Effects of hearing loss on vocal behavior are species-specific. To study the impact of auditory feedback on feline vocal behavior, vocalizations of normal-hearing, hearing-impaired (white) and congenitally deaf (white) cats were analyzed at around weaning age. Eleven animals were placed in a soundproof booth for 30 min at different ages, from the first to the beginning of the fourth postnatal month, every 2 weeks of life. In total, 13,874 vocalizations were analyzed using an automated procedure. Firstly, vocalizations were detected and segmented, with voiced and unvoiced vocalizations being differentiated. The voiced isolation calls (‘meow’) were further analyzed. These vocalizations showed developmental changes affecting several parameters in hearing controls, whereas the developmental sequence was delayed in congenitally deaf cats. In hearing-impaired and deaf animals, we observed differences both in vocal behavior (loudness and duration) and in the calls’ acoustic structure . The fundamental frequency decreased with age in all groups, most likely due to maturation of the vocal apparatus. In deaf cats, however, other aspects of the acoustic structure of the vocalizations did not fully mature. The harmonic ratio  was higher and more variable in deaf cats than in the other study groups. Auditory feedback thus affects the acoustic structure of vocalizations and their ontogenetic development. The study suggests that both the vocal apparatus and its neuronal motor control are subject to maturational processes, whereas the latter is additionally dependent on auditory feedback in cats. Congenital deafness affects the development of the auditory system  shares some common features with vowels of human language and has been suggested to be similar in structure and function to the cry human infants generate under social isolation Newman , 2007. TIn the present behavioral experiments, four normal-hearing mongrel cats, four hearing-impaired white cats and three congenitally deaf cats (CDCs) were used. All hearing-impaired and deaf animals were drawn from a colony of white cats. Hearing status was assessed during a screening procedure at the age of 4 weeks postnatally . To prevent exploration, the available space for the animal was further limited by an acoustically transparent cage (dimensions 45 × 30 × 30 cm). The calibrated microphone (Bruel & Kjaer 4165 condenser ½” microphone with a Bruel & Kjaer 2209 amplifier) was positioned in front of the cage at a distance of 50 cm. The situation reliably induced spontaneous vocalizations that were recorded continuously for 30 min, sampled at 44.1 kHz and stored on a computer. The data were analyzed offline using custom-tailored software programmed in MATLAB (©Mathworks) , with a total of 13–15 recording sessions in each animal group. The data were subsequently pooled into four age categories: 1 month (days 30–35 p.n.), 1.5 months (days 36–42 p.n.), 2 months (days 44–58 p.n.) and 3 months (days 64–103 p.n.). This corresponds to the developmental timeline of the auditory brainstem . Individual spectrograms were then aligned to the onset of vocalization and normalized to the maximum amplitude within the individual spectrogram. The normalized aligned spectrograms were averaged. Onset alignment enabled us to track the changing frequency components from onset of vocalization onwards. Overall representation of frequency content of vocalization (power spectra) was additionally evaluated using the Welch method (FFT length of 1024). The power spectra, too, were first normalized to the maximum power and subsequently averaged. To assure that the onset part of the vocalization was well represented in the average spectrogram, vocalizations that contained multiple unvoiced components  were excluded from the construction of average spectrograms, as were vocalizations shorter than 100 ms or longer than 1,500 ms.For further quantitative analysis, 10 ms non-overlapping frames were used to differentiate voiced and voiceless segments  of voiced vocalizations was determined from the autocorrelation function; to avoid confusion with first harmonics, the robustness of the results was increased by limiting the frequency range to around ±30 % of the expected maximum frequency of F0  and t tests (two-tailed uncorrected). The significance level was 5 % in all cases.The data’s normality was tested using the Stephens–Pearson test (α = 10 %) and the data were compared using The present data were obtained from three groups of animals: hearing controls from a colony of normal-hearing cats, animals from the white-cat colony with well-preserved hearing  and congenitally deaf cats from the white-cat colony . The hearing screening . For the vocalization identification and segmentation procedure, a subset of 3,508 vocalizations  was manually post-processed and compared with the results of the automatic segmentation procedure. The mean error rate per processed recording was 2.73 %, with the maximum value observed at 5.8 %. As a rule, more vocalizations were identified by the automatic procedure than manually, the reason being inappropriate ‘splitting’ of some vocalizations into two calls during automatic segmentation. Because of these low error rates, we used the automatic procedure for further processing.t test, hearing vs. deaf: p = 0.442; hearing vs. impaired: p = 0.0096; impaired vs. deaf: p = 0.0025). This relation was similar across all age groups. The voiced vocalizations were identified as isolation calls based on the overall acoustic structure  in all experimental groups  were mainly purr-like vocalizations and combined calls, starting as an unvoiced call and continuing as an isolation call Fig. . The durCongenitally deaf cats also produced isolation calls Fig. . Manual The spectrograms of the vocalizations of each animal were amplitude-normalized to maximum, aligned to the onset of vocalization and averaged for each age group Fig. . These aFurthermore, in the assessment of the vocalizations’ harmonic structure, the mean power spectra of the entire call Fig.  confirmeTo statistically confirm the results of Figs. p = 0.557; hearing vs. deaf: p = 0.172; impaired vs. deaf: p = 0.09). This was still evident when comparisons were performed within one age group . However, the deaf animals tended to exhibit the longest vocalizations .First, the general properties of the vocalizations were analyzed groupwise for all ages pooled, with means calculated per session; these were then pooled irrespective of age, resulting in a grand mean. The animals vocalized quite often in the soundproof booth: they generated 200–600 vocalizations per session Fig. . This hip = 0.184; hearing vs. deaf: p = 0.0011; impaired vs. deaf: p = 0.0002). This is interesting since, combined with the tendency for longer duration in deaf cats, the effects on the overall energy of the calls are even greater.The sound-pressure level (SPL) of the vocalizations was additionally analyzed; however, these results have to be considered with caution, as the absolute SPL picked up by the microphone critically depends on the relative position of the animal and the microphone. As the animals were unrestrained within the cage, this position was variable. Call loudness was significantly higher in deaf cats Fig. . The vocp > 0.05). However, the developmental pattern of vocalization duration differed between the groups. Hearing animals showed increasing call duration with increasing age . The difference, however, was not significant at 2 months (p = 0.103) and reversed at 3 months, when hearing cats had longer vocalizations than deaf cats (p < 0.001).Next, the developmental pattern of the call properties was analyzed. This involved generating age-related statistics within groups. The number of vocalizations did not differ between the three investigated groups at any age  and the faintest in hearing-impaired animals (p < 0.001), whereas those of both the hearing and hearing-impaired animals first increased and then decreased with increasing age  spectral structure than in hearing controls. This indicates a graded effect of mild to profound hearing loss based on the degree of auditory feedback.Consequently cat vocalizations are not solely produced by automated neuronal programs but partially depend on feedback and show developmental plasticity. A prominent finding was the greater variability of the spectral properties of vocalizations in deaf cats. Therefore, auditory feedback is used to control and stabilize the vocalizations during development in hearing cats.Previous studies on feline vocalizations documented about 10 different vocalizations generated in different behavioral contexts Moelk . The preThe few first weeks of postnatal life are important for the kitten-–mother bond. While kittens already vocalize in the first days of life, we started with data collection from the age of 1 month to prevent the kittens undergoing a lengthy separation from their mother in the first weeks after birth.In the present study, the investigated vocalization was referred to as the isolation call (‘meow’). This was due to the context in which the vocalization has been elicited: the animal being unrestrained but socially isolated. However, the behavioral role of this call may change throughout early development in the cat  and non-auditory development of the animals. The white cats, on the other hand, may have a different genetic and social background, which are potential confounding factors. To rule out these effects, we also investigated mildly hearing impaired animals of the same white-cat colony. In all grand mean comparisons Figs.  and 6, tThe most equivocal measure in the present study is loudness. Obviously, this measure depends on the exact position and orientation of the cat’s articulatory organs. As this is impossible to precisely control in the present setting, the data may be influenced by a difference in position and have to be considered with caution. We assume, however, that these variations were similar between the groups.Finally, the behavioral importance of vocalizations will be also dependent on the behavioral reaction of the mothers; as the mothers of the deaf cats were also deaf, some of the observations might rather be attributable to the hearing state of the mother than to the hearing of the kitten. Nonetheless, previous studies (see below) reported louder, longer and more variable vocalizations in deafened kittens  but should less affect the harmonic ratio. Indeed, no systematic effect of age on the harmonic ratio was observable in hearing cats Fig. . The harIt is interesting that the deaf cats showed the largest difference in F0 compared with hearing animals in the youngest group but not at later ages when the F0 has decreased in hearing cats. One possible explanation of lower F0 in deaf cats is that it is moved into the range where somatosensory receptors can represent vibrations in a phase-locked manner and be used to control vocalizations, particularly at the youngest ages when F0 is highest. Somatosensory receptors can detect vibrations with high precision and reliable phase-locking at least up to 1 kHz .


Genetic tests for hereditary hearing loss inform clinical management of patients and can provide the first step in the development of therapeutics. However, comprehensive genetic tests for deafness genes by Sanger sequencing is extremely expensive and time-consuming. Next-generation sequencing (NGS) technology is advantageous for genetic diagnosis of heterogeneous diseases that involve numerous causative genes.GJB2 mutations (the gene most often associated with inherited hearing loss), mitochondrial m.1555A>G or 3243A>G mutations, enlarged vestibular aqueduct, or auditory neuropathy. Clinical features of subjects were obtained from medical records. Genomic DNA was subjected to a custom-designed SureSelect Target Enrichment System to capture coding exons and proximal flanking intronic sequences of 84 genes responsible for nonsyndromic or syndromic hearing loss, and DNA was sequenced by Illumina GAIIx (paired-end read). The sequences were mapped and quality-checked using the programs BWA, Novoalign, Picard, and GATK, and analyzed by Avadis NGS.Genomic DNA samples from 58 subjects with hearing loss from 15 unrelated Japanese families were subjected to NGS to identify the genetic causes of hearing loss. Subjects did not have pathogenic ACTG1, DFNA5, POU4F3, SLC26A5, SIX1, MYO7A, CDH23, PCDH15, and USH2A, suggesting that a variety of genes underlie early-childhood hearing loss in Japanese patients. Mutations in Usher syndrome-related genes were detected in three families, including one double heterozygous mutation of CDH23 and PCDH15.Candidate genes were identified in 7 of the 15 families. These genes were Targeted NGS analysis revealed a diverse spectrum of rare deafness genes in Japanese subjects and underscores implications for efficient genetic testing. GJB2 mutations are the most common cause of childhood hearing loss worldwide  symbols represent bone conduction thresholds of the left ear; arrows pointing to the bottom left represent scale-out hearing level of the right ear; arrows pointing to the bottom right represent scale-out hearing level of the left ear.Click here for file


Signalling plays an important role in facilitating and maintaining affiliative or cooperative interactions in social animals. Social grooming in primates is an example of an interaction that requires coordination between partners but little is known about communicative behaviours facilitating this activity. In this study, we analysed the communication of wild chimpanzees of Budongo Forest, Uganda, as they entered and maintained a naturally occurring cooperative interaction: social grooming. We found that lip-smacking, a distinct multimodal oral gesture produced during grooming, coordinated this activity. Lip-smacking at the beginning of grooming bouts was significantly more often followed by longer and reciprocated bouts than silent grooming initiations. Lip-smacks were more likely to be produced when the risk of termination of the interaction by the recipient was high, for instance when grooming vulnerable body parts. Groomers were also more likely to produce lip-smacks during face-to-face grooming where the visual aspect of the signal could be perceived. Data are consistent with the hypothesis that chimpanzee lip-smacks function to coordinate and prolong social grooming, suggesting that this oral signal is an example of a communicative behaviour facilitating cooperative behaviour in chimpanzees. Advertising behavioural intentions or subsequent behaviour is essential in initiating and maintaining friendly or cooperative interactions in animals25In many primate species, lip-smacking, defined as the rapid closing and opening of the mouth and lips81091416171920232426To maximise the benefits of grooming, signals advertising grooming commitment and encouraging partners to engage in this interaction or preventing them from terminating it, would be beneficial. To date, however, little is known about the role of communication in instigating and maintaining grooming. Lip-smacking is a good candidate to function in this manner, since it is often associated with grooming. This especially applies to chimpanzees, which, in contrast to monkey species, produce lip-smacks almost exclusively in grooming contexts. In chimpanzees lip-smacks are produced by the groomer33The aim of our study was to investigate the role of lip-smacking in coordinating dyadic grooming bouts in chimpanzees. We hypothesised that lip-smacks produced by the groomer function to initiate and prolong grooming bouts and to facilitate grooming reciprocation within the bout. As such, we predicted that grooming bouts would be longer if lip-smacks were produced at the start of a grooming interaction compared to bouts where these signals were not used. We also predicted that the groomee would be more likely to reciprocate grooming if lip-smacks were given by the groomer, and that groomers would be more likely to produce lip-smacks when engaging in higher-risk grooming of vulnerable body parts, to signal benign intent and to prevent early termination of the grooming bout by the recipient e.g.,35. SinceLip-smacks occurred in 65% of grooming bouts. In 54% of grooming bouts lip-smacks were produced during the first 10 s of the bout. Lip-smacking occurred in 41% of all 10 s samples taken from all groomers (47% in reciprocated and 35% in unreciprocated grooming bouts). Although there was some variation among the focal males in terms of the rate with which they produced lip-smacks , all of them produced lip-smacks .Grooming bouts with lip-smacks given by the groomer in the first 10 s of the bout were significantly longer than those in which no lip-smacks occurred during the same period . Therefore, the lack of stable friendship patterns may have reduced the chances of seeing a clear pattern between the occurrence of lip-smack signals and the level of affiliation between the partners. It is also possible that, during times of social instability, it pays individuals to use affiliative signals to neutral social partners or lower ranking males to cultivate potential future allies. Alternatively, lip-smacking may be a flexible short-term affiliative signal employed between both preferred and neutral social partners in a similar way as pant hoot chorusingOverall, our data suggest that chimpanzees flexibly modulate the production of lip-smacks, which in turn seems to influence the social nature of grooming bouts by making them longer and reciprocated. In this respect, lip-smacking might be interpreted as a signal that facilitates cooperative acts in chimpanzees, with similarities to how language facilitates and coordinates joint activities in humans. Indeed, on a proximate level, lip-smacking, although unvoiced, requires some control over the supra-laryngeal parts of the vocal tract and rhythmic facial expressions77In conclusion, our study suggests that chimpanzee lip-smacking functions to maintain and prolong grooming bouts, as well as to facilitate within-bout reciprocity. Lip-smacks seem to encourage the recipient to engage in this activity and this coordination of the activities of the interactants is critical to the occurrence of long, reciprocal and thus socially valuable grooming bouts. Lip-smacking might therefore be an example of an oral signal that facilities cooperative behaviour in chimpanzees.2. Study subjects were adults  of the start and end of a grooming bout, the identity of the individual initiating a grooming bout and the identity of the partner. To minimise the problem of temporal non-independence of the data-points, we recorded details on grooming and lip-smacking from the first 10 s of every minute during the bout . During Our data set comprised of a total of 1385 10 s samples taken from 192 grooming bouts (of which 56 were reciprocated and 136 unreciprocated). We omitted in the analyses the 10 s samples (N = 347 of 1385) where two individuals were grooming each other simultaneously since it was difficult to reliably establish whether both or only one grooming partner was lip-smacking. Of the remaining 1038 10 s samples (501 from reciprocated and 537 from unreciprocated grooming bouts) that were entered into analyses, 748 of these samples from 129 bouts were provided by focal males when grooming other adult or late adolescent males, 199 samples from 35 bouts by focal males when grooming females or early adolescent males, and 91 samples from 28 bouts by non-focal individuals .PSPs were identified only for adult and late adolescent males. PSPs were established on the basis of three different dyadic association measuresDominance status was established only for adult and late adolescent males, using the Elo-rating procedureLinear Mixed-Effect Models (LMM) and Generalised Linear Mixed-Effect Models (GLMM) were used in statistical analyses. In the majority of the analyses the first 10 s within every minute of a grooming bout was set as a single data-point. In our models the identities of both the provider and receiver of grooming for every data-point were put as random effects. Since grooming bouts often generated more than one data-point, we put the identity of a grooming bout as an additional random effect. All statistical analyses were conducted using STATA 12.0 software .To investigate whether grooming bouts in which there was a lip-smack at their onset were longer than those in which there was not, we created a LMM in which we put as the dependent variable grooming bout duration (in seconds) and as the independent variables whether or not (0/1) lip-smacking occurred during the first 10 s of the grooming bout and whether or not (0/1) the grooming bout was reciprocated. In this analysis a grooming bout was a single data-point and data from all grooming bouts were included (N = 192). Groomer ID and groomee ID were put as random effects.To investigate whether lip-smacking at the onset of a grooming bout predicted whether or not a grooming bout was reciprocated, we created another model in which as the dependent variable we used the type of a grooming bout (reciprocated (1) and unreciprocated (0)) whereas as the independent variable we put whether or not (0/1) there was a lip-smack during the first 10 s of that bout. We also inserted into the model as the potential confounding independent variables, whether or not the groomee was a PSP to the groomer (0/1) and the dominance distance between the two grooming partners (in Elo-rating scores). Since we had data on affiliative and dominance relationships only for adult and late adolescent males, we conducted this analysis only for this age/sex category. In this analysis, a grooming bout was a single data-point (N = 129 grooming bouts). Groomer ID and Groomee ID were put as random effects.To test which factors affect the probability of the groomer producing lip-smacks, we created a GLMM in which we put as the dependent variable whether or not (0/1) the groomer lip-smacked in the first 10 s of every minute, and as the independent variables the dominance status distance between the grooming partners (in Elo-rating scores), whether or not (0/1) the groomee was a PSP to the groomer, whether or not (0/1) the groomer was in a direct visual contact with the groomee, as well as whether or not (0/1) vulnerable body parts were being groomed. Again, since we had data on PSPs and dominance statuses only for adult and late adolescent males, only samples from grooming bouts between these males were analysed in this model (N = 748 10 s samples from 129 grooming bouts). The ID of the groomer, the ID of the groomee and the ID of the grooming bout were put as random effects.How to cite this article: Fedurek, P. et al. Chimpanzee lip-smacking facilitates cooperative behaviour. Sci. Rep.5, 13460; doi: 10.1038/srep13460 (2015).


TCs) were previously shown by our group to form a tandem with stem/progenitor cells in cardiac stem cell (CSC) niches, fulfilling various roles in cardiac renewal. Among these, the ability to ‘nurse’ CSCs in situ, both through direct physical contact (junctions) as well as at a distance, by paracrine signalling or through extracellular vesicles containing mRNA. We employed electron microscopy to identify junctions (such as gap or adherens junctions) in a co‐culture of cardiac TCs and CSCs. Gap junctions were observed between TCs, which formed networks, however, not between TCs and CSCs. Instead, we show that TCs and CSCs interact in culture forming heterocellular adherens junctions, as well as non‐classical junctions such as puncta adherentia and stromal synapses. The stromal synapse formed between TCs and CSCs  was frequently associated with the presence of electron‐dense nanostructures (on average about 15 nm in length) connecting the two opposing membranes. The average width of the synaptic cleft was 30 nm, whereas the average length of the intercellular contact was 5 μm. Recent studies have shown that stem cells fail to adequately engraft and survive in the hostile environment of the injured myocardium, possibly as a result of the absence of the pro‐regenerative components of the secretome (paracrine factors) and/or of neighbouring support cells. Herein, we emphasize the similarities between the junctions described in co‐culture and the junctions identified between TCs and CSCs in situ. Reproducing a CSC niche in culture may represent a viable alternative to mono‐cellular therapies.Telocytes ( Different cell types have been investigated and even considered for regeneration of the injured heart in animal models and humans, of which notably: embryonic stem cells(ESCs), adipose‐derived stem cells, mesenchymal stem cells, bone marrow‐derived cells and CSCs in situin cultureAmong the cell types recently suggested to be involved in cardiac homoeostasis and regeneration are TCs in culture.In this work, we demonstrate that TCs form ‘atypical’ junctions with stem cells not only in tissue but also Cardiac TCs were isolated from the hearts of 3‐month‐old Wistar rats and cultured as described previously Cells cultured for 24 and 48 hrs, were fixed and embedded in epoxy resin (Agar 100) as described previously Telocytes were identified in culture Fig.  based onTelopodes originating from different TCs frequently formed homocellular networks, establishing both adherens (AJ) and gap junctions (GJ) at the site of contact Fig.  and thisstromal synapses (Figs Telocytes formed junctions with adjacent CSCs Figs , 4, 5A, ses Figs  and 4. TPuncta adhaerentia type junction was infrequently found connecting TCs and CSCs (Fig. via different types of extracellular vesicles (Fig. The SCs Fig. C. Telocyles Fig. A, howeveIntercellular contacts between TCs and CSCs were also analysed in tissue samples for comparison. The stromal synapse associated with electron‐dense nanostructures Figs B and 6 win culture as they do in tissue and form the same types of cell‐to‐cell junctions between themselves, as well as with CSCs, supporting the idea of an active involvement in the homoeostasis of cardiac tissue. This study suggests that replicating the architecture of CSC niches in vitro may be possible.Here, we confirm that TCs display a similar morphology In tissue, TCs may be identified as a result of their peculiar morphology, with Tp forming vast cellular networks, and these characteristics were exhaustively discussed in a previous review from our group in culture. This may have been the result of the relatively short culture times, but more likely represents a characteristic of the partnership between the two types of cells. Similarly, embryonic fibroblast feeder cells were not found to form GJs with human ESCs and mouse ESCs Telocytes formed homocellular networks by means of GJs, however, no such junctions could be seen between TCs and CSCs Puncta adhaerentia, consisting of cadherin–catenin clusters, are found at cell–cell contacts often during early stages of AJ assembly Telocytes formed both homocellular, as well as heterocellular AJs with CSCs. Classic AJs formed between stem and support cells in epithelial and stromal niches were previously shown to control the geometry of division, by facilitating the proper positioning of centrosomes Additionally, previous studies from our group The exact role of TCs in cardiac tissue regeneration remains an elusive, but desirable target, with potential applications in stem cell therapy. Future in‐depth studies of the molecular components of stromal synapse and the dynamics of these components in normal and diseased tissues will shed a new light on the signalling mechanisms between stem and support cells Fig. . This, iThe authors confirm that there are no conflicts of interest.


The New Jersey Safe Schools Program (NJSS) offers courses required for secondary school vocational–career–technical education teachers to become school-sponsored structured learning experience supervisors. The “Federal Wage and Hour and Child Labor Laws, Regulations and Hazardous Order Course” (FWH) was originally conducted in-person by U.S. Department of Labor-Wage and Hour Division from 2005 to Summer 2013, and then NJSS began conducting this course in-person (October 2013–April 2015). Staring in March 2015, this course was conducted online; beta-/pilot tests were conducted in Winter 2014–2015. Starting in May 2015, this course was offered exclusively online. This paper analyzes data from the in-person and online versions of the FWH, including overall course evaluation data comparing two versions with similar questions/constructs.The New Jersey Safe Schools Program modifications to FWH included adding information regarding the Fair Labor Standards Act’s Section 14(c) and supplemental case studies. The online version included information/resources provided during the in-person training plus assessments to supplement each module; the online version was split into modules to allow participants scheduling flexibility. Participants were given multiple possible attempts to achieve a minimum passing grade of 70%, excluding two ungraded activities (crossword puzzles simply completed). Descriptive statistics evaluated user satisfaction online compared to the in-person version of FWH and performance on aforementioned online assessments replacing in-person discussions/interactions.Between October 2013 and April 2015, 160 participants completed the training in person; 156 had complete data. Between April and November 2015, 78 participants completed the training online; 74 participants had complete data. Other enrolled participants were in progress (not done as of 12/23/2015). Overall satisfaction was similarly high for in-person and online versions of FWH; over 95% of responding participants recommended this course to colleagues. Course evaluations for in-person participants indicated 83% felt the course objectives were completely met, whereas 95% of the responding online cohort felt course objectives were completely met. Further analyses examined performance of online assessments regarding number of attempts and scores achieved and performance on highlighted questions in certain module lessons.Data suggested the online format as a viable alternative to an in-person version of this training and provided NJSS and agency partners with ideas on how modifications/improvements can be made. There are currently 507 secondary schools in the State of New Jersey (NJ) where high school students are able to participate in vocational-career-technical CTE programs across 16 different career clusters . The U.STraditional classroom-based in-person training courses are administered without the predominant use of online technology. Information is given orally or in writing by a certified instructor, who sometimes may demonstrate where to find relevant information on agency websites, e.g., USDOL. The online training format differs as the content is delivered through the use of online platforms on multiple types of hardware – desktop and laptop computers, tablets, and smartphones – without any face-to-face interaction. For adult worker S&H, online learning (e-learning) has been conducted in the construction industry with an emphasis on fall prevention, equipment use, and work practices in Taiwan  and in tCurrently, a fast growing trend in education is online learning. Notably, online college enrollment has grown almost 30% since 2010 . PotentiRecently, the United States Department of Education (USDOE) conducted an evaluation of 51 online learning studies, with results suggesting that students who took classes online performed better, on average, than those who took the classes in traditional classroom settings . ResultsAcross several different fields, current literature has suggested, in terms of performance, no statistically significant difference between online learning and classroom-based learning , 12, thoThis paper’s first objective was to describe the creation/conversion process of a training originally conducted in person with agency staff [2005–2013 school years  in April, 2015.n = 5) and Central NJ (n = 2). They completed the course, coursework was reviewed, and course evaluations were examined.The final online version of the course was first completed by eight NJ teachers/supervisors representing seven different school districts (one participant did not identify representing school district). These teachers had initially signed up for the March, 2015 in-person version of the training, but, there was an inclement weather/schools closed day throughout NJ. These eight teachers were from school districts in Northern NJ  aggregated. The consistent pattern of Lesson 2, Lesson 5, and Lesson 8 having the highest average score upon further stratification by gender suggests that the lesson content, module layout, and assessment type were more effective than other modules. The three lessons with the top average score for males were Lesson 8 (99.4%), Lesson 6 (96.9%), and Lesson 9 (96.0%), of which only one lesson (Lesson 8) was consistent with the top three lesson score averages of both females and males aggregated. However, the number of males (n = 20) compared to females (n = 54) is to be noted. Both females and males had relatively similar average attempts of 1.8 and 1.7, respectively, across the lessons of this training. Both females and males had the highest average number of attempts of 2.7 and 2.6, respectively, in Lesson 9, which suggested possible lesson difficulties or decreased attention while trying to finish the last lesson of the course. The lowest average number of attempts was observed in Lesson 2 (1.3) for females and in Lesson 7 (1.3) for males.Table Table S1 in Supplementary Material presents the number and correct percent of question answers by the highest score among attempts and lesson number, stratified by the gender. In other words, as an expansion of Table It should be also noted how across gender stratums only one question out of the four lessons had an overall failing score (≤60%): question one from Lesson 4 with a score of 28.4%. The lowest overall question score for each individual lesson is as follows: 73.0% for question 9 in Lesson 1, 67.1% for question 7 in Lesson 3, 28.4% for question 1 in Lesson 4, and 69.4% for question 10 in Lesson 7.n = 3) was small relative to both Central NJ (n = 28) and Northern NJ (n = 43). The lessons with the top average score for Northern NJ were Lesson 5 (100%), Lesson 8 (99.3%), and Lesson 2 (97.7%), which were consistent with the top three lesson score averages of females and aggregated participants. The lessons with the top average score for Central NJ were Lesson 2 (100.00%), Lesson 5 (99.1%), and Lesson 8 (99.0%), which were consistent with the top three lesson score averages of Northern NJ, females, and participants aggregated. It should be noted how the top scoring lessons within these regional stratifications follow the aforementioned trend noted within the aggregate and gender stratified results. The lessons with the top average score for Southern NJ were Lesson 5 (100%), Lesson 8 (100%), and Lesson 7 (98.3%), two of which (Lessons 5 and 8) were consistent with Northern NJ and Central NJ, females, and participants aggregated. The three NJ regions had similar average number of attempts between 1 and 3, across lessons: Northern NJ with average number of attempts of 1.6, Central NJ with average number of attempts of 2.0, and Southern NJ with average number of attempts of 2.0. Lesson 9 had the highest average of attempts per individual lesson for Central NJ and Southern NJ, while Lesson 3 had the highest average number of attempts per individual lesson for Northern NJ. Lesson 5 and Lesson 7 (each 1.4) had the lowest average number of attempts for Northern NJ, whereas Lesson 2 (1.3) and Lesson 5 (1.0) had the lowest average number of attempts for Central NJ and Southern NJ, respectively.Table Table S1 in Supplementary Material presents the number and the correct percent of question answers by the highest score among attempts and lesson number, stratified by region of NJ. Thus, as an expansion of Table n = 3) from Southern NJ. Data from course evaluations  for each lesson as well as the highest frequency of failing question scores (≤60%). This may be attributed in part to the low study subsample . About one-quarter (24.6%) of participants indicated that they were able to complete it in less than 4 h, and the remaining participants (26.7%) indicated that it took them 6 h or more to complete.Results from in-person evaluations revealed similar results for course satisfaction Table . A majorOnline participants (74 out of 78) were relatively more likely to recommend this course to others than in-person participants (156 out of 160), although both groups were highly satisfied with the course  from the State of NJ New teachers who will be tasked with supervising school-sponsored structure learning experiences across career clusters and program pathways and who have not yet completed the NJDOE/NJSS SLE supervisor curriculum will continue to take the online version of this specific NJ SLE training in the future, starting again in February, 2016. In the future, it may also be possible for teachers in other states to complete and benefit from the course. Data to date suggested that an online format can be a viable alternative to the in-person version of this training on federal laws concerning wages and hours worked, child labor, and hazardous occupations within agriculture and non-agricultural industries. This is because available course evaluation clearly suggested that participants in both formats were satisfied with integral components of the course, such as course content and course organization. In addition, the online course format also has produced more data, as written activities replaced in-person anecdotes/discussion, generating rich feedback for NJSS to continue to amend and improve upon the course. Moreover, this evaluation provided NJSS and agency partners with ideas – beyond incorporating changes to federal laws and newly available agency resources – on how course delivery modifications and content improvements can be made to this online training, and evidence for potentially changing the number of NJ approved PDUs awarded to teachers for this course from 6.0 to 5.0 PDUs.The New Jersey Safe Schools Program has Institutional Review Board (IRB) approval from Rutgers Biomedical and Health Sciences, New Brunswick/Piscataway, New Jersey (formerly UMDNJ) for training evaluation analyses and state law based injury surveillance data analyses .DS, AA, AP, LM, and SK contributed to the conception and design of this effort. DS, AP, and LM led data management and analyses, and the interpretation of results; AA assisted the manuscript. DS, AA, AP, LM, and SK contributed to drafting and revising the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Direction-specific interactions of sound waves with the head, torso, and pinna provide unique spectral-shape cues that are used for the localization of sounds in the vertical plane, whereas horizontal sound localization is based primarily on the processing of binaural acoustic differences in arrival time  and sound level . Because the binaural sound-localization cues are absent in listeners with total single-sided deafness (SSD), their ability to localize sound is heavily impaired. However, some studies have reported that SSD listeners are able, to some extent, to localize sound sources in azimuth, although the underlying mechanisms used for localization are unclear. To investigate whether SSD listeners rely on monaural pinna-induced spectral-shape cues of their hearing ear for directional hearing, we investigated localization performance for low-pass filtered , high-pass filtered , and broadband  noises in the two-dimensional frontal hemifield. We tested whether localization performance of SSD listeners further deteriorated when the pinna cavities of their hearing ear were filled with a mold that disrupted their spectral-shape cues. To remove the potential use of perceived sound level as an invalid azimuth cue, we randomly varied stimulus presentation levels over a broad range (45–65 dB SPL). Several listeners with SSD could localize HP and BB sound sources in the horizontal plane, but inter-subject variability was considerable. Localization performance of these listeners strongly reduced after diminishing of their spectral pinna-cues. We further show that inter-subject variability of SSD can be explained to a large extent by the severity of high-frequency hearing loss in their hearing ear. Listeners with total single-sided deafness (SSD) lack the ability to localize sounds on the basis of interaural differences in time (ITD) and sound level (ILD). As a result, SSD listeners encounter significant problems with the processing of auditory information in daily life  and 15 control listeners  participated in the present study. Table Nineteen listeners with complete SSD  monaural hearing; (ii) monaural hearing with a custom-made mold, fabricated from rubber casting material , inserted in the pinna of the better-hearing ear without obstructing the ear canal.All control listeners were tested under normal hearing conditions, and after altering their pinna-cues with custom-made molds in both pinna.Listeners were asked to localize (i) low-pass ; (ii) high-pass , and (iii) broadband  filtered Gaussian white noises. Spectral cues are minimal for LP noises  at a sampling rate of 50 kHz, and were delivered through a BB loudspeaker, moved by a computer-controlled motorized system at a distance of 1.15 m from the listener's head. Stimulus coordinates for BB and HP stimuli ranged from −85° to +85° in azimuth and from −30° to +30° in elevation. LP stimuli were presented at 0° in elevation.2) black plastic plate positioned in front (40 cm) of the listener's eyes. Listeners were asked to point the laser dot as fast and as accurately as possible in the perceived sound direction after stimulus exposure. Listeners were observed continuously by the experimenter with an infrared camera, but did not receive any feedback about their performance during the experiments.For a detailed description of the setup see Bremen et al. . BrieflyThe experimental session started with a brief visual calibration experiment to establish the off-line mapping of the coil signals onto known target locations. After this, listeners performed a brief practice session containing 10 trials to become familiar with the head-movement response procedure.During the sound-localization experiments the listener first fixated on an LED that was located at 0° azimuth and 0° elevation and then triggered the start of the trial by pressing a button. Between 150 and 300 ms the LED disappeared, and 200 ms later the sound stimulus was presented. After stimulus exposure the listener had to direct the head-fixed laser pointer as fast and accurately as possible, by making a rapid head movement toward the apparent sound direction.We analyzed the azimuth (α) responses separately for each stimulus condition  and for each listener. We determined the best linear fit (based on the mean-squared error criterion) of the stimulus-response relationship (pooled across presentation levels and elevation angles for HP and BB noises):RESP is the response azimuth (in degrees), αSTIM is the stimulus azimuth (in degrees), b is the response bias (in degrees), and g the response gain (dimensionless). We also computed Pearson's correlation coefficient between fit and data, as well as the coefficient of determination (r2). To dissociate the potential contribution of the proximal sound level, L, from that of the actual stimulus location, we performed a partial correlation analysis:where αp as the dimensionless azimuth coefficient and q as the dimensionless proximal sound-level coefficient; each determines to what extent sound-source azimuth or proximal sound level explains the observed responses. Variables αRESP, αSTIM and L were transformed into their (dimensionless) z-scores with x the variable to be z-transformed, μx its mean, and σx its standard deviation  responses to BB and HP noises the best linear fits of the stimulus-response relationships were also determined.RESP and εSTIM are the response elevation and stimulus elevation in degrees, b is the response bias (in degrees) and g the response gain (dimensionless).εn = 19) for frequencies up to 4 kHz. At 8 kHz 11 SSD listeners demonstrated normal hearing. The other SSD listeners demonstrated thresholds ≥20 dB HL, with six listeners demonstrating thresholds ≥40 dB HL  in the functioning ear were confirmed in all listeners with SSD n = 9 for fren = 15), two older listeners (age 56 and 61 years) suffered from a symmetric hearing loss at 8 kHz (thresholds ≥ 40 dB HL). All other control listeners demonstrated normal hearing thresholds between 500 Hz and 4 kHz, and thresholds of 40 dB HL, or better, for 8 kHz.In the group of control listeners . Listener P3 with SSD demonstrated good localization performance for BB and HP stimuli . In contrast, listener P12 with SSD demonstrated poor sound-localization abilities. This listener perceived the stimuli mainly at the hearing side, which resulted in a considerable leftward bias (b = 80° for BB stimuli), and small coefficients of determination (r2 < 0.10) for all stimuli and conditions. The hearing thresholds at 8 kHz in the better ear are listed in Table Figure g > 0.44 vs. g < 0.2).Figure n = 15), all SSD listeners with 8 kHz thresholds below 40 dB HL (n = 13), and SSD listeners with 8 kHz thresholds higher than 40 dB HL (n = 6), for BB, LP and HP stimuli. If the right ear was the deaf ear, data are presented without modification. If the left ear was the deaf ear, data of left and right ears were swapped before pooling the data. The figure demonstrates that listeners without high-frequency hearing loss outperformed listeners with 8 kHz thresholds higher than 40 dB HL, for BB and HP sounds. The figure hints at the possibility that SSD listeners with 8 kHz thresholds below 40 dB HL were able to use spectral pinna-cues, as they could localize the BB and HP stimuli in azimuth, but were not able to localize the LP sounds. Listeners with 8 kHz thresholds higher than 40 dB HL were equally poor in localization of BB, LP, and HP stimuli.Figure Figure Figure r = 0.83, p < 0.01). The SSD listeners with 8 kHz thresholds below 40 dB HL demonstrated higher azimuth and elevation gains than the SSD listeners with thresholds above 40 dB HL. The latter group of listeners had both gains close to zero, indicating poor directional hearing performance in both azimuth and elevation. The r2 were also small . The far majority of control listeners had azimuth and elevation gains that were close to the ideal value of one. The two older control listeners with high-frequency hearing loss demonstrated small elevation gains, confirming earlier reports of deteriorated vertical sound localization performance in the elderly . Note that the SSD listeners with a relatively low high-frequency hearing loss demonstrated a clear deterioration in their sound localization performance in both directions. Molds in the pinnae of control listeners only affected their elevation performance. This deterioration of sound localization abilities in elevation, after altering the pinna-cues with custom-made molds in both pinna, has been reported previously . Good high-frequency hearing in the only hearing ear appears to be an important requirement for adequate sound localization performance. The variation in localization performance is not explained by the onset of unilateral deafness . In addition we also included the data of 9 listeners with SSD from the study of Van Wanrooij and Van Opstal  and for the proximal sound level (q in Equation 2) for the BB stimuli, for SSD listeners (circles) and control listeners (crosses). These partial correlation coefficients reveal the relative contributions of the actual target azimuth and the perceived sound level at the hearing ear to their azimuth localization responses. For SSD listeners with an 8 kHz threshold below 40 dB, the contribution of proximal sound level varied systematically with the azimuth coefficient. Responses were more influenced by sound level when the  estimate of azimuth was poor. Indeed, those SSD listeners typically perceived louder sounds on their hearing side. A similar effect of sound level on localization performance in cochlear-implant listeners has been reported by Majdak et al.  device. These devices transmit sounds presented at the deaf side to the hearing ear. Currently, the two most commonly applied CROS interventions are the wireless conventional CROS hearing aid, and the percutaneous bone-conduction hearing device (Bosman et al., In several countries cochlear implantation has become a treatment option (Arndt et al., The present study emphasizes the importance of a precise evaluation of the monaural hearing abilities of listeners with SSD, especially at the higher frequencies for which the spectral-shape cues become unambiguous for sound localization. Some SSD listeners were using monaural pinna-induced spectral-shape cues of their hearing ear, for localization of BB noises in both azimuth and elevation. Because spectral cues are minimal for LP noises (Middlebrooks, The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Bisphenol A (BPA) and triclosan (TCS) are two nonpersistent chemicals that have been frequently measured in spot urine samples from the general population but less so in pregnant women; however, data are limited on the free (bioactive) and conjugated forms of these phenols.The Maternal-Infant Research on Environmental Chemicals (MIREC) Study addressed these data gaps by utilizing stored maternal urine samples from a large multicenter cohort study of Canadian pregnant women.Concentrations of free and conjugated forms of BPA and TCS were measured in about 1,890 first-trimester urine samples by ultra performance liquid chromatograpy–tandem mass spectrometry using isotope dilution.The glucuronides of BPA and TCS were the predominant forms of these chemicals measured , whereas the free forms were detected in 43% and 80% of samples, respectively. The geometric mean urinary concentrations for glucuronides of BPA and TCS were 0.80 μg/L  and 12.30 μg/L , respectively. Significant predictors of BPA included maternal age < 25 vs. ≥ 35 years, current smoking, low vs. high household income, and low vs. high education. For TCS, urinary concentrations were significantly higher in women ≥ 25 years of age, never vs. current smokers, and women with high household income and high education.The results from this study represent the largest national-level data on urinary concentrations of free and conjugated forms of BPA and TCS in pregnant women and suggest that maternal characteristics predicting elevated urinary concentrations of these phenols largely act in opposite directions.http://dx.doi.org/10.1289/ehp.1408187Arbuckle TE, Marro L, Davis K, Fisher M, Ayotte P, Bélanger P, Dumas P, LeBlanc A, Bérubé R, Gaudreau É, Provencher G, Faustman EM, Vigoren E, Ettinger AS, Dellarco M, MacPherson S, Fraser WD. 2015. Exposure to free and conjugated forms of bisphenol A and triclosan among pregnant women in the MIREC cohort. Environ Health Perspect 123:277–284;  Leaching of bisphenol A (BPA) has been reported from food cans and polycarbonate bottles, paper receipts, and dental sealants and fillings . BPA doeExposure to BPA is widespread, with > 90% of the populations of the United States and Canada having detectable urinary concentrations . EvidencTriclosan (TCS) is an antibacterial compound used in some cosmetic products, toothpaste, treated textiles, and food contact materials, such as cutting boards and countertops . TCS mayTriclosan is highly lipid soluble and rapidly absorbed from the gut, and it has a urinary elimination half-life of 11 hr, with an estimated 0.5% present in the unconjugated form within 24 hr of exposure and the majority of the compound as the glucuronide . TCS wasGiven the unique vulnerability of pregnant women and their fetuses and the possibility that these chemicals may be endocrine disruptors, it is important to examine the extent of exposure in this population, especially during the critical exposure window of the first trimester. Because the free form of these phenols may be more toxicologically active than the conjugated forms, measurement of urinary concentrations of free BPA and TCS may provide a superior metric of the biologically effective dose.a) measure the extent of exposure to free and conjugated forms of BPA and TCS during pregnancy among a population of Canadian women; and b) identify predictors of elevated body burdens of these chemicals. This research offered a unique opportunity to efficiently examine these issues in a large diverse population using stored biological specimens and data collected in this prospective pregnancy cohort study.The objectives of the current study were to Participants. The Maternal-Infant Research on Environmental Chemicals (MIREC) Study is a national-level pregnancy cohort of approximately 2,000 women recruited in the first trimester of pregnancy from 10 cities across Canada between 2008 and 2011  was used as a surrogate matrix for urine during the process. Water was poured into polypropylene cups and transferred to polypropylene storage tubes using the same material as for urine samples. Water samples were analyzed following pentafluorobenzyl bromide derivatization by gas chromatography–tandem mass spectrophotometry (GC-MS/MS) using the method previously developed in our laboratory and described by Analytical methods. The liquid chromatography (LC)-MS/MS methods for the analysis of free and conjugated forms of BPA and TCS in urine were described previously by 13C12-BPA and 13C12-TCS, were derivatized with dansyl chloride directly in 1 mL of urine. A liquid–liquid extraction with hexane was subsequently performed and the organic phase evaporated prior to reconstitution in a solution of acetonitrile:H2O . The LC-MS/MS  was operated in electrospray positive and multiple reaction monitoring mode. Chromatographic separation was achieved on an Acquity UPLC HSS T3, 1.8 μm, 50 × 2.1 mm analytical column (Waters) using a mobile phase gradient with 0.1% aqueous formic acid solution and acetonitrile.6d, BPADS-6d, BPAG-6d, TCSS-3d, and TCSG-3d were extracted from 1.5 mL of urine by solid phase extraction using a weak anion exchange phase . Analytes were eluted from the cartridge using a solution of 1% ammonium hydroxide (NH4OH) in methanol. The extracts were evaporated to dryness and reconstituted in a solution of 25% methanol in water. The same LC-MS/MS instrument and analytical column were used as for the free species, but the MS/MS was operated in the electrospray-negative and multiple reaction monitoring mode. A mobile phase gradient from aqueous NH4OH (2%) to an NH4OH–methanol solution (0.1%) was used to obtain proper chromatographic resolution of conjugated compounds.The conjugated metabolites BPAS, BPADS, BPAG, TCSS (TCS sulfate), and TCSG (TCS glucuronide) and their isotope-labeled standards BPAS-Laboratory quality control (QC). Several QC samples, reagents blanks, and urine blanks were incorporated into each batch of samples. In-house reference materials were prepared by spiking human urine to yield low (0.18 μg BPA/L and 0.9 μg TCS/L) and high (1.5 μg BPA/L and 7.5 μg TCS/L) concentrations. For conjugated species, human urine was spiked to obtained reference materials at three different concentration levels: low (0.2 μg/L), medium , and high . The intraday precision varied from 2.5% to 7.7%, and the interday precision ranged from 4.3% to 13% depending on the analyte. The accuracy was –3.7% for free BPA and –1.0% for free TCS. The accuracy for conjugated forms ranged from –2.1% to 13.3% depending on the analyte. Detailed quality assurance/QC procedures were described by Statistical analysis. Two different approaches were used to calculate summary statistics for the biomonitoring results that were below the limits of detection (LODs). The first approach used values generated by the laboratory instrument, and observations that were reported as zero were replaced by one-half the next smallest value (other than zero) for that contaminant. In the second approach, censoring methods were used by applying survival analysis techniques to left-censored data that have been demonstrated by other authors  equivalents. Total BPA or TCS was calculated by summing the free and conjugated forms, and the most conservative LOD of the components was assigned to the total to determine the percentage below the LOD.p < 0.05), separate treatment regression lines need to be estimated and then compared , pairwise comparisons were carried out using the Scheffé correction for multiple comparisons to determine significant group differences. This correction ensures that the overall false-positive rate from multiple comparisons is < 0.05.We calculated GM urinary concentrations for each level of potential predictive variables for all analytes that had at least 50% of the data above the LOD in all groups  and R (R Development Core Team). For the censoring methods, we used functions from the R libraries NADA and SURVIVAL. Unless otherwise indicated, a 5% significance level (α = 0.05) was implemented throughout.Participant characteristics. Our participants tended to be well-educated, > 30 years of age, and born in Canada, and they had never smoked, had an underweight or normal body mass index (BMI) prior to pregnancy, and had a parity of 0 or 1 .BPA. Detectable urinary concentrations of free BPA, BPAG, and BPAS were found in 43.24%, 94.75%, and 23.43% of the samples, respectively  and the log of total BPA  (data not shown). The log of free BPA was positively correlated with the log of total BPA  . As log total urinary BPA increased, free BPA also increased and the ratio of free to total BPA decreased  .The log of BPAG was positively correlated with the log of free BPA  and total TCS . Similarly, free TCS was correlated with total TCS .All of the log-transformed TCS metabolites were highly correlated with each other (data not shown). TCSG was correlated with free TCS  and TCSG  in womenp-values for the censoring methods were similar to machine readings, but were generally more conservative (data not shown).Comparing the two methods for dealing with values < LOD, the median from the Kaplan-Meier was similar to the median based on machine readings, and the GM from the censored MLE compared well with the GM based on machine readings  have been measured in several cohorts of pregnant women in the United States, with medians ranging from 1.0 to 2.0 μg/L , somewhaOnly one study has reported urinary concentrations of free BPA in pregnant women ; howeverA few studies have measured the conjugated forms of BPA in adult populations e.g., . In a stThere are a number of possible explanations for differences between studies in levels or proportions of free and total BPA, including disparities in laboratory methods and their sensitivity, differences in the study populations, hydrolysis of the conjugates, and contamination of the samples. Although free BPA has been measured in several studies  is comparable to that reported following oral ingestion of TCS in 10 volunteers, where 0.9% of the TCS excreted was reportedly in the free form 24–48 hr after exposure .Several studies have measured total TCS in urine from pregnant women . Median urinary concentrations were considerably higher in studies conducted in Puerto Rico  and FranData on major predictors of TCS exposure are limited. In MIREC, urinary TCS concentrations were significantly elevated in women with a university degree (compared with those with less education), > $100,000 compared with ≤ $100,000 household income, ≥ 25 compared with < 25 years of age, and never smokers compared with former or current smokers. In the U.S. general adult population, TCS concentrations were significantly higher in households with income of ≥ $20,000; they appeared to peak in the third decade of life and then decline slowly thereafter . In PuerA major concern in conducting biomonitoring studies is potential contamination of biospecimens via materials and processes , resultiConsideration of the ability of a single spot urine sample to predict an individual’s exposure over a period of time is especially important for short-lived chemicals such as BPA and TCS. Previous studies have indicated that although the intraclass correlation coefficient (ICC) for BPA is low (< 0.25) across pregnancy , the ICCThe present study is one of only a few to measure the extent of exposure to free and conjugated forms of BPA and TCS, especially during the critical period of pregnancy. We observed median concentrations that were lower than the estimated biomonitoring equivalents  for total BPA  can vary by age and education  and poss(846 KB) PDFClick here for additional data file.


We aimed to evaluate the association between hearing impairment and the prevalence of chronic kidney disease (CKD) in the largest population-based cross-sectional study to date.2. The participants were also evaluated for the presence of other contributing factors related to kidney dysfunction. We divided the participants at the 40-dB threshold into hearing-impairment and no-hearing-impairment groups, using the average threshold of all six frequencies  for both ears.This cross-sectional study was based on the Korean National Health and Nutritional Examination Survey (KNHANES). It included 5226 participants ≥19 years of age whose estimated glomerular filtration rate (eGFR) and hearing threshold had been measured. We diagnosed CKD as an eGFR <60 mL/min/1.73 mp-value < 0.001) in participants with an eGFR <60 mL/min/1.73 m2 than in those with an eGFR ≥60 mL/min/1.73 m2 after adjustments for age, sex, smoking, alcohol, body mass index, diabetes mellitus, hypertension, dyslipidemia, and microalbuminuria. Among the risk parameters of CKD associated with hearing impairment, linear regression analysis adjusted for age and sex determined that each increase of serum creatinine or blood pressure was positively associated with an increase in hearing threshold .The odds of hearing impairment was 1.25 times higher (95 % confidence interval: 1.12–1.64, The odds of hearing impairment were greater with lower eGFR than with normal eGFR. Individuals with CKD were more likely to also have hearing impairment. We recommend screening the hearing of patients with CKD to provide earlier identification of hearing impairment and earlier intervention, thereby preventing progression of hearing impairment and providing appropriate treatment strategies. The association between chronic kidney disease (CKD) and hearing impairment was first reported more than 80 years ago by Alport , who desThe kidney and stria vascularis of the cochlea share physiologic, ultrastructural, and antigenic similarities that could explain the link between CKD and hearing impairment . The simWe conducted a large cross-sectional analysis on the associations between hearing impairment with CKD in a representative sample of the adult Korean population from the Korean National Health and Nutritional Examination Survey (KNHANES). The aim of this study was to confirm whether CKD could be an independent contributing factor for hearing impairment and then to identify which CKD-related risk parameters were important contributing factors for hearing impairment among people with nonsyndromal CKD.The KNHANES is a cross-sectional and nationally representative survey composed of a health questionnaire survey, health examination, and nutrition survey. The KNHANES V was conducted from 2010 to 2012 using a stratified, multistage probability sampling design, and we used recent data from the third year (2012) of the KNHANES V in this study. In the 2012 survey, 10,069 individuals participated in health interviews, health examination surveys, and nutritional surveys. A trained otologist performed face-to-face interviews with each patient to collect a comprehensive medical history that included information about the presence of syndromes related to kidney disease and hearing. We analyzed the data of participants aged ≥19 years. Participants with any of the following conditions were excluded: (1) history of treatments for renal failure or syndromes associated with kidney dysfunction ; (2) history of chronic otitis media; (3) external- or middle-ear problems on physical examination; (4) history of noise-induced hearing impairment; or (5) incomplete hearing results. Using the questionnaire for quantitative evaluation , 101 patients with noise-induced hearing impairment were excluded, as noise-induced damage of the cochlea could create bias when evaluating the independent role of CKD for hearing. Exposure to occupational noise was determined according to whether the participant had worked in a location with loud machines for ≥3 months. A loud noise was defined as whether the participant had to raise his or her voice to hold a conversation.A total of 5226 participants who underwent audiometric testing were included in this cross-sectional analysis. The Korea Centers for Disease Control and Prevention approved the study protocol, and written informed consent was obtained from all participants before the study began.Ear examinations were performed by ear, nose, and throat residents using a 4-mm 08 endoscope (Xion GmbH) and the ML 150 vision system (JRMed Trade). The air-conduction hearing threshold was measured by well-trained examiners in a double-walled soundproof booth using an automatic audiometer  at 500, 1000, 2000, 3000, 4000, and 6000 Hz. In this study, the pure-tone audiogram average was calculated as the average threshold of all six frequencies. We included the 6000-Hz threshold to detect early presbycusis .2) was calculated by dividing weight by the height squared. Smoking history was positive if subjects had smoked >5 pack-years during their entire lives. Alcohol drinking was indicated by the consumption of at least 30 g/day over the last year. Seoul  and six other metropolitan cities were grouped as urban areas, while the remaining regions were grouped as rural areas. Educational status was divided into having or not having attended college. Histories of explosive or occupational noise exposure were considered positive or negative according to the subjects’ recall. An explosive noise was defined as a sudden loud noise such as an explosion or gunshot. Exposure to occupational noise was determined according to whether the participant had worked in a location with loud machines for ≥3 months. A loud noise was defined as the participant having to raise his or her voice to hold a conversation. Subjects were asked if they had ever been diagnosed with diabetes, hypertension, dyslipidemia, thyroid dysfunction, or chronic renal failure by a doctor.Information on age, sex, residence area , educational status, smoking exposure, alcohol drinking, occupation, and other variables in the KNHANES were obtained through a self-reported survey. Body mass index , total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, blood urea nitrogen (BUN), and creatinine concentrations were measured using a Hitachi 7600–110 chemistry analyzer . Fasting blood sugar was measured using an automated analyzer with an enzymatic assay . Urinary albumin was measured using a turbidimetric immunoassay . To compare the amount of albumin in each sample against the concentration of creatinine in the spot urine sample, the albumin/creatinine ratio (ACR) was used; microalbuminuria was defined as ACR ≥3.5 mg/mmol  or ACR ≥2.5 mg/mmol . The estimated glomerular filtration rate (eGFR) was determined using the CKD-EPI formula: eGFR (mL/min/1.73 m2) = 141 × minα × max −1.209 × 0.993Age × 1.018, if female  [2.Blood samples were taken after an 8-hour fast during the period in which this survey was performed. The glycosylated hemoglobin  . We focut-test was used to compare continuous variables and the chi-square test was used to compare categorical variables.We used the stratification variables and sampling weights designated by the Korean Centers for Disease Control and Prevention. Continuous data are expressed as means and standard deviation, while categorical data are expressed as percentages. An independent P < 0.05 were considered statistically significant. Statistical analyses were performed using SPSS version 18.0 .Demographic characteristics were analyzed according to the hearing threshold (<40 or ≥40). Logistic regression analysis was applied to examine whether hearing impairment was associated with CKD and was fitted with increasing degrees of adjustment: a model adjusted for age (Model 1) and a model adjusted for age, sex, smoking, alcohol, BMI, DM, HTN, dyslipidemia, and microalbuminuria (Model 2). Linear regression was used to assess the associations between risk parameters related with kidney dysfunction and hearing impairment with adjustment for age and sex. For the multicollinearity diagnosis among variables, variance inflation factors (VIF) were calculated for each variable. The VIF provided an index that could measure how much the variance of an estimated regression coefficient increased due to collinearity. The odds ratios (OR) and 95 % confidence intervals (95 % CI) of the associated factors for reduced eGFR were determined. Values of p-value < 0.05 for all) than those without. The risk parameters  for kidney dysfunction were worse in the hearing-impairment group than in the no-hearing-impairment group. Regarding eGFR, the key parameter for CKD, 10.1 % of patients with no hearing impairment and 47.1 % of patients with hearing impairments had reduced eGFR (<60 mL/min/1.73 m2). The ratios of microalbuminuria between the groups were significantly different; however, the occurrences in both groups were not high (1.3 % vs. 2.9 %).The participants’ general and anthropometric characteristics and clinical and laboratory parameters stratified by hearing impairment status are shown in Table To evaluate which frequencies would be influenced by low eGFR, we divided the frequencies into three groups . All of the three groups had meaningful negative correlations with eGFR values Fig. . Further2) in each group also was analyzed using logistic regression analysis with adjustment models . In Model 2, after adjustment for age, sex, smoking, alcohol, BMI, and parameters of chronic disease, the OR was also significant only for the parameter of eGFR .The prevalence of CKD  was 1.25 times higher in participants with an eGFR <60 mL/min/1.73 m2 than in those with an eGFR ≥60 mL/min/1.73 m2 after adjustment for age, sex, smoking, alcohol, BMI, DM, HTN, Dyslipidemia, and microalbuminuria. We observed in this study that CKD (defined as an eGFR <60 mL/min/1.73 m2) could affect hearing impairment independently. Blood pressure and serum creatinine were found to be positively correlated with hearing impairment among kidney dysfunction–related risk parameters.In this study, we used KNHANES data to investigate the potential association between CKD and hearing impairment in Korean adults. An eGFR <60 mL/min/1.73 mRecent research in a large Australian population reported a relatively high prevalence of hearing impairment associated with moderate CKD, and more than half of the participants with CKD had any level of hearing impairment . Others +-K+-ATPase activity resulted in hearing impairment [2), blood pressure treatment should be considered to preserve hearing function. Such a finding in this large-population study, performed by adjusting related important contributing factors, may be meaningful, compared with other small studies.The pathophysiology between hearing impairment and CKD remains unclear. Various factors can contribute to cochlear dysfunction in patients with CKD, and a combination and interaction of several factors could result in reduced cochlear functioning in patients with CKD. The cochlea and the kidney show noteworthy similarities such as membranous structures, the central role of ciliated epithelial cells, and tubular organization . One respairment . We testOtotoxic medication, including furosemide, can affect ionic gradients between the endolymph and perilymph, resulting in edema of the epithelium of the stria vascularis . It is fTaken together, these observations support the hypothesis of a close correlation between kidney dysfunction and cochlea impairment. The cochlea appears to be most influenced independently by CKD, which can translate into hearing impairment in adults. Although prospective studies are required to assess this association between CKD and hearing impairment, our results have considerable validity in that we confirmed this correlation in the largest population of participants to date after adjustment for other contributing factors and analyzed the study using the questionnaire and a laboratory assessment. We recommend that screening the hearing of patients with CKD to provide earlier identification of hearing impairment and earlier intervention, thereby preventing progression of hearing impairment and providing appropriate treatment strategies.The odds of hearing impairment were greater with lower eGFR than with normal eGFR. Our conclusion is that CKD combined with hypertension may be an important factor in disturbing hearing thresholds. Further studies may well reveal the common pathophysiology between kidney dysfunction and cochlear impairment.


Middle ear ossicle malformations are an uncommon event. Among them, the congenital absence of the stapes is a very rare condition that is seldom described in the literature. We report the cases of two women, aged 19 and 22 , who presented with a long history of conductive deafness. An exploratory tympanotomy was performed and the absence of the stapes superstructure and an abnormal position of the facial nerve could be observed. A bone anchored hearing aid (BAHA) was implanted in both patients with good results. It is believed that stapes agenesis is related to an abnormal development of the facial nerve, which by the 5th to 6th week of gestation would interpose between the otic capsule and the stapes blastema, preventing these structures from contacting. A long history of nonprogressive hearing loss from birth or early childhood is the key to reach a diagnosis. Several treatment options have been described. The authors opted for a hearing aid due to the high risk of facial nerve lesion, with good functional results. The congenital anomalies of ossicular chain of the middle ear are an uncommon event that can present in a variety of ways –4. AmongWe report the case of two sisters of Brazilian origin, aged 19 and 22, who presented with a long history of nonprogressive hearing loss. There was no familiar history suggestive of congenital otologic anomalies. Both patients were products of uneventful pregnancies and deliveries with no history of previous infections, trauma, or related illnesses. The external ear and eardrum were normal on both sides. The audiogram showed bilateral conductive hearing loss for all studied frequencies (250 Hz–4000 Hz) with an average air-bone gap of 60 dB in both patients. Rinne tests were negative bilaterally and the result of Weber's test was central. With an initial suspicion of otosclerosis, a right exploratory tympanostomy was performed to examine the ossicular chain. After careful inspection of the middle ear, we noted the complete absence of the stapes and the stapedial tendon with an abnormal positioning of the facial nerve, which appeared anterior and inferior to the tympanic cavity, above a hypoplastic oval window . Due to Due to the similarity of both cases, high resolution temporal bone CT scans were performed, showing the bilateral absence of the stapes superstrucure and the abnormal positioning of the facial nerve and the incus .Ossicular chain reconstruction was discarded, due to the location of the facial nerve and the high risk of lesion during the procedures. The authors decided to implant a right bone anchored hearing aid (BAHA) in both patients with good functional results. The patients were then referred to the genetic department of our hospital, but no genetic syndrome could be identified.CAS is a very infrequent condition, seldom described in the literature, first described by Mcaskile and Sullivan in 1955 , 8. It hA long history of nonprogressive conductive hearing loss since birth or early childhood, with no history of trauma or infection, is highly suggestive of a congenital middle ear malformation. Differential diagnosis should include otosclerosis and acquired traumatic or infectious disruption of the ossicular chain. Also, in the congenital cases, the hearing loss is usually more severe than in the acquired cases , 7, 8. ADue to the lack of published series, management of the CAS remains controversial . Several


Age-related hearing impairment (ARHI) is becoming a more significant issue as geriatric population increases. Sarcopenia in older people is known to have a diverse health problem in various circumstances in recent studies. We assessed whether the decrease in muscle mass is related to ARHI. We used the 2010 data of the Korea National Health and Nutrition Examination Survey (KNHANES) to examine the associations between sarcopenia and ARHI. A total number of participants was 1,622 including 746 males and 876 females aged 60 years or older. Muscle mass was assessed as an appendicular skeletal muscle mass, and hearing loss was defined as the pure-tone averages (PTA) of test frequencies 0.5, 1, 2, 4 kHz at a threshold of 40 dB or higher in worse hearing side of the ear. Among 1,622 participants, 298 men and 256 women had hearing loss. Appendicular muscle mass (ASM), expressed as kg, was categorized in tertiles. In female population, after adjusting for age, smoking, drinking, amount of exercise, total body fat, education level, income level, and tinnitus, the odds ratio (OR) for hearing loss was 1.57  = 0.92–2.68) in the middle tertile and 1.79 (1.03–3.08) in the lowest tertile, compared with the highest tertile. P for trend in this model was 0.036. Controlling further for hypertension, diabetes mellitus, chronic kidney disease, and three types of noise exposure did not change the association. Larger muscle mass is associated with lower prevalence of hearing loss in elderly Korean females. Age-related hearing impairment (ARHI), also known as presbycusis, is the most common sensory dysfunction in adults . This coThere have been some studies on the risk factors of hearing impairment such as Body Mass Index (BMI), waist circumference, adipose tissue volume and environmental risk factors as smoking, noise and alcohol consumption. Having a high BMI has been regarded as a risk factor for hearing impairment , 6, and In recent studies, sarcopenia in older people turned out to have an adverse health effect in various circumstances. Sarcopenia is an age-associated loss of muscle mass. Among the people aged 60–70 years, 5–13% of them have low muscle mass, and this prevalence increases to almost 50% in people over 80 years old , 10. TheBoth BMI and waist circumference are well known predictors of developing metabolic impairments. Obesity is also known as a risk factor for ARHI, whereas the audiometric association with sarcopenia in elderly population remains unexamined. Given the fact that both sarcopenia and obesity are associated with metabolic impairments, we assumed that the association between sarcopenia and ARHI has clinical significance.We supposed that Sarcopenia is related to the larger prevalence of age-related hearing impairment, and we used the Korea National Health and Nutrition Examination Survey (KNHANES) Ⅴ (2010–2012) data to support the hypothesis.The Korea National Health and Nutrition Examination Survey (KNHANES) Ⅴ (2010–2012) is the fifth cross-sectional survey for the South Korean population conducted by the Korean Ministry of Health and Welfare since 1998. This survey comprises a health interview survey, a health examination survey, and a nutrition survey. Each year’s survey of KNHANES is independent and represents the South Korean population based on its rolling sample design. We used 2010 data of KNHANES which is comprised of 8,958 South Koreans. We excluded people who were younger than 60 years old . Among people aged 60 years or older, we excluded people who only tested for either hearing loss or muscle mass (n = 381). We also excluded 13 people who had liver diseases such as hepatitis B, hepatitis C, and liver cirrhosis, and 12 people who had chronic kidney diseases with GFR<30, because these diseases can affect people’s body composition. A total of 1,622 subjects  were included in the analysis.Muscle mass in each subjects was assessed as appendicular muscle mass (ASM) measured by dual-energy x-ray absorptiometry (DXA) . ASM (kg) was derived as the sum of the lean soft tissue mass of the upper and lower appendixes (arms and legs), after the method of Heymsfield et al. .Muscle mass was categorized in tertiles: t1 (subjects whose ASM<18.6 in men and <12.64 in women), t2 (subjects whose ASM is 18.6–20.85 in men and 12.64–14.24 in women), and t3 (subjects whose ASM≥20.85 in men and ≥14.24 in women).Pure-tone audiometry, the gold standard for hearing loss evaluation, was conducted by a SA 203 audiometer  in the study population. The test was conducted in a soundproof booth and the instruction was given by certain otolaryngologists trained to operate the audiometry. Subjects used supra-auricular headphones in a soundproof booth, and only air conduction was measured. Study subjects pushed a button when they heard a tone. Test frequencies were 0.5, 1, 2, 3, 4, and 6 kHz.Hearing loss was defined as the pure-tone averages (PTA) of test frequencies at 0.5, 1, 2, 4 kHz at a threshold of 40 dB or higher in worse hearing side of the ear. In Information on the study population including BMI, waist circumference, total body fat percentage, smoking, alcohol consumption, exercise level, education level, household income, noise exposure, stress level, and medical conditions were obtained from the result of KNHANES. BMI was calculated based on the data of height which was measured to the nearest 0.1cm with a stadiometer  and weight which was measured using an electronic scale  with an accuracy of 0.1kg. Waist circumference was measured using a measuring tape  to the nearest 0.1cm.2, and the GFR was calculated based on the formula of the Modification of Diet in Renal Disease Study.Hypertension was defined as a systolic blood pressure (BP)≥140mmHg, a diastolic BP≥90mmHg, or treatment with antihypertensive agents. Blood pressure was measured three times with a mercury sphygmomanometer , and the average of the second and the last measurements became each subject’s final blood pressure. Diabetes mellitus (type 2) was defined by a fasting plasma glucose level≥126mg/dL, treatment with oral hypoglycemic agents or insulin, or diagnosis by a physician. Chronic kidney disease was defined as estimated glomerular filtration rate (GFR)<60mL/min/1.73mFor certain items such as smoking, alcohol consumption, exercise level, education level, household income, noise exposure, stress level, and medical conditions such as melancholy, tinnitus, and dizziness, all participants were asked to fill out the self-reported questionnaires. In terms of smoking, the participants were categorized into current smokers and the others. A participant was considered as a ‘drinker’ if he/she drank more than once per month over the past year. A strenuous physical activity done for at least 20 minutes at a time and more than three times a week was considered as a regular exercise. Education level was categorized into two groups: longer than 9 years of education (finished beyond middle school) and the others. The lowest quartile of household income was considered as low income. ‘Noise at work’ was defined as the work experience of more than 3 months at the place where people have to speak up to communicate with one another. ‘Noise outside work’ was defined as the experience of more than 5 hours per week outside workplace where people have to speak up to communicate with one another. ‘Noise at any given moment’ was defined as the exposure to a very loud noise such as gunfire noise. The subjects answered ‘yes’ if they think they experience a lot of stress in everyday life. Depressed mood for more than 2 weeks that affects everyday life in the past year was considered as melancholy. In terms of tinnitus and dizziness, the subjects answered ‘yes’ to each question if they have experienced each condition in the past year.2 tests were done, respectively. The association between ASM and PTAs of 0.5, 1, 2, 4 kHz was presented as Pearson’s correlation, and age-adjusted linear regression was used to verify the relationship between body composition variables and PTAs. Multiple logistic regression analyses were also performed to estimate the association between ASM and hearing loss. And we presented as OR (95% CI) after adjusting for certain factors in each model using hierarchical analysis . All statistical analyses were stratified by gender and performed using the SAS survey procedure  to reflect the sampling weights and complex sampling design analyses of KNHANES. All data of P < 0.05 were accepted as statistically significant results.The data in Characteristics of the subjects are shown in Association with ASM and the maximum dB in males and females is shown in 2 . The correlation between hearing impairment and other factors were statistically insignificant in males.Age-adjusted linear regression analysis between hearing loss and anthropometric parameters is shown in When ASM was categorized in tertiles, smaller amount of muscle mass was significantly associated with hearing impairment in females after adjusting for age (p for trend = 0.0084; In this cross-sectional study, larger muscle mass was associated independently with lower prevalence of hearing impairment in elderly Korean females, and this association was statistically significant even after adjusting for age, smoking, drinking, exercise level, total body fat, education level, income level, tinnitus, hypertension, diabetes mellitus, chronic kidney disease, and noise exposure. This is the first time to reveal the relationship between sarcopenia and ARHI. These findings may suggest possible lifestyle changes to prevent ARHI and even help people delay the onset of ARHI.Low level of physical activity is one of the causes of sarcopenia, along with inflammation, changes in endocrine function, chronic disease, and nutritional deficiencies . Even thIn recent studies, physical exercise appears to induce neuroprotection by improving synapse density and neuronal complexity, and it also appears to restore hippocampal neurogenesis after irradiation in rodents , 17. BraOxidative stress may also be a potential explanation for the association of sarcopenia with ARHI. Cochlea is sensitive to the changes in arterial blood supply as the stria vascularis is highly vascularized . InterruIn our study, larger muscle mass was associated independently with lower prevalence of hearing loss in elderly Korean females. This association was not shown in Korean males. In general, men tend to have greater muscle mass and physical performance compared with women , 34. WheStrengths of our study include its nationally representative data and validated methods to quantify sarcopenia and hearing loss. Our study subjects were comprised of the general population of South Korea and they represented the whole South Korean population as KNHANES is a rolling sample design. Also, KNHANES used the pure-tone audiometric test to whole study population which is a gold standard of hearing tests, and degree of sarcopenia was represented as ASM which was measured by DXA.There are several limitations in our study. It is difficult to evaluate causal relations between muscle mass and age-related hearing impairment because our study is a cross-sectional study. Therefore, our study invites future prospective studies on sarcopenia and hearing loss, which will allow us to have insight on causal relationship of them. Second, our study used KNHANES data which used self-reported questionnaires to examine respondents’ lifestyle, which can have the recall bias.Sarcopenia is underdiagnosed and undertreated even though it is very common among elderly people . Paying


Periplaneta americana L., was significantly enhanced in the presence of odor.The main features of grooming behavior are amazingly similar among arthropods and land vertebrates and serve the same needs. A particular pattern of cleaning movements in cockroaches shows cephalo-caudal progression. Grooming sequences become longer after adaptation to the new setting. Novelty related changes in grooming are recognized as a form of displacement behavior. Statistical analysis of behavior revealed that antennal grooming in American cockroach,  Self-grooming is an innate behavior that is represented across wide range of animal species, such as land vertebrates and arthropods. Despite only far relatedness between these groups, the main features of grooming behavior are amazingly similar and serve the same needs, namely, body cleaning and disease prevention, distribution of some substances across body surface, and as a displacement behavior in stressful conditions –3.Body cleaning is the most obvious purpose of grooming behavior; in insects it can be elicited by small particles, irritants, and weak mechanical stimulation –9. Care Grooming of various body parts is organized in particular sequences, usually with cephalocaudal progression , althougRobust orientation toward the odor source is a vital trait for the insects of various species. Volatiles activating odorant receptors inside olfactory sensilla are eliminated by odor-specific as well as nonspecific enzymes , 23, terPheromone-degrading enzymes were found on the surface of antennae, wings, and legs of some moth species , 29, but Periplaneta americana L. were transferred from the stock colony to the cage and were reared under 12 : 12 LD regime at 28 ± 1°C. After the last molt cockroaches were transferred into the experimental setup, they were kept for at least two weeks prior to tests. The experiments started in the first half of a dark phase, a period of maximum locomotor activity [Male last-instar nymphs ofactivity  and pheractivity . Water a2 and 79% N2) was blown through the cartridge with flow rate 3.5 mL/min. Two consecutive video recording sessions lasting 30 min started after a 10 min acclimation period and followed each other with a 10 min break. After an experiment the test chamber was removed and the cockroach was not returned to the cage. This scheme was chosen to avoid marking of the cockroaches, since it was shown to affect grooming in insects [The setup consisted of 3 components: a transparent plastic cage (30 × 45 × 30 cm) containing food and water, a shelter (17.0 × 17.0 × 5.5 cm) that was constantly dark inside, and an exchangeable test chamber (20 × 20 × 8 cm); the latter 2 were separated from the cage with plastic doors (modified from [ insects . Three sSeries 1 (Blank). Cartridge was loaded by control stimulus in both sessions (n = 8). Series 2 (Odor). Cartridge was loaded by control stimulus in the first session and with eucalyptol in the second session (n = 13). Series 3.  Cartridge was loaded with control stimulus, only one session was run (n = 5). For statistical evaluations purposes the data of this series was pooled with those from the first session of Series 1 and Series 2, so the sample size became n = 26.Video files were processed manually frame by frame. Following grooming events were counted and timed separately: cleaning of antennal flagellum by mouthparts; scraping of antennal base by foreleg; cleaning of forelegs and midlegs/hindlegs (pooled together) by mouthparts; scraping of cerci and genitalia by hindlegs. To evaluate locomotor activity, the top-view projection of the test chamber was divided into 4 quadrants and the number of visited quadrants was calculated.http://vassarstats.net/). Results of these tests are available in Supplementary Table available online at http://dx.doi.org/10.1155/2014/329514. Kolmogorov-Smirnov goodness of fit test (http://contchart.com/goodness-of-fit.aspx) was performed before running Student's t-test  or Mann-Whitney and Wilcoxon signed-ranks test (in other cases).Obtained values were transferred to MS Excel for further data processing and statistical evaluations. Differences (session 2 − session 1) were calculated for every cockroach from each series for each groomed body part Tables . Two-facThe background grooming frequencies were calculated from the first sessions of Series 1, 2, and 3 . Since cGrooming sequences mostly started from antennal grooming followed by scraping antennal bases with ipsilateral foreleg and passing this foreleg through mouthparts and facultatively ended with midleg or/and hindleg grooming. 74.6 ± 4.8% of grooming events was comprised in sequences which lasted 75.0 ± 4.9% of all grooming time.P < 0.01, Student's t-test), antennal bases (P < 0.05 Mann-Whitney test), and forelegs , but not midlegs and hindlegs  as well as for the time spent for grooming  (P < 0.05) in comparison with control. There was no difference across consecutive sessions for the number of observed flagellum groomings and for the time spent for grooming. Total time spent on antennal maintenance  reveals significant influence of the both factors: odor presence (P < 0.05) and consecutive sessions (P < 0.05). Duration of a single cleaning event of antennal flagellum raised at the second session (P < 0.01) and differences were significant for both series ; there was no difference between Series 1 and 2 in duration of antennal cleaning events. Antennal bases were groomed more often and took more time in the second sessions (P < 0.01), but the odor did not significantly affect this behavior (P > 0.05) (Tables P < 0.01) and longer (P < 0.05) cleaning events were observed in the second session; also the time spent for grooming increased (P < 0.01) (Tables Preexisting differences between Series 1 and 2 were found for the frequencies in grooming of antennae (ey test) . The fir) Tables , and 4. ) Tables , and 4.P < 0.01 for the time and P < 0.05 for number of elements) than in the first ones. The changes in sequence parameters look similar for both series , but the small difference between two control consecutive sessions was statistically significant . Sessions of the Series 2 did not differ (P > 0.1).Proportion of grooming events comprised in sequences was similar for all samples , but uniform decrease was found for both series . Percentage of antennal grooming is also similar: 35% [Our data on earlier , namely,lar: 35% , 36% in lar: 35%  was obvilar: 35% .Importance of grooming behavior for the maintenance and acuity of sensory organs was suggested for some insects in different circumstances –38, but Grooming is a complex multipurpose behavior which is reflected in its “microstructure” . Exposur Varroa mites [Importance of chemosensory-dependent grooming should not be overlooked in applied studies, since excessive grooming affects target behavior of beneficial insects . Sugar doa mites , so appl P. americana cockroaches clean more often their main olfactory organs, the antennal flagella, in presence of eucalyptol, the plant odor, underpinning the importance of grooming behavior in olfactory reception.Novelty stress disturbs grooming pattern by shortening grooming sequences and duration of cleaning events, similarly with phenomena known for birds and mammals. These data raise a question on origin and evolution of physiological mechanisms involved in manifestations of displacement behavior throughout the main lineages of animal taxa. Males ofhttp://vassarstats.net was used to obtain P values.Two-way ANOVA with Repeated Measures on One Factor was performed for the number of grooming events, time spent for the particular kind of grooming during the whole session, and duration of a single grooming event (Sessions 1-2 as repeated measures and Series 1-2 as independent factors). Online calculator