This study aimed to explore perceptions and experiences about how owning a hearing dog can influence the functioning and the autonomy of people with hearing loss. Three adults participated in a semi-structured interview. The interviews were video recorded, transcribed, and coded. A procedure combining qualitative content analysis and interpretative phenomenological analysis was used. The study shows how specific aspects of hearing dogs are associated with increased autonomy and sense of security among owners. The attentive dog-owner pairing, the outstanding training and the companion role of the hearing dog are the main elements supporting the high satisfaction related by all the participants. In regard of the location context , ongoing challenges for owners are reflected in the lack of visibility of this rehabilitation means and its poor recognition from the society, resulting in the constant burden to explain the dog’s work to others. For adults with hearing loss, the hearing dog is a relevant way of offering both the benefits of functional assistance and the psychosocial support of a pet. The association between owning a hearing dog and improved overall well-being suggests that this form of rehabilitation should be considered as a pertinent option by hearing health professionals. The consequences of auditory impairment are numerous and important. It can lead to communication and speech-understanding issues, but also to weariness, anxiety, social isolation, psychological distress, and depression ,2,3. FurPerson-centered rehabilitation should not only focus on reducing activity limitations and participation restrictions, but also take into account the emotional and psychological needs of the individual . RehabilThe benefits of service dogs can have far-reaching implications for individuals with disabilities, for society and for the economy by promoting independence, learning, and employability . Most inSignificant positive effects of the hearing dog in different components of functioning have been found. It has been repeatedly shown to perform well for the tasks it is trained to do, consisting primarily in alerting ,18,19. IIn contrast to these findings, Singh et al.  found noImproved social integration, in which the hearing dog acts as a catalyst for social interactions, have been discussed: owners reported being less inclined to avoid interactions with other individuals after the placement of the dog . Thus, tAlthough most interrogated owners reported being satisfied with the acquisition of their hearing dog, some negative aspects were also brought up. Among those are the dog’s behavior problems, the lack of training, the lack of post-pairing follow-up, the additional training necessary for certain tasks specific to the owner, the making of travel arrangements, the access to public places, the responsibilities associated with maintenance and care of the dog, the distraction of the dog by people, and the failure to alert to sound ,16,17. TCharter of human rights and freedoms protects any person with a disability who uses a guide or assistance dog. The hearing dog being recognized by this authority since 2007, any person accompanied by such a dog has the right to access without discrimination public places, public transport and taxis, workplaces and places of leisure  so if I lost something I had the visual support next to it”.The three participants interviewed learned about the existence of hearing dogs and the If there is an alarm, [the dog] will perceive the alarm through the movie [at the cinema]”. P1 and P3 mentioned that their dog could warn them when their child cries or calls them from another room in the house. The dogs were also trained to respond to basic commands  in French, English, and gestures. P1 also trained her dog for new tasks related to her workplace. All participants mentioned that their dog had been trained for tasks adapted to their needs and that there was nothing more the dog could do for them. More so, they claim that hearing dogs do a lot more than what they are trained to do. Nearly half of the statements about the experience of owning a hearing dog concerned non-auditory aspects. Among these, companionship is the most mentioned element, followed by the reduction of anxiety , improvement of physical health and motivation and reduction of loneliness. On this matter, it is impressive to note that a participant’s dog saved her twice by warning her of a heart attack in progress.The main task performed by the participants’ hearing dogs was alerting to repetitive sounds . P1 also mentioned the dog’s ability to perceive alarms in background noise: “It kind of freed my family. Completely also my entourage. This makes me feel less handicapped. You feel a lot more normal” (P3). They also all reported a higher sense of security since acquiring the dog, particularly in relation to the detection of alarms. Having a hearing dog would also increase social interactions by promoting conversations with strangers or other dog owners. Thus, concerning participation, we obtained testimonials demonstrating either an augmentation in participation  or no effect on participation. Only P1 experienced serious participation restrictions at her workplace because of her dog. On this subject, difficulties regarding access to buildings with the dog was a massive concern reported by all respondents: “[It was] more than a huge difficulty in Quebec. […] being refused to go somewhere because it is not a Mira dog […]” (P3). In terms of recognition and visibility, all expressed feelings of not being understood by Quebec government’s instances, which seem, from the perspective of the participants, to recognize very little of the rights of people with hearing disabilities and even less of those who benefit from a hearing dog. Thus, the recognition, but also the lack of knowledge on the part of the Quebecers  is a significant issue for the participants.The three participants mentioned an increase in their autonomy since acquiring their dog: “She is extraordinary. Extraordinary is not even the word. I don’t even know how to explain it” (P3). P1 and P3 also mentioned that those around them were satisfied with the arrival of the dog in the household. In the particular case of P2, pairing failure was a difficult event to go through: “Well, I was definitely disappointed, you know. But I have a kind of small mourning, there you know, but after that I came back”. P2 had to return the dog due to cleanliness problems, but also because he found the dog was not performing well all the time. It was sleeping a lot and sometimes did not alert its owner when it was supposed to. Although he returned his initial dog, P2 appreciated the experience enough to request another one; he is currently on the Foundation’s waiting list.Owning a service dog sure comes with its own set of responsibilities. The most irritating commitment reported was the permanent and constant duty to take care of the dog. Otherwise, cleanliness and behavior  were also mentioned a lot. When asked about the negative aspects of owning such dog, respondents all mentioned having been bothered several times by strangers touching or talking to the dog while it was at work. They must therefore constantly explain the role of the dog and educate people on how to behave around an assistance dog. Access restriction to certain buildings was also discussed. Being authorized to have only one assistance dog at a time (even once the dog is retired), the limited space for the dog on the plane and the costs related to the animal have all been named once on the negative side of the experience. Despite these negative aspects, respondents reported having complete confidence in their dog’s abilities and expressed overall satisfaction with the daily support provided by their dog: “The opinion that hearing aids or cochlear implants are essential tools to improve communication for a person with hearing loss is unanimous among the interviewees. Although the connectivity options available are appreciated, it turns out that these aids are not flawless: some sounds remain inaudible, and adaptation can be difficult. Hearing assistance technology such as EAS can be used as a complementary rehabilitation means to help alert the individual of inaudible sounds. P1 and P3 were unhappy with this system and stopped using it altogether because of its many inconveniences, even if it meant having to find other ways to get by or relying more on their support network. On the other hand, P2 reported understanding the usefulness of the EAS and continues to use it to this day, although he finds it to be a rather artificial technology. He sees a complementary utility in it, the dog being for him mainly a tool to counter social isolation.Foundation’s role. What was mentioned the most was the attention paid to pairing the animal with the right beneficiary and the quality of the post-pairing follow-up. The good coordination of services was appreciated by all. However, it should be mentioned that one of the participants experienced a difficult pairing and did not like how the situation was handled by the Foundation. Accessibility during training is another point which did not make consensus among participants. The Foundation’s personnel are said to be trilingual , but P2 was disappointed with the poor quality of certain staff members’ French. Additionally, sign language was used, but only in ASL, while the man would have been more comfortable with the Quebec sign language (LSQ). P1 and P3 had less profound hearing loss and did not have communication and accessibility difficulties during training. Another negative point mentioned by all is the website’s lack of user-friendliness.Participants were asked to describe their experience in Oakville and their general satisfaction with the To close this topic, all participants mentioned that the Foundation requires a high level of care for the animal . Although, this is not perceived as a negative element for beneficiaries who want the good of their animal. To conclude the interview, all respondents expressed a keen interest in the installation of a Foundation training center in Quebec to ease access to the service and promote the recognition of hearing dogs in the province.This project focused on service dogs specializing in hearing loss and aimed to describe the experience of owners of this little-known means of hearing assistance. This research is founded on three semi-structured interviews conducted with adults who currently own or have owned a hearing dog. The small sample size was predictable since only a few individuals currently own a hearing dog in Quebec. There may be several reasons why there are few hearing dogs in this province, including the lack of a local provider, inadequate promotion and accessibility or the rare recommendation of this assistive help as a means of auditory rehabilitation. Maintenance costs associated with caring for this type of help could also be one of these reasons, as participants mentioned that food and veterinary services are expensive and that they do not receive the same monetary compensation from the government as owners of blind dogs. Nevertheless, the explorative aim of this research is satisfied by the extent of the sample size presented. We propose to discuss the benefits and disadvantages of owning a hearing dog, but also the participants’ satisfaction, social participation, and sense of security since acquiring the dog. Interpreting the results in a multidimensional approach will allow us to observe the relevance of the hearing dog as rehabilitation means for adults with hearing loss.The perceptions and attitudes reported by the respondents are consistent with the frequent consequences of hearing loss mentioned in the literature such as social isolation, anxiety, frustration, or the feeling of being a burden ,2,3. ParIn regard to the results, the most salient benefits of owning a hearing dog can be summed up in four words: usefulness, safety, motivation, and companionship. These findings are consistent with much of the previous literature on the subject. Almost no effect on social interactions or social participation was reported by the individuals interviewed, except for the increase in autonomy to carry out certain activities more easily. This point stays a controversy element in literature as some have not found any effect of hearing dogs on social interactions ,14.The three interviewees demonstrated a high level of satisfaction. Their main expectations were met and exceeded. The dogs were trained in an exemplary and adapted manner, resulting in a strong trust from the owner and leading to an increased feeling of security both at home and during daily activities. On the other hand, various benefits reported were not related to the auditory sphere , these results being consistent with the previous literature ,19,20. WSocial acceptance and recognition were the most discussed topics during the interviews. The participants repeatedly noted the lack of knowledge about assistance dogs among Quebecers. Furthermore, access to businesses and buildings appears to be the biggest issue for the owners. These situations seem particularly problematic in Quebec, compared to the other places visited by the individuals interviewed. According to them, the existence and visibility of Mira foundation’s dogs could be part of the explanation since almost exclusively these dogs seem to be recognized by the general public. Constantly having to explain and justify the presence of the dog appears to weigh heavily on the daily life of the beneficiaries, even if the benefits of the dog remain greater. It should be remembered that in Quebec and Canada, no specific law makes it possible to circumscribe the use of hearing assistance dogs , which sWe reiterate here that the main role of the hearing dog is in some ways comparable to that of an EAS . So why get a dog instead of a technological system? According to the participants, dogs have particularities the EAS has not. First, they can alert the beneficiary when someone calls him/her by her name. This point was mentioned many times by our respondents as a facilitator in social contexts, where they often miss many communication cues. Second, the dog is seen as less constraining for the rest of the household since it does not bark or disturb anyone when alerting its owner. Finally, and most importantly, the dog can follow the beneficiary in his/her daily activities outside of the house. This represents a huge advantage when taken from a social participation and inclusion viewpoint, but also from a security perspective. However, it is worth mentioning that recent technological devices, such as intelligent watches, are now capable of alerting the owner of certain signals outside of its house . This could potentially provide an alternative means of being alerted for individuals with hearing loss.Although the current acquisition process available to Quebecers requires doing business with an organization located in Ontario, we found that the service is nevertheless accessible from a financial and linguistic point of view for people from Quebec. It should be noted, however, that the participants were either retired, on sick leave or business owners, which eliminated or reduced the constraints related to employment . The individualized pairing process, the training offered to dogs and beneficiaries, as well as the post-pairing follow-up remain the strengths of the Foundation. It is a generally positive portrait that emerges from the process carried out to obtain a hearing assistance dog through the Lions Foundation of Canada.Nevertheless, all participants were interested and in favor of the establishment of a system for acquiring hearing dogs in Quebec province. Two main reasons motivate this point of view. First, respondents believe that a Quebec service point would facilitate the acquisition process, but also the post-pairing follow-up. For instance, veterinary care could be provided for free or at a reduced rate and the staff would be more comfortable in Quebec French and LSQ. The other reason resides in the idea that a more local branch would allow the Quebec population to be more aware of the existence and role of hearing dogs. Participants did not feel that their dogs were recognized in the same way as dogs from other specialties or foundations. Having a distribution service located in Quebec would allow them to obtain better civic and organizational recognition, which would improve their inclusion in everyday society. This shows that local implementation entails more than practical benefits for owners. In regard of their own context and particular needs, other jurisdictions could also benefit from local hearing dog providers for the previous reasons.It is important to acknowledge that our study has certain limitations that should be considered when interpreting the results. We did not systematically control for variables such as personality or prior experience with hearing assistive technologies, which could have affected the participants’ experiences. However, we did consider these factors when interpreting the data and recognized the potential impact they may have had on the results. We did not include a measure of QoL in our study, as it is typically assessed using a specific questionnaire or similar tool. While it would be interesting to examine thoroughly the impact of this intervention on QoL, it was beyond the explorative scope of our current study. Finally, it is also important to recognize that our sample size was relatively small, which may have limited the generalizability of our findings. In order to fully understand the potential value of this hearing assistance for the Quebec population, the main findings put forward by the three participants will have to be re-evaluated on a larger scale within the population to better demonstrate the relevance of this aid in this context. Despite these limitations, we believe that our study provides valuable insights.The results of our study suggest that the acquisition process for hearing dogs was perceived positively due to the ease of the process, the availability of supportive resources, and the provision of free travel accommodations. These factors were identified as particularly beneficial to the participants. Once the pairing with the hearing dog was completed, participants mentioned their dog was trained to perform tasks tailored to their specific needs. Additionally, the participants seemed to feel that the hearing dogs were able to do a lot more for them than what was originally intended. Although the primary function of hearing dogs is to alert its owner to repetitive sounds, a significant portion of the collected data focused on non-auditory benefits. Among those, the most frequently mentioned was companionship, followed by reduced anxiety , improved physical health, increased motivation, and reduced feelings of loneliness. According to the three participants in our study, the dogs enabled them to have more independence and feel safer in their daily lives, but also facilitated conversations with strangers and other dog owners. These findings indicate that the impact of hearing dogs may extend beyond the auditory realm and imply elements that are directly related to well-being , suggestBased on the interviews conducted, it appears that social understanding and lack of institutional recognition of hearing dogs were major concerns for the participants. Access to businesses and buildings was also identified as a particularly significant issue. This lack of recognition can create barriers for individuals with hearing loss who rely on these animals for support and highlights the need for more widespread awareness and acceptance of these dogs within institutions and the broader community. There is a need to increase awareness and understanding of the role and value of assistance dogs among the public. By doing so, we can help to ensure that individuals with hearing loss and their dogs have equal access to spaces and opportunities. One potential way to address this issue is through the establishment of a Foundation training center in Quebec, which could provide a resource for education and support for individuals with hearing loss and their hearing dogs.Overall, the hearing dog is a relevant way to combine the benefits of technical assistance with the psychosocial and emotional support of a pet. Considering that rehabilitation options that take emotional and psychological needs into account are among the most effective to increase the user’s QoL , hearing


While age-related hearing loss is common, disparities in care exist by race, ethnicity, and socioeconomic position. HEARS is a hearing care intervention that incorporates over-the-counter hearing technology, partnering older adults with peer mentors via a community health worker (CHW) model to address disparities. Through a randomized controlled trial, 8 older adult peer mentors delivered the HEARS intervention. We aimed to understand the CHWs’ perspectives on their role through photovoice. Participants took photos illustrating the intervention, their role, and hearing health. Semi-structured interviews (n=4) and a focus group were conducted. CHWs responded positively to the intervention and were satisfied with their role. A notable theme was the value of serving as a peer mentor. Participants valued the opportunity for generativity, learning as older adults, and associated social benefits. Our findings demonstrate an opportunity to engage older adults in the evaluation process, expanding access to hearing care, and in peer mentorship.


The threshold equalizing noise (TEN(HL)) is a clinically administered test to detect cochlear “dead regions” , using a “pass/fail” criterion based on the degree of elevation of a masked threshold in a tone-detection task. With sensorineural hearing loss, some elevation of the masked threshold is commonly observed but usually insufficient to create a “fail” diagnosis. The experiment reported here investigated whether the gray area between pass and fail contained information that correlated with factors such as age or cumulative high-level noise exposure (>100 dBA sound pressure levels), possibly indicative of damage to cochlear structures other than the more commonly implicated outer hair cells.One hundred and twelve participants  who underwent audiometric screening for a sensorineural hearing loss, classified as either normal or mild, were recruited. Their age range was 32 to 74 years. They were administered the TEN test at four frequencies, 0.75, 1, 3, and 4 kHz, and at two sensation levels, 12 and 24 dB above their pure-tone absolute threshold at each frequency. The test frequencies were chosen to lie either distinctly away from, or within, the 2 to 6 kHz region where noise-induced hearing loss is first clinically observed as a notch in the audiogram. Cumulative noise exposure was assessed by the Noise Exposure Structured Interview (NESI). Elements of the NESI also permitted participant stratification by music experience.Across all frequencies and testing levels, a strong positive correlation was observed between elevation of TEN threshold and absolute threshold. These correlations were little-changed even after noise exposure and music experience were factored out. The correlations were observed even within the range of “normal” hearing (absolute thresholds ≤15 dB HL).Using a clinical test, sensorineural hearing deficits were observable even within the range of clinically “normal” hearing. Results from the TEN test residing between “pass” and “fail” are dominated by processes not related to IHCs. The TEN test for IHC-related function should therefore only be considered for its originally designed function, to generate a binary decision, either pass or fail. Degradation of the mammalian auditory system has been shown to be caused by a variety of factors such as age, genetics, oto-toxic pharmaceuticals, and noise exposure . ElevatiHearing deficits can be observed even before the audiogram shows a loss of sensitivity beyond the range of “normal.” Clinically, this can take the form of measured difficulties with speech perception in noise  of lesion. Such identification  are available clinically. One clinical tool that offers a differential diagnosis of the likely cause of dysfunction is the threshold equalizing noise (TEN) test . In the As originally developed, the TEN test results in a binary decision: pass or fail at each frequency tested. However, anecdotal reports observe some elevation of the detection threshold in individuals with hearing impairment. Some of this elevation was expected, as described in the original version of the TEN test : damage The primary hypothesis behind this report is that elevation of the TEN threshold, insufficient to be classed as a “fail,” may indicate the onset of a “sick,” rather than a “dead” region. Identification of such could provide an early warning, such as behaviour modification, well before the perceptual consequences of a dead region become apparent. In this sick region, we would expect to see the balance between the OHC and IHC contributions gradually shifting, but possibly with a dependence varying with frequency. For example, noise-induced damage in humans is typically first observed clinically as a notch in the audiogram between 3 and 6 kHz . If the This study reports results from the use of the TEN test during the screening of a participant pool for the “battery” project mentioned above. In particular, the experiment had certain similarities to that reported by Another test that is possible to administer clinically in a short period of time is the “Temporal Fine Structure—Adaptive Frequency” (TFS-AF) test, a test of acuity to binaurally presented temporal fine structure . It adapOur recruitment sought older participants, because the previous reports using low-SL testing had selected younger people (group means < 35 years) with, at most, mild losses. Since it arose from a screening process, our recruitment was less targeted and less selective than that of Participants were recruited for screening whose self-reported lifestyle of noise exposure might have caused sensorineural damage. Since the initial recruitment was on the basis of lifestyle and not reported hearing difficulties, it was expected that some would have normal or near-normal hearing. For the purposes of the experiment described here, the detection of subclinical losses, these participants were retained.The selection criteria for passing this screening were that participants were as follows: greater than 18 years of age, fluent speakers of English since birth, in generally good health for their age, physically able to travel for testing, and available for multiple sessions, if successful.Clinically, it was intended that they:(a) had no underlying neurological problems or history of head trauma,(b) had never worn hearing aids in the past ,(c) were audiometrically likely to benefit from a hearing aid, that is, they had a mild-moderate high-frequency hearing loss , here mo(d) had no history of major middle ear dysfunction, and an intact tympanic membrane,(e) had a negligible conductive component to the hearing loss (≤10 dB).For the experiment reported here, condition (c) was only enforced to set an upper limit to their hearing loss to select participants to go forward to further testing  a moderate hearing loss in the better ear, defined as a minimum of 41 dB HL, based on the average of the pure-tone air conduction (AC) hearing threshold levels from 0.5 to 6 kHz, including half octaves . This ex(g) a threshold elevation above 15 dB HL at 0.75 and/or 1 kHz, the reasoning for this will be detailed later,(h) no episodes of noise exposure exceeding 100 dBA.Routine audiological screening was performed bilaterally, consisting of otoscopic examination of the external auditory meatus, tympanometry, bone conduction, and AC audiometry. In addition, all participants were interviewed by the experimenter so as to complete the NESI . The NESOf the initial 167 participants tested, a total of 51 were excluded from further consideration due to violating one or more of conditions (f), (g), and (h) earlier. These numbered 11, 24, and 18 participants, respectively.Criterion (g), having low-frequency thresholds within the “normal” range was used as a proxy to select for participants who we expected to have had well-within-normal hearing at birth, and our observations were of the hearing status after some postnatally acquired hearing loss. In addition, measures of hearing ability at low frequencies could also act as a within-participant statistical control for any differential effects of noise exposure across frequency, such as that expected to primarily affect the 3 to 6 kHz region. The choice of 15 dB HL, rather than the more common 20 dB HL, will also be explained later.Of the remaining 116 participants, 74 were female. The group mean age was 51.8 years with a median of 53 years, and with a range of 21 to 91 years. All participants were paid an honorarium for their attendance, as well as travel expenses. The remaining testing, described later, except for those undergoing TFS-AF, was performed unilaterally on the better ear, as defined by the AC audiometry.The study received ethical approval from the NRES Committee North West—Greater Manchester Central (REC number 16/NW/0260).The tones in the TEN test are usually presented continuously, as per manual audiometry. However, some pilot trials showed that participants with tinnitus performed more reliably when the probe tone was pulsed, rather than the usual continuous-presentation method. The TEN test was administered by replay off a CD player , routed through an audiometer , and delivered via a single earpiece of a TDH39 headphone . The level of the target tone was adjusted in 2-dB steps and presentation controlled via manual audiometry. The AC absolute threshold (to the pulsed tones) was obtained at four frequencies, 0.75, 1, 3, and 4 kHz, and the TEN threshold measured with noise densities of 12 and 24 dB SL relative to these absolute thresholds. The TEN thresholds were transformed to calculate the elevation of the tone threshold relative to the TEN noise level. In line with The design of the TEN spectral shape , and hearing protection usage (if any).The NESI  was thenThe sound level of these activities (units of dBA) was estimated by the participants based on recall of the vocal effort required to hold a conversation in each activity. For instance, an activity with estimated noise of 99 dBA would require the participant to shout from 4 feet (1.2 m) to hold a conversation. The calculation procedure to estimate the cumulative exposure is detailed in o. However, thresholds obtained from listeners with normal hearing range between 1100 and 1700 Hz  performed the TFS-AF test. The IPD was set to 180 1700 Hz , so the  1700 Hz . It shou 1700 Hz , and so Both correlational analyses and mixed-effects modeling were employed. Pearson correlations were used to explore contradictory claims about the relationship between NESI and Age , 2019 anMixed models were performed separately for the absolute thresholds and STRs. In these models, absolute threshold, and STRs were entered as dependent variables; frequencies, presentation level (12 or 24 dB SL), hearing, noise, and music group were evaluated as fixed effects.These analyses were performed to generate evidence for the hypotheses mentioned in the Introduction by asking: (1) is a gradual shift in balance between OHC-related  and IHC-related deficits (observable within a “dead” region) as the STR becomes elevated demonstrable in the data, and, (2) does amount of noise exposure, as measured by NESI100 ) dominate elevated STRs?ggplot2 .Figure The degree of hearing impairment of each participant was calculated as the mean AC threshold obtained by manual audiometry, averaged over the same test frequencies as used in the TEN test, 0.75, 1, 3, and 4 kHz. Participants with a mean exceeding 20 dB HL  were classified as having “mild” hearing impairment, the remainder had “normal” hearing. Their distribution is shown in Figure The noise exposure interviews of (1) conventionally, as cumulative exposure for all exposures where the sound level was estimated to exceed 80 dBA, which corresponds to all exposures recorded by the NESI, and is likely to capture exposures from PMPs. We refer to this measure as “NESI80.”(2) cumulative exposure for exposures where the sound level was estimated to exceed 100 dBA, more in line with the exposures recorded by Stone and colleagues. This calculation of the NESI will be referred to in figures and Tables as “NESI100.”In the statistical analyses to be presented, the pattern of the results when modeling with the NESI8 0 scores was very similar to that for the NESI100 scores, and so NESI80 scores will not be considered further except to address the secondary hypothesis from the Introduction that the pattern of results should vary depending on whether NESI80 or NESI100 was used as the noise-exposure metric. However, to capture music experience, the NESI80 data set was required.The cumulative units of exposure were cube-root transformed to obtain a distribution approximately Gaussian. This scaling was also used in The distribution of NESI100 scores is shown in Figure The low-exposed group comprised 43 participants , with a mean PTA of 11.3 dB HL, and age range 38 to 74 years (mean of 54.9 years). The medium-exposed group comprised 23 participants , with a mean PTA of 16.3 dB HL, and age range 32 to 69 years (mean of 51.8 years). The high-exposed group comprised 46 participants , with a mean PTA of 11.9 dB HL, and age range 33 to 68 years (mean of 49.9 years).The NESI data were further processed to produce the cumulative number of hours spent in practising a musical instrument, including in choirs. Since these data were only originally captured for exposures exceeding 80 dBA then some musicianship may have been under-quantified, if the preferred instrument was very quiet, for example, lute or acoustic guitar. Our measure therefore should be regarded as a proxy measure, hence its stratification into categories for the purpose of analysis of the data. Fifty-two participants  had no music experience, 60  had some experience or were expert musicians.The distribution of hours of music experience is shown in Figure Figure (1/3)]. The subranges of scores were from 0.12 to 2.82 for the low group ; from 2.86 to 4.07 for the medium group ; from 4.14 to 8.01 for the high group . The data show a modest negative Pearson correlation of cumulative noise exposure with age (r(110) = –0.277, p = 0.0031). This finding will be discussed later.The overall range of exposure scores was between 0.12 and 8.01 , Hearing group , and their interaction ; the participants were entered as random intercepts. The biggest differences in absolute threshold between the Hearing groups were at 3 and 4 kHz. This result was trivial due to the criteria used for the allocation to Hearing group. Of more interest is that the absolute thresholds were similar between the groups when grouped by either noise exposure or music experience; this indicates no effect of these two factors on absolute thresholds.The absolute threshold data were best explained by a linear mixed model . The effect of controlling individually for absolute threshold, NESI100, and music experience, produced two further significant correlations, again at 24 dB SL, and 3 kHz. STR correlated with Age, when controlling for Absolute Threshold, for both the full  and restricted  range of absolute threshold .There were no significant correlations between STR and Age except at 24 dB SL at 3 kHz and only when considering data with Absolute Threshold ≤ 15 dB HL , 12 dB SL at 3 kHz and 24 dB SL at 4 kHz. This extends the findings of The lower two lines of the two halves of Table Figure 2 = 0.61, marginal R2 = 0.19), using fixed effects of frequency , and hearing group , and their interaction ; with by-participant adjustments to the intercept and by-participant adjustments  to frequency. Homogeneity of variance for the participants over frequency was assumed. The STR threshold increased significantly at 3 kHz relative to the other frequencies. The mild HL group had significantly higher STRs than normal-hearing groups at 3 and 4 kHz. There were no effects of noise exposure or music experience.Comparing the data between the low and high noise-exposure groups, the same structure of linear mixed model was used as for the absolute threshold data to best explain the STR data .Our primary hypothesis, outlined in the Introduction, was that, independent of absolute threshold, the STR should be correlated with measures of noise exposure. There was no correlation found between STR and the general noise exposure metric, NESI80 . Consequently a Bonferroni-adjusted significance level of 0.00625  was used. The dependent variable was the STR for each combination of presentation level and frequency. The predictors were absolute threshold at the same frequency, age, NESI100, music experience, and the interaction term between NESI and age since these two were significantly correlated  STR @ 12 dB SL & 3000 Hz, and(2) STR @ 24 dB SL & 3000 Hz.NESI100 and music experience were not significant predictors in any of the eight models. Table negative: STR is modeled as improving with age, an unlikely result unless some other, unmeasured, factor, such as lifestyle, is adding heterogeneity to the participant pool. Unlike with the correlations performed using the full span of absolute thresholds in the data set . Since Age was significantly correlated with both NESI scores and TFS-AF thresholds  = –0.316, p = 0.003), after controlling for Age, the correlation between TFS-AF thresholds and NESI scores was insignificant: r(84) = 0.021, p = 0.850. This test therefore found no evidence of putative noise-induced damage to the IHC-pathway outside of the classic 2 to 6 kHz region where noise-related damage is first observed in the human audiogram.The Pearson correlation of TFS-AF thresholds as a function of NESI was not significant (r(82) = 0.110, p = 0.681, n = 84) or at 24 dB SL .The Spearman correlation of TFS-AF thresholds as a function of STR at 1 kHz (the frequency closest to the bulk of the TFS-AF thresholds) was not significant either at 12 dB SL  = 4.079, p = 0.002]. The significant predictors of TFS-AF were age  and music experience . We have replicated the link of TFS-AF thresholds with age that has been shown before (A multiple linear regression model was run on TFS-AF using PTA (derived as the average of the audiometric thresholds at 1 and 2 kHz), Age, NESI and music experience as well as the interaction term between NESI and age as predictors. The model was significant [Rn before , but notThe motivation for this work was that a quickly administered clinical test, the TEN test, could provide more information about the frequency-specific patency of the hearing system than just that provided by the audiogram. A noise-exposure measure was also included to address the modern concerns that noise, specifically recreational, rather than industrial, in origin (in high- and middle-income countries), is the main driver of “modern” noise-induced hearing loss (NIHL), especially in young adults .The data presented here do not support the hypothesis of a linkage between elevation of the STR in the 3 to 4 kHz region in the TEN test and high-level (>100 dBA) noise exposure. The data do show another association that expands our understanding of the gradual decline of the human auditory system over the course of the lifespan. Our data provide a strong link between a form of hearing deficit, the elevation of STR, and absolute thresholds, even when absolute thresholds at a wide range of individual frequencies were clinically “normal.” This is in agreement with the data shown by We do not attribute the observed elevation of STR within the range of “normal” absolute thresholds to the effect of altered auditory filter shape. Neural tuning curves in animals have been successfully modeled as a parallel combination of a low-sensitivity, wide-bandwidth linear filter (the “tail filter”) and a high-sensitivity, sharply tuned filter (the “tip filter”) whose output is nonlinear with level, but whose sharp tuning is invariant with level . The comThe slope of the elevation of the STR as a function of absolute threshold was 1/5 at 1 kHz and 1/7 at 4 kHz, units of dB/dB HL as plotted in Figure Similar observations have been made with measures of Distortion Product OAEs: strength of the emission has been shown to correlate with pure-tone absolute threshold, even for values of absolute threshold below 20 dB HL at the test frequency . Howeverr = 0.52, p < 0.01) among 126 young participants aged 18–36 years, barely overlapping with ours (p < 1–10), but the correlation among the older participants was insignificant .Figure ith ours . Expandiith ours  recruiteOne explanation is that younger and older populations are not following the same life course : the younger population are acquiring a cumulative exposure faster than their antecedents, a manifestation of the more recent increased access to high-sound levels . This efThe lack of effect of the NESI exposure tool to reveal a link of cumulative noise exposure with absolute threshold is notable, especially since accumulated dose forms the basis of predicted damage in medico-legal cases . There wA recent review of the evidence for noise-induced synaptopathy in humans suggests that one reason for the many contradictory findings may be the variability in the populations studied . This would support the postulate of A group of 112 participants ranging in age from 32 to 74 years were selected from a larger pool by screening for clinically normal hearing at 0.75 and 1 kHz, and questioned about lifetime noise exposure at high-sound levels by use of the NESI. They performed the TEN test at four frequencies, and at two levels, 12 and 24 dB, above absolute threshold. The selection by normal hearing at low frequencies, as well as lack of a conductive component to their hearing thresholds was intended to select for people with postnatally acquired hearing damage, if any.Correlational analyses showed the following:(1) A strong contribution of aging to the elevation of absolute threshold, the classical definition of presbyacusis.(2) No link between degree of noise exposure and elevation of absolute threshold.(3) Across a wide range of center frequencies, the elevation of the TEN threshold into the “sick” region between “pass” and “fail” was (a) almost entirely driven by the elevation in absolute threshold, and (b) occurred even when the absolute threshold was within a “normal” range, even when drawn more stringently (≤15 dB HL) than the clinically conventional ≤20 dB HL. Although some elevation of TEN threshold has previously been reported at high testing levels, such high testing levels were rarely used here due to the selection criteria and thresholds encountered.We conclude that an elevation of the TEN threshold less than the “fail” criterion:(1) appears to reflect a general degradation of multiple cochlear mechanisms, primarily related to OHC dysfunction because of its strong dependence on elevation of absolute threshold, rather than with dysfunction in the IHC-pathway.(2) occurs at a rate of 1 dB for every 7 dB of absolute threshold elevation above 0 dB HL.(3) is not sensitive enough to indicate putative effects of noise damage, and therefore should not be used as such in a clinical setting other than as a pass/fail decision tool.These data, derived from a clinical rather than a laboratory tool, do not support the previous findings in much younger cohorts by The data presented add further support to This work was funded by the UK Medical Research Council , reference MR/M023486/1 and by the NIHR Manchester Biomedical Research Centre .


Borrelia burgdorferi is an important tickborne human pathogen comprising several strains based on nucleotide sequence of the outer surface protein C (ospC) gene. Detection and characterization of different ospC genotypes is vital for research on B. burgdorferi and the risk it poses to humans. Here we present a novel, multiplex assay based on Luminex xMAP technology for the detection of B. burgdorferi ospC genotypes. The assay has five major steps: amplification of the ospC gene, hydrolyzation of surplus primers and nucleotides, incorporation of biotinylated nucleotides into the template DNA, hybridization to Luminex microspheres, and detection of fluorescent signals corresponding to each ospC genotype. We validated the protocol by comparing results obtained from our method against results from an established ospC genotyping method. This protocol can be used for the characterization of ospC genotypes in B. burgdorferi infected ticks, reservoir hosts, and/or clinical samples. Borrelia burgdorferi sensu stricto (hereafter B. burgdorferi) is the tick-borne bacterium that causes human Lyme disease in the United States [Ixodes scapularis) populations in the northeastern United States [ospC allele group, or genotype, is composed of members with less than 2% nucleotide sequence difference within a genotype and more than 8% nucleotide sequence difference between genotypes [B. burgdorferi in host and tick samples is commonplace, ospC genotyping of those B. burgdorferi samples is seldom done. This has potential public health significance since there is evidence suggesting that only a subset of ospC genotypes  cause the overwhelming majority of disseminated infections in humans [B. burgdorferi ospC genotypes in natural tick and host populations, and determining which ospC genotypes routinely infect humans, can improve research on the basic biology of this pathogen and its associated public health risks.d States . The gend States –5. An osenotypes . While mn humans –8. IdentospC genotypes is the reverse line blotting (RLB) assay [ospC genotypes. However, RLB cannot readily be adapted to a high-throughput process. Recently, a method based on next generation sequencing (NGS) was developed for ospC genotyping [ospC genotyping protocol using Luminex multiplex technology in a 96-well plate format that is straightforward, provides same-day results, and offers improved efficiency compared to RLB (An established method to detect different B) assay –5, 9, 10notyping . The NGSd to RLB .ospC nucleotide sequences.Luminex xMAP technology is powered by spectrally distinct MagPlex-TAG microspheres (beads) that are coupled with a unique 24 bp oligonucleotide sequence (anti-TAG) . The couospC-specific probes previously used in the RLB method for DNA hybridization [ospC primer is paired with a corresponding bead type. The step-by-step protocol is described in the ospC genotypes are amplified via nested PCR  value is generated for each ospC genotype.The dization –5 were rsted PCR  and the ospC K genotype sequence  assay is relatively simple, scalable, and efficient. It is appropriate for studies that require identification of ospC genotypes in B. burgdorferi positive samples. Due to the multiplex platform, the protocol can be modified as needed to include additional analytes for detection of yet to be discovered, novel ospC gene sequences.The assay was designed using general recommendations from the Luminex xMAP Cookbook  with subsequence . The K pK primer . The optospC genotype C is a chimera of other genotypes. We were not able to identify a primer/probe sequence to distinguish the chimera from the donor sequences. This is a known shortcoming of hybridization based genotyping approaches, including RLB [ospC genotypes for which there is a specific primer. However, the presence of novel ospC genotypes can be inferred where an amplicon hybridizes with the “ALL” primer but fails to hybridize with any of the known nucleotide sequences  and analyzed using the developed assay. The ospC genotypes infecting the ticks used in the present study had previously been determined using the RLB method [ospC genotypes and directly compare results between this assay and RLB.To validate the LOG assay against a biologically relevant sample set, cks N = 12 and anaospC infections, including the identification of 12 different ospC genotypes in a single tick sample. The average number of ospC genotypes per tick was 3.51 and 3.30 for LOG and RLB, respectively.The LOG assay detected multiple To compare the genotypes detected in each sample by both methods, 152 pairwise comparisons were performed . GenotypospC genotypes G and M are overrepresented, and that ospC genotypes V and W are underrepresented by LOG. However, after correcting for multiple comparisons, only ospC genotype V was detected at significantly different frequencies between the two methods . One explanation could be that ospC genotype V is not being properly detected by our new method. However, during the optimization process using synthetic double-stranded ospC gene fragments , ospC genotype V was routinely detected in single and mixed infections  were also coinfected with ospC genotypes F, M, or O as confirmed by both methods. Utilizing the RLB results from all 152 samples, a positive association was found between genotypes V and F  and genotypes V and O . Since the V probe used in RLB shares the same sequence with the V primer in LOG, we suspect that cross-reactivity of the V probe with other genotypes may occur during DNA hybridization in RLB but is less likely to be detected in our developed assay that requires hybridization and extension of the target to generate a positive signal. It is also important to consider that every sample went through two separate nested PCR reactions: one for RLB and one for LOG. Therefore, any differences between methods may be attributable to DNA degradation by repeated freeze/thaw cycles and different nested PCR efficiencies.All 19 genotypes for which we have specifically designed primers were detected using the LOG assay and at relatively similar frequencies compared to RLB . It appefections . AnotherB. burgdorferi infected tick samples that had been previously genotyped by RLB. The new ospC genotyping protocol can detect multiple ospC infections and most of the pairwise results  were either an exact match or additional genotypes were detected by LOG compared to RLB. All 19 genotypes designed for our assay were detected and more genotypes can be added as needed going forward. The LOG assay has demonstrated utility and offers another tool for researching B. burgdorferi.The LOG assay performed comparably well using S1 File(PDF)Click here for additional data file.S2 FileospC K reference sequence (Genbank accession number AY275214).The Qiu et al. K probe and our modified K primer sequences are shown in (DOCX)Click here for additional data file.S3 FileospC genotype are highlighted in yellow. The ospC A-U reference sequences were obtained from Di et al. [ospC V and W accession numbers are FJ649656 and FJ649657, respectively.The nested PCR first round primer hybridization sites are highlighted in blue and the second round primer hybridization sites are highlighted in red. Overlapping regions for the first and second round forward primers are highlighted in purple. The hybridization site for the ASPE “ALL” primer is highlighted in green. The ASPE primer hybridization sites for each i et al. . The osp(DOCX)Click here for additional data file.S4 File(XLSX)Click here for additional data file.S1 FigospC genotype K was amplified from a plasmid stock and diluted to approximately 7 ng/ul. Duplicate samples of the genotype were detected using two ASPE primer mixes: one with the original Qiu et al. K primer and the other with the modified K primer. Average MFI values with standard deviation as the error bars are shown for each treatment.(DOCX)Click here for additional data file.S1 TableFor LOG, a representative run of 90 samples was chosen to leave room on the 96-well plate for negative and positive controls, as detailed in the step-by-step protocol.(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 TableospC genotype standards (ospC gBlocks and a K plasmid stock) were amplified, diluted to approximately 5 ng/ul in samples representing single and mixed B. burgdorferi ospC genotype infections, and then analyzed using LOG. As described in the step-by-step protocol, a ratio to NTC value of ≥ 3 was considered positive.(DOCX)Click here for additional data file.


In 2015 a new regulation and guidelines for the universal newborn hearing screening by AABR measurement have been implemented in Hungary. The aim of our study was to analyse (1) the past 5 years of data from our diagnostic centre about the incidence and types of congenital hearing losses, and (2) the first experiences with the National Newborn Hearing Screening Registry, started in 2019, and (3) the influence of the screening on the pediatric cochlear implant program.1269 children referred to our diagnostic centre between 2017 and 2021 were investigated. A third AABR measurement and full audiological evaluation were performed. Furthermore, one-year period data of the screening registry, and the number of implanted children at or under the age of 3 were analysed using the national databases.Altogether 276 newborns (22% of the referred cases after the two-stage screening) had hearing loss, 134 (49%) out of them was conductive origin, almost twice frequent in male as in female. Permanent sensorineural hearing impairment was found in 142 (51%), 58 (40%) of them had bilateral, severe to profound hearing loss, occurring more frequently in male as in female. The national digital registration of the screening data within 12 months concerned 68%. The number of early cochlear implantation in one year increased from 1 to 23 children in the past 15 years.A third AABR after the two-stage screening increased the efficiency and filtered the 78% false-positive cases. The audiological diagnostics verified and typed the hearing losses ensuring the early intervention. The early recognition of neonatal hearing impairment is very important. The development of the hearing system begins in a very early phase of the fetal life and even the intrauterin hearing initiates the sensation , 2. The The newborn hearing screening can ensure the discovery the congenital hearing losses –10. The The aim of this study was to analyse the types and severity of the hearing losses diagnosed in babies referred to an audiological centre over a 5-year period. The patients were followed up regarding the intervention as well. It is known, that children who receive appropriate intervention, hearing aid under 6 months of age have a much better outcome in speech development, speech understanding, and even at school they have larger vocabularies. , 5. AccoThe appropriate data collection would help to give precise information about the epidemiology of the congenital hearing loss, the judgement of the screening coverage, the calculation of the devices  and special education required for the rehabilitation and the cost-effectiveness of the screening and intervention. Therefore, we devised a national data collection, with a registry, that was improved in Hungary by informatics development. The National Newborn Hearing Screening Registry is able to collect and summarize the screening results from the hospitals, and the diagnostic results from the verification centres. According to these data, all newborn babies can be followed up and with a feedback system all undiagnosed cases can be attended. Our goal is to provide full access to newborn hearing screening and ensure for all children the early recognition of hearing loss and give early intervention.Cochlear implantation is a chance for patients with profound hearing loss or deafness to create normal hearing . CochleaIn the neonatal departments and neonatal intensive care units of the hospitals on the day 0–4 all the babies  are tested with AABR. The acoustic stimulation is a 35 dB CE-Chirp stimulus, with both ears tested. Children who do not pass the first screening are measured within the first month of age in the same hospital at an outpatient neonatal department . The chiThe referred babies are guided to regional verification centres. In our clinical audiologyThe objective measurements are done in the natural sleeping of the patient in the first months of life. No anesthesia is used. This needs strong cooperation of the parents and sometimes a long waiting time for the medical staff. In cases of hearing loss further investigations are performed to explore the causes: genetic testing for connexin 26 with DNA sequence analysis of GJB2, imaging with CT, MRI, serology and searching for potential syndromes.The audiological results of a 5-year period, 01 January 2017 to 31 December 2021 were analysed using the clinical database. According to our audiological investigations, the types and severity of the hearing losses were characterized. In cases of outer ear malformation with ear canal atresia permanent conductive hearing loss (atresia on charts) was diagnosed. Based on the physical investigation and with a B-type tympanogram the diagnosis was the conductive hearing loss caused by otitis media with effusion (OME). Without any pathological finding in the conductive system, with A-type tympanogram the diagnosis was sensorineural hearing loss (SNHL) and the average estimated hearing threshold regarding the objective threshold levels of 500, 1000, 2000, 4000 Hz was calculated. Mild hearing loss is diagnosed between 35 and 40 dB, moderate hearing loss 40–60 dB, severe between 60 and 80 dB and profound with more than 80 dB, respectively. The incidence of congenital hearing loss in male and female were compared.The National Newborn Hearing Screening Register started in 2019. From the neonatal departments and NICUs there is an equipment-to equipment data transfer, supplied by different types of AABR informatically connected to the register with a special informatics solution. Analysing one-year (September 2020–August 2021) data according to the register we could calculate the efficacy of data collection. We compared the registered number of measurements with the number of births in Hungary in the same period.As an assessment of early intervention, the number of children implanted at or under the age of 3 is presented according to the data of the National Health Insurance Fund.Based on audiological diagnosis the OME cases were followed up, children with ear canal atresia were given bone conductive hearing aid, the unilateral sensorineural hearing loss cases were observed, and in profound cases cochlear implantation was offered. In cases of bilateral sensorineural hearing loss depending on the severity hearing aid was given. Mild cases mostly do not need intervention, these cases were followed up, biannually they have audiological investigations. For moderate to profound hearing losses hearing aids were given. In cases of profound hearing loss—after providing a high-power hearing aid for some months—CI is offered.In our clinic, in the past 5 years, 1269 children referred from newborn hearing screening were investigated. According to the AABR—as a third stage measurement—22% of the cases has been verified as hearing impairment, which means that 276 newborns had congenital hearing loss, the other 78% proved to be false-positive result after the two-stage screening at the hospitals. The verified hearing loss ratio increased year by year, from 15.9 to 26.7%.According to the physical investigation and tympanometry of 134 newborn babies, 49% of the positive cases had conductive hearing loss. With B-type tympanogram of 121 children, 44% had a transient problem, caused by otitis media with effusion. Counting the incidence within our cases 80 boys, 66% of the OME cases and 41 girls, 34% of the cases were found (Table Sensorineural hearing losses were diagnosed in 142 infants, that was 51% of the verified cases, 28% unilateral and 72% bilateral impairment were among them. (Fig. The National Newborn Hearing Screening Registry collects data from 63 hospitals in 49 cities all over the country, where the screening takes place with 106 pieces of AABR equipment at neonatal divisions and at intensive care units. (Fig. The number of cochlear implantation in Hungary at or under the age of 3 years was only 1 case in 2005, and it has been growing to 23 cases in 2020. (Fig. The early recognition and intervention of neonatal hearing impairment give the chance for all affected children for normal speech and language development. Different screening protocols exist for detecting congenital hearing loss. These protocols vary in the measuring methods, in the number and timing of measurements –18. AfteThe newborns who failed at the screening are sent to one of the five verification centres. Fig.  AnalysinAccording to our guidelines, the screening should be performed up to 1 month of age, then we should give a diagnosis up to 3 months of age and we begin the rehabilitation, for example we give hearing aid up to 6 months of age for the children, if it’s possible. Children with bilateral hearing loss of moderate, severe or profound severity were supplied with hearing aid at their 4–10 months of age. The mild cases mostly do not need any correction in this early phase , but theUnexpectedly, we found a significant gender difference in the incidence of congenital hearing impairments. Several studies have now shown equal AOM prevalence in males and females  and manyWith starting the National Newborn Hearing Screening Register the goal was to find all children who need help due to hearing impairment. During the international data collection by the EUSCREEN project , 40, a lAlthough we have the guidelines for newborn hearing screening, the appropriate amount of equipment, educated health professionals, national data collection, we can always, even nowadays, find late-diagnosed children, who could not get appropriate interventions at the time that could have improved their language development. The cause of these “late” cases can be delayed diagnosis or delayed intervention. Behind these delays could stand lack, not enough or not appropriate information, fear, worry, concern, denial, lack of awareness, low education of the parents or bad social circumstances. In some cases, children are not sent to the centres, in other cases the parents do not want the appropriate intervention, for example, the cochlear implant. We face these problems however we have the universal newborn hearing screening and financial support, as the intervention is totally financed by the government health insurance. What we need is a more accurate follow-up system and we should give more information, even in the healthcare system to the specialists and even to the public, for example in the social media and education. The parents’ and professionals’ awareness of the possibility of full rehabilitation and their knowledge about the consequences of the delayed diagnosis have to be enhanced.


C. elegans can sense innocuous temperature, and navigate themselves towards memorize past cultivation temperature (Tc) of their preference. For this thermotaxis, AFD thermosensory neuron is pivotal, which stereotypically responds to warming by increasing intracellular Ca2+ level in a manner dependent on the remembered past Tc. We aimed to reveal how AFD encodes the information of temperature into neural activities. cGMP synthesis in AFD is crucial for thermosensation in AFD and thermotaxis behavior. Here we characterized the dynamic change of cGMP level in AFD by imaging animals expressing a fluorescence resonance energy transfer (FRET)-based cGMP probe specifically in AFD and found that cGMP dynamically responded to both warming and cooling in a manner dependent on past Tc. Moreover, we characterized mutant animals that lack guanylyl cyclases (GCYs) or phosphodiesterases (PDEs), which synthesize and hydrolyze cGMP, respectively, and uncovered how GCYs and PDEs contribute to cGMP and Ca2+ dynamics in AFD and to thermotaxis behavior.Living organisms including bacteria, plants and animals sense ambient temperature so that they can avoid noxious temperature or adapt to new environmental temperature. A nematode  C. elegans senses not only noxiously high and low temperature [C. elegans can sense temperature, memorize past cultivation temperature (Tc), and compare the present temperature with the memorized past temperature. AFD is a major thermosensory neuron responsible for thermotaxis of C. elegans [2+ level in a manner dependent on past Tc [in vitro [c and compare the present temperature with the memorized past temperature.Thermosensation is universal from bacteria to plants and animals, and enables organisms to react to noxious temperature or adapt to new environmental temperature , 2. C. eperature –5 but alperature . Thermot elegans , 8, whic past Tc –14. Thisin vitro , indicatFor AFD thermosensation, three guanylyl cyclases (GCYs), GCY-8, GCY-18 and GCY-23, which are specifically expressed in AFD, and cyclic nucleotide-gated (CNG) channels TAX-4 and TAX-2, which are far more sensitive to cGMP than to cAMP, are essential –19: AFD C. elegans. It was indeed reported recently that cGMP level in AFD increases in response to warming in a manner dependent on past Tc similarly to Ca2+ dynamics [c, which is consistent with the previous report [Genetic experiments indicated that cGMP should transduce temperature information to Ca2+ dynamics in AFD neurons of dynamics . We furtdynamics  and by as report .gcy double mutants, in which only one gcy gene out of the three is intact, exhibit differential abnormality in thermotaxis behavior [c, respectively [gcy double mutants and uncovered differential contributions of these GCYs to cGMP dynamics in AFD.The three GCYs, GCY-8, GCY-18 and GCY-23, expressed in AFD redundantly but differentially contribute to thermotaxis and thermosensation of AFD. Three of behavior  and AFD behavior , 20. GCYC. elegans, four PDEs  are supposed to hydrolyze both cGMP and cAMP, whereas PDE-4 and PDE-6 are supposed to be cAMP-specific [2+ in AFD and uncovered complex co-operation among the PDEs to properly form AFD Ca2+ dynamics and thermotaxis behavior.cGMP is hydrolyzed by phophodiesterases (PDEs). Of six PDEs in specific –28. Thesspecific . We invec as described previously using FlincG, a GFP-based cGMP probe [2+ [c gcy-8 gcyperature  Figs. Th2+ in AFD and thermotaxis behavior. First, we analyzed thermotaxis of pde mutants. Among single mutants for pde-1, pde-2, pde-3 and pde-5, product of which hydrolyze cGMP, pde-5 mutants were significantly defective in thermotaxis when cultivated at both 17°C and 23°C C. elegany neuron . We alsos report . We furtbehavior  Figs. Moior Figs .2+ level already start decreasing at the temperature range where cGMP level was constant, which was more prominent in soma rather than in sensory endings  [2+ influx and/or decrease Ca2+ level despite high cGMP level exist in AFD, which is consistent with our previous finding that SLO K+ channels contribute to decrement of Ca2+ level in AFD after its increment by warming [2+, and the resulting K+ efflux may prevent Ca2+ influx possibly by inactivating voltage-dependent Ca2+ channels. Interestingly, all the three gcy double mutants cultivated at 17°C showed prolonged Ca2+ signal at soma  , 39, thi warming ; both SL at soma , left, sshutdown , althoug so much , upper.2+ increase stereotypically in AFD in response to warming according to past Tc, however, it remains elusive how the past Tc is memorized as the onset temperature for cGMP and Ca2+ increment. Interestingly, when GCY-18 are ectopically expressed in chemosensory neurons, they confer fixed onset temperature for Ca2+ increment regardless of animals’ Tc [2+ increment in AFD of gcy-23 gcy-8 double mutants, in which only GCY-18 is expressed out of the three GCYs essential for the AFD thermosensation, was changeable according to Tc  plates seeded with E. Coli OP50 strain (Caenorhabditis Genetics Center (CGC), Twin Cities, MN, USA) as described [escribed . N2 (Briescribed . StrainsPopulation thermotaxis (TTX) assays were performed as described previously . Brieflyi: number of animals in each section i (i = 1 to 8), N: total number of animals on the test plate.Ngcy-37p::cGi-500::unc-54 3’UTR was a gift from Dr. de Bono [unc-54 3’UTR were fused by MultiSite Gateway Technology .pAF-EXPR-26  de Bono . cGi-500pde-1b, pde-2a and pde-5 cDNAs were PCR amplified from DupLEX-A Yeast Two-Hybrid cDNA library C. elegans (adult)  and cloned into AgeI-NotI site of pIA138 gcy-8p::VN173::unc-54 3’UTR.Plasmids used in this study are listed in 2+ imaging with GCaMP3 and tagRFP), (R–R0) / R0, was plotted, where R0 is average of R from t = 1 to t = 31.cGMP and Calcium imaging was performed as described elsewhere , 40. BriExpression of PDE-5::GFP was observed with LSM880 confocal microscope .The error bars in histograms for thermotaxis assays indicate the standard error of mean (SEM). In the boxplots, the bottom and top of boxes represent the first and third quartiles, and the band inside the box represents the median. The ends of the lower and upper whiskers represent the lowest datum still within the 1.5 interquartile range (IQR), which is equal to the difference between the third and first quartiles, of the lower quartile, and the highest datum still within the 1.5 IQR of the upper quartile, respectively. For multiple-comparison tests, one-way analyses of variance (ANOVAs) were performed, followed by Tukey-Kramer test or Dunnett test. Statistic analyses were done by R programming language.S1 Fignd cooling were made bold and bold-italic, respectively.Individual traces of fluorescence ratio (CFP/YFP) change shown in (TIF)Click here for additional data file.S2 Figgcy double mutant animals indicated that express GCaMP3 Ca2+ indicator and tagRFP in AFD were cultivated at 17°C (left) or 23°C (right) and subjected to imaging analysis with temperature stimuli indicated (orange line). Warming and cooling was at the rate of 1°C/20 sec. n = 4 to 6. Individual (gray) and average fluorescence ratio (GCaMP/RFP) change at AFD sensory ending is shown. B. Temperature at which Ca2+ level started increasing in response to warming was extracted using a MATLAB command ‘findchangepts’ and plotted. p values were indicated, or *** indicates p < 0.001 .Wild type and (TIF)Click here for additional data file.S3 Figgcy double mutant animals indicated that express GCaMP3 Ca2+ indicator and tagRFP in AFD were cultivated at 17°C (left) or 23°C (right) and subjected to imaging analysis with temperature stimuli indicated (orange line). Warming and cooling was at the rate of 1°C/20 sec. n = 4 to 6. Individual (gray) and average fluorescence ratio (GCaMP/RFP) change at AFD soma is shown. B. Temperature at which Ca2+ level started increasing in response to warming was extracted using a MATLAB command ‘findchangepts’ and plotted. p values were indicated, or *** indicates p < 0.001 .Wild type and (TIF)Click here for additional data file.S4 FigA. Mean values of YFP fluorescence intensity between t = 0 and t = 31, while temperature was kept constantly at 23°C, were plotted for 23°C-cultivated animals of indicated genotype expressing cGi-500 cGMP indicator specifically in AFD thermosensory neurons used in (TIF)Click here for additional data file.S5 Figpde-1, pde-2 and pde-1; pde-2 animals and pde-1; pde-2 animals that express PDE-1 or PDE-2 specifically in AFD were cultivated at 17°C or 23°C and then subjected to thermotaxis assay. n = 8 for N2 and pde-1; pde-2. n = 4 for others. The error bars in histograms represent the standard error of mean (SEM). The thermotaxis indices of strains marked with distinct alphabets differ significantly (p < 0.05) according to the Tukey-Kramer test. B. Wild type and mutant animals lacking pde gene(s) indicated that express cGi-500 cGMP indicator in AFD were cultivated at 23°C and subjected to imaging analysis. Warming and cooling was at the rate of 1°C/6 sec. Individual (gray) and average (blue) fluorescence ratio (CFP/YFP) change at AFD sensory ending is shown. C-D. Wild type and mutant animals lacking pde gene(s) indicated that express GCaMP3 Ca2+ indicator and tagRFP in AFD were cultivated at 23°C and subjected to imaging analysis. Warming and cooling was at the rate of 1°C/20 sec. Individual (gray) and average (pea green or green) fluorescence ratio (GCaMP/RFP) change at AFD sensory ending (C) and soma (D) is shown.A. Wild type, (TIF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file. This paper was transferred from another journal. As a result, its full editorial history  may not be present.27 Oct 2022AttachmentcGMP_rebuttal_221025_IM_IA_IM.docxSubmitted filename: Click here for additional data file. 15 Nov 2022cGMP dynamics that underlies thermosensation in temperature-sensing neuron regulates thermotaxis behavior in C. elegansPONE-D-22-29567Dear Dr. Mori,We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. 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Matsuyama for critical reading; and K. Ikegami, Y. Murakami, for technical assistance. Some strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). This work was supported by JSPS KAKENHI Grant Numbers JP 16H01272, JP 16H02516 and JP 18H04693 (to IM)."We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. 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Given its broad usage, TCS is present ubiquitously in the environment. Given its strong lipophilicity and accumulation ability in organisms, it is potentially harmful to biohealth. Several reports suggest the toxicity of this compound, which is inserted in the class of endocrine disrupting chemicals (EDCs). In September 2016, TCS was banned by the U.S. Food and Drug Administration (FDA) and the European Union in soap products. Despite these problems, its application in personal care products within certain limits is still allowed. Today, it is still unclear whether TCS is truly toxic to mammals and the adverse effects of continuous, long-term, and low concentration exposure remain unknown. Indeed, some recent reports suggest the use of TCS as a repositioned drug for cancer treatment and cutaneous leishmaniasis. In this scenario it is necessary to investigate the advantages and disadvantages of TCS, to understand whether its use is advisable or not. This review intends to highlight the pros and cons that are associated with the use of TCS in humans.Triclosan (TCS), a broad-spectrum antimicrobial agent, has been widely used in personal care products, medical products, plastic cutting boards, and food storage containers. Colgate Total Triclosan (TCS), along with its congener triclocarban (TCC)  1], is , is 1], ® toothpaste, containing 10 mM TCS, was the number two leading toothpaste in sales in 2016 [TCS CAS#3380-34-5), also named Irgasan DP300, CH3565, GP41-353, FAT 80′023, Irgacare MP and Irgasan , was fir0-34-5, a in 2016 . In toot in 2016 ,13. TCS  in 2016 ,15. More in 2016 .® [w/w) [® when it is used in plastics and clothing, and Biofresh® when it is used in acrylic fibers [® has been also reported to be used to coat door handles in hospitals [The worldwide production of TCS is very high. It has been recently estimated that it reaches 1500 tons per year and that 132 million liters of TCS-containing products are used annually in the USA alone . The pro® ,56. In t® [w/w) ,58. TCS c fibers . Microbaospitals . Given iospitals   with an essential oil-containing mouth-rinse. The use of antibacterial toothpastes without the mouth-rinse lowered the viability of adhering bacteria Lactobacilli, Streptococcus sobrinus ATCC 33478, and S. mutans ATCC 10449. It was noticed that the combination of TCS-containing toothpaste with an essential oil-containing mouth-rinse yielded a reduction in the prevalence of S. mutans from 30% to 5%, determining a drastic shift in the composition of the oral microbiome. However, the TCS-containing toothpaste alone produced major increases in the prevalence of adherent S. oralis/S. mitis, S. sanguinis, and S. mutans. Thus, the authors hypothesize that the non-polar TCS could increase the hydrophobicity of the S. mutans cell surface, facilitating the removal of S. mutans by hydrophobic oils, such as oil-containing mouth-rinses [Pseudomonas aeruginosa biofilms [P. aeruginosa to transiently survive aminoglycosides including as tobramycin. TCS was shown to enhance tobramycin effectiveness in vivo in a mouse wound model. The association of TCS with tobramycin may be considered a new antibiofilm strategy as it enhances the susceptibility of P. aeruginosa biofilms to aminoglycosides [Escherichia coli strains on Foley catheters [®, showed that the growth of S. mutans is inhibited by this mouth rinse on toothbrush bristles that were used by children [TCS has shown interesting activity against biofilms . The stubiofilms . This acycosides . The comatheters . Antisepchildren .Leishmania donovani with a half minimal inhibitory concentration (IC50) of 30 µM. Studies in silico suggested that this action was due to the interaction of TCS with L. donovani enoyl-acyl carrier protein reductase [Mycobacterium tuberculosis while at higher concentrations it determined the disruption of bacterial protein synthesis. It is noteworthy that, although less active than the antitubercular drug isoniazide, TCS doesn’t require any activation and is able to directly affect the function of InhA [Plasmodium falciparum and P. vivax dihydrofolate reductases, classic targets for the blood stage of the parasite. It inhibits blood-stage Plasmodium, via its action against dihydrofolate reductase, and the liver stage of the parasite, via the inhibition of ENR [In vitro and ex vivo drug assays on promastigotes and amastigotes showed that TCS possesses antileishmanial activity against eductase . TCS has of InhA . Severaln of ENR . Severaln of ENR ,87.®, Colgate-Palmolive, New York, NY, USA) contains 0.3% TCS along with 0.24% sodium fluoride [Streptococci, Actinomyces, hydrogen-sulphide (H2S)-producing bacteria, Fusobacteria, and Veillonella) in diverse oral sites in comparison with the other two dentifrice formulations as seen 12 h after brushing [® toothpaste determined a greater uptake of TCS to enamel and buccal epithelial cells than from a dentifrice containing TCS alone. Moreover, Colgate Total® provided superior protection against plaque and gingivitis, caries, oral malodor, exhibited superior stain removal, and provided protection against the progression of periodontal disease [Porphyromonas gingivalis [Toothpastes containing TCS have been shown to reduce plaque and gingivitis ,89. Mostfluoride ,91,92. Afluoride . It was brushing . In anot disease . Another disease . Tobacco disease . TCS actngivalis . Recentlngivalis , has beengivalis .Ka = 8.1 at 20 °C). The high octanol-water Partition coefficient (log Kow 4.76) and high hydrophobicity leads to bioaccumulation potential [® toothpaste users compared to non-users.Exposure to TCS may occur mainly through oral intake and dermal contact . Moreoveotential . The halotential . Recentlotential . As an ootential . TCS is otential . The shootential . A studyotential  in AsianTCS is transformed biologically into several compounds , includiSimilar to the parent compound, TCS transformation products have themselves demonstrated undesirable environmental and health characteristics. MTCS has been demonstrated to be more persistent than TCS and, similar to TCS, possesses endocrine disrupting capabilities ,115. TCSp-dioxin  [p-dioxin  has been also suggested [In photochemical reactions and incineration processes TCS could be converted to the more toxic compound 2,8-dichlorodibenzo-,8-DCDD) . Photode,8-DCDD) . The phouggested . There auggested .®) and other two, containing cetylpyridinium chloride and chlorhexidine digluconate  by the Somatic Mutation and Recombination Test in Drosophila melanogaster [Despite the numerous and several years of studies about TCS, it remains a controversial antibacterial regarding its toxicity. Rodrigues et al. (2007)  comparednogaster , employinogaster  demonstrnogaster . Howevernogaster  to invesnogaster .TCS is often considered a potential thyroid disruptor. However, its hormone disruptive potential has mainly been studied in animals and requires confirmation in human studies . The advThe endocrine system is important for male reproductive development because androgens, such as testosterone, promote the maturation of male secondary characteristics and the process of spermatogenesis. Male reproductive health, specifically testosterone and sperm count were declined, which is correlated with an increase in a variety of EDCs . In vitrCYP19A1 or P450arom), human 3 beta hydroxysteroid dehydrogenase type 1 (3β-HSD), and human 17 beta hydroxysteroid dehydrogenase type 1 (17β-HSD) [TCS is suspected of having endocrine-disrupting properties, but few human studies have examined the developmental effects of prenatal TCS exposure. Some epidemiological studies report that gestational urinary TCS concentrations during pregnancy are associated with decreased birth weight and length, head circumference, and gestational age ,155. Mor17β-HSD) . There a17β-HSD)  conclude17β-HSD)  suggeste17β-HSD)  of 15 st17β-HSD) , althoug17β-HSD) ,165,166.17β-HSD) . A recen17β-HSD) . Prenata17β-HSD) . Wang et17β-HSD) . Berger 17β-HSD) . However17β-HSD) ,173. Rec17β-HSD) . A recen17β-HSD) . A recen17β-HSD) . A succe17β-HSD) .Rattus norvegicus) and their male offspring were studied. The lactating mother rats were dosed with 0 mg, 3 mg, and 5 mg/kg/day of TCS until 28 days after the day of delivery so that the pups consumed TCS through breastfeeding. It was noticed that the mothers passed the detrimental effects to their untreated male pups as shown by reduced androgen synthesis and subsequently decreased sperm count [A recent study was carried out to evaluate the influence of TCS on the male reproductive system of postnatal pups. Pregnant female Wistar rats (rm count .Danio rerio) embryos highlighted that TCS caused heart edema and slow heartbeat [Danio rerio embryos confirmed that the exposure to TCS during neurodevelopment, especially during axonogenesis, is toxic and suggests a possible mechanism, through which TCS-induced ectopic expression of proneural genes may affect sequential neural development and impair glial cell function, resulting in CNS structural abnormalities during neurogenesis such as reduced axon growth and synaptic density [Exposure to TCS may adversely affect neurodevelopment and may exert adverse effects on central nervous system (CNS) functions, mainly through the induction of apoptosis and oxidative stress . Neurotoeartbeat , delayedeartbeat , and impeartbeat  at conce density . In vivo density . Success density . A recen density . TCS has density . Some ofSeveral researches have revealed to some extent that TCS exposure could induce nephrotoxicity. TCS administration induced function alterations, oxidative stress, pro-inflammatory, and fibrotic response in mouse kidneys. The lipid accumulation related to lipotoxicity and the decreased fatty acid oxidation may play a causative role in TCS-induced nephrotoxicity . TCS hasPangasianodon hypophthalmus, TCS induced immunosuppression and reduced the survivability of this fish during the challenge to a fish pathogenic bacterium Edwardsiella tarda [Over the last decade, researchers have shown that endocrine disrupting chemicals, such as TCS, can affect the development, function, and lifespan of immune cells. TCS may attenuate immunity against infections or cause a hyperactivity of immune responses such as allergies and autoimmune diseases. Exposure to TCS has been related to food and aeroallergy and asthma exacerbation in humans, both in adults  and chilla tarda . The topla tarda . The assla tarda .Proteobacteria in adults, as well as in infants with high urinal concentrations of TCS [®  system, was used to simulate the human gut microbial community in vitro, and to look at the community in the three different regions of the gut, the ascending colon, transverse colon, and descending colon regions, as well as in both the lumenal and mucosal phases. It was found that treatment with TCS significantly impacted the community structure in terms of reduced population, diversity, and metabolite production, most notably in the ascending colon region. However, after a two-week recovery period, most of the population levels, community structure, and diversity levels were recovered for all the colon regions [Lactobacillus plantarum ST-III increases the diversity of the gut microbiota in zebrafish, thus lowering the toxicity of chronic exposure to TCS. Additionally, a probiotic-rich diet reduced the risk of lipid-metabolism disorders such as increased triglyceride and total cholesterol levels [The understanding that humans readily absorb TCS and are able to pass TCS through breastmilk has led to a need to explore the impact of TCS on the human gut microbiome. Moreover, the extensive use of TCS over the past several decades , along ws of TCS . A recen regions . The inv regions . Finally regions . Interesl levels .P. aeruginosa energetics. At consumer-relevant, non-cytotoxic doses, TCS inhibits the functions of both mitochondria and mast cells, a ubiquitous cell type [2+ dynamics in mast cells [2+ mobilization in human Jurkat T-cells [socs3b, and simultaneously activated the downstream IL-6/STAT3 signaling pathway [Several mechanisms of action have been proposed for TCS, most of which are related to the different effects, either positive or toxic, that are exerted by this compound . TCS at ell type . Recentlst cells , and it  T-cells . The mec T-cells . Szychow T-cells  demonstr T-cells . Kim et  T-cells  demonstr pathway . The hep pathway  recently pathway . A study pathway . Besides pathway . The rol pathway . Spontan pathway .Caenorhabditis elegans and human mesenchymal stem cells (hMSCs) [Gambusia affinis) that were exposed to TCS showed that the up-regulations Sirt1 and Sirt2 in Sirtuin family could activate the Nrf2/ARE signaling pathway and the down-regulations of miR-34b and the up-regulations of miR-34a, miR-144, and miR153a-5p modulated the Nrf2/ARE signaling pathway [Catla catla [Drosophila melanogaster showed that TCS caused a redox imbalance with an increase on the catalase activity and a decrease on reactive oxygen species (ROS) level [Labeo rohita showed that TCS caused biochemical and transcriptomic alterations that resulted in oxidative stress, an impairment of metabolic processes, and dysfunction of the liver, kidney, and digestive system. A total of 96 h exposure to 0.06–0.097 mg/L TCS significantly declined the levels of glucose, triglycerides, urea, and uric acid and the activity of alkaline phosphatase, GOT and GPT [In this subparagraph, studies regarding the possible mechanisms of toxicity in animal models are summarized. Yoon et al. (2017) showed that TCS induces oxidative stress and intracellular accumulation of toxins, at least in part, by disrupting the SKN-1/Nrf2-mediated oxidative stress response, specifically by inhibiting the nuclear localization of SKN-1/Nrf2 and the expression of its target genes, in both a model organism, the nematode  (hMSCs) . A recen pathway . Sublethla catla . StudiesS) level . Liu et S) level . Studies and GPT .−7 M of TCS resulted in increased migration and invasion as measured by xCELLigence technology for all three cell lines. The authors suggested a mechanism involving epithelial to-mesenchymal transition (EMT), as the reduction in levels of E-cadherin mRNA and of E-cadherin protein was observed [DNMT3A and DNMT3B in the SCC-15 cell line after 24 h treatment [Although TCS is considered to be an EDCs, it is not considered a carcinogen by the International Agency for Research on Cancer (IARC) . Indeed,observed . A recenobserved . Finallyreatment . SummingProvidencia rettgeri group, namely P. rettgeri MB-IIT strain, could be advantageously used to degrade TCS that was present in the wastewater [Glyceria maxima showed that the amounts of TCS in plant shoots were 1.4–2.5 times higher than that in roots [3O4) nanoparticles [Due to the potential ecological and adverse health effects of TCS, several strategies are addressed to find new technologies for specific and effective removal of TCS from water, thus minimizing its threats in the environment. TCS mitigation from environmental matrices was obtained by using various oxidoreductases, including laccases, and peroxidases including soybean peroxidase, versatile manganese peroxidase, and horseradish peroxidase . Photo-dstewater . A recenin roots . Li (202in roots  reportedin roots . A recenarticles .Leishmania and affecting 12 million people in 98 countries. TCS has shown preclinical efficacy in a murine Leishmania amazonensis model, suggesting that the parasite plasma membrane may be a possible target of the drug, leading to an irreversible lethal antiparasitic effect [Perkinsus spp, TCS exhibited a high activity through the change in lipid content distributions and an increased oxidative stress [L. panamensis and Trypanosoma cruzi and showed an interesting potential as leads for the further development of drugs [The drug repositioning process significantly saves the development time of new drugs. It only costs researchers approximately 3–12 years to develop an old drug for new treatments compared to the time required to develop a new drug that is approximately 12–17 years . Repositc effect . Otero ec effect  reportede stress . Finallyof drugs . All togTCS is a synthetic, chlorinated phenolic antimicrobial agent that is commonly used in commercial and healthcare products. As a broad-spectrum antibacterial agent, TCS has been ubiquitously added to different personal hygiene products, such as antimicrobial soaps, deodorants, mouthwashes, toothpastes, disinfectants, and cosmetics, typically at concentrations of 0.1–0.3% and also to other products such as bedding, fabrics, clothes, toys, trash bags, household cleaning products, kitchen utensils, and seafoods. Besides, there is strong evidence for TCS’s disrupting effects on the endocrine system, especially on reproductive hormones. TCS belongs to EDCs, which adversely affects the endocrine system leading to compromised functions of hormones. Epidemiological studies report a link between TCS exposure and several adverse health outcomes including alterations in thyroid function and an increased risk for hypersensitivity diseases suggesting an immunomodulatory role for the synthetic chemical. Even though TCS possesses wide biological properties that have been exploited in several applications, there are many important considerations that need attention and represent severe issues that should be taken into account and diminish its indiscriminate and widespread use. Amongst them, the ascertained rise of bacterial resistance, the high impact on gut microbiota, immune and endocrine system, different organs, with related negative effects on human health, and, ultimately, its environmental accumulation as a polluting substance represent critical aspects. On the contrary, under a scientific research point of view, it is desirable to study TCS as repositioning drug, considering some interesting preliminary studies regarding its potential anticancer and antiparasitic properties. This review provides the main information and highlights advantages and disadvantages regarding the different TCS applications, inviting to a more rationale and limited use and unveil the TCS potential as repositioning drug.