Lip reading (LR) is unconsciously practiced as we communicate and has currently been widely used in the assessment of hearing impaired people. The hearing challenged individual is able “to read” lip position and thus interpret the speech sounds of the speaker; however, it is very likely that the best lip reader can only catch 50% of the words uttered.30 individuals of both gender, with age ranging from 27 to 89 years, carriers of moderate bilateral sensorineural hearing loss. The assessment encompassed speech recognition test of monosyllable words in four situations: without hearing aid (HA) and LR; without HA and with LR; with HA and without LR; and with HA and LR.we noticed an improvement in the percentage of correct answers in 93.5% of the patients with HA and LR when compared to those patients in the other situations.Lip reading is an important communication strategy for those with hearing impairment, and it can support the hearing aid fitting process. Communication difficulty is considered the most important consequence of hearing impairment. Hearing impaired individuals try to minimize this difficulty by using some mechanisms in order to have a better understanding of what is being said. Thus, they send the message back to the person they are talking to, in a much easier way. These mechanisms are called by Speri (2000) “communication strategies”According to Boéchat (1992), communication strategies are a set of given attitudes that work as facilitating agents for the message to be easily received, both in a visual and hearing wayThe same author organized communication strategies in groups, according to their nature and she classified them into cognitive, interventional, mechanic, palliative, remedial, waiving and simulative ones.Among the cognitive strategies, which aim to rescue the content of the message, there is lip reading (LR).Besides using communication strategies as facilitating agents for communication effectiveness, the use of Lip Reading becomes essential for this purpose. According to Kozlowski (1997), the visual processing of speech is used even among listeners, as part of speech perceptionFor Demorest, Bernstein (1992) lip reading is the most prevalent expression within the cognitive type of strategy, where individuals use several clues to understand speech, as for example, paying attention to facial expressions, recognition of gesture clues, paying attention to environmental clues and othersThe use of lip reading is unconsciously done when we communicate watching for facial expression, gestures, change of posture and clues that show us ways to decode the information, and it is being currently used when assessing hearing impaired individuals.The hearing challenged individual is able “to read” lip positions and thus interpret the speech sounds of the speaker; however, it is very likely that the best lip reader can only catch 50% of the words uttered, since many phonemes have an invisible articulation and others have the same articulation.For Russo (1999), it is necessary to select, indicate and adapt hearing aids together with global audiologic rehabilitation programs, in order to minimize the psychosocial reactions of older people to the aspects mentioned above, helping this hearing impaired person and his/her relativesTherefore, this study aimed to research lip reading benefits during the hearing aid fitting process in adults.This study was done at the Speech Therapy Sector at the Otorhinolaryngology Department at Marilia Medical School (FAMEMA), and approved by the Research Ethics Committee under protocol # 322/04. Thirty one patients were selected with ages ranging between 27 and 89 years (average= 65.6 median = 71), whose threshold tonal audiometry indicated symmetric bilateral hearing impairment, both sensorineural and moderate, being these two the inclusion factors. All the patients were being submitted to hearing aid selection and indication process for the first time, and they have never had a hearing aid test previously, exclusion factors.Data collection was done through the analysis of medical records. We selected medical records from patients meeting the inclusion and exclusion factors described above.-Without HA and without LR (1st situation);-Without HA with LR (2nd situation);-With HA without LR (3rd situation);-With HA with LR (4th situation);We collected data relative to a speech perception test of monosyllable words  in 4 situations, which were presented as followsThis specific procedure is used as routine for hearing aid selection on this institution.The different tests were applied orally by the same investigator, with an intensity of around 70/75 dB, in a well-lit room and with minimum noise. The distance between the subject and the investigator was 1 meter.Davis, Silvermann's classification (1970) was used to determine the degree of hearing impairmentResults were analyzed in a descriptive manner, according to statistical orientation.On Situation number 1 presented a higher degree of difficulty for the hearing impaired individual, since the patient was without HA and without LR.When comparing situations 1 and 2, without HA, without and with LR, it can be noticed that 100% of the patients (31 patients) scored higher using LR. The same happened when we compare the two following situations, 3 and 4, with HA, with and without LR, since 100% scored better using HA.Results such as these, are also similar between situations 1 and 3, without LR, with and without HA, it was confirmed that 100% of patients scored better using HA.However, in situations 2 and 4, where patients used lip reading both with and without using HA, 2 patients obtained the same score in both situations.And comparing the last situation with the rest, 93.5% improved using HA and LR.We will now establish comparison relationships between the different situations.When we compare the first two situations, where almost all individuals did not have HA, 97% of them obtained an improved rate on speech recognition when they did LR.Therefore, we can observe that using LR only, most hearing impaired patients can benefit from this communication strategy.Now, regarding the two following situations, 3 and 4, where all individual used HA, on situation 3 without LR and on situation 4 with LR, 100% of patients assessed obtained a better percentage on getting the words right when doing LR.This type of data is similar to those reported by Schartz et al. (2004), who stated that watching a speaker's lips helps the listener to hear better, and as a consequence, to understand betterBetween situations 1 and 3, where they did not have the help of LR, we can notice a better score on 100% of individuals assessed, when they used HA.Silva et al. (2002) described that the use of hearing aids, is important to improve the cognitive functions in older hearing impaired patientsThe same thing did not happen between situations 2 and 4, where almost every individual did LR, situation 2 was without HA and situation 4 with HA, since 6.5% obtained the same result, both with and without HA.Situations 2 and 3 called our attention, since between them 16% of individuals scored higher on the second one, where they did not have HA but did LR.It is likely that this fact happened because the test with HA was done with hearing impaired patients who had never used or tested HA previously, but knew how to do LR as a communication strategy, being the hearing stimulus on a second level.This could also be justified by the conclusion reached by the study done by Boéchat (1992), where it is said that those individuals dealing better with hearing impairment, used the communication strategies more often 2.Blamey et al. (1989) described that when hearing does not offer proper sensorial information on speech, the visual and tactile aspects can be used as additional or alternative sensorial channels, and therefore, they should be used with the aim of increasing the communication potential of those with hearing loss Hull (1992) talked about the importance of introducing older patients to hearing difficulties on a hearing rehabilitation program, where LR training would be emphasized, making up for the communicative difficulties found when they only use HA In a similar study, authors concluded that HA and LR together, offer a significant improvement in recognizing consonants. Whereas LR offers information regarding the point of articulation, HA helps with its point and manner, and with information on vowelsOther studies such as this one, were not found on compulsory literature, however, on a paper done by Mello et al. (2004), the authors reported that 100% of assessed individuals used the cognitive strategies, LR among them, both with HA and without HAMarques et al. (2004) showed on their study, that after a hearing rehabilitation program, aiming to assess the capability to integrate visual and hearing clues, through phoneme training, only one subject did not improve the speech perception of monosyllable words.With this study, we can state the importance of LR as a mechanism to facilitate speech understanding and keeping a successful conversation. From a general point of view, LR brings benefits to patient's well-being with more self-esteem and, therefore, a better social life.An important factor we should highlight is the fact that this study was carried out in a quiet and lit setting, which makes easier the use of these strategies. When the patient is in a noisy place where noise is louder than speech, the HA system may not be effective and the strategies play a significant role for communication.From the results obtained with this study one can conclude that:Most individuals obtained a better score in the speech perception test when they used lip reading, both with and without HA.The most beneficial situation for hearing impaired individuals was situation number 4, where they had HA and LR.Lip reading is an important communication strategy for hearing impaired individuals and its recommendation helps the HA fitting process.


Since 1999, more than 100 “Maisons des Adolescents” (MDAs) - “House of Adolescents” - have been developed in France. These integrated youth-friendly facilities enabled young persons to gain access to specific care. The various medical programs of MDAs depend on the priorities of local communities rather than on official regulations. Most MDAs offer the following essential services: a “Health and Prevention Space” open daily; multidisciplinary consultations; consult liaison for youths hospitalized in medical units; a home visiting service; outpatient clinic including art workshops; refresher courses for school work; peer and parent support groups. The MDAs from the start addressed an age group (young people aged 11-21 years) rather than an illness. They thus provide primary prevention for young persons according to the World Health Organization definition of health as “a state of complete physical, mental and social well-being.” The success of the MDA network is already widely acknowledged by users, professionals, and policymakers.No significant relationships.


The high prevalence of a connexin 26 gene mutation, and its easy identification have made the diagnosis possible. The most frequent gene mutation is called 35delG. The purpose of this study was to evaluate the prevalence of 35delG mutation in children submitted to cochlear implantation who had severe and profound hearing loss previously diagnosed as idiopathic.The study was done at the Cochlear Implantation Clinic of the Otolaryngology Department and at the Laboratório Genética Humana-CBMEG, UNICAMP-SP. 32 children with severe to profound sensorineural hearing loss were evaluated. The detection of the 35delG mutation was made by a allele - specific PCR, using primers and polymerase chain reaction.69% had a normal exam, 12% were homozygous for the mutation, 19% of the cases were heterozygous. The 35delG mutation in heterozygousity is not a cause of hearing loss.The data confirm the high prevalence of the 35delG mutation in nonsyndromic bilateral profound sensorineural hearing loss. It was also possible to diagnose the cause of hearing loss as genetic in a significant percentage of patients. That stresses the importance of the molecular investigation in those cases formerly classified as idiopathic. In developed countries, it is estimated that 60% of the cases of prelingual deafness have genetic causes, and the remaining 40% have the most varied etiologies . Moreover, 1/1000 children become deaf after learning to speak, before coming of adult age; between 30-50 years of age, 0.3% of the population report hearing loss above 65 decibels (dB) and 2.3% of the population have it between 60 and 70 years of age. The prevalence keeps growing and continues to increase, reaching 50% in 80 year old patients .In Brazil, environmental factors (67%) are among the major causes of hearing loss, mostly congenital rubella and neonatal anoxia (33.5%), followed by those of unknown etiology (18.5%) and those of recessive autosomal heritage (15.5%). With the major environmental factors, high deaf risk neonates are those with a 2-5% chance of developing hearing loss if they suffer one or more of the following factors: asphyxia or anoxia, with Apgar score below 7,1; bacterial meningitis, specially caused by Haemophilus influenza; prenatal congenital infections ; head and neck malformations; high bilirubin levels (levels that require transfusion); family history of hearing loss; neonatal ICU stay for more than 48 hours; birth weight below 1.5 kilogram and use of ototoxic drugs by the pregnant mother or during the neonatal period .Cases of late hearing loss, that being, postlingual, may also occur after exposure to ototoxic drugs, family history of hearing loss, and in some cases it may be a sudden hearing loss of unknown origin, thus being considered idiopathic.Although genetic therapy is still far from practical use, cochlear implants are a feasible resource, and one available for bilateral severe/or profound hearing loss for those who do not benefit from individual sound amplification devices (hearing aids).In those cases of patients who lost hearing before learning to speak , age is one of the most important factors. In very young patients, results are better than in older children. Usually the cause of this early hearing loss is congenital, of genetic origin, without physical injury to the hearing apparatus, and this favors the cochlear implant.The 35delG mutation on the connexin 26 gene is not rare; and quite the contrary, its presence in heterozygosis may be found in up to 3% of the individuals of some populations. Notwithstanding, in Italy, its prevalence is around 1:32, in Portugal it is of 1:40, and in Spain it is 1:45. If we group together these three European populations, of which a great part of the Brazilian population descend, we can see that the average frequence of heterozygosis for the 35delG mutation is of 1:42; considering a random union of heterozygosis and the 25% chance of affected descendents, we know that in these regions 1 in 5,069 children would be born deaf because of the 35deIG mutation homozygosis .Recent progresses in molecular biology, with the finding of different genes involved in hearing loss, allow the possibility of identifying the hearing loss etiology. The high prevalence of mutations on the GJB2 gene and the ease of study allow the diagnosis of many a patient, and have suggested that these individuals would be potential candidates to receive a cochlear implant.The most frequent mutation in this gene is the one called 35delG, the one most involved in cases of hearing loss of genetic etiology.This mutation is the loss of a guanine base in the DNA sequence of the gene on position 35. It is very important to search for the 35delG mutation in the hearing loss etiology, since 2 to 4% of the individuals are carriers of this mutation, in other words, they are heterozygotes. The 35delG mutation corresponds from 75 to 80% of the possible mutations found in this gene .It is necessary for the individual to inherit two mutated genes, one from the father and the other from the mother, for deafness to set in. This way, it is impossible for connexin 26 to be codified by the altered GJB2 gene. When the patient presents a 35delG mutation in heterozygosis, it means that there is mutation in only one of the alleles, and thus it is possible for the other allele to codify the protein. This implies in a smaller number of codified connexin 26.The connexin 26 protein is indispensable to the normal functioning of the inner ear, and the gene alterations responsible for its coding is the main cause of hereditary, non-syndromical prelingual deafness, identified by the 35delG mutation study through the “genetic hearing loss test”.Some studies have concluded that the cochlear implant in patients with GJB2 gene-related hearing loss allows them to acquire voice recognition equal to or better when compared to prelingual deaf children, with deafness of unclear origin, or even with congenital hearing loss, such as those caused by cytomegalovirus . This is so, because when the damage occurs to the gene, its expression does not interfere in the placement of the implant, in other words, the cochlea is not involved, such is the case of traumatic or infectious hearing loss, from meningitis or ototoxicity, which damages the cochlear physical structure, often times preventing a larger number of electrodes to be inserted because of cochlear calcification, for instance. Genetic damage happens to the connexons structure, impairing intercellular communication, and this does not harm the cochlear physical structure, favoring the implant.The feasibility and the benefits of tracking the connexin 26 gene mutations are being reflected on public health. The use of molecular tests, together with audiometric tests will help in the early detection of hearing loss, which is extremely important in managing these patients, specially in cases of progressive hearing loss, since language development in its critical period makes the children learn to communicate before the hearing loss becomes more severe. Moreover, today it is possible to make a predictive diagnosis in those individuals affected by mutations in the connexin 26 gene, even before hearing loss sets in. The consequences of this prediction in both the social and family environment are deafness prevention or to help reduce costs assigned to the special education of such individuals, its medical management and professional decision .The goal of the present work was to check the incidence of 35delG mutation in children in need of and who received cochlear implants and had their hearing loss diagnosed as supposedly idiopathic.This study was carried out in the otorhinolaryngology outpatient ward of the University Hospital - UNICAMP-SP, cochlear implant and human genetics lab departments CBMEG, UNICAMP-SP.32 children candidates to cochlear implantation, with sensorineural, severe to profound, supposedly idiopathic, bilateral hearing loss. The major inclusion criteria of these children in the cochlear implant department are the non-benefit of individual sound amplification device (hearing aids). These individuals were submitted to a multiprofessional assessment, including otorhinolaryngologists, speech therapists, psychologists, social workers, nurses and geneticists.The 35delG mutation tracking was carried out based on peripheral blood DNA colected in a purple lid Vacutainer. In order to detect the 35delG mutation we used the allele specific PCR method (AS-PCR), using primers and the aforementioned polymerase chain reaction - modification patented by CBMEG - UNICAMP. . This method easily differentiates the normal allele from the mutant and through two reactions it is possible to distinguish normal homozygotes, 35delG homozygote and heterozygote carriers of the 35delG mutation.The molecular analysis results were correlated with the hearing loss etiology presented by the patients analyzed.In the present study, 32 children with idiopathic hearing loss underwent GJB2 gene 35delG mutation tracking. The mutation was found in 4 homozygotes of the total children assessed (12% of the cases), this being the hearing loss cause. The mutation was found in heterozygosis in 6 children (19% of the cases); It was not possible to diagnose the hearing loss in these patients. The 35delG heterozygosis mutation does not diagnose the hearing loss cause, it only proves the patient is a carrier of the mutation and, as shown in the literature, heterozygote patients can always hear. In the graph below, we can see the results obtained from the genetic study.With the progress of research in this area, it is clear the importance of gjb2 gene mutation studies due to the easy detection of mutations in the conexine 26, specially mutation on 35delg - considered the most frequent mutation of any gene already studied in caucasoids. This is the first gene indicated for molecular analysis in families with sensorineural hearing loss .In a prior study carried out in a sample of the Brazilian population, mutations to the GJB2 gene were found in 22% of the families with non-syndromical sensorineural hearing loss, once again pointing out that molecular analysis of this gene in patients with non-syndromical hearing loss should be the first step in determining hearing loss causes in our country . This is particularly true for cases of family hearing loss, among which the frequence of mutations in this gene was found to be of 50%, but also for sporadic cases, among which the frequence was a bit higher than 11% (approximately 1:9) . In Brazil, the prevalence of 0.97% of 35delG mutation carriers was found to be approximately 1:103 heterozigotes, in a survey carried out with 620 newborns in the region of Campinas-SP .Our results are in agreement with studies carried out and described in the literature for different populations. The relative distribution of the 35delG mutation for non-syndromical hearing loss in these populations varied from 0%  to 70% , showing the genetic heterogeneity among the different countries, despite the fact that some of these studies were based on a small number of patients and the investigation criteria regarding mutation tracking methods were different as well.Data obtained in the present study confirmed the high prevalence of GJB2 gene 35delG mutation in cases of profound and bilateral non-syndromical sensorineural hearing loss. It is also possible to use genetics to diagnose the cause of hearing loss in a significant number of children.The results stress the importance of molecular studies in patients with supposedly idiopathic hearing loss, since this test allows us to clarify the hearing loss etiology when there is a positive result.


Mucormycosis is a life-threatening fungal infection caused by mucormycetes, fungi of the Mucorales order.4The endoscopic appearance of gastric mucormycosis is usually a large ulcer with necrosis, eventually presenting an adherent, thick, green exudate.,4,Clinical diagnosis of GI mucormycosis is challenging. It can present an array of nonspecific symptoms, including discomfort, diarrhea, fever, gastrointestinal bleeding, necrosis, perforation, and as a necrotizing enterocolitis in premature neonates.5The reported images belong to a 48-year-old man who died from septicemia seven days after a road traffic accident. At autopsy, apart from findings of traumatic injuries, the stomach showed multiple rounds to oval lesions with sizes ranging from 0.5cm X 0.5 cm to 3cm X 2 cm, over the pylorus with elevated margins and necrotic base . On the The microscopic examination showed that the ulcers were extending to the muscularis propria. They were invading underlying arteries and veins in the submucosa, indicating angioinvasion . Gomori In conclusion, gastric mucormycosis cases usually go unnoticed clinically unless it becomes symptomatic or diagnosed incidentally on endoscopy. Thus, such cases are being diagnosed at autopsy.


Congenital fixation of the malleus and incus is an uncommon cause of conductive hearing loss. Congenital malleus bar is a term coined by Nomura et al.A 16 year-old boy presented with right-sided hearing loss that had been present since childhood. He had no history of ear disease such as otitis media. Otoendoscopic examination showed that the external auditory canal and tympanic membrane were normal. However, a white band-like mass mimicking myringosclerosis was noted in the right tympanic membrane . His resThe many different types of congenital ossicular anomalies can be broadly divided into major and minor ear anomalies. Minor anomalies are restricted to the middle ear, whereas major anomalies can involve the middle ear, external meatus, and auricle.The pathogenesis of malleus fixation is not completely understood. It may be caused by tympanosclerosis, chronic infection, trauma, otosclerosis, or Paget's disease, or it may be congenital or idiopathic.Drilling of the bar may induce tinnitus and sensorineural hearing loss by direct transmission of sound energy into the cochlea. Unlike drilling, curettage avoids the risk of tinnitus and sensorineural hearing loss. In the present study, we removed the malleus bar by using a microcurette at the junction between the bony annulus and malleus bar. Before removing the bar, dexamethasone was injected intraoperatively to prevent noise trauma. Intraoperative dexamethasone administration can also protect the inner ear.Congenital malleus bar is rare. In this case, partial removal of the bar was performed using a microcurette and intraoperative dexamethasone injection. This method prevented sensorineural hearing loss and tinnitus.The authors declare no conflicts of interest.


Beauveria bassiana (strain SB010) was exposed to spaceflight (ChangZheng 5 space shuttle during 5 May 2020 to 8 May 2020) as a part of the Key Research and Development Program of Guangdong Province, China, in collaboration with the China Space Program. The study revealed significant differences between the secondary metabolite profiles of the wild isolate (SB010) and the spaceflight-exposed isolate  of B. bassiana. Some of the secondary metabolites/toxins, including enniatin A2, brevianamide F, macrosporin, aphidicolin, and diacetoxyscirpenol, were only produced by the spaceflight-exposed isolate . The study revealed increased insecticidal activities for of crude protein extracts of B. bassiana spaceflight mutants  against Megalurothrips usitatus 5 days post application when compared crude protein extracts of the wild isolate (SB010). The data obtained support the idea of using space mutation as a tool for development/screening of fungal strains producing higher quantities of secondary metabolites, ultimately leading to increased toxicity/virulence against the target insect host.Studies on microorganism response spaceflight date back to 1960. However, nothing conclusive is known concerning the effects of spaceflight on virulence and environmental tolerance of entomopathogenic fungi; thus, this area of research remains open to further exploration. In this study, the entomopathogenic fungus  An increase in space exploration has resulted in an increase in studies on understanding the changes in physiology of living organisms under spaceflight conditions . SpaceflStreptomycetes, Escherichia coli, and Bacillus) and are mostly limited to one or a few metabolites per study. These studies have suggested altered secondary metabolite production levels, but the specific responses have been unique to each species. Furthermore, previous studies have been either limited to already-known metabolites or have focused on microorganisms that are already well-known metabolite producers. However, space vehicles hold diverse species whose behaviors are unstudied and could have responses under microgravity beyond prediction. Additionally, understanding microbes at a global metabolomics level could provide more comprehensive knowledge about the overall responses exhibited under microgravity.Metabolism is the sum of all reactions in a living cell required for maintenance, development, and division. Microbial metabolism is comprised of primary metabolites (the intracellular molecules that enable growth and proliferation) and secondary metabolites (predominantly extracellular molecules that facilitate a microbe’s interaction and adaptation with its environment) ,11. SecoBeauveria  are one of the most common insect pathogenic fungal species. Beauveria species are known to cause widespread epizootics of insect populations because of their saprophytic behavior [Filamentous fungi are a major group of microorganisms critical to the production of different commercial enzymes, biopesticides, and organic compounds ,20. Sevebehavior . Studiesbehavior ,24,25,26Beauveria bassiana (SB010) was sent to the Taingong space station for exposure to spaceflight conditions on the ChangZheng 5 space shuttle during 5 May 2020 to 8 May 2020 as a part of the Key Research and Development Program of Guangdong Province, China, in collaboration with the China Space Program. The aims of this study were to (i) extract and characterize the changes in mycelial protein extract/secondary metabolites profiles of different B. bassiana spaceflight mutants and (ii) undertake toxicity assays of mycelial extract/secondary metabolites B. bassiana spaceflight mutants against Megalurothrips usitatus (Thysanoptera: Thripidae).In this study, the entomopathogenic fungus B. bassiana. The total protein concentration of ethyl acetate extract differed significantly among all four isolates . The highest protein contents were obtained from the spaceflight mutant BHT021, with a mean value of 1.87 ± 0.035 mg/mL. For the wild isolate (SB010), the concentration of protein observed was 1.26 ± 0.03 mg/mL. The lowest protein content was produced by the space mutant BHT098, with a mean value of 1.05 ± 0.05 mg/mL (Secondary metabolite production was quantified from the ethyl acetate extracts of wild isolate (SB010) and spaceflight mutants  of 05 mg/mL .As determined by LC-MS analysis, several differences were observed between the metabolite profiles of the four isolates . A sectiB. bassiana wild isolate (SB010) and spaceflight mutants  showed marked variations for functional groups of secondary metabolites (−1 (due to overlap of O-H or N-H stretching) along with sharp peaks at 2924.24 cm−1 , methylene functional group), 1647.51 cm−1 (carbonyl stretch amide linkage), and 641.42 cm−1 (aromatics).The FTIR analysis of ethyl acetate extracts of abolites . The FTIB. bassiana spaceflight mutant (BHT021), the O-H or N-H stretching was observed at 3308.91 cm−1. The methylene (C-H) peak was observed at 2955.88 cm−1, whereas amide linkage peaks were observed at 1649.24 and 1623.96 cm−1, respectively. Furthermore, four sharp peaks of ester (C-O) functional group were observed at 1385.16, 1329.16, 1298.90, and 1241.14 cm−1, respectively. Five strong peaks representing the presence of aromatics were observed between 766.32–559.74 cm−1.In the case of B. bassiana spaceflight mutant (SB030) showed a broad peak at 3311.40 cm−1 (due to overlap of O-H or N-H stretching) along with peaks at 2924.24 cm−1 , methylene functional group), 1648.98 cm−1 (carbonyl stretch amide linkage). Furthermore, four sharp peaks of ester (C-O) functional group were observed at 1384.34, 1330.16, 1306.46 and 1241.58 cm−1, respectively. Five strong peaks representing the presence of aromatics were observed between 766.66–559.74 cm−1 (The FTIR profiles of .74 cm−1 .B. bassiana spaceflight mutant (BHT098), the O-H or N-H stretching was observed at 34.44.52 cm−1. The methylene (C-H) peak was observed at 2924.49 cm−1, whereas amide linkage peaks were observed at 1647.75 cm−1, respectively. Furthermore, a single sharp peak of ester (C-O) functional group was observed at 138,400 cm−1. Two strong peaks representing the presence of aromatics were observed at 637.34 and 562.01 cm−1 (In the case of .01 cm−1 .B. bassiana spaceflight mutants when compared to the FTIR profile of B. bassiana wild isolate (SB010).The results showed the presence of additional ester (C-O) and aromatic functional groups in ethyl acetate extracts of the B. bassiana wild isolate SB010 through NMR analysis revealed bands of high resonance in allylic, pyrrolidine ring and NeCH (2.0–2.6 ppm and 2.7–3.5 ppm), and ether linkage (3.5–4.3 ppm) regions. The NMR spectrograms of metabolic compounds produced by B. bassiana space mutants  showed high-resonance bands in aliphatic (0.9–1.4 ppm), allylic, pyrrolidine ring (2.0–2.6 ppm), NeCH (2.1–3.5 ppm), and ether linkage (3.5–4.3 ppm) regions (The characterization of protein extracts of  regions .B. bassiana wild isolate (SB010) and space mutants  applied at different concentrations against M. usitatus adults differed significantly among different isolates and their concentrations . The different concentrations of the crude protein extracts of B. bassiana spaceflight mutants (BHT021 and BHT030) had higher efficacy against M. usitatus adults 5 days post application compared to the wild isolate (SB010) (M. usitatus adults following treatment with different concentration of the crude protein extracts of spaceflight mutants (BHT098) were lower than the mortality values observed for the crude protein extracts of wild isolate (SB010).The insecticidal activities of crude protein extracts from  (SB010) . The obsM. usitatus treated with mycelial extracts of space mutants (BHT012 and BHT030) were sparse, disorganized, and vacuolated, while some of them were even completely lost. The nuclei of the blank group were flat, but after treatment with mycelial extracts of space mutants (BHT021 and BHT030), the fat body cells of M. usitatus adults were severely damaged, and their nuclei appeared expanded, folded, and detached, and the lipid droplet membrane structure became transparent and vacuolated  ,28. The m et al. , who obso et al.  observed X and Z . Similarcontrols . All theB. bassiana in response to spaceflight exposure can be strain, growth media, and secondary metabolism production pathway, depicting an inconsistent trend. These changes are induced by the indirect physical influences of spaceflight (such as variations in fluid dynamics or the extracellular transport of metabolites) [B. bassiana space flight mutants went through same sort of extracellular stress, the amount and number of secondary metabolites produced by each mutant were different. In short, the changes in secondary metabolites production in B. bassiana following spaceflight exposure were induced by the variations in micro-environments around the fungal cells [Based on already-reported data and the results of this study as well as complexity of regulatory mechanisms of secondary metabolites production, the observed changes in secondary metabolite profiles of bolites) . Furtherbolites) . Therefoal cells . Howeveral cells , and theB. bassiana wild isolate SB010 and space mutants , applied at different concentrations against M. usitatus adults, showed significantly different rates of insect mortality. The crude protein extracts of B. bassiana space mutants BHT021 and BHT030 (used at 100 μg/mL) induced higher mortality of M. usitatus adults when compared with the mortality caused by B. bassiana wild isolate SB010. On the other hand, M. usitatus mortality observed in response to treatment with crude protein extracts of B. bassiana space mutant BHT098 was lower than mortality caused by B. bassiana wild isolate SB010. Our results are similar to the already-reported previous studies on changes to microbial pathogens  induced by spaceflight [M. usitatus adults showed nuclei enlargement, folding of nuclear membrane, and detachment of microvilli from midgut cells. The secretion of secondary metabolites is important for pathogenic processes, and variations in secondary metabolite profiles of spaceflight mutants compared to the wild isolate may have led to variations in biological activities of crude protein extracts against M. usitatus. These data further explain that increased secondary metabolism can increase the virulence of microorganisms [The examination of the biological activities of fungal crude protein extracts from ceflight . The traB. bassiana, which can serve as a baseline information for the studies on the effects of microgravity on insect pathogenic fungi. However, the specific reasons or mechanisms regulating the above-mentioned changes are unclear. Therefore, further studies on corresponding gene expression/regulation and characterization of micro-environments around the fungal cells should be emphasized in near future for identification as well as application of secondary metabolites produced by B. bassiana.The results obtained from our studies provide substantial evidence that spaceflight exposure can alter the secondary metabolites production and biological activities of M. usitatus adults were reared on cowpea pods following the method of Espinosa et al. [M. usitatus adult females were used in subsequent studies.a et al.  and Du ea et al.  for multBeauveria bassiana isolate SB010  was used for the study. The fugal isolate was cultured on potato dextrose agar (PDA) plates for 15 d followed by preparation of a basal conidial concentration (1 × 108 conidia/mL) using the method of Ali et al. [i et al. .B. bassiana conidial suspension (1 × 106 conidia/mL) grown on PDA broth was individually loaded into polypropylene (PE) centrifuge tubes, which were then sealed with parafilm M . Four PE centrifuge tubes were exposed to simulated microgravity in a 3D rotating experimental device  for 72 h at Shenzhou Space Biology Science and Technology Corporation, Ltd., Beijing, China.One milliliter of B. bassiana conidial suspension were exposed to spaceflight conditions by the following method. The PE tube samples were pooled and placed into experimental boxes. The experimental boxes were placed in the ChangZheng 5 space shuttle. The samples were then flown to space within the shuttle during 5 May 2020 to 8 May 2020. The samples stayed in higher earth orbit  for 67 h and faced the Van Allen radiation belt (high-energy particle radiation belt) several times during the spaceflight. The sample box was retrieved from the returning space capsule and opened after 10 days, with the PE tubes being stored at −20 °C until further use.Polypropylene centrifuge tubes (4 tubes) having B. bassiana conidial suspension exposed to spaceflight conditions were cultured on PDA plates (15 plates each) following the method of Zhao et al. [B. bassiana conidial suspensions were selected and named as BHT021, BHT030, and BHT098.The fugal isolate was cultured on PDA plates for 15 d followed by preparation of a basal conidial concentration (1 × 108 conidia/mL) using the method of Ali et al. [Aliquots (200 µL) of o et al. . The fasi et al. .2PO 0.39 g, Na2HPO4·12H2O 1.42 g, NH4NO3 0.70 g, and KCl 1.00 g), added to Erlenmeyer flasks (250 mL), was inoculated with 5 mL of conidial suspension (1 × 108 conidia/mL) of wild isolate (SB010) and spaceflight mutants  of B. bassiana followed by incubation at 150 rpm and 27 °C for 5 days. After 5 days of growth, cultures were centrifuged in an Eppendorf 5804R centrifuge  at 10,000 rpm, 4 °C for 10 min, and the resultant supernatant was extracted with ethyl acetate (1: 1 v/v ratio) following Wu et al. [Sterilized growth medium, 100 mL,  analysis of obtained protein extracts was carried out by the method of Wu et al.  using LC−1 over a frequency range of 400 to 4000 cm−1. The liquid sample was loaded directly, and the spectra were recorded at room temperature.Fourier-transformed infrared spectroscopy analysis was performed by using MIR8035 FTIR spectrometer . All measurements were made at a resolution of 4 cmNuclear magnetic resonance (NMR) was performed using a Bruker advance III-HD 600 NMR spectrometer  by following the method of Wu et al. .B. bassiana against M. usitatus adult females was studied by following the method Du et al. [2O with 0.05% Tween-80 served as control. Adult females of M. usitatus  were inoculated to treated bean pods with camel hair brush, and bean pods were placed in a treated centrifuge tube. Each centrifuge tube was sealed with a cotton plug to prevent the thrips from escaping and was placed at 26 ± 1 °C, 70 ± 5% R.H., and 16:8 L:D photoperiod. The insects were observed on a daily basis for 5 days to record mortality data as outlined by Du et al. [The concentration mortality response of crude protein extracts of wild isolate (SB010) and spaceflight mutants  of u et al. . Brieflyu et al. . The treM. usitatus midgut were directly monitored at 5 days post treatment under a JEM1011 transmission electron microscope  following Du et al. [M. usitatus adults were sampled at 5 d post treatment and were dissected under stereo microscope  to obtain midgut samples. The samples were fixed overnight in 2.5% glutaraldehyde + 2% paraformaldehyde solution at 4 °C followed by rinsing with PBS buffer (0.1 M). The samples were then stained overnight with 1% uranyl acetate at 4 °C followed by dehydration with gradient concentrations of ethanol. The dehydrated tissues were embedded in silica gel blocks, and sections were cut using automatic microwave tissue processing instrument  and cryo-ultramicrotome .Changes in the appearance of the infected u et al. . The trep < 0.05). Mortality (%) data were arcsine transformed before further analysis. The mortality (%) data were subjected to ANOVA-2, and significance between means was also tested by Tukey’s HSD test (p < 0.05). SAS 9.2 was used for all statistical analysis [Data regarding total protein concentration were subjected to ANOVA-1. Means were compared by Tukey’s HSD test (analysis .


Modern medicine offers a wide spectrum of different hearing devices, and bone conduction implants can be found among them.The presentation of the outcomes of the implantation of a new active bone conduction hearing implant – the Osia®, and its comparison with the well-known passive transcutaneous system – the Baha® Attract.Eight adult patients with bilateral mixed hearing loss were randomly divided into two groups. Group 1 was implanted with the Osia®, and group 2 was implanted with the Baha® Attract. The details of the surgery were analyzed, along with the functional and audiological results.In all the cases, the surgery was successful, and the healing uneventful. In both groups, it was observed that pure tone audiometry and speech audiometry in free field improved significantly after the implantation (mean gain in pure tone audiometry for the Osia group 42.8 dB SPL and for the Baha group 38.8 dB SPL). In the Osia group, the results after the surgery were much better than with the Baha® 5 Power processor on the Softband. The patients implanted with the Osia® evaluated the quality of their hearing as being superior to those implanted with the Baha® Attract. There was an evident improvement in the abbreviated profile of hearing aid benefit questionnaire and in the speech, spatial and qualities of hearing scale for both systems. In the abbreviated profile of hearing aid benefit, changes were more evident in the Osia group .Implantation of the Osia® is an effective treatment option for the patients with bilateral mixed hearing loss. The surgery is safe but more complex and time-consuming than the Baha® Attract implantation. The preliminary audiological results as well as the overall quality of life indicate that the Osia® is a better solution than the Baha® Attract. However, future studies should be carried out to make further observations in a larger group of patients, and with longer follow-up. Modern medicine offers a wide spectrum of different hearing devices for hearing impaired patients – both hearing aids and many implantable systems. Currently, four types of hearing implants are available: cochlear, bone conduction, middle ear and auditory brainstem implants.4Bone conduction implants (BCIs) or bone-anchored hearing aids (BAHAs) are well-established solutions used in treatment of patients with unilateral and bilateral, mixed, and conductive hearing loss, as well as those with single-sided deafness. The first implantation was described by Tjellström and Granström in 1977The most popular systems are known as passive bone conduction systems, which consist of an implantable part being gradually osseointegrated with a bone, an external sound processor with a transducer, and a connecting part – the skin penetrating abutment (in percutaneous solutions) or system of magnets (in transcutaneous devices). Both systems are very helpful in the above-mentioned patients, however, they are not ideal and have some disadvantages and limitations.The percutaneous BAHA is a skin penetrating solution, which enables direct and high-quality sound transmission. However, it breaks the continuity of the skin barrier, so it requires lifelong daily hygienic care. What is more, it causes the risk of local skin complications including infections, skin overgrowth and sometimes even implant loss.The transcutaneous BAHA, in contrast to the previous one, uses magnetic attraction force, and leaves no permanent skin defect. Thus there are no hygienic problems, and the aesthetic effect is good. However, the quality of sound can be limited due to the sound attenuation by the intact skin between the magnets.In recent years, a few systems, known as active bone conduction systems, have become available.The aim of this study was to present the outcomes of the implantation of the new Osia® system , and to compare it with the well-known Baha® Attract .The prospective study was conducted between June and December 2019. Eight adult patients with bilateral mixed hearing loss, who were candidates for the bone conduction device implantation, were enrolled and randomly divided into two groups. Group 1 (n = 4) was implanted with the Osia®, and Group 2 (n = 4) was implanted with the Baha® Attract. The detailed descriptions of the patients are presented in The new active piezoelectric bone conduction system evaluated in this study is composed of implantable parts  and an external sound processor .Figure 11Surgery and healing: type of anesthesia, time of a surgery, length of incision, implant position, soft tissue reduction, bone polishing, bleeding from bone at the site of implantation, or any additional problems and difficulties during a surgery, as well as healing process, pain and numbness.2Audiological results:a)Pure tone audiometry, performed (1) with headphones (air and bone conduction), and (2) in free field: first, before the surgery with the Baha® 5 Power sound processor on the Softband, and then, after the surgery with an implanted device (the Baha® Attract or the Osia® system); free field thresholds were measured using warble tones presented from a loudspeaker that was positioned 1 m in front of a patient,b)Speech audiometry (a Polish monosyllabic word test), performed (1) with headphones, and (2) in free field: first, before the surgery, with the Baha® 5 Power sound processor on the Softband, and then, after surgery with implanted device (the Baha® Attract or the Osia® system), both in quiet and noise ,c)Direct bone conduction through an implanted device (BC in situ),d)Patients’ subjective evaluation of the quality of sounds after the implantation of a device – 4 parameters were evaluated according to a scale ranging from 1 (the worst) to 5 points (the best): sound loudness, sound distinction, hearing of one’s own voice and reverberation.3The quality of life benefits: the APHAB (the Abbreviated Profile of Hearing Aid Benefit) and the SSQ  questionnaires.The following parameters were evaluated, and might be presented as three categories:The audiological evaluation as well as the assessment of patients’ quality of life was performed 3 months after the surgery. All the audiological tests were performed with the Otometrics Madsen Astera in a soundproof room. The investigation was approved by the local Ethics Committee [decision number 42/19].In all the cases, the surgery was successful, and the healing uneventful. However, the Osia® surgery required general anesthesia, and was much more complex, in comparison to the Baha® Attract, as a longer skin incision and more bone work were needed . Three months after the surgery, the operated area was free of pain in all the cases, but in 1 Osia patient and 1 Baha patient, skin sensitivity was still limited.The details are presented in In both groups, significant improvement was observed in terms of pure tone audiometry in free field with an implanted device, in comparison to unaided hearing. The mean gain in PTA for the Osia group was 42.8 ± 4.9 dB SPL, and for the Baha group 38.8 ± 8.5 dB SPL. In both groups, the obtained results were comparable to those with the Baha® 5 Power processor on the Softband .Figure 3In comparison to the unaided situation, it was observed that speech understanding in free field, both in quiet and noise, improved significantly after the implantation of a device in both groups. For silent and loud speech (50 dB and 80 dB), both in quiet and noise, the results for the Osia and the Baha group were similar. Nevertheless, for medium speech (65 dB), the results in quiet were better for the Baha group, whereas in noise, they were better for the Osia group. Generally, after the surgery, the results for the Osia group were better, in comparison to those with Baha® 5 Power processor on the Softband. The results for the Baha group after the implantation and with the Softband were similar .Figure 4In the Osia group, for most sound frequencies, the results of the Bone Conduction (BC) in situ measurements with an implanted device were better, in comparison to the preoperative levels. In the Baha group, the respective results were similar or even worse after the surgery .Figure 5The patients evaluated four aspects of the quality of their hearing: sound loudness, sound distinctness, hearing of their own voice, and reverberation. The subjects implanted with the Osia® marked three out of four above-mentioned aspects of hearing higher, in comparison to the patients implanted with the Baha® Attract. In the case of the remaining one aspect, the results were similar for both groups. The details are presented in The number of hearing problems evaluated by the APHAB scale in different acoustical situations was significantly reduced after the implantation, for both systems. The changes were more evident in the Osia group . The details are presented in Currently, the place of bone conduction devices in hearing restoration is well-established, and the benefits of these solutions in well-qualified cases are not in dispute. However, the available systems are not flawless, and the choice of an optimal option is not usually simple.Our study showed that an Osia® surgery is safe, but more complex and much more time-consuming than a Baha® Attract surgery.What is more, we found that there is an evident audiological benefit after an Osia® implantation, and a significant improvement in pure tone audiometry and speech audiometry, as well in quiet as in noise, when compared to the unaided conditions. Moreover, the audiological gain of our patients after the Osia® implantation was compared to the performance with the Baha® 5 Power on the Softband before the surgery and we found an evident improvement in speech audiometry, both in quiet and noise. In our research, the advantages of the Osia® compared to the Baha® Attract were also discussed. We did not find any evident differences between both groups, in terms of free field pure tone audiometry or speech audiometry improvements. However, there were some differences in the age and preoperative hearing status before the surgery in our randomized groups. In comparison to the Baha group, the mean age in the Osia group was higher, and the initial hearing performance was worse. Thus, a similar hearing improvement for both devices indicates that the Osia® is a better choice. Moreover, the patients’ subjective evaluation of the quality of their hearing with both devices confirms this observation. Additional audiological evaluation of our patients showed better results of bone conduction measured with the Osia® device (BC in situ) than, the one measured preoperatively with bone headphones, which resulted from the direct stimulation of the bone without attenuation by the skin. In the Baha group, such difference was not observed, which is obvious. Moreover, according to our study, the subjective evaluation of four aspects of the quality of hearing indicates that the Osia® is a better solution than the Baha® Attract.We also observed an evident improvement in quality of life after the Osia® implantation, measured by the APHAB and the SSQ scale. The gain in the reduction of hearing problems in different acoustical situations, evaluated by the first questionnaire, was more evident in the Osia group than in the Baha group, and the results of the second questionnaire were slightly better for the Baha group.Our studies have some limitations, such as a small number of implanted patients, short time of observation, and the differences in age and preoperative hearing status before the surgery in our randomized groups.The Osia®, a new active bone conduction implant, is an effective treatment option for the patients with bilateral mixed hearing loss. The surgery is safe, but more complex and time-consuming than the Baha® Attract implantation. In comparison to the Baha® Attract, the early audiological and quality of life results, observed in our small groups of patients, indicate that the Osia® is a better solution. The exceptionally noteworthy results are: significant improvement in speech understanding after the Osia® implantation, in comparison to the Baha® 5 Power on the Softband, the subjective patients’ evaluation of the quality of hearing, and the obtained gain in the reduction of hearing problems in different acoustic situations. Nevertheless, further observations should be carried out in groups of cases, and with longer follow-up.The study was not supported by Cochlear Ltd.The authors declare no conflicts of interest.


As a result, HGTree v2.0 has a set of expanded data results of 6 361 199 putative horizontally transferred genes integrated with additional functional information such as the KEGG pathway, virulence factors and antimicrobial resistance. Furthermore, various visualization tools in the HGTree v2.0 database website provide intuitive biological insights, allowing the users to investigate their genomes of interest.HGTree is a database that provides horizontal gene transfer (HGT) event information on 2472 prokaryote genomes using the tree-reconciliation method. HGTree was constructed in 2015, and a large number of prokaryotic genomes have been additionally published since then. To cope with the rapid rise of prokaryotic genome data, we present HGTree v2.0 ( Genetic materials are often inherited from parents to their offspring vertically, but for prokaryotes such as bacteria or archaea , the inheritance can be demonstrated horizontally during species interactions, from one adjacent organism to another, for their evolutionary benefit . The asehttps://hgtree.snu.ac.kr)   in 2015.u.ac.kr)  or DarkHu.ac.kr)  utilize u.ac.kr) ,10. and u.ac.kr) . As a reSince 2015, the amount of prokaryotic genome data has increased dramatically, and so has the requirement for a more comprehensive HGT database with more reliable and efficient processes to detect HGT events. After the publication of HGTree, new methods to detect HGT events have been continuously developed. ShadowCaster hybridized the implicit method with the support-vector-machine (SVM) method , and tooTherefore, we present a newly updated HGTree database in response to these demands, coping with the growing interest in HGT. The updated HGTree (referred to as HGTree v2.0) includes approximately eight times larger genomes than the previous version, yielding approximately over six times more putative HGT event results. The newly revised HGT events detection procedure ensured increased detection reliability, and the detected events are presented in the HGTree v2.0 website equipped with more user-friendly interfaces, including the modified user query processor, which allows users to navigate their genomes.https://www.ncbi.nlm.nih.gov/assembly; May 5, 2021) (https://gtdb.ecogenomic.org/)  . A totalic.org/) .–6 and the minimum identity score was 98%. To retrieve 16S rRNA, RNAmmer (ver. 1.2)   was usedSaccharomyces cerevisiae for the outgroup.For each detected orthology group and its corresponding 16S rRNA group, CLUSTAL Omega (ver. 1.2.3)  was usedRanger-DTL 2.0  was usedhttp://www.mgc.ac.cn/VFs/main.htm) (The 20 536 genomes were scanned against PfamScan (ver. 1.6)  under thain.htm)  to classain.htm)  was condain.htm) ,30 was uain.htm)  .–6, but users can choose their parameters too. The best-hit results will be assigned to HGTree v2.0 database orthology groups to generate new orthology groups that include sequences of the query genome. Third, gene trees and corresponding species trees are generated through ClustalO and FastTree2. Finally, Ranger-DTL2 is performed to detect putative horizontally transferred genes from the input genome. In the same way as we built the HGTree v2.0 database, if users choose ‘Strict Mode’ in the second step, Ranger-DTL will run twice, once with default transfer cost and the secondly time with transfer cost of ‘3’ and ‘4’. Users will be given a final text file stating the ratio of HGT-related gene events and a list of them, separated into two sections: ‘Donated Genes’ and ‘Received Genes’. Virulence factors, antimicrobial resistance, gene names, product names and the genomes of the donors and recipients will also be included in the text file. The workflows of both database construction and the user query processor are illustrated in Figure We also modified the user query processor of the previous version, and the steps are as follows: first, Prokka ver. 1.11)  and Barr.11  and http://mariadb.org/) management system was used. The web-based user interface was generated with HTML5, django, CSS and JavaScript. DataTable (ver. 1.11.5) (https://datatables.net/) and jQuery (ver. 2.1.1) (http://jquery.com) were used to implement the user interface widgets. To generate circular phylogenetic trees, jsPhyloSVG-1.55 (The MariaDB (ver. 10.5.8) , constructed by aligning the same number of orthologous gene groups, were used to calculate all optimal gene tree rootings to additionally produce 1 500 857 trees, which results in 4 529 646 pairs of trees altogether. The first version of Ranger-DTL  did not Actinobacteriota and Bacteriodota had a higher ratio of predicted HGT events relative to their total gene ratios. Based on this result, we also calculated the frequency of HGT events among each taxon, whether the events occurred within or between taxa  to detect 3 028 789 orthologous groups and, by using them, we predicted 6 361 199 HGT-related genes   clusters were annotated to putative horizontally transferred genes. For a further analysis of bacterial horizontally transferred genes, the current update also focused on other functional annotations such as the KEGG  pathway,In addition, the metadata of each genome were also retrieved from the NCBI database, which includes the ‘Country’, ‘Isolation Source’ and ‘Collection Date’. The metadata are displayed on the website together with the world map  and will provide further geographical insights to the users.In the current update, we revised the website interface so that users can interpret results more thoroughly and quickly. The ‘Browse’ section provides all putative HGT events and the genes of 20 536 genomes. It also displays other information such as the genomes’ taxonomy, isolation source, geographical location and the ratio of HGT-related genes in the genome (the HGT index). Users can choose a genome in the database to see its HGT-related genes  and can also view charts illustrating the ratio of other genomes’ genera that share the same transferred genes. Furthermore, users can inspect the external links connecting to the website page of Pfam, COG and KEGG database, virulence factor and antimicrobial resistance information by clicking ‘Detail’. Like version 1, the HGT relationship plot related to each genome's HGT events and their corresponding phylogenetic trees are concisely provided, along with the summary of HGT-related genomes in graphs and the world map. Lastly, with the Jbrowse program , gene clUnlike the ‘Browse’ section, the ‘Search’ section only shows HGT events in the selected genomes. Within this section, we utilized the program ‘Circos’  to visuaIn our future research, we intend to concentrate on constructing a database that can additionally accommodate data of other types of microorganisms that can demonstrate HGT, such as viruses, fungi and other eukaryotes. Future works will also involve finding a method to predict HGT-related genes at the strain level since the major drawback of our approach was that we employed 16S rRNA sequences when constructing the species phylogenetic trees. 16S rRNA gives similar  sequences for organisms which are more closely related ; thus, mIn summary, the HGTree database was updated with a more extensive number of prokaryotic datasets and predicted a much larger number of HGT-related genes than it could do before. Changes in the prediction procedures allowed more reliable detection of putative HGT-related genes. The website interfaces are now more user-friendly with various types of visualization equipment.http://hgtree2.snu.ac.kr.The data are available at


Pontastacus leptodactylus  (Decapoda:Astacidea:Astacidae) constitutes an ecologically and economically highly important species. In the present study, the mitochondrial genome of the freshwater crayfish P. leptodactylus from Greece is analyzed for the first time, using 15 newly designed primer pairs based on available sequences of closely related species. The analyzed coding part of the mitochondrial genome of P. leptodactylus consists of 15,050 base pairs including 13 protein-coding genes (PCGs), 2 ribosomal RNA gene (rRNAs), and 22 transfer RNA genes (tRNAs). These newly designed primers may be particularly useful in future studies for analyzing different mitochondrial DNA segments. Based on the entire mitochondrial genome sequence, compared to other haplotypes from related species belonging in the same family (Astacidae) available in the GenBank database, a phylogenetic tree was constructed depicting the phylogenetic relationships of P. leptodactylus. Based on the results, the genetic distance between Astacus astacus and P. leptodactylus is smaller than the genetic distance between Austropotamobius pallipes and Austropotamobius torrentium, despite the fact that the latter two are classified within the same genus, questioning the phylogenetic position of A. astacus as a different genus than P. leptodactylus. In addition, the sample from Greece seems genetically distant compared with a conspecific haplotype available in the GenBank database, possibly implying a genetic distinction of P. leptodactylus from Greece. Pontastacus leptodactylus  (Decapoda:Astacidea:Astacidae), apart from representing a keystone species operating as a benthic scavenger, is, in parallel, a food product of high economic value. Although it is characterized as least concern in IUCN [The narrow-clawed crayfish  in IUCN . Further in IUCN . The nar in IUCN ,6, as it in IUCN . AdditioAstacus astacus, P. leptodactylus, Austropotamobius torrentium) and one alien (Pacifastacus leniusculus) species. Previous studies concerning its geographic spread report that P. leptodactylus is native to the Evros region in Greece [P. leptodactylus have been carried out in new habitats where it had not been recorded in the past. Lakes Vegoritida and Volvi in north Greece are included in the above category, as P. leptodactylus was recently recorded there, probably reflecting recent translocation events [Four freshwater crayfish species are found in Greece. More specifically, there are three indigenous , 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and the control region (D-loop), which is usually an AT-rich noncoding sequence [tellites ,23,24,25tellites . The lac studies . HoweverCherax genus [DNA barcoding, especially when cytochrome oxidase 1 (cox1) is used as a reference , has raiax genus ,31. The ax genus . One majax genus . Therefoax genus .P. leptodactylus species complex will provide information to fill the existing gap and reveal more details regarding the species status after and before translocations, as well as shedding light on the investigation of existence of specific species and subspecies within this genus.Hence, taken all together, it is clear that a more robust analysis, including more mtDNA concatenated sequences or even the whole mitogenome, will provide a clearer picture regarding the phylogenetic position of the narrow-clawed crayfish inhabiting Greece. Therefore, the aim of the present study was firstly to analyze the entire mitochondrial genome of the specific species collected from Greece for the first time, in an attempt to further clarify its systematic classification. The development of a low cost and easy-to-use tool for the whole mitogenome analysis will be the first step towards future studies regarding the reconstruction of the phylogeny of the family Astacidae. More specifically, the phylogenetic investigation of P. leptodactylus from 5 crayfish individuals caught in Lake Vegoritida, northwestern Greece. The samples were immediately embedded in liquid nitrogen prior to DNA extraction. Forty (40) μg muscle tissue were obtained for DNA isolation, performed using the kit Nucleospin tissue  following the recommendations of the manufacturer. The concentration and the purity of the DNA were evaluated using a Q5000 microvolume spectrophotometer . Fifteen primer sets were designed using the Primer 3 software  [A. leptodactylus and A. astacus, respectively. Genomic DNA was extracted from the abdominal muscle tissue sample of Estonia)  accordinThe 15 mtDNA segments were amplified in 15 separate PCRs for each individual. Each 20 μL PCR reaction contained 10 μL FastGene Taq 2X Ready Mix , 0.6 μL of each primer (10 μM), 1 μL of DNA (50 ng/μL), and ultrapure water up to the final volume. The conditions of each reaction were 95 °C for 3 min, 94 °C for 30 s, annealing temperature  for 40 sThe nucleotide composition and the relative synonymous codon usage (RSCU) were determined using MEGAΧ . AT skewP. leptodactylus was conducted according to the sequence with accession number KX279349, which is deposited in the GenBank database. The preliminary annotation was conducted using MITOS, which revealed the locations of the protein-coding and rRNA genes. The start and stop codons were identified using ORF finder and Blastn of NCBI [Finally, using the MEGAX software , the res of NCBI .P. leptodactylus sample were amplified . More specifically, the base composition is found to be T: 39.1%, C: 11.3%, A: 32.3%, and G: 17.3%. Additionally, a negative AT skew (−0.095) and a positive GC skew (0.210) are observed.The base composition of the coding mitogenome of P. leptodactylus. For the tRNA and rRNA region, the length is found to be 1427 bp and 2139 bp, corresponding to 9.48% and 14.57%, respectively, of the whole coding mitogenome of narrow-clawed crayfish. The length of the 13 PCGs ranges between 159 bp (atp8) and 1734 bp (nad5), while the length of 22 tRNAs ranges between 62 bp and 70 bp. Eleven of the PCGs are initiated by a canonical ATN codon while the other two  are initiated by near-cognate GTG codon. Furthermore, ten of them terminate with a typical TAA stop codon while the other three  terminate with TTA, TAG, and ATT, respectively. The rRNA genes  are located between trnN and trnV and between trnV and trnL1, and they are 814 bp and 1379 bp in length, respectively. The D-loop region is located between trnE and trnQ and it is estimated to be between 1500 and 2000 bp in length.The PCG region is calculated to be 11,153 bp in length, which corresponds to 74.11% of the whole coding mitogenome of The average codon frequencies of the PCGs were calculated. From the results, it is shown that the RSCU values of the preference codons are all greater than 1 . FurtherP. leptodactylus sample from Greece (OQ131122) in comparison with other species  are depicted in the maximum likelihood dendrogram of P. leniusculus, A. pallipes, A. torrentium, A. leptodactylus, and A. astacus, respectively. After alignment, these haplotypes were trimmed and eventually only sequences of the same length were compared. Further, branch lengths were included in the phylogenetic tree to evaluate the differences in the lengths of the branches.The phylogenetic relationships of P. leptodactylus is a species with no clear systematic classification due to its various different morphological characteristics. Its most prevalent classification at the genus level is P. leptodactylus [dactylus . HoweverEven among the same basin, in the Caspian Sea the genetic structure of seven populations revealed significant haplotype diversity and nucleotide diversity . FurtherP. leptodactylus with other freshwater crayfish species, such as A. astacus [A. torrentium [P. leptodactylus with other freshwater species sharing common habitats can probably led to inbreeding and potential hybrids [P. leptodactylus to survive in the places where it has been transferred and established populations. From all the above, it is clear that these data from the mitochondrial genome sequence will provide useful information regarding translocations events for future biogeography analysis.The translocations that took place during the last years to new habitats were carried out by humans, where the narrow-clawed crayfish established populations. In most of these cases, there is a co-occurrence of  astacus ,43 and Arrentium ,9. There hybrids ,23. TherP. leptodactylus in Greece, as well as representing the first step towards the investigation of its phylogenetic position based on the entire mitochondrial genome. The complete coding mitogenome of P. leptodactylus was submitted to GenBank, and given the Accession number OQ131122. Furthermore, we report 15 primer pairs that can be used for both P. leptodactylus and A. astacus for further analyses of different mitochondrial segments. The majority of P. leptodactylus genetic studies, so far, are mainly focused on genetic markers such as allozyme, single mitochondrial genes, and microsatellites [The present study, to the best of our knowledge, represents the first attempt to analyze the mitochondrial genome of tellites ,24,25 anP. leptodactylus from Greece is genetically closer to A. astacus, and, indeed, has a smaller genetic distance compared to that of A. torrentium and A. pallipes, which belong to the same genus. In addition, the genetic distance between the analyzed sample and the unique sequence of P. leptodactylus submitted to GenBank (KX279349.1) is quite high, even though the individuals belong to the same species. The above observation suggests that the years after its translocation and population establishment, together with potential introgression with other species, has probably exerted an impact on the mitochondrial genome of wild P. leptodactylus individuals, leading to a distinct genetic identity after local adaptation. Of course, further research at population genetic level, including other Greek habitats where crayfish occur naturally or have been translocated, is necessary to draw conclusions regarding the genetic profile of the species and the potential existence of founder effects or cryptic species. Determination and enlightening of stock complexity at different locations is crucial for efficient species management strategies and stock assessment for aquaculture development [Astacus and Pontastacus genera, which is based either on partial genes or on morphological data. Instead, complete mitochondrial-genome-based analysis seems to question the assignment of Astacus and Pontastacus in different genera, as well as raises concerns regarding the genetic distance of introduced populations in comparison with native ones.We observed that elopment ,45. FurtP. leptodactylus in Greece is of major importance for three main reasons: firstly, for a more efficient management of bloodstocks towards restocking purposes when populations decline; secondly, as a useful tool for aquaculture, as it can be used for genetic diversity, bloodstock management, and species identification [Genetic analysis of the populations inhabiting Greece could lead to proper management in cases where the natural populations decline, omitting negative impacts in the ecosystem. Freshwater habitats, due to the existence of natural barriers, are characterized by restricted gene flow. Thus, the limited gene flow in combination with the co-occurrence of different freshwater crayfish species could result in different genetic structures as the years pass by. The improper management of translocations and restocking could lead to reduced genetic diversity and increased inbreeding levels. From all the above, it can be assumed that the genetic characterization of fication ; and thiP. leptodactylus from Greece. It should be mentioned at this point that although one P. leptodactylus mitochondrial complete genome haplotype was already available in GenBank database, to the best of our knowledge, there was no study discussing this submission. Thus, this is the first study that describes the P. leptodactylus mitochondrial genome as well. The analyzed mitogenome of P. leptodactylus consists of 15,050 base pairs including 13 protein-coding genes, 2 rRNAs, and 22 tRNAs. From the phylogenetic relationships between the species studied, it is revealed that P. leptodactylus and A. astacus are closer than A. torrentium and A. pallipes, although the latter are classified in the same genera. Furthermore, high genetic distance is observed between the sample from Greece with the sequence existing in the GenBank. Hence, this partial mitogenome, together with the primer sets we describe, can operate as important and valuable information to support ongoing comparative genomic, phylogenomics, and molecular-based breeding studies for aquaculture, conservation, and biodiversity-related studies and can be improved upon over time, with the generation of additional data. To conclude, in addition to providing a fully annotated coding mitogenome of P. leptodactylus depicting its phylogenetic position, we developed a useful set of primers for further comprehensive analysis.Here, we provide the first analysis of mitogenome of


Diabetes mellitus (DM) is a chronic degenerative disease that impairs normal insulin production and use. DM chronic auditory complications may include spiral ganglion atrophy, degeneration of the vestibulocochlear nerve myelin sheath, reduction of the number of spiral lamina nerve fibers, and thickening of the capillary walls of the stria vascularis and small arteries.This paper aims to verify the hearing thresholds of individuals with type 1 DM.Sixty patients were enrolled in this trial and divided into case and control groups featuring diabetic and non-diabetic subjects respectively. All individuals were interviewed and underwent physical examination, ENT examination, and audiometric tests.Statistically significant difference was observed in hearing thresholds of case group subjects at 250, 500, 10,000, 11,200, 12,500, 14,000 and 16,000 Hz for both ears and ear average. Case group subjects had higher likelihood of having hypacusis at any frequency regardless of ear than controls.Statistically significant differences were seen in the audiological findings of case group subjects when compared to controls. Thorough audiological examination including high frequency audiometry is required for subjects with diabetes mellitus type 1. Diabetes mellitus (DM) is a chronic disease derived from the inadequate production of insulin in the pancreas or from the ineffective use of available insulin. It is characterized by increased blood sugar levelsDM is a relevant chronic degenerative disorder. Prevalence rates vary regionally. It has been estimated that by 2025 there will be an astounding 300 million diabetic individuals in the world, twice as many as in 2000There are four main types of DM: type 1 -results from the autoimmune destruction of the pancreatic beta-cells; type 2: insulin metabolism or secretion disorder; secondary diabetes related to genetic predisposition, drug use, unknown cause; gestational diabetesDM type 2 accounts for 80% to 90% of all cases and is closely linked to obesity. DM type 1 accounts for the remaining 10% to 20% and affects 8.4:100,000 people in Brazil. Eye, kidney, cranial nerve, peripheral nerve, ear, and vascular system disorders reside among the chronic complications of diabetes mellitus. In the auditory system, DM may lead to spiral ganglion atrophy, degeneration of the myelin sheath of the vestibulocochlear nerve, reduction on the number of nerve fibers in the spiral lamina, and thickening of the capillary walls of the stria vascularis and small arteries inside the ear canalThere is some disagreement in regards to the pathological changes DM introduces to the auditory system. Various types of auditory involvement have been reported in diabetic subjects. One of them is the gradual onset of bilateral sensorineural hearing loss, involving mainly higher frequencies in elderly patients. This is similar to presbycusis, but with greater losses than expected for the age range. Other authors have reported sudden onset early sensorineural hearing loss affecting low and mid-range frequenciesThere is no consensus in the literature as to the incidence of hearing loss related to DM, as reports range from 0 to 93%Current research indicates that DM may cause hearing loss, but a firm cause-effect correlation has not been described yet. It is known that a series of variables may favor the association of both conditions, but more studies are required to establish the true role these factors play. As seen above, diabetes and hearing loss may be interdependent components, or even components of a genetic syndromeDM may be incurable, but it is a manageable condition. Therapy aims to avoid the complications consequent to the chronic evolution of DM, such as vascular, neurological, and metabolic disordersMetabolic disorders are a relevant diagnostic component in the inner ear involvement secondary to DM. More studies are required to assess the auditory signs and symptoms present in individuals with insulin-dependent DMGiven this scenario, the purpose of this paper is to verify the hearing thresholds of individuals with DM type 1.A lot has been discussed on the association between diabetes and hearing loss since it was first considered in 1857 by Jordão, but a clear relationship between these factors is yet to be established. In general terms, the literature presents contradictory information. The Framingham trial pointed to an association between glucose levels and hearing loss in women. In 1997, the central and peripheral auditory pathways of DM type 2 patients were analyzed, and it was found that the cochlear receptor is the main structure affected in diabetic patients and that central auditory pathways showed no involvementToday the association between DM and hearing loss is being given a lot of attention. Complaints related to the auditory and vestibular systems and metabolic disorders affecting glycides and lipids have been pointed out as the main etiologic factors related to hearing loss, tinnitus, and dizzinessHigher mean hearing thresholds have been reported (more hearing loss) in diabetic patients with neuropathy when compared to diabetic patients without neuropathy in all frequencies from 250 Hz to 8.000 Hz. DM-related hearing loss follows a similar pattern to that of presbycusis given the linear distribution between frequenciesin situ” (when routine workup cannot diagnose diabetes) hearing loss is usually fluctuating, as characterized in hydrops secondary to altered sodium/ potassium gradients and reduced endocochlear potentials. As the disease progresses, microangiopathy and diabetic neuropathy assist in the progression of dysacusisIt has been reported that in patients with “diabetes Dysacusis manifests gradually and progressively. It is usually a bilateral sensorineural condition that involves mainly higher frequencies. Sudden unilateral hearing loss may also occur. Diabetic angiopathy and neuropathy, either combined or separately, have been singled out as the cause for vestibulocochlear disorders as they affect the vessels in the inner ear and stria vascularisIt has been reported that insulin resistance and hyperinsulinemia increase the rate of triglyceride productionIt is known that in order for the inner ear to function properly there has to be a good balance between insulin and glucose levels. DM patients have glucose in their blood, but it cannot enter the cells of the inner ear because of the lack of insulin thus producing functional disordersThe main proposed mechanisms are the interference on nutrient transportation through thickened capillary walls, flow reductions due to narrowed vessels, and secondary degeneration of the vestibulocochlear nerve causing neuropathyAngiopathy occurs mainly in the stria vascularis and on the spiral ligament. Studies in diabetic rats suggest that hearing impairment is caused primarily by the reduction on the number of spiral ganglion cells and secondarily by edema in the stria vascularisSome authors, however, insist that hypacusis progression occurs due to central auditory pathway involvement and not because of cochlear angiopathy progressionSpiral ganglion neuron atrophy and demyelination of the vestibulocochlear nerve have been described in four diabetic patients. It was shown that demyelination is also the early injury to the peripheral nerves of extremities on diabetes and that traces of myelin metabolic disorder may have a relevant role in the pathogenesis of diabetic neuropathy. The following were observed through an optical microscope: auditory nerve demyelination by degeneration of the myelin sheath with minor axon alterations and perineurium fibrosis; severe atrophy in the spiral ganglion with cell loss in the basal and middle turn of the cochlea, and reduction on the number of nerve fibers of the spiral lamina. Other findings included reduction on the number of ganglion cells in the ventral and dorsal cochlear nuclei, minor loss of ganglion cells in the superior olivary nucleus, inferior colliculus, and medial geniculate body. No changes directly connected to DM were observed in the temporal lobe auditory centersOther authors claim that neuropathy is the primary injury in hearing loss and that PAS-positive material found on vessel walls is non-specific, as it is also found in other conditions. Twenty patients with peripheral diabetic neuropathy underwent audiometric testing and had their test results to those of 32 controls without DM. The thresholds of individuals with peripheral neuropathy were always worse than those of the controls in all frequencies. The thresholds of patients above the age of 60 were also worse in both groups. Typically, subjects were found to have progressive sensorineural hearing loss, and patients above 60 had more intense involvementGenetic syndromes have also been considered by other authors, once more cases of hypacusis have been observed in diabetic individuals with diabetic mothers in an inheritance pattern connected to mitochondrial DNAleu)This association was first alluded to in a case of DM concurrent with sensorineural hearing loss with onset at 30 years of age in maternal relatives of a German familyIn a prospective trial, Newkirk et al.The possible association with DNA mutations was first considered by Luft et al.This association was confirmed in 1986, when Petty et al.Statistics reveal that the association between diabetes and hearing loss of maternal inheritance accounts for 1.5% of all cases of diabetes in the Netherlands and Japan•Insulin-dependent diabetes: at onset diabetes may not be of the insulin-dependent type, but it usually progresses to become insulin-dependent as mitochondrial disorders alter insulin secretion by the pancreas.•Inheritance is maternal only.•Patients are thin.•Onset occurs before 40 years of age.•Patients have sensorineural hearing loss, initially at higher frequencies; the condition is progressive and recruiting, suggestive of strictly cochlear involvementThe clinical characteristics of DM related to deafness of maternal inheritance are well established:Diabetic angiopathy has been characterized by endothelial proliferation, intimal accumulation of glycoproteins, and thickening of the capillary and small vessel basement membranes. Internal auditory artery wall fibrotic thickening and lumen stenosis were also observed. Accumulation of PAS-positive substance in the internal auditory artery walls, modiolus vessels, and stria vascularis capillaries was also seenFukushima et al.Glucose metabolism significantly affects the inner ear. Both low and high sugar levels may alter inner ear function. Patients with glucose metabolism disorders may have auditory, vestibular, or mixed symptoms as seen in diabetesThe inner ear presents intense metabolic activity, but has no energy storage capability. Therefore, minor sugar level changes affect inner ear functionThe presence of severe peripheral neuropathy or retinopathy seems to increase the risk of hearing loss. However, there is no association between the duration and severity of diabetes and hearing lossSome groups are at a greater risk of having hearing loss for various reasons and predisposing factors. Among such groups are the individuals exposed to noise, patients taking ototoxic drugs, and subjects with metabolic disordersAuditory complaints vary significantly, and may range from fluctuating hypacusis to sensorineural hearing loss. Tinnitus and aural fullness may also occurIt has been found that high frequency hearing loss (similar to presbycusis) may be present in diabetic patients before they reach the age of 60. After this age, the incidence of hearing loss is not significantly different for diabetic and non-diabetic subjectsThe relevance of aerobic glucose metabolism in maintaining endolymphatic potential has been documentedHypoglycemia affects the active transportation of sodium and potassium and thus produces hydro electrolytic imbalances in the endolymph. Endolymph is similar to the intracellular medium in its make-up, as it is rich in potassium and poor in sodium. Perilymph, on the other hand, is rich in sodium and poor in potassium, and is similar to the extracellular medium. As fluid are separated by a permeable membrane, potassium tends to shift from the endolymph to the perilymph, while sodium tends to go the opposite way. This passive spontaneous mechanism would lead to high sodium levels in the endolymph and to more water shifting into this compartment, causing the onset of endolymphatic hydrops and the ensuing clinical repercussions: vertigo, tinnitus, hypacusis, and aural fullnessThe diagnosis of metabolic disorder is very relevant for ENTs and their patients, once inner ear involvement may be exacerbated of popular drugs for labyrinthine conditions such as cinnarizine and flunarizine are used, as they are vasoactive drugs that increase the consumption of glucose by nerve cells and thus strengthen metabolic disordersIn DM patients the histological alterations observed are microangiopathy and peripheral neuropathy. They impair terminal blood flow and glucose supplyin vivo and in vitro studies present data consistent with this assumption, and further suggest that the synthesis of the basement membrane and tissue regeneration are increased in diabetes while basement membrane degradation is reduced or impairedA literature review mentioned that the capillary basement membrane is made up of collagen proteins commonly found in connective and scar tissues. The synthesis of these proteins is up-regulated in response to a variety of stimuli and injuries. The authors stated that capillary basement membrane thickening associated with diabetes is considered to be a proliferative response of capillary cells to injury. Others reported that thickening can be assigned to repeated episodes of endothelial cell death and regeneration . They concluded that a wide range of The authors of a study done on the temporal bones of 32 individuals of various agesThe study confirmed non-generalized angiopathy, as certain capillary systems of the inner are preferentially involved, as is the case of the stria vascularis. Nonetheless, the authors were unable to clearly establish the chemical nature of the precipitate, as numerous substances are dyed by this method. They reported that the findings are typical of diabetes, but not specific to the disease, observing that atherosclerosis is reduced peripherally, while diabetic angiopathy is intensified in the vicinity of small vesselsSixty individuals of both genders were enrolled in this study and divided into two groups.Group 1: the case group was made up of thirty individuals with DM type 1, 17 (57%) males and 13 (43%) females with ages ranging from 18 to 55 years .Group 2: the control group included 30 non-diabetic subjects. Clinical examination and workup  were done for all individuals. None of them had a family history of DM. Seventeen (57%) were males and 13 (43%) were females with ages ranging from 18 to 55 years .Audiological tests were done to rule out individuals from either of the groups with malformations, chronic otitis media, ear wax plug, or blood in their ears.This study was approved by the Research Ethics Committee and given permit number 009/2005. Enrolled individuals signed an informed consent form, as per the guidelines and standards in effect for research using human beings.Members from both groups were first interviewed to gather relevant data on indicators over ear symptoms. Controls were also interviewed to find our whether they had cases of DM in their families.a)Welch Allyn model 20200 USA® to rule out disorders that could affect the study results.ENT assessment – otoscopic examination with device b)Conventional audiological assessment – pure-tone air conduction audiometry from 250 to 8000 Hz, and bone conduction audiometry from 500 Hz to 4000 Hz, followed by speech recognition threshold (SRT) and speech detection threshold (SDT) tests were carried out in a soundproof booth.® Interacoustics audiometer and TDH-39® earphones with thresholds in dBNA were used in the tests.An AC40c)® digital earphones with thresholds in dBNA were used in the tests.High-frequency audiological assessment -the audiometer mentioned above was used to test auditory thresholds for frequencies ranging from 9000 Hz to 16000 Hz; KOOS HV/PROThe following procedures were then carried out:High-frequency audiometry looks into frequencies ranging from 9000 to 18000 Hz and is an important test in the early detection of hearing loss at the base of the cochlear duct. Auditory involvement can be detected before the conventional frequency range is compromised, and patients taking ototoxic drugs can be monitored to prevent spiral organ deteriorationThe equipment was calibrated based on standard ISO 8253. The criteria developed by Davis and SilvermanThe results in this study are presented as mean, median, minimum, maximum values, and standard deviations (quantitative variables) or frequencies and percentages .The quantitative variables used to compare the case and control groups were treated using Student's t-test for independent variables and the Mann-Whitney U test.p < 0.05. The data sets were treated with software Statistica v.8.0.The qualitative dichotomic variables of both groups were compared using Fisher's exact test. Statistical significance was attributed when All tests had 0.05 or 5% as the threshold to test the null hypothesis. Significant values were marked with an asterisk.Following are the frequencies in which statistically significant difference was found with increased audiometric thresholds in case group subjects when compared to controls: 250, 500, 10000, 11200, 12500, 14000 and 16000 Hz for both ears and mean values for both ears; and only right ears and mean values for both ears at 9.000 Hz.The analysis described below considered hearing loss occurred when subjects had auditory involvement at any frequency or ear.The tested null hypothesis stated that the hypacusis probabilities were the same for case and control group members, versus the alternate hypothesis in which probabilities are different.p-values.The case group had a statistically higher probability of hypacusis at any frequency, regardless of tested ears or frequencies, with 23 individuals versus 12 controls.p < 0.001), as did another study from 1981 in which significant differences were found only at 2KHzTay et al.In our study, the case group had a higher probability of having hearing loss. Statistically significant differences between case and control groups were seen at 250, 500, 10000, 11200, 12500, 14000 and 16000 Hz or both ears and mean values for both ears; and only right ears and mean values for both ears at 9.000 Hz.Patients had sensorineural hearing loss, predominantly at higher frequencies. Thirty-three (37.5%) of the tested 88 ears had hearing loss; 24 (72.7%) ears had mild hearing loss and nine (27.3%) had moderate hearing loss. Twelve (27.3%) subjects had bilateral hearing lossMitochondrial deafness is a bilateral symmetrical sensorineural type of hearing loss. The degree of impairment and age of onset vary, and it may or not be associated with syndromic conditions as DMp = 0.008) higher probabilities of hypacusis were seen in the case group, with 23 patients with hearing loss against the 12 in the same condition found in the control group.Auditory acuity may be compromised in patients with dysfunctional glucose metabolism and insulin disorders regardless of their age. Marchiori & GibrinSalvenelli et al.Alvarenga et al.Maia & CamposMaia & CamposAudiological examination indicated the presence of statistically significant differences between case and control groups at 250, 500, 10000, 11200, 12500, 14000 and 16000 Hz or both ears and mean values for both ears; and only right ears and mean values for both ears at 9.000 Hz.Case group subjects were statistically more likely to have hypacusis in either ear and at all frequencies.Statistically significant differences were found between case and control groups through audiological examination. Diabetes mellitus type 1 patients must be thoroughly examined from the audiological standpoint, as they are at risk of developing hearing loss. It is recommended that high frequency audiometry be included in the patients' clinical examination protocol.