At this time of the COVID-19 pandemic, potentially effective treatments are currently under urgent investigation. Benefits of chloroquine and hydroxychloroquine for the treatment of COVID-19 infection have been proposed and clinical trials are underway. Chloroquine and hydroxychloroquine, typically used for the treatment of malaria and autoimmune diseases, have been considered for off-label use in several countries. In the literature, there are reports of ototoxic effects of the drugs causing damage to the inner ear structures, which then result in hearing loss, tinnitus, and/or imbalance. This mini-review represents a summary of the findings from a systematic search regarding ototoxicity of chloroquine and hydroxychloroquine in the published literature. The characteristics of sensorineural hearing loss and/or tinnitus after chloroquine or hydroxychloroquine treatment can be temporary but reports of persistent auditory and vestibular dysfunction exist. These are not frequent, but the impact can be substantial. Additionally, abnormal cochleovestibular development in the newborn was also reported after chloroquine treatment in pregnant women. The suggested dose of chloroquine for COVID-19 infection is considerably higher than the usual dosage for malaria treatment; therefore, it is plausible that the ototoxic effects will be greater. There are potential implications from this review for survivors of COVID-19 treated with chloroquine or hydroxychloroquine. Patient reports of hearing loss, tinnitus, or imbalance should be noted. Those with troublesome hearing loss, tinnitus and/or imbalance are encouraged to be referred for hearing evaluation and interventions once they are stable. Clinical trials of chloroquine or hydroxychloroquine should also consider including audiological monitoring in the protocol. Three out of ten cases had temporary sensorineural hearing loss after chloroquine treatment that improved after cessation of the medication , 19. A pWhile there was no difference in hearing thresholds between children who were and were not exposed to chloroquine during gestation , there wSix case reports, describing ototoxic effects associated with hydroxychloroquine, were identified and are displayed in The manifestation of sensorineural hearing loss and/or tinnitus and/or imbalance after chloroquine and hydroxychloroquine can be either temporary or permanent. Most of the studies on this topic were case series or case reports with only a few observational studies. Information from a definitive large study with good methodology is still lacking. Ototoxicity after chloroquine use tends to be more sudden, while hydroxychloroquine is more likely to cause ototoxicity after prolonged use. This could be due to different drug efficacy and equivalent dosage. Furthermore, hearing loss in these patients could be associated with other possible causes rather than chloroquine and hydroxychloroquine including autoimmune disease e.g., systemic lupus erythematosus , sudden in vitro model (The suggested dose of chloroquine for patients diagnosed with COVID-19 infection (1 g daily for 10 days) is substantially higher compared with the usual dosage of chloroquine for malaria treatment (1 g daily for 3 days) . There iro model , and 600Due to the potentially substantial number of the world's population who may take chloroquine or hydroxychloroquine, there is the prospect of a significant number of people being affected with ototoxic side effects. It is therefore vital to build awareness about the presentation and impact of the symptoms of drug-induced ototoxicity. Patient reports of hearing loss, tinnitus, or imbalance should be noted. Those with troublesome hearing loss or tinnitus are encouraged to be referred for hearing evaluation, including extended high frequencies audiometry at 8–16 kHz where possible, once they are stable. Available options of audiological interventions for those with bothersome hearing impairment or tinnitus are counseling, hearing aids, and tinnitus therapy. The possibility of exacerbation of pre-existing hearing loss and/or tinnitus should be considered. Synergistic adverse auditory effects when other ototoxic medication is administered with chloroquine or hydroxychloroquine, such as aminoglycoside antibiotics and azithromycin, is a further risk . Severe Although chloroquine and hydroxychloroquine are generally considered safe in pregnant women, the use of chloroquine during pregnancy in the first trimester should be contemplated with particular caution since there are reports of abnormal cochleovestibular development in newborns. Hydroxychloroquine has a safer clinical profile in pregnancy, thus is a more suitable option than chloroquine .Recent publications have brought attention to the possible benefit of chloroquine and hydroxychloroquine in COVID-19 treatment. It is important to build awareness about the possibility of ototoxicity in survivors of COVID-19 treated with these drugs. Patient reports of hearing loss, tinnitus, or imbalance should be noted. Those with troublesome hearing loss or tinnitus are encouraged to be referred for hearing evaluation and interventions once they are stable. Clinical trials of chloroquine or hydroxychloroquine should also consider including audiological monitoring in the protocol.All authors planned and structured the paper. PP undertook the review and wrote the first draft. DB, PP, and AK jointly revised the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To gauge the prevalence of hearing loss in school children in Bishkek, Kyrgyzstan, and refer pupils with positive results for further diagnostic testing.According to WHO data, hearing disorders are common in school-age children. Screening for hearing loss is an important preventative tool, helping to avoid further complications. Expenditure that supports early child development can reduce future outlay on health care and social services; it can eliminate disability problems, education deficits, and social maladaptation in later adult life.Pure-tone air-conduction hearing thresholds were obtained at 0.5–8 kHz. The results of the hearing screening examination were regarded as positive if pure-tone thresholds were higher than 20 dB HL in one or both ears at one or more of the test frequencies. Data were also obtained from follow-up visits of children who failed the initial screening.This study included 452 children aged 7–13 years old. Based on audiograms, screening showed that 123 (27.2%) of the children had hearing impairment. The study has important implications for clinical practice and health policy. There is a need for systematic monitoring of hearing status among children of this age, and parents and educators need to be made aware of the significance of hearing loss. Pure-tone audiometry is the gold standard for hearing screening programs for school-age children , in which the hearing threshold for 500 and/or 1000 Hz was above 20 dB HL, while the threshold for other frequencies did not exceed 20 dB HL.High-frequency hearing loss (HFHL), in which the hearing threshold for 4000 and/or 8000 Hz was above 20 dB HL, while the hearing threshold for other frequencies did not exceed 20 dB HL.Other: abnormal screening results in which the hearing threshold was greater than 20 dB HL at two or more frequencies.A positive  test result was taken to be an air conduction threshold value higher than 20 dB HL at one or more frequencies in at least one ear , while the other 329 children (72.8%) had audiometric thresholds below the 20 dB criterion. There were 80 children (65% of 123 children with positive outcome) who had unilateral impairment and 43 children (35%) who had bilateral impairment. The data divided into age groups are presented in Table 2 = 9.98; P = 0.002. Positive results were found more often in the younger children (32.2%) than in the older children (18.4%).There was a statistically significant difference in the frequency of positive results between the younger and older children: χ2 = 0.44; P = 0.505. Positive results in one ear were more frequent than in both ears, regardless of age.There was no statistically significant difference in the laterality of positive results between the younger and older children: χConsidering ears which had hearing thresholds above the 25 dB criterion, there were 166 in total, including 78 ears with HFHL (47%), 12 ears with LFHL (7.2%), and 76 ears with other type of hearing loss (HL). The data divided into age groups are presented in Table P < 0.001) and in the older group (P = 0.041). The rate of HFHL was significantly higher in younger children than in older children (P = 0.040). The rate of LFHL was significantly higher in the older children than in the younger group (P = 0.039). The ratios of other types of HL were similar in both age groups (P = 0.323).HFHL was more frequent than LFHL both in the younger group . In nine cases, the ear canal was blocked with ear wax. After removing it, hearing was found normal. In five cases, otitis media was diagnosed and appropriate antibiotic treatment was implemented. Tympanoplasty was ordered in two children due to perforation of eardrum. In two children, hyperplasia of the pharyngeal tonsil was found and adenotomy was recommended. Two children had a recent infection of the respiratory system which could have affected hearing and the doctor decided to wait for a complete recovery. One child had otosclerosis which had been diagnosed earlier and was not declared by the parents before hearing screening. Hearing impairment was not confirmed in six children with positive outcome of hearing screening .Three selected audiograms of children with follow-up data are presented below. The first child was diagnosed with an eardrum perforation in the left ear. The ENT specialist ordered a laboratory test to identify any bacterial infection, antibiotic drops were prescribed, and tonal audiometry was ordered. A second visit took place one month later. There was no bacterial infection in the left ear, but the eardrum was still perforated. Tympanoplasty in the left ear was ordered Figure . The secThe primary purpose of this study was to investigate the rate of hearing disorders in school-age children in Bishkek and, if tested positive, refer them for detailed diagnosis. An additional goal of the screening program was to alert parents and the school more generally to children’s hearing problems. These activities were part of a general effort to improve the state of medical care in Kyrgyzstan using information technology. The effectiveness of using an asynchronous model of screening for hearing impairment in children from primary school was also evaluated. The telemedical model of screening was aimed at improving the hearing health services for children in Kyrgyzstan where long-distance travel to regional or city centers is often difficult and sometimes impossible.et al., et al.  was more common than bilateral losses, in line with results reported by Skarzyński et al. . A unilaet al. . On this et al., .In the current study, we were only able to collect follow-up data from 27 children who had a positive result. The low follow-up rate among students referred from the school suggests a low level of support for the program from caregivers. This could be because of lack of knowledge about hearing disorders or problems with traveling to the medical center. Engagement of parents and school personnel with the school screening program is essential if the prevalence of treatable ear diseases and associated hearing disorders in children is to be reduced.The data from the present study suggest that it is possible to use a telemedicine model to assess the hearing status of school-age children and to provide long-distance expert assistance. It is necessary to train local medical staff to perform hearing screening. However, health care personnel involved with hearing services in less modern locations need consistent training, oversight, and feedback by experienced audiologists in order to provide quality services. Hearing screening opens up the possibility of detecting hearing problems and then directing the children to further specialist diagnostic evaluation and intervention.The present study was confined to the capital city of Kyrgyzstan, and it is difficult to generalize the findings to the whole pediatric population of the country. Moreover, only air conduction thresholds were measured; there was no bone conduction, otoscopy, tympanometry, or otoacoustic emission measurements.


Vaccinium species, tea leaves, grape seeds, and tannin-rich medicinal herbs. Despite a vast amount of in vitro and cell-based evidence of immunomodulatory there are still only a few animal and clinical studies. Most of the reports, regardless of the used model, indicated the efficiency of these phytochemicals from cranberries and other Vaccinium species and tea extracts (green or black). Other sources such as grape seeds and traditional medicinal plants, were seldom. In conclusion, the potential of flavan-3-ols and their derivatives in prevention and alleviation of periodontal disease is remarkable but clinical evidence is urgently needed for issuing credible dietary recommendation and complementary treatments.This paper continues the systematic review on proanthocyanidins and flavan-3-ols in the prevention and treatment of periodontal disease and covers the immunomodulatory effects, and animal- and clinical studies, while the other part discussed the direct antibacterial properties. Inflammation as a major response of the periodontal tissues attacked by pathogenic microbes can significantly exacerbate the condition. However, the bidirectional activity of phytochemicals that simultaneously inhibit bacterial proliferation and proinflammatory signaling can provide a substantial alleviation of both cause and symptoms. The modulatory effects on various aspects of inflammatory and overall immune response are covered, including confirmed and postulated mechanisms of action, structure activity relationships and molecular targets. Further, the clinical relevance of flavan-3-ols and available outcomes from clinical studies is analyzed and discussed. Among the numerous natural sources of flavan-3-ols and proanthocyanidins the most promising are, similarly to antibacterial properties, constituents of various foods, such as fruits of  In the previous paper, we demonstrated that both free flavan-3-ols and oligomeric proanthocyanidins are very promising constituents for combating various bacteria involved in periodontitis pathogenesis . Here, uAccording to the latest concept of periodontitis etiopathology, the development of the disease requires the co-existence of dental plaque that accumulates on the teeth surface and the patient’s immune-inflammatory response . PeriodoSearch strategy, as well as inclusion, exclusion criteria, and data organization are described in our previous review , in whicThe search terms included all combinations of the following key words: ‘periodontitis’ OR ‘periodontal diseases’ OR ‘gingivitis’ OR ‘gingival diseases’ AND ‘proanthocyanidins’ OR ‘condensed tannins’ OR ‘flavan-3-ols’ OR ‘catechin’ OR ‘epicatechin’ AND ‘anti-bacterial’ OR ‘antiadhesive’ OR ‘anti-inflammatory’, respectively. All titles with abstracts were imported into a citation manager program “Mendeley” , and all duplicates were removed. References of imported articles were also screened for other relevant studies. Two investigators (N.-H.I. and K.-R.P.) independently reviewed the titles and abstracts of the imported references to determine whether they met the inclusion and exclusion criteria .Finally, 31 studies were reviewed in , f, fAggregVaccinium macrocarpon) juice [Among the reviewed studies, eleven reported the influence of proanthocyanidins or flavan-3-ols on periodontitis in animal models. Three of them used grape seed proanthocyanidin extract ,53,54; fn) juice  and one n) juice . Toker en) juice  to rats P. gingivalis induced periodontitis in the animal model. The mice orally inoculated with P. gingivalis in PBS, received sterile food and drunk water with 0.02% epigallocatechin-3-gallate solution in the period from 8 weeks to 15 weeks. It was found that epigallocatechin-3-gallate has significantly reduced alveolar bone resorption as well as decreased the high expressions  of inflammatory cytokines and other mediators both in serum and in gingival tissue  and mitogen-activated protein kinase (MAPK) pathways. Despite these promising results there is still much less studies using animal models (11) and only a few clinical studies (3). In conclusion, the potential of flavan-3-ols and their derivatives in prevention and alleviation of periodontitis is remarkable but clinical evidence is urgently needed for issuing credible dietary recommendation and complementary treatments.Among the numerous in vitro studies (36) on the immunomodulatory effect of proanthocyanidins or flavan-3-ols on the host cells, most concern the tea leaves extract and its compounds- catechins with presence of the galloyl moiety as the most active, as well as of A-type proanthocyanidins from fruits of


Mycoplasma hyopneumoniae (M. hyopneumoniae), a representative pathogen causing swine enzootic pneumonia, generally infects piglets vertically. However, it is difficult to ascertain the M. hyopneumoniae infection state of sows due to limited detection methods. This report investigated sow herd stability by applying nested PCR to laryngeal swabs of suckling pigs, which is reportedly the most sensitive method.M. hyopneumoniae was detected in 14 farms (63.6%) and 127 piglets (6.5%). The prevalence of sows likely to transmit M. hyopneumoniae in herds (11.1%) was calculated. In addition, there was a significant difference in detection rates among farms depending on herd size, gilt replacement rate, acclimation method, and antibiotic usage, suggesting various parameters that influence sow stability.The results demonstrated that laryngeal swabs from suckling pigs have provided useful information regarding vertical transmission from sows in South Korean farm conditions. This result demonstrated that farms with larger herd sizes, higher gilt replacement rates, and a practice of naturally exposing gilts for acclimation had higher detection rates in weaning piglets, indicating an unstable sow infection state. M. hyopneumoniae [M. hyopneumoniae adheres to the ciliated epithelium of the respiratory tract of the pig. After attachment, the infected pigs show chronic coughing and increased susceptibility to other respiratory infections. Due to these symptoms, the pigs have reduced weight gain [Mycoplasma, such as M. hyorhinis or M. hyosynoviae, are also able to infect pigs. M. hyorhinis commonly causes polyserositis and arthritis in nursery pigs. M. hyosynoviae causes arthritis in grower-finishers [M. hyopneumoniae due to decreased performance, increased treatment and vaccination costs [Swine enzootic pneumonia (EP), a chronic respiratory disease in pigs of all ages characterized by dry coughing, growth retardation, and poor feeding efficiency, is mainly caused by eumoniae . M. hyopght gain . In someinishers . Howeveron costs .M. hyopneumoniae is one of the most common diseases on pig farms worldwide. Several abattoir surveys have demonstrated that a high prevalence of enzootic pneumonia is associated with lung lesions in pigs [M. hyopneumoniae were mainly detected in pigs 14–22 weeks old [Swine enzootic pneumonia caused by infection with  in pigs –7. In Soeeks old . In anoteeks old .M. hyopneumoniae from an infected sow to her piglets is recognized as the typical pathway of enzootic pneumonia. Infected sows are considered a risk factor for M. hyopneumoniae infection in the preweaning period [M. hyopneumoniae shedding through direct contact with vertically infected piglets is the initial infection source in susceptible pig populations during the nursery and growth-finishing periods [M. hyopneumoniae [Vertical transmission of g period . M. hyop periods . “Sow heeumoniae –13.M. hyopneumoniae has been considered the gold standard. However, it is not used for routine diagnosis due to the highly time-consuming process. The organism can also be detected by immunofluorescence testing, but the test has limited sensitivity. Although serology can be used to show the presence of M. hyopneumoniae at a herd level, it is not proper for diagnosis on individual animals [For conclusive diagnosis, culturing of  animals . Current animals .M. hyopneumoniae infection compared to the more commonly used investigation of nasal swabs and tracheobronchial lavage fluid in eight-week-old pigs [M. hyopneumoniae using laryngeal swabs of suckling pigs. Although M. hyopneumoniae is barely detected in adult pigs, the detection rate of weaning pigs that were vertically infected by sows was comparably high. This is the first report of infection and virus shedding among sows in South Korean field conditions, and it was conducted by testing the laryngeal samples of weaning piglets with nested PCR.A recent study indicated that laryngeal swabs investigated with the PCR method are superior in detecting early old pigs . PCR resTwenty-two pig farms in South Korea were selected for testing. To investigate the test, swine veterinarians visited 22 farms under the contract. Without consideration of specific geological area, the veterinarians selected 22 farms according to pig population from four representative swine producing provinces of South Korea ; eight farms in Chungcheong-do (28%); six farms in Jeolla-do (25%); five farms in Gyeongsang-do (23%)). Accordingly, the farms were distributed in balance across the nation.The farms had a higher number of marketed pigs per sow per year (MSY) than the average South Korean pig farm and were managed with a three-way cross breeding program . The average MSY in South Korea was reportedly 17.9 in 2018 based on the Korean pork producer association. The average MSY of 22 farms joined in this study was 21.8. Only three farms  had less MSY than 20. Farm no.15 and 17 had unexpected events including porcine reproductive and respiratory syndrome (PRRS) and porcine epidemic diarrhea (PED) outbreak a year before the experiment. Farm no.16 had a sudden change of gilt source. Their relatively low MSY might result from such internal factors.M. hyopneumoniae for acclimation was selected on three farms, and vaccination with commercial products (Ingelvac® MycoFLEX and Zoetis Respisure ONE®) was used on 14 farms. While investigating, antibiotics  were used. In the case of antibiotics, there were three categories associated with the treatment. The sows and growing pigs were inoculated with antibiotics on randomly selected farms: only growing pigs on seven farms, only sows on five farms, and both on six farms. In the case of gilt introduction, seven farms introduced gilts from their own female pigs, and 15 farms brought gilts from grandparent farms (GP). Their protocols are usually used in South Korean swine production systems.All farms had farrow-to-finish production systems and weaned their piglets at three to four weeks of age, while each farm had individual acclimation, antibiotics, and gilt replacement protocols Table . The expM. hyopneumoniae from randomly collected sera were detected by ELISA (BioCheck), and PCR results of necropsied lung tissue showed the presence of M. hyopneumoniae. Including M. hyopneumoniae, type 1 and type 2 PRRSV, porcine circovirus type 2d (PCV2d) and bacteria  were also isolated from some cases. Concretely, 6 farms reported the presence of type 1 PRRSV, and 7 farms reported type 2 PRRSV. Only 3 farms had two PRRSVs simultaneously. PCV2d was the only porcine circovirus type on the farms and was detected on 5 farms. Salmonella and Streptococcus were isolated from one farm.The veterinarians reported signs of porcine respiratory disease complex (PRDC) in growing pigs based on previous tests, which were performed to monitor the herd disease state at least 1 month prior to this experiment. The antibodies against Sampling was conducted as previously described . BrieflyIn most farms, there were 15 ~ 20 farrowing sows in each batch, and most of them were selected for sampling. There were newborn piglets every week because all farms had the same weekly production system. The test was conducted 1 day per week in three consecutive weeks, and the date was selected by veterinarians during October and November 2018.M. hyopneumoniae strain SNU98703 culture supernatant) was tested for nested PCR sensitivity. The reaction detected M. hyopneumoniae DNA to a dilution of 10− 8, approximately 1 fg, as in a previous report [Laryngeal swab samples were processed for DNA isolation with a Viral gene spin RNA/DNA extraction kit  on the sampling day, and nested PCR was performed with isolated DNA as previously described . The poss report . A pristP < 0.05 was considered significant.The chi-square test and Fisher’s exact test were used to compare the prevalence of infected piglets with the following parameters: herd size, gilt replacement rate, acclimation procedures, and antibiotic usage. A value of M. hyopneumoniae DNA was detected by nested PCR in 14 farms (63.6%) and 127 piglets . After sequencing PCR products, 16S ribosomal RNA of M. hyopneumoniae was confirmed. The prevalence of sows likely to transmit M. hyopneumoniae (11.1%) was calculated as the ratio, specifically, the number of sows with M. hyopneumoniae-positive piglets divided by the number of total sows. The mean prevalence of 13% at the first sampling time was reduced to 2.2% at the final sampling time.M. hyopneumoniae DNA was detected in 48 piglets (5%) from 11 farms with ≤550 sows and 79 piglets (8%) from 11 farms with > 550 sows. Farms with > 550 sows had a significantly higher detection rate   from 13 farms that replaced ≤40% of gilts and 85 piglets (8.6%) of 9 farms replacing > 40%. Farms replacing > 40% of gilts had a significantly higher detection rate   from 13 farms introducing serologically M. hyopneumoniae-positive gilts and 51 piglets (6.3%) from nine farms introducing serologically M. hyopneumoniae-negative gilts  from seven farms with self-replacement of gilts and in 84 piglets (6.4%) from 15 farms with gilts brought from grandparent farms  from three farms that naturally exposed gilts to M. hyopneumoniae. M. hyopneumoniae DNA was detected in 67 piglets (5.4%) from 14 farms that vaccinated gilts. Finally, M. hyopneumoniae DNA was detected in 15 piglets (3.3%) from five farms without acclimation treatment. Farms that naturally exposed gilts to M. hyopneumoniae had a significantly higher detection rate than the others   from 11 farms using antibiotics in the sow herds and 46 piglets (4.7%) from 11 farms that did not   [M. hyopneumoniae during the early stages of infection is critical but technically challenging. The colonization of pigs by M. hyopneumoniae starts from tracheal cilia as the initial postexposure infection site. Therefore, direct access into the trachea cilia could be useful for sensitive detection of M. hyopneumoniae. And the tracheobronchial lavage fluid may be regarded as the best sample to detect the infected microorganism in this case [M. hyopneumoniae DNA detection among oral, nasal, and tracheobronchial samples as well as antibody detection in serum samples [M. hyopneumoniae infection or colonization, it may function as a vestibular site that may incur lower airways. Consequently, the larynx could be the place where M. hyopneumoniae is concentrated, and it is a preferred sampling site for favorable access and proximity to the lower airways [M. hyopneumoniae detection are practical and reliable.x (PRDC) . PRDC isx (PRDC) , 19. To his case . However samples , 21, 22. airways , 23. As M. hyopneumoniae DNA was detected by nested PCR in 14 farms (63.6%) and 127 piglets . The prevalence (11.1%) of sows likely to transmit M. hyopneumoniae in herds was calculated as the ratio of total sows to sows with pathogen-positive piglets. Compared to previous reports, it was similar to the results obtained in other countries. Using laryngeal swabs, 7% of weanling pigs and more than 50% of sow herds tested positive for M. hyopneumoniae in the USA. In Germany, 18.7% of weanling pigs and 75% of sow herds were PCR positive [As a result, positive .There was a significant difference depending on herd size and gilt replacement rate. Farms with more than 550 sows had a significantly higher detection rate than those with lower sow numbers. Farms where more than 40% gilts were replaced also had a significantly higher detection rate than those where gilts were less replaced. In general, larger farms tend to replace gilts at a higher rate. Larger farms or high gilt replacement rates may support the transmission of pathogens due to management factors such as biosecurity, vaccination practices, and herd density . HoweverM. hyopneumoniae detection in piglets among farms with regard to the serological status of introduced gilts. Similarly, the detection rate in farms with self-replacing gilts was not significantly different from the rate in farms with gilts brought from grandparent farms. With regard to the long shedding period, sows exposed to M. hyopneumoniae at the age of 50–100 days are expected to develop immunity early enough so that the possibility of transmission is reduced at their first farrowing [There was no significant difference in the prevalence of arrowing .M. hyopneumoniae had a significantly higher detection rate in piglets than farms with vaccinations or without acclimation. Considering that experimentally infected pigs shed M. hyopneumoniae for up to 200 days [Notably, farms where gilts were naturally exposed to 200 days , naturalM. hyopneumoniae in the farms where antibiotics were actively used. Farms with severe enzootic pneumonia outbreaks tend to use antibiotics more frequently as a control measure. In addition, the symptoms may reappear after cessation of treatment or development of antimicrobial resistance [Amoxicillin, enrofloxacin, tilmicosin, ceftiofur, penicillin, streptomycin, gentamicin, tulathromycin, lincomycin, and spectinomycin were used on most farms. Treatments were applied to sows, piglets, or both. Interestingly, the detection rate was significantly higher in farms that used antibiotics in the sow herds than in those that did not use antibiotics. The detection rate was also significantly higher in farms using antibiotics in suckling pigs than in farms where antibiotics were not used. Although antibiotics have been demonstrated to be effective at controlling enzootic pneumonia , only lisistance .M. hyopneumoniae is not consistent in the different farrowing batches. In addition, a minimum of three sampling time points is necessary to assess sow herd stability. The necessity of multiple samplings has already been suggested [As a result of subpopulation in a batch, there was a great deal of variability in the prevalence between farrowing batches within a farm. The mean prevalence at the first sampling time was 13%. In the second sampling time, a mean prevalence of 4.9% was reported. For the third sampling time, the mean prevalence was 2.2%. These results suggest that the time of infection and shedding of uggested .M. hyopneumoniae prevalence in weaning-age or approximately weaning-age piglets among farms. The results demonstrated that laryngeal swabs of suckling pigs provided useful information with regard to sow herd stability in farms with enzootic pneumonia. In addition, it provided some insight into influencing parameters such as herd size, gilt replacement, acclimation, and antibiotic usage.In summary, this study investigated the difference in


Medical topic therapies can relieve symptoms associated with lichen planus of external auditory canal. Further, bone‐anchored hearing devices represent an optimal solution for hearing restoration in otic lichen planus. Medical topic therapies can relieve symptoms associated with lichen planus of external auditory canal. Further, bone‐anchored hearing devices represent an optimal solution for hearing restoration in otic lichen planus. Otic LP, indeed, presents two main problems: the stenosis of the external auditory canal and an associated conductive hearing loss.We report a case of oticus LP with mixed hearing loss; the clinical presentation, diagnostic issues, and treatment are discussed. Moreover, we propose a new approach for the treatment of this disease.2PM, a 78‐year‐old woman, was admitted to our audiologic service for bilateral ear discharge, ear pruritus, and pain and progressive hearing loss, developed 5 years before, but rapidly worsened in the last 12 months. In the last 6 months, the patient had been using bone conduction hearing aid spectacles with very limited benefit. The otomicroscopy and the otoendoscopy showed an easy to blood, totally stenotic external auditory canal (EAC) bilaterally. Tympanic membrane was not visible, bilaterally. Tonal audiometry revealed a severe‐to‐profound mixed bilateral hearing loss, with an air conduction pure tone average 500‐1000‐2000‐4000 Hz (PTA) of 96.25 dB on the left side and 103.75 dB on the right side and a bone conduction PTA of 50 dB on the left side and 45 dB on the right side .Further, in order to improve hearing results, we proposed a bone‐anchored hearing device (BAHD) implantation surgery. We did not propose a cochlear implant procedure for the age of the patient and for the middle ear involvement by the inflammatory disease. After a complete preoperatory counseling and a test with the bone conduction softband device, the patient gave her informed consent to the procedure. The left side was chosen because of patient subjective preference. During the device test on softband, indeed, no significant objective differences were noticeable between left and right sides at free field audiometry or open‐set perception evaluation. We implanted the device abutment, and after one month, we activated and fitted the device.Considering the patient's bone conductive hearing threshold, we proposed the most powerful hearing processor that was available on the market, indicated for a bone conductive PTA better than 65 dB.2.2After 3 months from the activation of the BAHD and two fitting sessions, the patient had a PTA of about 40 dB HL at free field audiometry and an open‐set disyllabic word recognition score of 70% in silence and 35% with background noise with device on  and monolateral in 10 cases .The diagnosis is clinical, with a coexistence of mucosal LP and conductive hearing loss associated with EAC stenosis.In the few cases reported in the scientific literature, medical and surgical therapies have been proposed for the otologic management. Martin et al in one case proposed a surgical treatment with the removal of the inflammatory tissue and the calibration of the external auditory canal and, after the three months of relapsing, a medical therapy with oral acitretin  followed by oral prednisolone (1mg/Kg/day) with a temporary clinical improvement of the otologic finding and the conductive hearing loss.The patient we report was treated with otologic drop achieving a good control of the local symptoms after twelve months of medical therapy.Regarding the hearing deficit associated with otic LP, a conductive or mixed hearing loss, with a variable degree of presentation, but frequently mild, is generally reported, due to the stenosis of the EAC and in some cases of the tympanic cavity. Sartori‐Valinotti et al reported the hearing loss to as the most frequent symptom in patients with otic LP, both conductive and mixed; they also report that four patients presenting a relevant hearing loss in their cohort received bilateral hearing aids.As we previously stated, some authors reported that patients may have benefit from the use of traditional hearing aidsIn the present article, we report a case with a rare otic localization of LP. In our case also, as previously reported in the scientific literature, topical therapies with antibiotics and corticosteroid eardrops proved to be effective to control the local symptoms.None declared.FL: drafted part of the manuscript and reviewed the literature. LB: drafted part of the manuscript and contributed to review the draft. SB: reviewed the draft. ADV: drafted part of the manuscript and helped reviewing the literature. FF: drafted part of the manuscript and contributed to review the draft. All authors: approved this final draft for consideration of publication.


ATP7B gene leading to impaired copper metabolism. Brain imaging, such as magnetic resonance (MR) and transcranial sonography (TCS) in WD patients, shows changes mostly in the basal ganglia. Heterozygotic carriers of one faulty ATP7B gene should not exhibit symptoms of WD, but one in three heterozygotes has copper metabolism abnormalities. This study examined heterozygote ATP7B mutation carriers using TCS to assess any basal ganglia changes compared with healthy controls.Wilson’s disease (WD) is an autosomal recessive disorder of Heterozygote carriers and healthy volunteers underwent the same standard MR and TCS imaging protocols. Heterozygotes were followed for 5 years and monitored for the development of neurological symptoms.p < 0.001). Bilateral substantia nigra (SN) hyperechogenicity was found in 8 heterozygotes and one control; another 3 heterozygotes had unilateral SN hyperechogenicity . Heterozygotes had larger SN area on both sides compared with controls .The study assessed 34 heterozygotes (21 women), with mean age of 43 years (range of 18 to 74 years) and 18 healthy controls (13 women), with mean age of 47 years (range of 20 to 73 years). Bilateral lenticular nucleus (LN) hyperechogenicity was found in 25 heterozygotes, but none of the controls (SN and LN hyperechogenicity were more frequent in heterozygotes than in controls, probably due to copper accumulation, but it remains unknown if this predisposes to brain neurodegeneration. Wilson’s disease (WD) is an autosomal recessive inherited disorder of hepatic copper metabolism that is caused by malfunction of a putative copper-transporting P-type ATPase, ATP7B. Cellular damage occurs due to copper deposition in affected tissues, principally the liver and the brain , 2. ClinATP7B gene—is high, at around 1 in 90 individuals or 1–2% of the general population [ATP7B gene should not exhibit symptoms of the disease, but copper metabolism abnormalities have been noted, in a range from 20% of cases [The number of WD heterozygotic carriers—having only one faulty copy of the pulation , 12. Howpulation . Heterozof cases , with upof cases , 16. Conof cases .ATP7B gene with TCS to assess any basal ganglia changes and to compare with healthy controls.The aim of this study was to examine heterozygote carriers of the ATP7B gene were mostly recruited from the relatives of WD patients admitted for family screening for presymptomatic WD. All included heterozygotes underwent the same assessments including medical history, neurological examination, routine laboratory screening , copper metabolism screening , DNA analysis, and brain imaging using standard MRI and TCS protocols. The methods used for copper metabolism and DNA analysis were as described previously [Heterozygote carriers of the eviously . HeterozThe control group consisted of healthy volunteers who had no abnormalities in routine laboratory tests and no neurological abnormalities. They underwent the same standard MR and TCS imaging examinations. Genetic sampling and copper metabolism were not conducted in this group.All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.In both groups, MRI was performed using a 1.5-T scanner . T1-weighted  and T2-weighted  images were acquired in axial planes with 5-mm slice thickness. Gradient echo T2* images were obtained as a single-echo sequence . The imaging results were assessed by a neuroradiologist. Changes in the basal ganglia, mostly in the GP, putamen, thalamus, and SN, were recorded.2 were considered hyperechogenic, those ≤ 0.23 cm2 were considered normal, and intermediate-sized areas were considered moderately hyperechogenic. The GP and putamen were assessed as one structure, the LN, as in the TCS protocols. The LN and thalamus were visualized in the third ventricular plane. The echogenicity of the LN and thalamus were classified as hyperechogenic if echogenicity was more intense than the surrounding white matter. The area of hyperechogenicity was measured by manually encircling the outer circumference of the hyperechogenic area. The width of the third ventricle was measured on the axial scanning plane, with width < 0.7 cm considered normal  by a sonographer (MS) who was blinded to the MRI results. The sonographer was also blinded to the genetic results at the time of ultrasound examination. The chosen ultrasound parameters were penetration depth of 14–16 cm and a dynamic range of 50 dB. Image contrast and brightness were adjusted to obtain the best image. SN echogenic size measurements were performed automatically on axial TCS scans after manually encircling the outer circumference of the SN echogenic area. For the ultrasound system used, SN echogenic areas of ≥ 0.25 cmp value of < 0.05 was considered to indicate statistical significance. Wilcoxon Test and Fisher’s Exact Test were used to compare heterozygotes and controls. Logistic regression models were used to investigate the relationship between copper metabolism and LN and SN hyperechogenicity.Statistical analysis was performed using SAS version 13.2. A ATP7B gene; 21 were women (61.7%), with a mean age of 43 years (range from 18 to 74 years). The mutations found in the study group were as follows: p.H1069Q (c. 3207C > A) in 28 cases, p.IVS162A > G (c.3557-2A > G) in 3 cases, p.Q1351X (c.4051C > T) in 2 cases, and p.A1135fs (3402 delC) in 1 case.The study group consisted of 34 heterozygote carriers of the Ceruloplasmin levels below the laboratory range were found in 11 cases, and mean level was 29.16 ± 9.02 mg/dL. Low copper plasma levels were observed only in 5 cases, and mean serum copper was 91.91 ± 30.10 μg/dL. In all cases, 24-h urine copper excretion levels were normal, with mean urinary excretion 23.72 ± 28.82 μg/24 h. Routine laboratory results did not reveal any abnormalities. MR changes were found in T2* sequences in 2 cases: in the first case, there was bilateral hypointensity in the GP and in the second case, there was bilateral hypointensity in the SN. Both had p.H1069Q mutations.p = 0.45 for sex and p = 0.38 for age). All controls had normal MRI results.The control group consisted of 18 healthy subjects; 13 were women (72.2%), with a mean age was 47 years (range from 20 to 73 years). There were no differences in sex and age distribution between heterozygote and control groups (p < 0.001). Unilateral changes were present in 5 control cases. Mean LN echogenicity area was larger for heterozygote carriers compared with controls (p = 0.001 for right and p = 0.002 for left). Depending on the type of mutation, mean LN echogenicity area was as follows: for p.H1069Q, 0.52 cm2 and 0.51cm2; for p.IVS162A > G, 1.05 cm2 and 1.12cm2; for p.Q1351X, 0.22 cm2 and 0 cm2; and for the only patient with p.A1135fs, 1.0 cm2 and 0.68 cm2, respectively, for the right and left side. Bilateral SN hyperechogenicity was found in 8 heterozygote carriers and one control, while unilateral SN hyperechogenicity was present in 3 heterozygote carriers and no controls . Heterozygote carriers had larger SN area on both sides compared with controls . Thalamic hyperechogenicity was found in 3 heterozygote carriers and no controls, but the difference between the groups was not significant (p = 0.19). The width of the third ventricle was within the normal range in all heterozygote carriers (< 0.7 cm) and in 15 of the control cases. Controls had wider third ventricles than heterozygote carriers (p = 0.016). There was no correlation between TCS LN or SN hyperechogenicity and ceruloplasmin level, copper plasma level, and urinary copper excretion . A second female heterozygote carrier complained of memory problems—this case had no SN hyperechogenicity but bilateral LN hyperechogenicity on TCS. None of the followed cases developed extrapyramidal symptoms.TCS has gathered great attention in different neurodegenerative disorders. It has become an additional diagnostic tool for idiopathic Parkinson’s disease (PD) as hyperechogenicity of the SN is observed in about 90% of patients in the very early clinical stages of PD . TCS seeLN hyperechogenicity is the most common TCS abnormality in WD, found in all patients with the neurological form of WD studied previously , 9. In tATP7B carriers, who had normal structural MR. In our study, almost three-quarters of heterozygotes carriers had bilateral LN hyperechogenicity, while bilateral LN hyperechogenicity was not found in the control group. There might be LN echogenicity area differences, depending on type of mutation—with the highest for p.IVS162A > G and lowest p.Q1351X. Since the groups were very small this is not necessarily true and no statistical analysis was performed.In the current study, we found hyperechogenicity of LN and SN in heterozygote Heterozygotes also had marked bilateral SN hyperechogenicity in about one-quarter of cases. Hyperechogenicity of SN is the imaging phenomenon typical for the PD, present in more than 90% of patients, but only in approximately 10% persons without PD . HyperecThalamic TCS hyperechogenicity was found only in 3 heterozygote carriers, with no differences between heterozygotes and controls. Thalamic hyperechogenicity has been previously described in WD patients , but theOne of the markers of neurodegeneration and brain atrophy is the width of the third ventricle, with many WD patients exhibiting dilatation of the third ventricle. Neurological disease severity, assessed with the United Wilson Disease Rating Scale, has been shown to correlate with the widths of the third ventricle on TCS , 9. In oATP7B gene raise questions regarding the nature of hyperechogenicity of deep brain nuclei, with the exact answer still unknown. TCS hyperechogenicity was first described in a small group of PD patients, showing changes in SN, which was later attributed to iron accumulation [TCS findings in healthy heterozygote carriers of the mulation . TCS stumulation , 24, WD mulation , 9, 25, mulation , 27 patimulation , 28. Hypmulation , 30. PosWe speculate that LN and SN hyperechogenicity in heterozygotes are also due to copper accumulation, but we failed to find a correlation with copper metabolism and TCS findings. Thus, it is possible that hyperechogenicity is influenced by not only metal ions themselves but also different ion compounds and binding partners. Experimental studies in an animal model of PD show that SN hyperechogenicity is caused by structural changes, such as gliosis, rather than by increased iron concentration .ATP7B mutation showing TCS basal ganglia changes in cases with normal MR results and neurological examination. We found that SN and LN hyperechogenicity were more common in heterozygote carriers than in controls and this is probably due to copper accumulation. It remains unknown if the SN and LN hyperechogenicity observed can predispose heterozygotes to brain neurodegeneration. In general population hyperechogenicity of the SN is a risk marker for PD, but in heterozygote, ATP7B carriers is not a risk factor for developing PD/parkinsonian syndrome over 5 years follow-up.To conclude, this is the first study, to our knowledge, of heterozygote carriers of an


In the article titled “Single-Nucleotide Polymorphisms in XPO5 are Associated with Noise-Induced Hearing Loss in a Chinese Population” , the inf


Macaca mulatta) the effect of observing group members groom on a subject's subsequent grooming behaviour and the potential modulation of contagion by relationship quality and social status. We recorded behaviour after subjects witnessed a grooming event and compared it to behaviour in a control condition with the same individuals in proximity but in the absence of a stimulus grooming event. Compared to the control condition, after observing others groom, females engaged in a grooming interaction sooner, and were more likely to be the initiator and to take on the active groomer role. Dominance rank of the focal individual and more weakly also of the stimulus individuals affected the latency to the next grooming interaction of the focal subject. Latency to the next grooming interaction decreased with increasing rank of the subject potentially reflecting lower social constraints faced by high ranking individuals in this highly despotic species. Relationship quality between the subject and the stimulus individuals had no effect on latency to grooming. Collectively, our findings provide evidence for visual contagion of affiliation in rhesus macaques. Future studies should explore the systematic variation in contagion of interactive behaviour in relation to a gradient of social tolerance.Social contagion of non-interactive behaviour is widespread among animals including humans. It is thought to facilitate behavioural synchronization and consequently group cohesion, coordination and opportunities for social learning. Contagion of interactive behaviour—particularly affiliation—has received much less attention. Here, we investigated in female rhesus macaques ( ContPan troglodytes) and common marmosets  when exposed to higher rates of aggressive vocalizations from neighbouring groups, increased their intragroup aggression  minus latency PG [s]). We then built a model with this delta latency (in seconds) as the response that increases with increasing contagion. Predictors were the standardized rank of the stimulus individual (selecting the higher of the two) and the standardized rank of the focal subject. Focal subject ID was a random factor (N = 106 observations). To test prediction 4 with data from groups DUS and TED, we again used a GLM to determine whether the effect of grooming contagion increased with increasing affiliative relationship strength between the bystander and the groomers. To do so, we built a model with the delta latency  to the next grooming interaction of the focal animal as the response. The sole test predictor was the DSI value between the focal bystander and the stimulus individuals (selecting the higher of the two). Focal subject ID was a random factor (N = 82 observations). Testing prediction 5, we used a Wilcoxon matched-pairs test to assess whether the proportion of next grooming interactions  for which the bystander was the initiator was higher in PG grooming bouts than in MC grooming bouts. The proportion was calculated based on the total number of grooming events of the focal bystander. Following Berthier & Semple [N = 13 individuals.Furthermore, we ran the following statistical tests in Statistica 13.3 . To investigate whether grooming was contagious, we used a Wilcoxon matched-pairs test to test whether an individual's proportion of ‘attracted’ PG–MC pairs was higher than that of ‘dispersed’ PG–MC pairs . Both predictors had relatively weak effects on grooming contagion; the higher the dominance rank of the higher ranking of the stimulus individuals the larger was the difference in latency between MC and PG, i.e. the shorter the latency in the PG observation after accounting for latency in the MC .The dominance rank model was significantly different from the null model with only the random effect .Relationship strength between the focal female and the individuals she witnessed grooming did not influence the degree of contagion; the relationship model was not different from the null model , the proportion of grooming bouts initiated by the bystander was significantly higher in PG observations, i.e. after observing grooming, compared to MC observations . Our results most closely resemble findings of visual contagion of grooming behaviour in another macaque species, i.e. the socially more tolerant Barbary macaque . In thisper se was not included in the best model explaining yawning contagion in bonobos [Extending research on social contagion of interactive behaviour, here we investigated the potential moderating effects of dominance rank and relationship quality on contagion propensity found for non-interactive behavioural contagion such as yawning or scratching ,21,40,50 bonobos  that dom bonobos ,51. And  bonobos . In our The latency to the next grooming bout a focal individual engaged in was also affected by the focal individual's own dominance rank: the higher ranking she was, the sooner she herself engaged in a grooming interaction. While this effect was stronger than the effect of the stimulus rank on the latency to groom, the significance of the effect was lower. This result is not simply due to an increased attractiveness of high ranking individuals as grooming partners, as our design directly compared the two conditions keeping the social environment constant. Instead, increased contagion in high ranking females could be driven by the enhanced social opportunity dominant individuals enjoy. As observing grooming also increased the degree of proactivity in terms of grooming initiation and grooming role (see below), subordinate individuals may simply be more constrained to act on this behavioural tendency.Societies of female rhesus macaques belong to the most despotic among macaques and exhibit steep hierarchies and minimal social tolerance . While aPongo pygmaeus), where contagion was more likely to occur if the relationship quality between the stimulus and the observer was low [Relationship quality, or bondedness, has been shown to enhance yawning contagion in several species, including geladas , bonobos was low . The lac was low ,55 (but  was low ). A pote was low  and cons was low . As withUltimately, social contagion may enhance behavioural synchrony which in turn facilitates coordination, cohesion and group performance ,57,58. OCollectively, our study adds to previous findings highlighting that social interactions do not occur in isolation but in group-living species may also affect the behaviour of others that are not directly involved. Thus, social behaviour needs to be explored beyond the primarily interacting individuals. A dyad or cluster of socially interacting individuals will affect others' social attention to update on the current social situation , may lea


OTOA. A missense variant in ESRRB was implicated for recessively inherited moderate to severe hearing loss. Two individuals were heterozygous for variants in MYO7A and CHD7, respectively, consistent with de novo variants or dominant inheritance with incomplete penetrance as the reason for their hearing loss. Our results indicate that similar to familial cases of deafness, variants in a large number of genes are responsible for moderate to severe hearing loss in sporadic individuals born to consanguineous couples.Hearing loss affects 380 million people worldwide due to environmental or genetic causes. Determining the cause of deafness in individuals without previous family history of hearing loss is challenging and has been relatively unexplored in Pakistan. We investigated the spectrum of genetic variants in hearing loss in a cohort of singleton affected individuals born to consanguineous parents. Twenty-one individuals with moderate to severe hearing loss were recruited. We performed whole-exome sequencing on DNA samples from the participants, which identified seventeen variants in ten known deafness genes and one novel candidate gene. All identified variants were homozygous except for two. Eleven of the variants were novel, including one multi-exonic homozygous deletion in  Nonsyndromic hearing loss is most frequently inherited as an autosomal recessive trait3 and the genetics of profound deafness in families has been well characterized4. However, few studies have explored the prevalence of sporadic, moderate to severe hearing loss in the broader population.Approximately 50% of hearing loss cases has a genetic etiology. Of these, 70% are nonsyndromic and the remaining 30% are syndromic6. The etiology of severe to profound deafness in sporadic cases has been explored in some populations and variants in GJB2 (OMIM 121011) and SCL26A4 (OMIM 605646) have been found to be the major contributors9. Worldwide, only a few comprehensive studies have been carried out on single affected individuals exhibiting moderate to severe hearing loss. These studies have shown that variants in GJB2, OTOG (OMIM 604487), STRC (OMIM 606440), TECTA (OMIM 602574) and SERPINB6 (OMIM 173321) are more frequent in individuals with moderate or moderate to severe hearing loss in Chinese, Korean, Japanese and American populations12. So far, no extensive study has been conducted to elucidate the genetic etiology of moderate to severe, sporadic hearing loss in Pakistan. We present the first report on the contribution of different deafness genes in the etiology of hearing loss of sporadic individuals born to consanguineous parents.De novo variants or variants with incomplete penetrance, recessively inherited X-linked variants and mitochondrial variants may occur in individuals with no family history of hearing lossThis study was approved by Institutional Review Board of School of Biological Sciences (SBS), University of the Punjab, Lahore, Pakistan. All the experiments were performed in accordance to the guidelines and regulations set by the relevant body. Twenty-one probands were identified from various schools for deaf and special education schools in Punjab. The selected participants were born to consanguineous parents. The syndromic associations with hearing loss in this cohort were excluded by observation of the clinical features in individuals and detailed questioning regarding phenotypes. Written informed consents were obtained from participants or their legal guardians.Audiometry was performed to measure average hearing thresholds for all the participants at frequencies of 0.5, 1, 2, 4 and 8 kHz under ambient noise conditions. Romberg and tandem gait tests were completed to identify vestibular defects.13. Libraries were sequenced on the Illumina HiSeq2500 platform to generate 125 bp paired end runs. Reads were aligned with BWA mem 0.7.8 to the 1,000 genomes phase 2 (GRCh37) human genome reference sequence. Variant calling was performed using GATK 3.3–0 joint calling with HaplotypeCaller. The data after final output was analyzed independently at BHCMG, USA and SBS, University of the Punjab, Lahore. The program wANNOVAR (https://wannovar.usc.edu/) was used for annotating the variant call files (VCF). Using either the PhenoDB Variant Analysis Tool14 or manually, the output data from wANNOVAR was filtered against the population frequencies in the 1,000 Genomes database, genome Aggregation Database (gnomAD) and the Exome Aggregation Consortium (ExAC) database. Variants were retained for further evaluation if they had an allele frequency of less than 0.01 in these public databases. Homozygous, hemizygous and compound heterozygous exonic and splice site variants were examined. Large deletions and copy number variations (CNV) were detected using ExomeDepth15 using read depth data from exome sequencing experiments. For these analyses, each test exome was compared to a matched, aggregate reference set of samples. CNV calls were annotated using AnnotSV16. Candidate CNVs were prioritized by minor allele frequency, exon number, Bayes factor (BF) and the ratio of observed/expected number of reads.Whole blood up to 10 ml was drawn from the participants and the DNA was extracted using a standard method involving sucrose lysis and salting out. Whole-exome sequencing was performed at the Baylor-Hopkins Center for Mendelian Genomics (BHCMG). Exome capture was performed using the Agilent SureSelect Human All ExonV5 kit using a low input library preparation protocolhttps://sites.google.com/site/revelgenomics/). The conservation of selected amino acid residues affected by variants was checked across vertebrate species. For this purpose, multiple alignments were carried out on the protein sequences obtained from UniProt (https://www.uniprot.org/) using ClustalO .The wANNOVAR files also included predicted pathogenic scores for these variants from Polyphen 2, Mutation Taster and SIFT along with the pathogenicity CADD scores indicating the probable impact of variation on the function of the encoded protein. In addition, REVEL pathogenicity scores for the variants were accessed  for better hearing ears ranged between 65 to 88 dB HL Fig. . Romberg17 twelve were pathogenic, three were likely pathogenic and two variants were classified as variants of uncertain significance. Most of the variants except for one variant in OTOF (OMIM 603681) were unique to only one individual. All variants observed during this study have been deposited in ClinVar . No candidate variants were obtained from four participants.Whole-exome sequencing identified candidate causative variants in eleven genes in seventeen individuals Table . Of the OTOF affected three individuals with deafness in this cohort. Two novel frameshift variants were identified for three unrelated participants. The variant c.4990_4991del (p.Tyr1497TyrfsTer10) was present in two individuals HLRBS13 and HLRBS14. Another novel frameshift variant c.2443delC (p.Gln815GlnfsTer1) was identified in individual SPK6. Variants in GJB2, SLC26A4 and OTOA (OMIM 607038) affected more than one individual. Among these, one missense variant c.158G > T (p.Cys53Phe) in GJB2 and one variant c.3188C > G (p.Pro1063Arg) in OTOA were identified for the first time. A large homozygous deletion including OTOA was identified in individual SPK7. Homozygous missense variants in RDX (OMIM 179410), CABP2 (OMIM 607314), and ESRRB (OMIM 602167) were found to contribute to hearing loss in three individuals. All these variants had high pathogenicity scores and the affected amino acids were conserved among different vertebrate species  were identified in two unrelated individuals. The characteristic symptoms of Barter’s syndrome such as polyhydramnios or salt loss during neonatal stages were ruled out in both individuals by detailed questioning from the mothers of the affected children. The variant c.35 T > C (p.Ile12Thr) has been reported to cause nonsyndromic deafness DFNB73 (OMIM 602522)18. Since the novel variant c.52G > C  affects an amino acid located in close proximity of the reported p.Ile12Thr variant, it is possible that p.Leu18Val is also a nonsyndromic deafness allele. However, some individuals with missense variants only manifest renal abnormalities as adults19. It is therefore possible that renal abnormalities could be manifested in future by the child with the p. variant in BSND.One known and one novel homozygous missense variant in exon 1 of CHD7 (OMIM 608892) was identified in individual HLGMSA01. The variant was predicted to be damaging by three of the four prediction tools. No characteristic features of CHARGE syndrome (OMIM 214800) such as unusual shape of external ear, facial asymmetry, small eyes, delayed growth and delay in development of motor skills were observed for the affected individual or reported by the parents and siblings at the time of sampling. Samples from parents and siblings were not available to establish the segregation of the variant and to determine if it was a de novo variant.A heterozygous variant c.1531G > A (p.Asp1711Asn) in MYO7A (OMIM 276903) was identified in HLMS32. The variant had high REVEL (0.75) and CADD scores (25.8), and was predicted to be damaging by three of the mentioned prediction tools.A heterozygous variant c.4313C > T  in BHLHE22 (OMIM 613483) was identified in individual HLMS7 who had moderate to severe hearing loss. This variant was predicted to be disease causing by various online prediction tools but was of uncertain significance according to ACMG guidelines. The variant had a high CADD score of 22.9 and a relatively low REVEL score of 0.38. The variant was rare as it had a low allele frequency of 0.00008 and 0.0001 in ExAC and gnomAD respectively, with no homozygous individuals in the control population. It had a GERP++ score of 3.18.A missense variant, c.187G > C (p.Gly63Arg) in 21 which increases the risk of prevalence of recessive disorders, including hearing loss. According to the World Health Organization, the Pakistani population has a high prevalence of recessive disorders (2.4%) as compared to the incidence worldwide (1.7%). In this study, we explored the genetics of moderate to severe hearing loss for the first time in Pakistan in single individuals born to unaffected parents who were cousins. It was suspected that variants in few genes like STRC, GJB2, SLC26A4, OTOG or TECTA may explain the hearing loss for the majority of individuals in our cohort as is the case in many other world populations for the individuals with moderate to severe deafness. However, the identification of variants in multiple genes associated with hearing loss in our cohort of sporadic cases suggests a similar genetic heterogeneity in sporadic and familial cases in Pakistan. The combined contribution of genes involved in profound deafness is 52% to moderate to severe hearing loss in this cohort. The phenotypic variability due to variants in the same genes implicate the involvement of extrinsic factors or modifiers affecting the severity of hearing loss.Consanguineous families have served as a rich resource for the identification of genetic causes of recessively inherited disorders. In Pakistan 40–60% marriages are among first cousinsOTOF were more frequent in our cohort as compared to the published data for different ethnicities or populations. Four other reports from Korea, Japan and China on the genetic predisposition of hearing loss in sporadic individuals included more than 60 participants with moderate to severe or profound deafness, in which they demonstrated SLC26A4 as the major contributor to hearing loss23.The variants in GJB2 related deafness accounts for 10% cases in our cohort, which is similar to the reported incidence of GJB2 variants (9.5%) obtained from screening of large consanguineous families11. However, a recent research from Pakistan on 40 individuals with profound deafness from Bannu and Kohat districts indicated that GJB2 variants caused deafness in 37% of non-familial cases9. The small sample size, difference of ethnic background and less severe hearing phenotype may explain this lower contribution of GJB2 variants in the present study.The ESRRB was identified for moderate to severe hearing loss in one participant of this study. ESRRB is an estrogen related receptor beta gene which is known to cause hearing loss at DFNB35 (OMIM 608565) locus. The encoded protein consists of two domains; DNA binding domain (DBD) and ligand binding domain (LBD). Seven of the previously identified variants affect the ligand binding domain of the protein. The variant identified in this study also affects the ligand binding domain and the amino acid at this position is conserved among vertebrate orthologues  in ues Fig. B. HoweveBHLHE22 was potentially implicated for moderate to severe hearing loss. BHLHE22 has a single coding exon which encodes a class E basic helix loop helix protein 22 (BHLHE22). It is a small protein of 381 amino acids which serves as a sequence specific DNA binding transcription factor and mediates cell differentiation and proliferation. Mutant murine models have demonstrated that BHLHE22 is necessary for retinogenesis24 and development of dorsal cochlear nuclei26. BHLHE22 has the highest expression in retina26 however, it is also expressed in cochlear hair cells, supporting cells and utricle in the inner ear .A missense variant c.187G > C (p.Gly63Arg) in BHLHE22 had a relatively low conservation score  and high pathogenicity scores . These scores may be explained by a previous study on transcriptional repressors, which suggested that nonconserved regions are vital for the DNA binding function of the proteins. They may also provide a drift during evolution for the correct folding and thus secondary structure of the respective protein27. Therefore, the identification of BHLHE22 variants in additional affected individuals or mice models will be useful to understand the role of this gene, if any, in hearing loss.The variant c.187G > C in MYO7A primarily cause autosomal recessive nonsyndromic hearing loss (DFNB2) (OMIM 600060) and Usher syndrome 1B (USH1B) (OMIM 27690)29. A heterozygous variant identified in individual HLMS32 suggests that hearing loss is probably nonsyndromic dominant as observed for DFNA11 (OMIM 601317) instead of USH1B or DFNB2 which are caused by biallelic variants of MYO7A. However, we cannot exclude the possibilities that either the individual is a carrier for DFNB2/USH1B variant or the variant may be benign, in spite of its prediction to be damaging.Majority of variants in 8. Few other studies on sporadic cases from Korea, China and Japan have reported the pathogenic variant detection rates of 20% (from 92 cases)30, 23.1% (from 13 cases)6, 32% (from 34 cases)31 and 45.4% (from 11 cases)5 using whole-exome sequencing. The mutation detection rate in these studies is lower as they screened the individuals for common variants of GJB2 and SLC26A4 prior to whole-exome sequencing. However, even after excluding the GJB2 cases, the identification rate for the present study still remains as high as 71%. This may perhaps be due to the fact that we specifically studied hearing loss in individuals born in consanguineous unions. This increased the possibility that the disorder was recessively inherited.The diagnostic rate for sporadic cases in this research was relatively high (80%) as compared to other studies. For instance, in a cohort of 63 simplex cases from China the successful diagnostic rate was of 12.7%Variants of uncertain significance were identified in multiple participants Table  while noOur study comprehensively evaluated the genetic cause of moderate to severe hearing loss in a cohort of sporadic individuals. Results show that a similar diversity of gene variants is responsible for sporadic deafness as seen for familial hearing loss. Therefore, such cohorts can serve as a rich source for the determination of genetic and molecular basis of hereditary deafness. These results also suggest that targeted sequencing of few common deafness genes prioritized according to the ethnicity, followed by whole-exome sequencing will be a simple and cost effective approach for the genetic diagnosis and management of isolated hearing loss.


Chemical exposure leading to ototoxicity is a fresh challenge for occupational healthcare in South Africa.The critical question is: ‘what is known about occupational ototoxic chemicals with or without noise exposure in South Africa?’This qualitative, mapping study was completed with published (peer-reviewed) and grey literature from 1979-2019. Data was analysed using the Preferred Reporting Items for Systematic reviews and Meta-Analyses: extension for Scoping Reviews and the Nursing and Allied Health Resources Section subcommittee on Mapping the Literature of Nursing and Allied Health (adapted). Numerical analysis of article type was completed, but the primary focus was on capturing patterns/trends using thematic analysis and ideology critique.African Journal of Disability, African Journal of Primary Health Care and Family Medicine, South African Medical Journal, The South African Journal of Communication Disorders [SAJCD] and Health SA Gesondheid) were included with the SAJCD containing one relevant item and seventeen other items were analysed. Research focusses on the mining sector (gold) in Gauteng, and ototoxic medication (tuberculosis and/or human immunodeficiency virus) take precedence. In KwaZulu-Natal, the focus is on commerce and industry across formal and informal sectors. There are no governmental policies that refer to chemical ototoxicity. Occupational hearing loss is configured exclusively on the meme that noise exposure is the only toxin.The Chemical exposures are only just beginning to be recognised as ototoxic in South Africa. Hearing conservation programmes should always serve the workers’ interests and never bow down to the econometric interests of employers. Every day, 6000 women and men die because of occupational fatalities. Of those who survive, 160 million live with disabling injuries or illnesses  commissioned a study on occupational health in the mining sector  exposures in the workplace. (p. 173)This is one of the most significant items in this World Bank’s discussion paper. Why? For hearing healthcare in Southern Africa, the World Bank’s recommendation represents a shift from relying on a ‘noise-only’ discourse to explaining occupational hearing loss. Critically, Osewe and Nkrumah have sounded alarm bells about workplace chemicals as contributing to hearing loss too. This positioning of chemical ototoxicity is a fresh challenge for occupational healthcare in less industrialised world regions such as sub-Saharan Africa.Among occupational healthcare professionals, it is widely known that chemicals affect hearing. It is well established that medications  are ototoxic as many illnesses, and diseases in the world are treated using medications with innate ototoxic properties, for example, aspirin  in the early 1990s, I worked with communities maimed by the British company, Thor Chemicals. This company is a case in point of how big business and government, enabled by a historicised geopolitical license, value the dollar over death and disability. In the 1970s, they started exporting toxic chemical waste (mainly mercury) to Cato Ridge (KZN) after disallowed to continue operations in England Carnie, . So, froCorporations with social responsibility on their minds and old and new democracies are bedfellows with the ‘unholy trinity’: the International Monetary Fund, World Bank and the World Trade Organisation Peet, . Big busThe aim of this study was to identify the coverage of the current body of knowledge of workers’ hearing when exposed to ototoxic chemicals with/without noise in literature focussed on South Africa. Topics that have attracted hearing healthcare researchers, and gaps or opportunities for further research will be identified.The selected design is a qualitative, literature mapping study because it is best suited to answer a research question that is about ‘what is happening’ in a particular subject or field of inquiry. Mapping studies, as a genre of evidence synthesis methods, have been used since the early 1990s by health science researchers. Perryman  reviewedTwo broad data sets were used: (1) published, peer-reviewed articles – obtained via two search strategies, detailed below, and (2) grey literature (including print/paper data) tagged by the type or authenticity of the data, see below. Databases used were limited to Google Scholar, PubMed/Medline and ScienceDirect/Scopus for a 40-year period (1979–2019) – selected for accessibility of electronic and print data.Theoretical or operational construct sampling was used because this form of purposive sampling is suited for the selection of data that represent important constructs about the phenomenon of interest Suri, . OperatiSCImago Journal and Country Rank that were likely to publish articles regarding occupational hearing loss owing to chemical ototoxicity with/without noise, focussed on South Africa. There are inherent limitations to this method, discussed later. However, words (or synonyms) in the journal titles had to be related to ‘occupation’ and/or ‘health’, ‘hearing’, ‘disability’, ‘communication’, ‘noise’, ‘pharmaceutical’, ‘chemical’ and/or ‘toxicology’. ‘African’ and/or ‘South(ern) Africa’ and synonyms were the terms selected to identify and include relevant journals.As the design is a mapping study but with principles adapted from other evidence synthesis methods like scoping and systematic literature reviews, two search strategies were used; the initial search (discussed in detail below) was performed by a research assistant. Firstly, we identified highly ranked journals via the international ScopusSCImago Journal and Country Rank is limiting by geopolitical intellectual space, I was concerned about a bias/risk of missing unindexed publications from/about South Africa. This then made it necessary to locate articles that were published but not necessarily peer-reviewed, namely, grey literature like items in paper (vs. electronic) versions, for example, unpublished university student dissertations. Across peer-reviewed published and grey literature items, I ensured that all were about South African workers, chemical (with/without) noise exposures and occupational hearing loss. Therefore, keywords searches were completed using medical subject headings (MeSH) or Boolean searches (as per database) for ‘chemicals’, ‘solvents’, ‘hearing loss’, ‘ototoxicity’, ‘audiologist’, ‘audiology’, ‘hearing healthcare’, ‘South Africa’, ‘Africa’, ‘industry’ and ‘occupational health and safety’.Secondly, a search strategy was adapted from the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Reviews (PRISMA-ScR)  chemical exposures with/without noise in the workplace and that (2) the South African workforce were considered for the time period stipulated above (1979–2019).A structured data collection schedule was developed and included the following items:booksgovernment documentsInternet sourcemiscellaneous.A determination of the number of citations and classification of the literature format types by:‘pre-’ if prior to the year range‘in-press’ for unpublished work in 2019‘unknown’ for undated items.Database sorting of format type, cited year (selected as 1979 onwards) and tagged as:When journals changed their titles/names, articles were listed under the most recent title of the journal.Development of the data collection tool and process: A data collection schedule was adapted from the protocols of the Nursing and Allied Health Resources Section (NAHRS) Subcommittee on Mapping the Literature of Nursing and Allied Health . The initial blind review for the selection of primary data assisted with data credibility too. Data dependability was checked via a detailed track record of the data collection process. As generalisability is not the goal of this type of qualitative research, transferability was what guided evaluations of the usefulness of data for other contexts or setting conducted via designing reflexive categories of analysis.This research received no specific grant from any funding agency in the public, commercial or not-for-profit sectors.In relation to stated aims and objectives, South African workers’ exposure to chemicals with/without noise is discussed under the following topics:current knowledgefocal research topicsknowledge gaps and research opportunities.SCImago Journal and Country Rank for South Africa only, and by the following subject areas: ‘Health Professions’, ‘Medicine’, Multidisciplinary’, ‘Neuroscience’ and ‘Pharmacology, Toxicology and Pharmaceutics’:The following journals were identified using the International Scopus (2018) African Journal of DisabilityAfrican Journal of Primary Health Care and Family MedicineSouth African Medical JournalThe South African Journal of Communication Disorders [Die Suid-Afrikaanse Tydskrif vir Kommunikasieafwykings]Health SA GesondheidSCImago Journal and Country Rank, in spite of the Occupational Health Southern Africa journal being accredited by the Department of Higher Education and Training in South Africa. Across all the journals, only one item per journal was included, which was the same as that identified in the data base searches  for a focus on chemical exposures with/without noise regarding South African workers. Notably, there were no occupational health-related journals ranked by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Reviews guidelines were used to electronically identify the initial data set of 2010 items. Inclusion criteria were applied – see the PRISMA-ScR flow chart , adaptedAWEH – Advancing Workers’ Engagement with Hearing)  taking precedence. In KZN there appears to be a clearer focus on commerce and industry – both in the formal and informal sectors. KwaZulu-Natal research and services are developed especially for workers exposed to solvents in combination with noise and other risk factors like ototoxic medication.The most critical gap noted is that there are no governmental policies that link chemical ototoxicity in any South African policy on occupational hearing loss, noise or otherwise. Occupational hearing loss is configured exclusively on the meme that noise exposure is the only toxin. For a more detailed analysis of this bias and/or omission in the law, see Manning and Pillay .Furthermore, hearing healthcare professional education programmes must consider the informal sector and workers’ exposure to a plethora of toxins. The notion of occupational hearing loss owing to ototoxicity must find its way onto the agendas of public and occupational health researchers and practitioners alike. Chemical exposures are only just being recognised as ototoxic in the mining (Osewe & Nkrumah, In response to the question, ‘what is known about occupational ototoxic chemicals with/without noise exposure in South Africa?’, this study has mapped the knowledge we are developing in this field in South Africa. However, it does indicate the need for a more systematic evaluation of the nature of studies, their methodological quality and a deeper evaluation of the contribution hearing healthcare researchers make in transforming how employers configure their employers’ healthcare.In a study, entitled ‘Hearing Conservation Programmes: For Employers/Workers?’, South African formulae for calculating workers’ compensation were evaluated for how one accounted for complexities like chemical ototoxic occupational hearing loss Pillay, . What wa


MYO15A as the only candidate pathogenic mutations cosegregated with the hearing loss in this family. These two variants were absent in 200 normal-hearing Chinese Hans and were classified as likely pathogenic and pathogenic, respectively, based on the ACMG guideline. Our study further expanded the mutation spectrum of MYO15A as the c.3658_3662del mutation is novel and confirmed that deaf patients with recessive MYO15A mutations have a good outcome for cochlear implantation.Hearing loss is a highly heterogeneous disorder, with more than 60% of congenital cases caused by genetic factors. This study is aimed at identifying the genetic cause of congenital hearing loss in a Chinese Han family. Auditory evaluation before and after cochlear implantation and targeted next-generation sequencing of 140 deafness-related genes were performed for the deaf proband. Compound heterozygous mutations c.3658_3662del (p. E1221Wfs∗23) and c.6177+1G>T were identified in  The Mng exons . Myosin ng exons –10. In hss DFNB3 . The pref ARNSHL –15.MYO15A were identified as the probable cause of the deafness.Here, we report a nonconsanguineous Chinese Han family with profound ARNSHL, in which compound heterozygous mutations in A Chinese Han recessive deafness family  was enroAudiometric assessments included otoscopic examination, pure tone audiometry (PTA), auditory brainstem response (ABR), and multiple steady-state responses (ASSR). Hearing level was assessed at 250, 500, 1000, 2000, 4000, and 8000 Hz. The hearing threshold was defined as the average of both sides. Inner-ear malformation and dysplasia of the auditory nerve related to the hearing loss were excluded by temporal bone Computerized Tomography (CT) scan and cranial Magnetic Resonance Imaging (MRI).GJB2, SLC26A4, and MT-RNR1 were excluded by Sanger sequencing. Targeted next-generation sequencing was then performed in the proband as previously reported [http://www.umd.be/HSF3/) was used to calculate the consensus values of potential splice sites.Blood samples were collected into an EDTA anticoagulant tube by venipuncture of the cubital vein. Extraction of genomic DNA was performed using a blood DNA extraction kit . As the first step, mutations in common deafness genes reported . A totalreported . Human SThe proband was a 14-year-old male from Zhejiang Province, China. He had congenital, bilateral, profound hearing impairment with a threshold above 95 dBHL as revealed by the PTA  and ABR MYO15A (NM_016239) were identified as the only candidate pathogenic mutations consistent with a presumably autosomal recessive inheritance. The mean depth of sequencing was 364.43X, and 98% of the targeted region was covered with at least 20X. Cosegregation of these two mutations with the hearing phenotype was confirmed within the family members (By targeted next-generation sequencing of 140 deafness-causative genes in the proband, compound heterozygous mutations c.3658_3662del and c.6177+1>T in  members . These t members . The c.6MYO15A mutations and recessive deafness DFNB3 was first discovered by Friedman et al. in Bali, Indonesia [MYO15A, all in a homozygous state, result in congenital, severe-to-profound hearing loss [MYO15A have been reported, mostly reported in consanguineous families from the Middle East [MYO15A were identified. Like many previously reported truncating mutations in MYO15A, the p.E1221Wfs∗23 variant is predicted to result in a truncated protein product without Motor, IQ, MyTH4, FERM, SH3, and PDZ domains (HCs in the cochlea play a critical role in converting mechanical sound waves into neural signals for hearing, and most of the hearing loss induced by gene mutation, noise, different ototoxic drugs, inflammation, or aging is caused by the HC malfunction –27. The ndonesia , in whicing loss . To datedle East –35. In t domains . The c.6MYO15A are associated with congenital, severe-to-profound deafness [MYO15A in the sensory HCs, the proband in our study had a marked improvement for hearing after cochlear implantation, showing a good prospective outcome for a similar procedure in other DFNB3 patients.Most recessive mutations in deafness , 33, 36,deafness . Both vaMYO15A may markedly benefit from cochlear implantation.The p.E1221Wfs∗23 and c.6177+1G>T compound heterozygous mutations in MYO15A are the probable cause of congenital, profound deafness in the Chinese Han family. Patients with recessive mutations in