: recurrence of disease after treatment.Resectable: a tumor or lesion that can be treated surgically, often because it is localized.Response rate (RR): percentage of patients whose cancer shrinks or disappears because of treatment.Unresectable: a tumor or lesion that cannot be removed surgically, as in the case of metastatic cancer.Gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) were originally identified as rare diseases occurring in the gastrointestinal tract and pancreas and displaying distinctive histopathological features from those of conventional gastroenteropancreatic epithelial cancers . Unlike The annual worldwide incidence of GEP-NENs has increased, with a fivefold increase over the past 30 years in the United States – from 1.09 to 5.25 cases per 100,000 persons – possibly due to improvements in endoscopic cancer screening . This inGEP-NETs are characterized by slow proliferation and initially occur as a localized disease. But treatment delays can result in the tumor's metastatic progression and death . GEP-NETDisease models are often used to gain new insights into the etiology and biology of human neoplasms and to develop novel treatments. These models include genetically engineered mouse models (GEMMs), cell lines and patient-derived xenograft (PDX) models. However, despite recent efforts, the establishment and application of GEP-NEN disease models have been limited, primarily due to the relatively small number of individuals affected by GEP-NENs. Considering the recent rise in incidence and the poor prognosis of these diseases, it is important to develop GEP-NEN disease models that more accurately reflect the biology of human GEP-NEN tissues in terms of diagnostic criteria and genetic alterations.In this Review, we provide an overview of the hallmark clinical features, diagnostic criteria, genetic background and currently available models of GEP-NENs. We also highlight the extent to which these models recapitulate human tumor biology and the major insights that have been gleaned from recent studies. We further discuss how emerging organoid models could provide a useful new platform for GEP-NEN disease modeling.As mentioned, GEP-NENs are largely divided into GEP-NETs and GEP-NECs, according to the classification criteria defined by the World Health Organization (WHO) . A GEP-NThe use of the Ki67 index as a disease marker, however, requires caution since it might not reflect the biology and the heterogeneity of the disease. The Ki67 index is highly influenced by the surrounding tumor microenvironment and by therapeutic interventions. Thus, it might reflect these biasing factors rather than the intrinsic properties of the tumor cells . Even inRevealing the mutational landscape of GEP-NENs is important for deepening our understanding of the biology of these diseases. Among recent insights, several mutations in cell-cycle-related genes have been discovered in GEP-NENs through advances in targeted gene sequencing.MEN1, which encodes menin –AKT pathway, which also regulates the cell cycle  or sporadic . To date, at least four familial GEP-NET syndromes have been reported: multiple endocrine neoplasia type 1 (MEN1), tuberous sclerosis complex (TSC), Von Hippel–Lindau (VHL) and neurofibromatosis type 1 (NF1) . MEN1 ises menin . The funll cycle . TSC is ll cycle . VHL is  pathway . Recentlton pump , and thepoptosis .MEN1 , PTEN , TSC1 and TSC2 , which encodes a key tumor suppressor that regulates the cell cycle  – a treatment strategy called PRRT , peptide receptor radionuclide therapy (PRRT), everolimus and sunitinib, has been reported to improve the survival of patients with unresectable GEP-NETs . Streptoled PRRT . This taled PRRT . Sunitinled PRRT .The response rates RRs;  for thesThe setting up of clinical trials to improve the therapeutic strategies available for patients with GEP-NECs is hampered by the low number of patients with this disease. Nevertheless, GEP-NECs have been treated using the treatment guidelines developed for small-cell lung cancer based on the histological similarities between these cancers. These strategies include using a combination chemotherapy regimen consisting of cisplatin or carboplatin with etoposide or irinotecan . In seveEstablishing experimental models that recapitulate GEP-NENs is important to increase our understanding of the biology of these diseases and to develop novel targeted therapies to treat them. Several disease models of GEP-NENs have been reported, and have contributed to drug development of the current standard of care. However, these disease models are limited by their insufficiency to recapitulate the genetic mutations and diagnostic features of the human neoplasms. An appropriate preclinical disease model would be one that mimics the genetic alterations that feature in GEP-NENs and fulfills the diagnostic criteria that can be applied for GEP-NEN drug screening and development. Here, we provide an overview of the current disease models for GEP-NENs, including emerging organoid models, and highlight their advantages and limitations, and some of the insights that have been gained from their characterization .Table 2GEMMs are a key tool in cancer research as they enable tumors to be evaluated in the presence of an intact immune system, and allow the causal relationship between genetic mutations and phenotypes to be explored. GEMMs can also be used in a variety of ways, such as for drug target validation and for testing novel therapies, for biomarker discovery, and for evaluating treatment toxicity .Men1 causes embryonic lethality at E11.5–E12.5, Men1 heterozygous knockout mice develop tumors in multiple organs and develop certain features found in patients with MEN1 -large T antigen (Tag)-based transgenic models. Genetic knockout models have been generated by engineering germline mutations in mouse strains . Whereasith MEN1 . GEMMs ttrinemia  and devetrinemia . The tumGEP-NETs . But theThe rat insulin promoter (RIP)-Tag mouse model is a representative model for SV40-Tag-based transgenic models, in which the RIP drives the expression of Tag specifically in islet cells, and β-cells become highly proliferative in response to SV40-Tag expression . The RIPTP53 and RB1 . CNDT2 is a cell line derived from a liver metastasis of a patient with a primary ileal NET. When initially transplanted into mice, this cell line developed as a tumor resembling the patient's tumor histology . Howeverin vivo applications, such as drug screening (including high-throughput screening), based on their relevance to patients' tumors, including stromal conservation  is an African rodent that tends to develop multi-centric gastric NETs as it ages (Sigmodon hispidus) and in a Mongolian gerbil (Meriones unguiculatus) model of gastric NETs induced by high doses of proton pump inhibitors  has been used to study genetics and development for over three decades  for cells to adhere to, and various growth factors to stimulate differentiation  have developed a colon cancer organoid biobank, which contains a diverse range of human colorectal tumor subtypes . We generganoids . There il cancer  Fig. 1)TP53 mutade novo pathway  have been derived to date, and the lack of GEP-NET organoids renders the current culture system insufficient for modeling GEP-NENs.TP53 and RB pathway mutations in GEP-NECs. These mutational differences might cause the different clinical features of these tumor subtypes and might influence the resulting treatment options, such as molecular targeted therapy for GEP-NETs and cytotoxic treatment for GEP-NECs. The establishment of these current standard therapies has arisen from studies conducted in GEP-NEN animal models, which were developed through extensive efforts. However, due to the small number of individuals affected by GEP-NENs, and the low success rates in the development of the disease models, the number of animal models remains limited. Additionally, some of the current animal models were validated under the previous WHO classification for GEP-NENs, when the role of genetic mutations was unknown and the terminology for GEP-NETs and GEP-NECs was less clear. Thus, very few of the existing GEP-NEN models have been confirmed clinically and genetically as being informative and accurate. Additional models of GEP-NENs are thus needed to advance this field and to gain new biological insights into the cellular and molecular etiology of these diseases. Modeling GEP-NENs in organoids could provide a new platform for this field and could also enable the reconstruction of the oncogenic steps that lead to GEP-NEN development through genetic engineering. Organoid models have the potential to contribute to the ongoing development of novel treatments and to the identification of the GEP-NEN stem cell of origin, which could lead to the eventual eradication of these diseases. Disease models that faithfully recapitulate GEP-NENs are indispensable for the translation of animal-model-based research to the clinic, to help overcome the limited treatment strategies for these diseases.GEP-NENs are heterogeneous diseases that have been understudied for several decades due to the small number of patients who develop these diseases. However, their increasing incidence, poor prognosis and limited treatment options has focused attention on these diseases, stimulating research in this field. Technological advances in clinical diagnosis and genetic analysis have furthered our understanding of the biology of these diseases in recent years. Data suggest that cell cycle dysfunction underlies the etiology of GEP-NENs and that different tumor subtypes feature different pathway mutations: mTOR pathway mutations in GEP-NETs, and


Cyclic Nucleotide Phosphodiesterase (PDE) comprise a family of 11 enzymes that hydrolyze cyclic nucleotides. PDEs aid in the regulation of secondary messengers, cyclic adenosine monophosphate (cAMP), and cyclic guanosine monophosphate (cGMP), through the degradation of phosphodiester bonds (Huang et al., Until recently, there was little knowledge about the expression of PDEs in the inner ear (Huang et al., in vitro (Kranz et al., The discovery of these various PDEs in the inner ear suggest that they play a crucial role in the auditory system Figure  and can The future of PDE research could thus have vast implications in the field of hearing loss. Future studies should focus on deciphering the molecular mechanisms of the PDE pathways in the cochlea as well as how increasing and decreasing cAMP/cGMP levels influence inner ear homeostasis. In particular, further studying PDE5, PDE4D, and PDE3B inhibitors and their exact ototoxic properties at varying concentrations could provide insight on whether these drugs have beneficial or harmful effects on the auditory system and hearing. Once these ototoxic properties are thoroughly understood, medicinal doses can be administered or reduced accordingly. If reduction is not an option, the consequent step would involve changing the biochemical properties and structures of these PDE inhibitors to maintain their therapeutic purposes while reducing their ototoxicity.All authors listed, have made substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Beauveria bassiana-28 ethyl acetate extracts on different larval stages and pupae of Culex quinquefasciatus mosquitoes. B. bassiana-28 ethyl acetate mycelial extracts produced mosquitocidal activity against larvae and pupae which was comparable to that of the commercial insecticide B. bassiana-22 extract. The LC50  values of B. bassiana-28 extracts for 1st to 4th instar larvae and pupae were 11.538, 6.953, 5.841, 3.581 and 9.041 mg/L respectively. Our results show that B. bassiana-28 ethyl acetate mycelial extract has strong insecticidal activity against larval and pupal stages of Cx. quinquefasciatus. Fourier transform infrared spectrum study of B. bassiana-28 extract shows peaks at 3226.91; 2927.94; 1593.13; 1404.18; 1224.18; 1247.94; 1078.21; 1018.41; 229.69; and 871.82 cm−1. Major spectral peaks were observed at 3226.91 cm−1, assigned to N–H stretching, 2927.94 cm−1 assigned to C–H bonding and 1595.13 cm−1 assigned to C–O stretching. Gas Chromatography-Mass Spectrometry studies of B. bassiana-28 ethyl acetate crude extract showed presence of six major compounds viz. N-hexadecanoic acids (13.6040%); Z,Z-9,12 octadecadienic acid (33.74%); 9-eicosyne (10.832%); heptacosane (5.148%); tetrateracontane (5.801%); and 7 hexyleicosane (5.723%). Histology of mosquito midgut tissue shows tissue lysis as a result of B.bassiana-28 extract exposure. The study shows that bioactive molecules obtained from B. bassiana-28 mycelial extract has insecticidal properties and can be used as alternative for mosquito control.Microbial-based pest control is an attractive alternative to chemical insecticides. The present study sought to evaluate the toxicity of the entomopathogenic fungus  Anophelinae, Culicinae and Toxorhynchitinae [Culex quinquefasciatus mosquitoes are responsible for lymphatic filariasis in tropical and subtropical regions [Mosquitoes are responsible for several vector-borne diseases . Mosquitchitinae . Female  regions ,4,5,6. H regions ,8,9. In  regions ,12,13,14 regions ,16.Chrysosporium [Metarhizium and Beauveria [Culicinomyces [Verticillium [Piper [B. bassiana, are well known as insect pathogens against agricultural lepidopteran pests [Beauveria spp., are highly specific to mosquitoes [B. bassiana is a cosmopolitan entomopathogenic fungus found in nature in soils. B. bassiana-derived insecticides have nowadays been registered and commercially developed worldwide as agricultural pest control methods [Beauveria bassiana conidia, blastospores and their secondary metabolites are used for controlling the mosquito population at a laboratory level [B. bassiana-28 extract in comparison with the commercially available microbial insecticide Beauveria bassiana-22 extract against different stages (Ist–IVth instar larvae) and pupae of Cx. quinquefasciatus mosquito and also performed larval histopathological studies at different time durations .Several studies on entomopathogenic fungi (EPF) and their metabolites have shown their control potential in various stages of mosquitoes ,18. Secoosporium , Metarhieauveria ,21; Culiinomyces , Verticiicillium  and Pipem [Piper  have beem [Piper . Entomopan pests ,26. Thessquitoes  and can  methods . Beauverry level . In the B. bassiana-28 fungal culture was isolated from dead cadavers of Spodoptera litura (tobacco cutworm or cotton leaf worm) insects in a cotton field in Dharmapuri, Tamil Nadu, India. B. bassiana-28 .siana-28  was sub-B. bassiana-22 was procured from Manidharma Biotech, Pvt Ltd. . Commercial Beauveria bassiana-22 targets several insect pests such as root weevils, plant hoppers, Japanese beetle, black vine weevil, spittlebug and white grubs and lepidopteron pests. The procured B. bassiana-22 was cultured on PDA, supplemented with ampicillin (3 mg/100 mL) and incubated for 10 days at 25 °C ± 2 °C. Commercially available powdered microbial insecticide B.bassiana-28 fungal strains based on morphological characteristics such as colony colour, aerial mycelial structures, pigment production and conidia stained with lacto-phenol cotton blue and viewed under light microscope  at 400× magnification.Morphological identification was carried out for isolated or unknown B. bassiana-28 broth was prepared for the mass culture of fungal mycelia as per the modified method of [B. bassiana-28 fungal conidia (1 × 107 per mL) were inoculated and grown in PDB. The flasks were incubated at 25 °C ± 2 °C for 25 days.ethod of . Four 10B. bassiana-28 and B. bassiana-22 was carried out in a 1000 mL Erlenmeyer flask containing 500 mL of PDB. The flasks were incubated under the optimized culture conditions (pH 7.0 at 27 °C) for 25 days. The fungal biomass was removed from the medium with help of Whatman No. 1 filter paper and washed more than five times with distilled water to remove the unwanted broth particles. Fungal biomass (100 g) was transferred to 500 mL glass beakers containing ethyl acetate (250 mL) which was mixed with the mycelium for cold extraction for 20 days at 25 °C ± 2 °C. After complete extraction the liquid portion was separated from the mycelium by filtering through Whatman No. 1 filter paper. Separated secondary metabolite ethyl acetate extracts were finally concentrated using a rotary vacuum evaporator  at 45 °C.Mass culturing of (TLC) was performed on commercial silica gel-H TLC plates  for principal components separation. The developed TLC plates were dried at room temperature. After air drying B.bassiana-28 extract were spotted at center of the plate with the help of a capillary tube. Then we prepared different solvent systems as mobile phases for thin layer chromatography because the biological molecules can be separated by different solvent system. The mobile phase solvent systems were chloroform:methanol in several ratios . After the running process the plates were observed under UV light at 350 nm. The retention factor (Rf) values were calculated using Equation (1) and based on the movement of samples in TLC plate (Thin layer chromatography LC plate .(1)Rf=DCx. quinquefasciatus egg rafts were obtained from the Institute of Vector Control Zoonoses, . The egg rafts were maintained in 2 L plastic jars containing tap water. The larvae were fed with dog biscuits and millet powder and yeast powder in 3:3:1 ratio. Larvae were kept at 27 °C ± 2 °C and 70–85% relative humidity with a 12:12 light and dark photoperiod.B. bassiana-28 and B. bassiana-22 mycelia extract were transferred individually to 250 mL round bottom flasks, then the ethyl acetate solvent was removed by using a rotary evaporator, . The bioassay has five testing concentrations and each concentration had three replicates of twenty larvae each. Test containers containing 20 mosquito larvae were stored in 150 mL plastic cups containing 99 mL of distilled water with the desired concentration . In the control 20 individuals were exposed to the same dose of dimethyl sulfoxide (DMSO) as negative control. After 24 h exposure, mortality (%) was calculated and corrected with control mortality using the Abbott formula [50 and LC90 values were calculated by probit analysis using the SPSS-16.0 software .Larval mortality bioassays were carried out according to the method suggested by the World Health Organization (WHO) , with sl formula . The larCx quinquefasciatus pupae from the laboratory maintained culture were used to examine the pupal toxicity of B. bassiana-28 and B. bassiana-22 extracts. Twenty pupae were transferred to 150 mL plastic containers containing 99 mL of distilled water. Five different concentrations of extracts  were separately dissolved in DMSO (1 mL) and the dissolved fungal extracts were added to the water in the bioassay vessels. Each concentration had three replicates and each replicate had twenty pupae. Mortality was calculated 24 h post treatment, and mortality in the treatments and control was corrected using Abbott’s formula [50 and LC90 values were calculated from the toxicity data by probit analysis using SPSS-16.0 software. formula . The LC5−1; and finally 300 °C, where it was held for 6 min. The mass detector conditions were 240 °C; ion source temperature at 240 °C; and ionization mode electron impact at 70 eV, a scan time 0.2 s and scan interval of 0.1 s. The fragments from 40 to 600 Da were collected. A Clarus 680 30 m × 0.25 mm ID × 250 μm silica column was used for GC analysis of the chemical constituents. This column was packed with elite-5MS (5% biphenyl 95% dimethylpolysiloxane). The chemical constituents were separated by using He at a constant flow of 1 mL/min as a carrier gas. The crude extracts (1 μL) were injected to the GC-MS instrument at 260 °C during the column running time. The temperature ramp was as follows: 60 °C (2 min); followed by 300 °C at the rate of 10 °C minCx. quinquefasciatus larvae were fixed in 3% formaldehyde solution for 2 h at 4 °C. The blocks were cooled at 27 °C for 3 h and cut to 8 μm thickness 1.3 mm ribbons with a microtome . Cross-sectioned larval gut was stained with haematoxylin and eosin stain. After air drying sections was viewed under a light microscope (Olympus-CH20i) at a magnification of 400×.The 6, 12 and 24 h post-treated and control 4th instar 50 and LC90 values it was found that B. bassiana-28 mycelium extract had insecticidal activity similar to that of the commercial microbial insecticide B. bassiana-22 on 1st to 4th instar of Cx. quinquefasciatus , eicosanoic acid (21.016%), octadecanoic acid (21.466%), tridecanoic acid (22.081%), pentadecanoic acid (22.136%), tetradecanoic acid (22.986%), octadecanoic acid (23.757%), eicosanoic acid (24.442%), heptadecanoic acid (25.117%), tridecanoic acid (25.778%), octadecanoic acid (26.468%), tridecanoic acid (27.143%), dodecanoic acid (27.888%), l-(+)-ascorbic acid 2,6-dihexadecanoate (28.749%) (N-hexadecanoic acid (13.6040%), Z,Z-9,12 dectadecadienoic acid (33.74%), 9-eicosyne (10.832%), heptacosane (5.148%), tetrateracontane (5.801%); and 7 hexyleicosane (5.723%)—may be involved in the insecticidal activity.Gas Chromatography-Mass Spectrometry results obtained from the 28.749%) . Six maj−1  analysis ; Table 4B. bassiana-28 extract. Gut tissue of lumen was mutually by way of a thin peritrophic membrane (Pm) in 6 h treatment as compared to control  were severely damaged by  control B, whereasiana-28 B,C also B. bassiana-28 and commercial B. bassiana-22 were found to produce toxicity in all stages like larval and pupal Cx. quinquefasciatus. Result clearly shows that B. bassiana-28 extract has strong mosquitocidal activity against 1st to 4th instar larvae of Cx. quinquefasciatus,  [Z, Z-9,12 dectadecadienoic acid (33.74%); 9-eicosyne (10.832%); heptacosane (5.148%); tetrateracontane (5.801%); and 7-hexyleicosane (5.723%) are also thought to be involved in insecticidal activity. In our results show the major compound in assessment with the standard was N-hexadecanoic acid, and based on the above we infer that N-hexadecanoic acid from B. bassiana-28 mycelial extract may be the principal metabolite which confers insecticidal activity to the extract.eratures . The FT-3.6040%) ; Z, Z-9,B. bassiana-28 mycelial extracts produced toxicity in larvae and pupae of Cx quinquefasciatus. In addition these extract produce considerable tissue damage to the midgut of mosquito larvae. Characterization of extracts showed that N-hexadecanoic acid is the main constituent of the extract which has insecticidal properties. In future work on control of mosquito larvae, N-hexadecanoic acid can be used as an alternative to chemical insecticides.


To evaluate the effect of hypoxia on the neonatal pig retina and brain, we analysed the retinal ganglion cells (RGCs) and neurons in the superior colliculus, as well as the response of astrocytes in both these central nervous system (CNS) structures.2: 10–15%), followed by a reoxygenation period of 240 minutes (FiO2: 21–35%). RGCs were quantified using Brn3a, a specific nuclear marker for these cells, and apoptosis was assessed through the appearance of active caspase-3. A morphometric analysis of the cytoskeleton of astrocytes (identified with GFAP) was performed in both the retina and superior colliculus.Newborn pigs were exposed to 120 minutes of hypoxia, induced by decreasing the inspiratory oxygen fraction (FiOHypoxia produced no significant change in the RGCs, although, it did induce a 37.63% increase in the number of active caspase-3 positive cells in the superior colliculus. This increase was particularly evident in the superficial layers of the superior colliculus, where 56.93% of the cells were positive for active caspase-3. In addition, hypoxia induced changes in the morphology of the astrocytes in the superior colliculus but not in the retina.Hypoxia in the neonatal pig does not affect the retina but it does affect more central structures in the brain, increasing the number of apoptotic cells in the superior colliculus and inducing changes in astrocyte morphology. This distinct sensibility to hypoxia may pave the way to design specific approaches to combat the effects of hypoxia in specific areas of the CNS. Neonatal hypoxic-ischemic brain injury is a prominent cause of neurological disability in neonates , 2. SincThe retina pertains to the central nervous system (CNS) and it is one of the most metabolically active tissues in the body . Its higWhile the brain represents 2% of our body weight, it consumes 20% of the body’s oxygen demand. Moreover, the immature foetal and neonatal brains are particularly vulnerable to severe alterations in oxygen tension and they may develop neurovascular malformations when oxygen levels are low , 18. HowGlial cells play crucial roles in regulating neuronal development and neuronal activity, and astrocytes in particular are susceptible to reductions in oxygen tension . Given tGiven that hypoxia has a negative effect on neuronal metabolism and that it may be detrimental to the function of neurons, and since astrocytes have the capacity to sustain normal neuronal activity and to regulate the development of the vasculature, here we evaluated whether low oxygen conditions induce changes in neurons and astrocytes in the retina and superior colliculus.This study was carried out in strict accordance with the recommendations for the Experimental Research Committee of the Cruces University Hospital, which is registered in the Official Register of Breeders, Suppliers and Users of animals for experimental and other scientific purposes in the Basque Country (Spain). All the experimental protocols met with the European (2010/63/UE) and Spanish (RD53/2013) guidelines for the protection of experimental animals and they were approved by the Ethics Committee for Animal Welfare of the Cruces University Hospital. On the same day as the experiment, animals were obtained from a local farm authorized by the Basque Country Regulatory Agency to supply animals for research. All animals were free of any disease and they were transported with a certificate of health.Sus scrofa, n = 8) aged 2–4 days old (1.7± 0.2 kg) were sedated with an intramuscular injection of ketamine (15 mg/kg) and diazepam (2 mg/kg). Before performing the surgical procedure, anaesthesia and analgesia were induced by intravenously administering fentanyl (5 μg/kg) and propofol (1.2 mg/kg), and this state was maintained by continuous intravenous infusion of fentanyl (titrated as necessary: 5–20 μg/kg/h), propofol (titrated as necessary: 2–3 mg/kg/h) and midazolam (titrated as necessary: 0.5–2 mg/Kg/h). In addition, the animals used as controls were paralysed by continuous intravenous infusion of vecuronium bromide (3 mg/kg/h). In all cases, an ear vein was catheterized to deliver the anaesthesia and analgesia. A tracheotomy was performed, and a tracheal tube (4.0 mm ID) was inserted and connected to a ventilator. Animals were then positive pressure ventilated and changing ventilator parameters were performed to maintain adequate blood gas values of arterial oxygen pressure (PaO2) 80–110 mmHg, adequate arterial pressure of carbon dioxide (PaCO2) 35–45 mmHg and a pH 7.30–7.45.The protocol to produce hypoxia used in the present study has been described extensively elsewhere . Neonata2, PaCO2, pH, Base Excess (EB), oxygen saturation and lactic acid . Animals were also monitored continuously by three-lead electrocardiogram during the experimental procedure.An arterial catheter was inserted into the femoral artery to monitor the mean arterial blood pressure (MABP) and heart rate (HR), and to obtain arterial blood samples for blood gas analysis: PaO2, the fraction or percentage of oxygen in the space being measured) to 0.1–0.15 while increasing the concentration of inhaled nitrogen gas. Control pigs were not subjected to reduced oxygen concentrations. Following hypoxia, the FiO2 levels were increased to 21–35% in order to maintain normoxia for 240 minutes. At the end of the experiments (4 hours after the onset of hypoxic injury), the animals were sacrificed with an intravenous overdose of potassium chloride (0.35 mg/kg).The hypoxia model is based on a swine model of neonatal asphyxia . Two expNeonates eyes were enucleated, and the cornea, crystalline lens and vitreous humour were removed. Each eyecup containing the retina was fixed overnight by immersion in 4% paraformaldehyde (PFA) in 0.1 M phosphate buffer  and after then washing in phosphate buffered saline , the retina was carefully separated from the rest of the eye. In addition, the superior colliculus was extracted from the brain and immediately fixed overnight in 4% PFA. Control and hypoxic tissues were cryoprotected for 24 hours at 4°C in 30% sucrose diluted in 0.1 M PB, and the tissue was then embedded in OCT medium to obtain cryosections (14 μm thick) that were stored at −20°C.Whole mount retinas were immunostained as described previously , with soCryostat sections of the retina and superior colliculus were immunostained as described previously . After w2 per retina (2 mm2 per area). The method to quantify RGCs was slightly modified from that used previously [To analyse the RGCs and astrocytes in the retina, confocal microscopy images of whole mount retinas were obtained at a resolution of 2048 x 2048 pixel with a 20X objective . A Z-stack was obtained of five images with a 4 μm separation and a total of 8 whole mount retinas were studied (4 control and 4 hypoxic). Four different areas of the retina were analysed, the peripheral and central areas of the dorsal and ventral retina . Thus, aeviously , employi2 of the superior colliculus (cells/mm2).The cells with active caspase-3 were quantified in the retina and superior colliculus using a fluorescent microscope  and the Zeiss Zen software, analysing 5 sections from the same central and peripheral areas of each retina, and 20 sections from the same rostro-caudal level of each superior colliculus from 4 control and 4 hypoxic animals. Five images from each section were acquired at 20X on a Zeiss Axiocam MRM  and the active caspase-3 cells were counted double blind by two experimented researchers, as described previously . In the ad hoc programme was developed in Matlab R2010b  to quantify the morphological changes to astrocytes. This programme estimates the local orientation of the GFAP-positive lines in an image, taking advantage of the SURF extraction algorithm [To analyse the cytoskeletal morphology of astrocytes in the retina, images were taken of whole mount retinas using the same microscope and criteria as those used for RGC quantification. The astrocytes in the pig retina are organized mainly like those in the human retina, running parallel to the RGC axons. A semi-automatic method to measure astrocyte organization was used to measure the morphological changes in astrocytes. The aim was to quantify the differences in the astrocyte network between the control and hypoxic retinas based on the local orientation of the astrocytic processes. An lgorithm  and its lgorithm , a well-To analyse the astrocytes in the superior colliculus, a Zeiss Axiocam MRM fluorescent microscope  and the Zeiss Zen software was used, obtaining 20 images (20X) at the same rostro-caudal level of the superior colliculus of the 4 control and 4 hypoxic animals. Since the astrocyte organization in the superior colliculus does not follow a well-established parallel pattern, the method used to analyse the astrocytes in this structure differed to that used in the retina. The morphology of the astrocyte cytoskeleton was analyzed using ImageJ (version 1.49), the image processing program developed at the National Institutes of Health (NIH). Using the “threshold color” tool, the region formed by colored pixels (labeled with the antibody against GFAP) was selected, and this area of the cytoskeleton of selected astrocytes was measured. Given the entire image area, we could calculate the proportion of the area occupied by astrocyte´s cytoskeleton in the control and hypoxic superior colliculus.RGC density, the number of active caspase-3 positive cells and the morphological data from the astrocyte’s cytoskeleton were described as the mean and standard error of mean, and these parameters were compared between control and hypoxic tissues. Statistical analyses were carried out using IBM SPSS Statistics software v. 21.0 and the homogeneity of the variances was assayed with Levene´s test (p < 0.05). To assess whether there were significant differences in the number of RGCs, the number of active caspase-3 positive cells and in astrocyte morphology between control and hypoxia conditions, a Student T-test was used. In addition, a Mann-Witney U test was used to verify the differences between the control and hypoxic groups. For both tests, the minimum value of significance was defined as p<0.05.2) was 30 ±2%, the PaO2 was 108 ±16 mmHg, PaCO2 was 42 ±6 mmHg, pH was 7.34 ±0.08, EB (base excess) was -2 ±3 mmol/l and lactic acid was 2 ±1 mmol/l. In the hypoxic animals, the decrease in oxygen levels produced a FiO2 of 12 ±2%, and a significant impairment of gas exchange, with a PaO2 of 31 ±8 mmHg, PaCO2 of 45 ±8 mmHg, pH of 6.84 ±0.05, EB of -24 ±2mmol/l and lactic acid of 17 ±2mmol/l. Moreover, the MABP  decreased significantly after hypoxia (32 ±5 vs. 78 ±12 mmHg), while the HR (heart rate) remained unchanged relative to the basal values (184 ±36 vs. 196 ±33 bpm). These parameters partially reverted during the re-oxygenation period to a FiO2 of 33 ±1%, a PaO2 of 92 ±17 mmHg, a PaCO2 of 45 ±5 mmHg, a pH of 7.22 ±0.12, an EB of -8 ±6 mmol/l and lactic acid of 4 ±4 mmol/l. However, there was no change in either the MABP (36 ±9 mmHg) or HR (193 ±26 bpm). In the control animals that were not subjected to hypoxia, these parameters remained at the basal values throughout the experiment. Finally, the hemoglobin levels (7.0 ±0.7 g/dl) remained constant throughout the experiment and all pigs remained alive throughout.At baseline, all animals displayed adequate gas exchange, whereby the fraction of inspired oxygen  cells/mm2 after hypoxia  cells/mm2 in the hypoxic animals, (p = 0.002).Numerous active caspase-3 positive cells were found in the inner nuclear layer (INL) and ganglion cell layer (GCL) of the retina, both in control and hypoxic animals. Indeed, hypoxia did not produce a significant change in the number of positive active caspase-3 cells in the GCL, with 34 (±3) active caspase-3 positive cells/mm in control retinas and 38 (±4) cells/mm in hypoxic retinas . By contad hoc routine to quantify the differences in the retinal astrocyte networks in function of the local orientation of their processes highlighted the differences in the distribution of retinal astrocyte processes. However, no significant differences between control and hypoxic retinas were found in the area analysed. While these results indicate there was no significant difference between these populations (p >95%), there does appear to be a difference in the degree of organisation .When we analysed the cytoskeletal morphology of astrocytes in the control and hypoxic neonatal pig retina, retinal astrocytes appeared to be more disorganised following hypoxia, with an increase in the lateral extension of processes . Using tActivation and hypertrophy of astrocytes was evident in the hypoxic superior colliculus , implyinExperimental studies in suitable animal models commonly provide insights into human neonatal situations. In the present study we have used the pig as an animal model because beside other primates, the porcine retina is the most similar to the human retina among the large mammals –44. NeonWe found that hypoxia produces significant neuronal apoptosis in the superior colliculus but not in the retina. Moreover, we found significant gliosis of the astrocytes in the superior colliculus but not in the retina. However, in the retina some changes in RGC density in the peripheral retina were found, although they were not significant. The changes described probably reflect the earliest events that result from the hypoxic insult and we cannot rule out that longer recovery times will produce more significant damage in the retina.Limiting oxygen in the retina contributes to RGC neurodegeneration, which results in a loss of vision and ultimately, blindness. Cell viability can be dramatically compromised by hypoxic stress , 47 and Although several mechanisms of cell death are activated by oxygen depletion in neural tissues , 55, chaHypoxia induces astrocyte activation in many neurological disorders –63. ReacThe distinct susceptibility to hypoxia between the retina and the superior colliculus could be explained by the heterogeneity in metabolic and molecular regulation between different areas of the CNS  that proThe lack of morphological changes in the retina after hypoxia only reflect early events, suggesting that the brain is more vulnerable to hypoxia at these early time points. However, after longer reperfusion times, retinal cells might also be sensitive to such insult. Moreover, given the limited sample size in this study some changes could remain unnoticed, with only major differences becoming evident, such as those in the superior colliculus that appears to be more sensitive to hypoxia. Consequently, a therapeutic window appears to exists in which a neuroprotective action protects the retinal neurons even though the brain has suffered damage.The results of the present study highlight the differences in the reaction of neurons and astrocytes in different parts of the CNS. As heterogeneous responses to hypoxia were detected in different brain regions, interesting opportunities may be open to design therapeutic and preventative treatments specific to certain areas and structures in the nervous system.S1 FigSchematic drawing of a pig retina where the four different areas of the retina analysed are represented as black squares: DP, dorsal periphery; DC, dorsal centre; VC, ventral centre; VC, central periphery.(TIF)Click here for additional data file.S1 FileFile that contains the individual-level data points behind means, and variance measures presented in the results, tables, and figures.(XLSX)Click here for additional data file.