Beauveria bassiana is a soil fungus that parasitizes a large number of arthropod species, including numerous crop pests, causing white muscardine disease and is therefore used as a biological insecticide. However, some insects, such as the cabbage aphid (Brevicoryne brassicae), defend themselves chemically by sequestering dietary pro-toxins (glucosinolates) from their Brassicales host plants. Glucosinolates are accumulated by cabbage aphids and activated to form toxic isothiocyanates when under attack. While isothiocyanate formation protects aphids against most attackers, B. bassiana is still able to infect the cabbage aphid under natural conditions. We therefore investigated how this fungus is able to circumvent the chemical defense system of the cabbage aphid. Here, we describe how B. bassiana infection activates the cabbage aphid defense system, but the resulting toxins are metabolized by B. bassiana via the mercapturic acid pathway, of which the first step is catalyzed by glutathione-S-transferases of low substrate specificity. This detoxification pathway enhances B. bassiana growth when isothiocyanates are present in natural concentrations, and so appears to be an important factor in fungal parasitization of these chemically defended aphids. Multitrophic interactions involving plants, herbivores, and herbivore enemies are ubiquitous in natural and agricultural ecosystems. In these contexts, plant chemical defenses are not only used by plants for protection against herbivores and pathogens, but also traverse trophic levels and can be exploited by some herbivores as defenses against their own predators, parasitoids, and pathogens , 2. ManyBrevicoryne brassicae (Hemiptera: Aphididae), a specialist of Brassicales plants that sequesters plant GSLs. These insects ingest the plant pro-toxins via a piercing-sucking feeding mode that minimizes tissue rupture and so mostly circumvents GSL activation by the plant defense system . Th. ThB. baant Fig. . We thenhid Fig. . Hence, B. bassiana metabolizes the major GSL hydrolysis products it encounters, B. bassiana cultures were incubated with either 4MSOB-ITC (1a), A-ITC (2a), 2PE-ITC (3a) or goitrin (4b) dissolved in 0.025% ethanol in potato dextrose broth (PDB) for 24 h, and the resulting metabolites were analyzed via non-targeted metabolomics using ultra high performance liquid chromatography coupled to quadrupole time of flight mass spectrometry (UHPLC-qTOFMS). B. bassiana incubated with 0.025% ethanol in PDB was used as a negative control for metabolomic comparisons. Volcano plots of extracted LC-MS/MS features from these non-targeted analyses indicated that the major metabolites of 4MSOB-ITC (1a) produced by B. bassiana were 4MSOB-ITC-GSH (1b) (the glutathione conjugate), 4MSOB-ITC-Cys-Glu (1c) (the cysteinyl-glutamate conjugate), 4MSOB-ITC-Cys-Gly (1d) (the cysteinyl-glycine conjugate), 4MSOB-ITC-Cys (1e) (the cysteine conjugate), and 4MSOB-ITC-NAC (1f) (the N-acetyl-cysteine conjugate)  and 4MSOOB (4-methylsulfonylbutyl)-ITC (6a)  used herein , 4MTB-ITC-Cys-Glu (5c), 4MSOOB-GSH (6b), and 4MSOOB-Cys (6e) , A-ITC-Cys-Glu (2c), A-ITC-Cys (2e), and A-ITC-NAC (2f) conjugates were detected as products of A-ITC (2a) metabolism , 2PE-ITC-Cys-Glu (3c), 2PE-ITC-Cys-Gly (3d), 2PE-ITC-Cys (3e), and 2PE-ITC-NAC (3f) conjugates were the main products of 2PE-ITC (3a) metabolism by B. bassiana , no signals corresponding to a potential GSH conjugate or its derivatives could be observed, and no signals were present that suggested goitrin (4b) metabolism  Fig. . Identifein Fig. ), were a6e) Fig. . Similarism Fig. ; and 2PEana Fig. . Howeverism Fig. . The strB. bassiana appears to metabolize most ITCs via the general mercapturic acid pathway. This pathway involves the conjugation of the toxin to glutathione (GSH), followed by sequential hydrolysis of the amino acid constituents of GSH to form Cys-Glu, Cys-Gly, and Cys conjugates, and finally acetylation to give the NAC conjugate  metabolism by B. bassiana in more detail, fungal cultures were incubated with 100 µM or 400 µM 4MSOB-ITC (1a) in PDB medium for 4 h, 12 h, and 24 h, and 4MSOB-ITC (1a) metabolites were analyzed via targeted metabolomics using UHPLC coupled to a triple-quadrupole MS. B. bassiana absorbed 4MSOB-ITC (1a) from the PDB medium into their hyphae and blastospores, and the concentrations of this compound in B. bassiana increased during the 24 h incubation  present in the surrounding PDB medium significantly decreased during this time and 4MSOB-ITC (1a) was barely detectable in the 100 µM samples after 24 h  via GSH conjugation followed by the mercapturic acid pathway, and 4MSOB-ITC conjugates were increasingly observed both in B. bassiana tissues and in the surrounding PDB medium  and 4MSOB-ITC-Cys (1e) represented over 70% of the 4MSOB-ITC conjugates formed  and 4MSOB-ITC-NAC (1f) comprised over 85% of the total 4MSOB-ITC conjugates present in the medium after 24 h  formed was converted into 4MSOB-ITC conjugates, mainly in the form of 4MSOB-ITC-Cys (1e), after five days of B. bassiana infection  are typically responsible for the first metabolic step of GSH conjugation to ITCs and other electrophiles. To elucidate the involvement of particular GSTs, all seventeen putative GST-encoding genes present in B. bassiana were cloned. Predicted BbGST proteins were classified into twelve subfamilies based on conserved domains within the amino acid sequence. Proteins were assigned to the Ure2p_like, GTT1, Sigma_like, EF1Bγ, N_3 , N_2 , N_1 , glutathionyl hydroquinone reductase (GHR), Zeta, etherase_LigE, and Kappa subfamilies as well as to a group of unclassified GSTs. They were named as BbGSTUre2p, BbGSTGTT1, BbGSTS, BbGSTEF1Bγ, BbGSTN3, BbGSTN2, BbGSTN1, BbGSTGHR, BbGSTZ, BbGSTe, BbGSTK, and BbGSTU , A-ITC (2a), and 2PE-ITC (3a)) for 4 h were compared to those from a control treatment using RT-qPCR. Consequently, we observed that expression of BbGSTUre2p2, BbGSTUre2p3, BbGSTN21, and BbGSTGHR was upregulated by all three ITCs  and 2PE-ITC (3a), but not by 50 µM 4MSOB-ITC (1a) , the expression of BbGSTU was induced after 4 h of incubation, and BbGSTe1 and BbGSTN1 transcripts were up-regulated after 12 h of incubation  Fig. . When B.ion Fig. . The resTCs Fig. , and the1a), A-ITC (2a), and 2PE-ITC (3a) were measured using in vitro incubations of His-tag purified heterologously produced enzymes, with the corresponding GSH-conjugate products being quantified by their absorption at 274 nm [1a) than A-ITC (2a) or 2PE-ITC (3a), including BbGSTUre2p1, BbGSTUre2p2, BbGSTUre2p3, BbGSTN21, and BbGSTN3 , and BbGSTe1 had higher activity with 2PE-ITC (3a) ; BbGSTU was more specific to A-ITC (2a); and BbGSTe1 was more specific to 2PE-ITC (3a)  with 4MSOB-ITC (1a) than A-ITC (2a) and 2PE-ITC (3a) (Table kcat/KM) with A-ITC (2a). BbGSTe1 had the highest Vmax and kcat/KM with 2PE-ITC (3a) of any of the enzymes, although its kcat/KM with 4MSOB-ITC (1a) was still higher than that for 2PE-ITC (3a)  Fig. . Based o3a) Fig. . In vitrB. bassiana detoxifies ITCs via conjugation to GSH, the growth of this fungus was still negatively impacted by these GSL hydrolysis products. The growth of B. bassiana was negatively correlated with the concentrations of 4MSOB-ITC (1a), A-ITC (2a), and 2PE-ITC (3a) on potato dextrose agar (PDA) plates, but was not affected by the goitrin (4b) treatment  and A-ITC (2a) than on media with 2PE-ITC (3a). B. bassiana growth was reduced by 50% at 4MSOB-ITC (1a) and A-ITC (2a) concentrations around 180 µM, while 2PE-ITC (3a) reduced B. bassiana growth by two-thirds even at 30 µM  and A-ITC (2a) treatment, but not after exposure to 2PE-ITC (3a), which reduced the contents of not only glycine, but also proline, alanine, and tryptophan, compared to the untreated control group  towards B. bassiana  exposure, but only helped the fungus very slightly in the presence of 2PE-ITC (3a)  Fig. . TherefoB. brassicae [Phyllotreta armoraciae [Harmonia axyridis [Chrysoperla carnea [Adalia bipunctata [B. bassiana, an entomopathogenic fungus often used in pest control. Using A. thaliana wild-type plants and myb28myb29 mutant plants , we determined that aliphatic GSLs negatively impact B. bassiana , 4MSOB-GSL (1), or 2PE-GSL (3), respectively, supported much lower B. bassiana growth than aphids fed on B. napus, which accumulated 2OH3But-GSL (4), a GSL that does not yield a stable ITC toxin after activation , 4MSOB-ITC (1a), and 2PE-ITC (3a), but not of the 2OH3But-GSL (4) product goitrin (4b)  Fig. , suggestal types . Our res feeding –50. For  species . In thisB. bassiana counteracts the toxicity of ITCs by metabolizing these GSL hydrolysis products. Using both non-targeted and targeted metabolomic analyses, we determined that this entomopathogen detoxifies dietary ITCs by conjugation to GSH, and can hence successfully infect even host insects that accumulate and activate GSLs. During the mercapturic acid pathway, ITCs are initially conjugated to GSH followed by the removal of Glu and Gly and N-acetylation leading to the formation of ITC-Cys-Glu, ITC-Cys-Gly, ITC-Cys, and ITC-NAC derivatives. These modifications profoundly alter the electrophilicity and polarity of ITCs, reducing their reactivity with other biological nucleophiles such as protein thiols and amines and enabling a rapid efflux from cells [Diadegma semiclausum, circumvents toxic 4MSOB-ITC (1a) in its caterpillar host P. xylostella by inducing GSL excretion [C. carnea, efficiently degrades ITCs ingested from caterpillar prey via conjugation to GSH in the mercapturic acid pathway followed by anal excretion [The fungus om cells . This paom cells  and evenom cells . Variousom cells . For exaxcretion . In contxcretion .S-transferases (GSTs) are major detoxification enzymes in both prokaryotes and eukaryotes that play a crucial role in the deactivation of several chemically reactive xenobiotics by conjugating the tripeptide GSH to electrophilic centers [Exophiala pisciphila) also induced genes from diverse classes of GSTs [Glutathione- centers . Three m centers . Moreove centers . However centers . In this centers –62 namel centers , 63–65; B. bassiana, there were differences in substrate preference for ITCs. For instance, BbGSTs from the Ure2p, N3, and N2 classes have higher activity toward the aliphatic methylsulfinyl-containing 4MSOB-ITC (1a) than other ITCs , an aliphatic ITC with an unsaturated C-C bond , a benzenic ITC, than 4MSOB-ITC (1a) and A-ITC (2a)  Fig. . In preve-chains , 67. Herr plants , and theP. xylostella were negatively correlated with the expression of its sulfatase gene and enzyme, which are responsible for GSL detoxification [BbGST genes might serve to minimize the physiological costs of detoxification in B. bassiana  concentrations (400 µM) that strongly reduce fungal growth  detoxification delayed larval development [B. bassiana to infect the cabbage aphid is supported by several lines of evidence. First, the only ITC metabolites detected in B. bassiana were intermediates and products of the mercapturic acid pathway (Fig. B. bassiana to specific ITCs (Fig. B. bassiana growth in the presence of ITCs (Fig. The detoxification of plant defense compounds by insect herbivores is often associated with energetic or metabolic costs that lead to reductions in herbivore performance. For example, growth and development of the crucifer-specialist herbivore fication . Thus, iana Fig. . For insted Fig.  at high wth Fig. . Detoxifwth Fig. . A similelopment , 68. In way Fig.  suggestiTCs Fig. , and neaTCs Fig. . Third, TCs Fig.  is consiB. bassiana, an entomopathogenic fungus frequently used in pest control. The fungus possesses a general detoxification mechanism for the major GSL hydrolysis products involving GST enzymes that may facilitate aphid infection. However, these enzymes are likely not effective at high ITC concentrations due to limitations in the supply of the GSH cosubstrate.In conclusion, our results demonstrate how plant GSL defense compounds sequestered by a specialized aphid herbivore can negatively affect Detailed materials and methods with references are described in the supplementary methods (Supplementary file Supplementary file 1Supplementary file 2Supplementary file 3Supplementary file 4Table S1Table S2Table S3Table S4Table S5Table S6Table S7Table S8Table S9Table S10Table S11


This study was to explore the relationship between fibrinogen and advanced colorectal adenoma among inpatients.From April 2015 to June 2022, 3738 participants (566 case subjects and 3172 control subjects) who underwent colonoscopies enrolled, and smooth curve fitting and logistic regression models were applied to explore the association between fibrinogen and advanced colorectal adenoma. In addition, sensitivity and subgroup analyses were performed to assess the stability of the results.Compared with lower fibrinogen quantile 1 (< 2.4 g/L), the adjusted OR values for fibrinogen and advanced colorectal adenoma in quantile 2 (2.4–2.75 g/L), quantile 3 (2.76–3.15 g/L), and quantile 4 (≥3.16 g/L) were 1.03 , 1.37 (95% CI: 1.01–1.85), and 1.43 (95% CI: 1.06–1.94), respectively. A linear relationship between fibrinogen and advanced colorectal adenoma was observed. Sensitivity and subgroup analyses showed stable results.Complements the evidence that fibrinogen was positively associated with advanced adenomas, suggesting that fibrinogen may play a role in the adenoma-carcinoma sequence. The adeSome coagulation indicators have been recognized as potential biomarkers for CRC –10. HoweFibrinogen is a glycoprotein produced in the liver . The intElectronic medical records generate a wealth of clinical data that can describe conditions in detail and are the hallmark of modern healthcare systems. Clinical data can be extracted and integrated, cleaned and transformed, and converted into a data format to create a database that can be used for disease management, early diagnosis, or treatment decisions .Studies targeting advanced colorectal adenoma are more critical to understand the adenoma-carcinoma sequence  and be b2.2.1.The present study was conducted among 3,738 consecutive inpatients who underwent colonoscopies from April 2015 to June 2022. Subjects with advanced colorectal adenoma who underwent pathological examination to confirm the diagnosis were considered eligible cases. Subjects with normal colonoscopies during the same period were considered controls. Each patient was analyzed only once.Exclusion criteria: history of colorectal surgery, an incomplete colonoscopy (no cecum reached), inadequate bowel preparation, younger than 18, incomplete medical records, history of any cancer treatment within 3 years, combined with malignancies, and fibrinogen data missing. A total of 3,738 subjects  were included. Shown in 2.2.For the current study, individuals with advanced colorectal adenoma, defined in another piece of literature , comprisPlasma fibrinogen was measured using a Mindray CX-9000 automatic coagulation analyzer  and its associated reagents. As soon as possible after collection, all blood samples were processed within 4 h. In the present study, the median time between colonoscopy and fibrinogen testing is 2  days.The baseline variables were extracted from electronic medical records, including age, sex, weight, marital status, past medical history, family history , drinking status, smoking status, and co-morbidities. Marital status is divided into three categories: married, single/divorced, and other. The history of smoking and drinking were classified into four categories, respectively: never, former, current, and NA (not recorded). A current drinker is defined as an individual who consumes any type of alcohol per week (self-reported) . An ex-s2.3.t-test or the Mann–Whitney U-test for categorical variables or continuous variables, respectively, as appropriate.Continuous variables are expressed as mean ± standard deviation or median (quartile 1–quartiles 3) values according to the normality of the distribution. Comparisons between groups were performed by the chi-squared test or Fisher exact test and Student’s We evaluated the relationship between fibrinogen and advanced colorectal adenoma using smooth curve fitting and logistic regression analysis. To evaluate the impact of fibrinogen, fibrinogen level was categorized into quartiles . We constructed three models: (i) crude model, no other covariates were adjusted; (ii) adjustment by age and sex; and (iii) adjustment by age, sex, hypertension, DM, APTT, PLT, ALP, and CREA. These potential confounders were chosen based on previous scientific literature, or a more than 10% change in effect estimates. Subgroup analyses were performed using a binary logistic regression model and then performed an interaction test. In the subgroup analyses, considering the small number of ex-smokers and ex-drinkers, we integrated ex-smokers and ex-drinkers with non-smokers and non-drinkers as a group, respectively. Multiple imputations (five replications), based on a chained equation approach method in the R mice procedure, were performed to maximize statistical power and reduce bias to account for missing data. In addition, sensitivity analysis was conducted using all complete cases.http://www.R-project.org, The R Foundation) and Free Statistics software version 1.7.1, p < 0.05 (two-tailed test) was considered statistically significant.All statistical analyses were conducted by packages R 3.3.2 . Requirements for informed consent were waived due to the retrospective nature.3.3.1.p < 0.05).A total of 3,738 subjects, including 566 subjects with advanced colorectal adenoma and 3,172 with normal colonoscopies, were included in this study. The subjects’ baseline characteristics were shown in 3.2.p = 0.834), 1.37 , and 1.43 , 1.36; 95% confidence interval (CI), 1.21–1.53]. The results were similar after adjusting for age and gender . After adjustment for sex, age, hypertension, DM, APTT, PLT, CREA, ALP, and ALB, the OR value was also stable . When fibrinogen was performed as a quartile for analysis, a positive association between them was shown even after adjustment for potential confounders. Compared to participants with low fibrinogen in quartile 1 (<2.40 g/L), the adjusted OR values for fibrinogen and advanced colorectal adenoma in quartile 2 (2.40–2.75 g/L), quartile 3 (2.76–3.15 g/L), and quartile 4 (≥3.16 g/L) were 1.03 , sex , smoking status, drinking status, history of cancer, DM, CHD, HLP, hypertension, ischemic cerebrovascular disease, significantly affected the relationship between fibrinogen and advanced colorectal adenoma . In subgp = 0.903), 1.39 , and 1.41 , 1.17 (1.02–1.35); Model II: OR (95% CI), 1.13 (0.98–1.37)]. When serum GGT as a categorized variable, compared with individuals with lower fibrinogen in quartile 1 (<2.40 g/L), the adjusted OR values for fibrinogen and advanced colorectal adenoma in quartile 2 (2.40–2.74 g/L), quartile 3 (2.75–3.15 g/L), and quartile 4 (≥3.16 g/L) were 1.02  Fibrinogen can bind growth factors , and theOur study also had several limitations. First, the present research had limitations inherent to retrospective studies. We collected the participant’s data from the laboratory information system and electronic medical records, there were missing data for some indicators. However, we performed a sensitivity analysis and found that the results were stable with or without multiple imputations of the data. Second, in a present retrospective study, we did not collect repeated measurements for fibrinogen, which may not represent their long-term levels and their association with advanced colorectal adenoma. Third, even though multivariable logistic regression, subgroup analyses, and sensitivity analysis were performed, residual confounding effects from unmeasured or unknown factors could not be completely excluded. Finally, the findings were analyzed in a single-center database from China, and the generalizability may be limited for populations with different demographic characteristics.Studying risk factors for advanced colorectal adenoma could help develop more comprehensive and non-invasive screening recommendations and possibly provide a clearer understanding of the mechanism of colon cancer. Studies on the epidemiology of advanced adenomas may be crucial in revealing why only a fraction of common adenomas progresses to CRC.5.A linear association between fibrinogen and advanced colorectal adenoma was found among inpatients. With an increase in plasma fibrinogen, the risk of advanced adenoma increased. These findings provide evidence that fibrinogen may be a potential high-risk factor for colorectal screening.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Ethics Review Board of Shijiazhuang Traditional Chinese Medicine Hospital (NO.20220919029). Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.HW and HuZ conceived and designed the study and wrote and revised the manuscript. HW, XC, PM, JL, ZW, TZ, and HaZ collected and analyzed the data. All authors contributed to the article and approved the submitted version.This work was supported by the Hebei Administration of Traditional Chinese Medicine (grant number 2023145).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


OTOF gene. In this study, the time course of changes of distortion product otoacoustic emissions (DPOAEs) has been analyzed in a cohort of patients in the light of tympanometry results.Attenuation of otoacoustic emissions over time has been reported for many patients with hearing impairment harboring mutations in the OTOF-related hearing impairment were retrospectively analyzed.The changes of DPOAEs in 16 patients with All but one subject showed DPOAEs bilaterally at the time of diagnosis. Three patients diagnosed as adults still had DPOAEs at ages of 27, 31, and 47 years, respectively. Follow-up was available for 7 children diagnosed at the age of 1 to 3 years, who still showed preservation of DPOAEs at ages of 5 to 16 years. The responses were absent or attenuated in amplitude at some follow-up appointments in association with type B or C tympanograms.OTOF-related hearing impairment. The previously reported loss of DPOAEs may have been caused in some children by increased middle ear impedance due to otitis media.DPOAEs are preserved much longer than expected in a cohort of patients with  OTOF gene (DFNB9) encoding otoferlin, a transmembrane protein involved in glutamate neurotransmitter release (Auditory neuropathy (AN) is a hearing disorder characterized by alteration of temporal coding of acoustic signals in the auditory fibers, which results in severe impairment of speech perception and the absence of auditory brainstem responses . In cont release , vesicle release , proper  release .OTOF gene account for 1.4% to 5% of cases of autosomal recessive nonsyndromic congenital hearing impairment in different populations  has not been addressed. OME, which occurs with high frequency in children aged 1 to 4 years, impacts on the amplitude of OAEs recorded in the ear canal, eventually leading to OAEs disappearance  in a group of 16 patients with hearing impairment due to mutations in the OTOF patients were extracted from their medical records maintained at the Audiology Service of “Santi Giovanni e Paolo” Hospital in Venice (in accordance with the 1964 Declaration of Helsinki).In this retrospective study, the DPOAE data from OTOF gene at each follow-up appointment throughout the last 20 years. Genetic and audiological data have been previously reported  with hearing impairment due to biallelic mutations in the reported , 2021 exThe details of genetic findings together with audiological and electrophysiological measures for all subjects are summarized in Table The degree of hearing impairment was defined by the pure-tone average (PTA) threshold levels measured at 0.5, 1, 2, and 4 kHz, and was classified as mild (PTA 21 to 40 dB HL), moderate (PTA 41 to 70 dB HL), severe (PTA 71 to 95 dB HL), and profound (PTA > 95 dB HL) . ProfounComputed tomography (CT) and magnetic resonance imaging (MRI) scans of the head and ear as well as growth and motor development were normal in all subjects. No patient had a history of sound exposure.All but 3 patients received unilateral cochlear implant in the right year, except for subject no. 15, who was implanted in the left ear because of persistent OME in the right ear. Nine patients attempted a trial with hearing aids, but only 4 subjects  underwent a course longer than 1 year. One patient (no. 3) was still wearing the hearing aid in the nonimplanted ear at the last follow-up appointment.2O in children and 0.3 to 1.4 mm H2O in adults  of 16 patients showed OME at least at one follow-up appointment, which was diagnosed based on otoscopic evaluation and tympanometry recording (Grason-Stadler GSI TympStar or Madsen Zodiac impedance audiometer). Tympanogram was indicated as type A, As, B, or C according to n adults .f2/f1 ratio was kept at 1.21. The f2 frequency was increased in 1/4 octave steps from 1.0 to 8.0 kHz (ILO-92 system) or to10.0 (ILO-292 system).DPOAEs were obtained using an ILO-92 system up to 2018  and an ILO-292 OAE  system thereafter. Primary tones were presented at 65/55 dB SPL and the f2 frequency, DPOAEs were considered present when the signal to noise ratio was higher than 3 to 6 dB  lost OAEs few months after birth. DPOAEs rates at the first and last follow-up appointments in the presence of normal tympanometry  are reported in Figure Four patients  with profound hearing loss have been following up since the age of 1 to 2 years. They had DPOAEs in both ears before cochlear implantation and still showed DPOAEs in the nonimplanted ear at the age of 4.8, 16, 12.8, and 7.3 years, respectively. It is interesting that subject no. 3 still had DPOAEs at the age of 4.8 in the nonimplanted ear where he was wearing a power hearing aid since the age of one.The children diagnosed after the age of 3  still had DPOAEs in the nonimplanted ear at age of 6.4, 10.2, and 8.3 years, respectively. It is of note that in subject no. 8, the DPOAEs responses were recorded at the age of 6.4 also in the implanted ear. Of these patients, subject no. 8 showed profound deafness, whereas subjects nos. 9 and 10 had, respectively, mild and moderate hearing loss.In all subjects who underwent an extended follow-up the hearing thresholds measured in the nonimplanted ear remained stable over time.The 3 patients  diagnosed as adults still had DPOAEs at ages of 27, 31, and 47 years, respectively. The degree of hearing impairment was mild in subjects nos. 1 and 2, and moderate in subject no. 16  at follow-up appointments during which tympanometry indicated the presence of increased middle ear impedance are displayed in Figure , 8 at fof2 frequencies, while subject no. 16 showed robust DPOAEs only at f2 frequencies lower than 2.8 kHz. This patient also had elevated hearing thresholds at high frequencies.DPOAEs collected in the 3 patients diagnosed as adults are shown in Figure In conclusion, the 10 patients diagnosed and/or followed up after the age of 3 years showed DPOAEs in both ears  or in the nonimplanted ear  at the last follow-up appointment (age range 4.8 to 47 years).OTOF-related hearing impairment. The disappearance of OAEs responses or their attenuation in amplitude in some children was associated with the coexistence of OME as indicated by recording type B or C tympanogram.The findings reported in this study indicate preservation of OAEs in a cohort of patients with OTOF gene , one lost DPOAEs by the age of one year, whereas the remaining two showed preservation of OAE responses up to the age of 12.8 and 16 years, respectively.Of the patients included in this study, subjects with profound deafness had inactivating mutations in both f2 frequencies ranging from 1500 to 5000 Hz in comparison with teens and adults. These differences are deemed to result from the developmental changes occurring in the outer and middle ear  had no DPOAEs at high OTOF patients suffering from recurrent episodes of OME could have been missed, particularly in those children for whom an extended follow-up was not available. We believe that, at least in some patients, the OAEs presence has been underestimated as the attenuation in amplitude could have resulted from an increased middle ear impedance due to OME rather than from OHC loss.In some children, DPOAEs disappeared or were markedly attenuated in amplitude in the presence of an increased middle ear impedance due to OME even in cases of a minimal decrease of the peak compliance in tympanometry. Of note, in one child (subject no. 3), DPOAE amplitudes appeared markedly increased at the age of 5 in comparison to previously recorded DPOAEs, probably because the episodes of OME had faded away. Based on these findings, it seems conceivable that in some of the previously published studies, the presence of DPOAEs in OTOF-related hearing impairment deals with the high noise levels often observed in highly active toddlers, which could mask positive OAE responses. An example is given by the DPOAEs obtained in subject no. 15, for whom only noisy recordings were available. These cases require an extended follow-up to select the data collected in the best recording condition.Another point to be addressed when investigating the changes of OAEs in OTOF gene received the bilateral cochlear implant by 1 to 2 years of age. This could have been a critical factor in preventing an extended follow-up of DPOAEs recordings in these patients. All but 2 subjects of our sample received unilateral cochlear implant, which allowed us to extend the DPOAE follow-up after cochlear implantation. It is interesting that low-amplitude DPOAEs were recorded also in the implanted ear in one subject (no. 8) at the appointments in which type A tympanogram was obtained. This finding points to preservation of OHCs, which survived to the trauma induced by the insertion of the cochlear implant array.As bilateral cochlear implantation is the standard of care for children with severe to profound hearing loss, it is likely that profoundly-deaf children harboring mutations in the When considering the hearing aid use, only 4 subjects used hearing aids for more than one year, whereas the remaining patients tended to use acoustic amplification discontinuously. It is interesting that subject no. 3, who was wearing one power hearing aid in the nonimplanted ear since the age of one, still had robust DPOAEs at the age of 5 years. Nevertheless, due to unavailability of an extended follow-up for the other patients, the effects of amplification on DPOAEs cannot be assessed.OTOF-related hearing impairment. Although other unidentified genetic or environmental modifiers may be responsible for the early OAE loss in a minority of patients, the disappearance or the decrease of OAE amplitude in some patients could have been induced by an increase of middle ear impedance due to OME or to high noise levels during OAEs recording. These findings are relevant not only from the point of view of the elucidation of the mechanisms underlying hair cell damage induced by otoferlin deficiency, but mainly because they indicate preservation of the cochlear amplifier, which is crucial for the full restoration of hearing through gene therapy administration.In conclusion, this study demonstrates that DPOAEs are preserved much longer than expected in a cohort of patients with We are greatly indebted to Drs. Alessandra Ferraboschi, Serena Scardillo, Marica Pistello who helped with data collection.


This article has been corrected.1In the Original Investigation titled “Baseline Features and Reasons for Nonparticipation in the Colonoscopy Versus Fecal Immunochemical Test in Reducing Mortality From Colorectal Cancer (CONFIRM) Study, a Colorectal Cancer Screening Trial,”


Lawsonia intracellularis, which infects the cells lining the ileum and infrequently cells in the large intestine, and it causes inflammation, hemorrhages, and thickening of the intestinal wall  prior to a combined L. intracellularis/PCV2 infection mitigated the clinical signs, lesions, and level of shedding of L. intracellularis infection. Prior PCV2 exposure also led to higher L. intracellularis antibody titers after the combined infection, which warrants further research into the cross-talk between the immune response to viral infections and L. intracellularis infection. The microbiota of the pig's ileum is known to influence the susceptibility to L. intracellularis, and the infection also adversely affects the gut microbiota . Particularly, an increase of pathobionts, including Collinsella, Campylobacter, Chlamydia, and Fusobacterium, was observed in the gut during L. intracellularis infection. Interestingly, the live attenuated L. intracellularis vaccine, in addition to decreasing the L. intracellularis shedding, significantly decreased the relative abundance of pathobionts, highlighting the role of the broader gut microbiome composition in the pathogenesis caused by L. intracellularis. Another article in this collection by Hankel et al., which is based on their analysis of fecal microbiota in pigs naturally infected with L. intracellularis that were varying in their vaccination status  and clinical conspicuity, found the influence of the microbiota on the severity of disease. The causal role of the microbiota in the pathogenesis of L. intracellularis infection is yet to be completely unraveled.Porcine proliferative enteropathy (PE) or proliferative hemorrhagic enteritis (PHE), or commonly “ileitis”, is a severe intestinal disease of domestic pigs and wild boars. It is caused by the bacterium nal wall . The disnal wall . L. intrhamsters . The etidinburgh . L. intrllularis . Now, thllularis , 5. The fections . A crossrom 2015 . The vaccrobiota . In thisL. intracellularis consists of a 1.46 Mb chromosome and three plasmids of 0.03, 0.04, and 0.19 Mb (NCBI Refseq IDs NC_020127 to NC_020130). A complete genome dataset of at least nine strains of L. intracellularis from around the world  are available in the public domain at the end of March 2023. Notably, the E40504 strain was isolated from a horse. The genome data have led to the development of several semi-quantitative and quantitative PCR assays for the diagnosis of L. intracellularis. Xiao et al. expressed the outer membrane protein (Omp2) of L. intracellularis from a synthetic gene based on sequence information. Monoclonal antibodies specific to Omp2 were produced and found to be efficient in detecting L. intracellularis in Western blots, immunohistochemistry, and immunofluorescence assays. Moreover, a comparison of the gene expression profiles of the pathogenic low-passage and non-pathogenic high-passage isolate of L. intracellularis revealed that there were only 401 genes expressed by the pathogenic bacteria , which is commonly found in pathogenic bacteria to secrete effector proteins into host cells. Through a yeast expression array expressing hypothetical genes from L. intracellularis, it has been identified that LI1035 interacts with the mitogen-activated protein kinase (MAPK) pathway and regulates actin organization in yeast . They found that L. intracellularis is endocytosed by both clathrin-dependent and -independent mechanisms, but the internalization of heat-killed bacteria is decreased in the clathrin-depleted cells. This suggests potential active mechanisms, involving a secretion apparatus, effected by L. intracellularis during endocytosis, as observed in many pathogenic intracellular bacteria. Given the wealth of knowledge on the genome of L. intracellularis, potential effector proteins or virulence factors remain to be characterized. With the current level of information on the biology of L. intracellularis, a promising outlook prevails over the efforts for a comprehensive understanding of the pathogenesis of this fastidious organism, which may require uncharacterized inciting factors to cause disease.The reference genome of rea CBNU0, and Chiammation . The stiin yeast . PereiraThe author confirms being the sole contributor of this work and has approved it for publication.


Early linguistic background, and in particular, access to language, lays the foundation of future reading skills in deaf and hard-of-hearing signers. The current study aims to estimate the impact of two factors – early access to sign and/or spoken language – on reading fluency in deaf and hard-of-hearing adult Russian Sign Language speakers.In the eye-tracking experiment, 26 deaf and 14 hard-of-hearing native Russian Sign Language speakers read 144 sentences from the Russian Sentence Corpus. Analysis of global eye-movement trajectories (scanpaths) was used to identify clusters of typical reading trajectories. The role of early access to sign and spoken language as well as vocabulary size as predictors of the more fluent reading pattern was tested.Hard-of-hearing signers with early access to sign language read more fluently than those who were exposed to sign language later in life or deaf signers without access to speech sounds. No association between early access to spoken language and reading fluency was found.Our results suggest a unique advantage for the hard-of-hearing individuals from having early access to both sign and spoken language and support the existing claims that early exposure to sign language is beneficial not only for deaf but also for hard-of-hearing children. Poorer The foundation of early language development is access to language. In deaf and hard-of-hearing people, access to language can take different paths, be that access to sign language, to spoken language, or both. The precise role of each route for reading proficiency is under debate. However, early acquisition of sign language might be not the only road to proficient reading. The first factor – early access to sign language – primarily depends on the hearing status of the child’s parents, since deaf parents tend to be signers, and hearing parents tend to either learn sign language together with their child  signers: in addition to the early access to sign language, we also consider the access to spoken language approximated by the degree of hearing loss as a factor that can potentially influence reading fluency. While early access to sign language is clearly beneficial for reading skills of deaf individuals, it is less clear what role early access to spoken and/or sign language plays for hard-of-hearing individuals with partial access to speech sounds.2.To investigate global reading fluency in DHH Russian signers, we focus not on the isolated measures related to individual word reading, such as fixation durations and skipping rates, but rather on the global trajectories of eye movements in reading the entire sentences . While tTo illustrate the concept of a scanpath, Utilizing the scanpath method to compare reading strategies in English-Russian bilingual and Russian-speaking monolingual speakers, In the present study , we expect that early exposure to any type of language (sign or spoken) should be associated with greater reading fluency in DHH signers. That is, we expect higher reading fluency in both deaf and hard-of-hearing signers who have deaf parents and, therefore, were exposed to sign language from birth, and in hard-of-hearing signers who were exposed to spoken language from birth. We hypothesize that these readers will adopt a more fluent pattern of reading compared to DHH readers with less exposure to language (sign or spoken).per se is not a direct index of reading skill or successful comprehension: A text can be skimmed fast but poorly understood  and 14 hard-of-hearing participants . The individual characteristics of each participant can be found in The study included 40 DHH signers: 26 participants with complete hearing loss  and had hereditary deafness, while 11 were born in hearing families and had hearing loss due to other causes see . Seven oThe aim of the present study is to establish whether reading fluency of DHH signers is correlated with their parents’ hearing status and the individual degree of hearing loss. The mapping from these predictors to the main factors of interest, early access to sign and spoken language, is as follows: parents’ hearing status maps directly onto early access to sign language, which may benefit both deaf and hard-of-hearing individuals. In contrast, early access to spoken language maps onto the degree of participant’s individual hearing loss with more severe loss leading to lesser spoken input the individual receives during infancy. We also hypothesize that the degree of hearing loss might interact with the parents’ hearing status: the individuals with some access to speech sounds and at least one hearing parent are likely to have more early access to spoken sounds compared to individuals with deaf primary caretakers.https://Ruscorpora.ru) and normed for acceptability. Sentences had different syntactic structures: narratives, exclamations, and interrogatives, as well as sentences with non-standard word order. Sentences spanned from five to twelve words (with the average sentence length of 9 words) and were selected for being syntactically and lexically accessible. The Russian Sentence Corpus has been successfully read by advanced L2 learners and heritage speakers of Russian :In addition, approximate vocabulary size of print Russian was measured for each participant using an online computerized adaptive testing tool . During 3.2.Stimuli were presented on the ASUS VG248QE monitor . Eye movements were recorded at the rate of 1,000 Hz with desktop eye-tracker EyeLink 1,000+ using a chinrest. Eye-to-camera distance was 60 cm, eye-to-screen distance was 90 cm.The experiment started with 9-dot camera calibration. After the calibration, a black dot was presented at the position of the first letter of the first word in the sentence. After the camera registered a fixation on the black dot, the sentence appeared. Participants were asked to read the sentence without signing (silent reading). If no fixation was registered on the black dot within 2 s, calibration was repeated. After having read the sentence, participants had to look at the red dot in the lower right corner of the screen. Fixation on the red dot triggered the next trial.If the sentence was followed by a question, then after a fixation on the red dot was detected, the question appeared in place of the sentence. The response options were presented below the question. To select an answer, participants had to click on the response. The experiment started with three practice sentences and continued with 6 blocks, 24 experimental sentences in each. Between blocks, participants could have a break followed by a recalibration. The order in which the sentences appeared was randomized.3.3.To answer the main research question of the study, i.e., whether more proficient sentence reading trajectories in DHH participants are associated with early exposure to language, sign and/or spoken, we followed the steps in analysis in To find out whether early exposure to language affects cluster placement in DHH participants, we used a generalized mixed-effects model with the cluster as a dependent variable and parents’ hearing status, participant’s degree of hearing loss as predictors. We additionally used participants’ vocabulary size, age, and gender as covariates, as these factors are known to affect reading . The modhttps://osf.io/je8du/. The readers are encouraged to reproduce our analysis and to apply any other analyses they see fit to the data set.The data and analysis code are openly available at: 4.Based on the eye-movement characteristics, the two clusters earlier identified via Gaussian mixture modeling were labeled as more fluent and less fluent reading clusters  can be found at: The studies involving humans were approved by the HSE Committee on Interuniversity Surveys and Ethical Assessment of Empirical Research. The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.AZ and AL contributed to conception and design of the study. AZ programmed the procedure and performed data collection and wrote the first draft of the manuscript. OP, AZ, and AL performed the statistical analysis. AL wrote the revised version of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.This article is an output of a research project implemented as part of the Basic Research Program at the National Research University Higher School of Economics (HSE University). AL was funded by the Deutsche Forschungsgemeinschaft , project number 317633480, SFB 1287. Funded by the Deutsche Forschungsgemeinschaft  – Projektnummer 491466077.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The IBMP concentration of grape and the expression patterns of VvOMTs in berry skin were monitored over two consecutive years. The results showed that the IBMP concentration of grape treated with DCIP was significantly increased at maturity in both years. The relative expression levels of VvOMT1 and VvOMT3 in berry skin were positively correlated with the IBMP accumulation. After DCIP incorporation, the relative expression level of VvOMT1 and particularly that of VvOMT3 were obviously up-regulated and closely mirrored the IBMP accumulation pattern in two consecutive years. Therefore, we speculate that DCIP may be a key intermediate involved in the biosynthesis of IBMP and plays an important role in regulating IBMP accumulation.The metabolic pathway of 3-alkyl-2-methoxypyrazines (MPs) in grape remains largely unclear except for the final step. In this study, the 2,5-dicarbonyl-3-isobutyl-piperazine (DCIP), which is proposed as the key intermediate of 3-isobutyl-2-methoxypyrazine (IBMP) biosynthesis, was incorporated into Cabernet Sauvignon clusters in  At −1) ,10. The −1) ,12 is thred wine ,8.−1 level [In view of the extremely low concentration of MPs in grapes and wines, one of the key points in this research field is the detection and quantitative methods developing. In recent years, many studies were involved in this field and great achievements were made, and many accurate and efficient techniques with higher sensitivity were developed . Among t−1 level ,15. And −1 level ,16,17,18Generally speaking, excessive levels of MPs are unacceptable for both the wine maker and consumer due to the fact that it will make the wine be regarded as poor quality, which is made from the unripe grape ,20. The Pseudomonas perolens, for instance; the IPMP biosynthesis begins in the condensation of valine with glycine to form the 2,5-dicarbonyl-3-isopropyl-piperazine (DCPP), and then the DCPP was converted into the 3-isopropyl-2-hydroxypyrazine (IPHP). Finally, the IPHP was methylated to IPMP.Unfortunately, the biosynthesis pathway of MPs remains largely unclear . To dateVvOMT1, VvOMT2, VvOMT3, and VvOMT4, had been identified in wine grapes [It is worth noting that the common point of the two hypotheses is both of them suggested that the final step of MPs’ biosynthesis is the HPs being methylated to MPs ,25,26, we grapes ,28,29. Te grapes , wherease grapes . Therefoe grapes ,31,32, isitu. In order to avoid the influence of year difference on the results of the field experiments, the IBMP content in grape berries and the relative expression levels of VvOMTs in berry skins were monitored during the experimental period over two consecutive years (2020–2021). The effect of DCIP on IBMP accumulation in grape berry and the relative VvOMT genes expression pattern of berry skins was evaluated. The results would further broaden the understanding of IBMP biosynthesis pathway in wine grape.Our previous study had explored the effect of the assumed precursor L-Leu and the possible key intermediate AMPA on IBMP biosynthesis in grape berry according to the “Amino acid amidation pathway” . Here, wsitu was purchased from Aozeal Certified Standards . Deuterium-labeled IBMP  used as internal standard was obtained from CDN Isotopes .Solvents and reagents were of analytical reagent grade or higher. Sodium fluoride  used as sample extraction buffer was purchased from Guangfu Technology and Development Co., Ltd. . 3-Isobutyl-2-methoxypyrazine  used as the standard was purchased from Sigma-Aldrich . 2,5-Dicarbonyl-3-isobutyl-piperazine  used as soaking incorporation solution in The experimental vineyard that belongs to the Shandong Academy of Agricultural Sciences located in Zhangqiu, Shandong, China . Eight-year-old Cabernet Sauvignon vines, which were grafted on the 1103P rootstock, were used to carry out the experiments over two consecutive years (2020 and 2021). The vineyard soil type was typical loam soil, and it was managed under the traditional management. The vines’ training method was single cordon with a vertical shoot positioning trellis system, row spacing is 2.5 m, and plant spacing is 1.0 m, approximate south-north row direction.situ, respectively at pre-veraison, i.e., 59 days after anthesis (DAA) in 2020 and 63 DAA in 2021. Three replicates were used for each treatment, with 60 clusters for each replicate. All treatments were arranged in randomized blocks in three rows.In this study, three treatments were carried out: normal growth grapes (Mock); deionized water (Control) and DCIP solutions (5 mM) were fed into the grape clusters in A soaking incorporation method that was developed by Hayasaka et al.  and ChasIn 2020, berry samples were collected at 59, 67, 72 (veraison), 88, 104, 121, 128, and 154 DAA (maturity). In 2021, berry samples were collected at 63, 69 (veraison), 76, 85, 93, 101, 118, and 153 DAA (maturity). All samples were collected randomly in triplicate at each sampling time point. Approximately 250 berries were collected per replicate and were immediately placed on ice in foam boxes that were returned to the laboratory within 2 h. Among them, 15 berries were randomly selected in each replicate for peeling berry skins using a scalpel and then frozen in liquid nitrogen immediately. The grape berry samples were stored at −40 °C, and the berry skin samples were stored at −80 °C prior to analysis.According to the method that was developed by Koch et al.  and modi−1 internal standard solution was added into the centrifuge tube. The samples were homogenized and then placed in high-speed refrigerated centrifuge  at 5000× g for 7 min at −4 °C. Next, 10.0 mL supernatant was placed into the headspace vial, which contained 3.0 g NaCl, and was put under room temperature to equilibrate for 1 h before HS-SPME-GC-MS.Approximately 30.0 g of frozen grape berries were weighed and recorded accurately. The number of grape berries in each replicate was also recorded, in order to express the IBMP content in grape berries both in nanograms per kilogram (fresh weight) and picograms per berry. The frozen berries were de-seeded with the scalpel when they had melted slightly; the flesh was ground into a fine powder in liquid nitrogen immediately and then was transferred to the centrifuge tube (50.0 mL). Then, 10 mL extraction buffer (2 mM NaF) containing 200 ng·LThe parameter setting detail of the CTC autosampler system and the GC-MS condition was same as the method we had established .VvOMT1, VvOMT3, and VvOMT4 were according to Lei et al. [VvOMT2 were as follows: the forward primer was 5′-TCCGAGAAGATGGCTATGAG-3′, and the reverse primer was 5′-CTGCAAAGTTGGAATCTTTAA-3′.Total RNA was extracted from berry skins using the General Plant Total RNA Extraction Kit . cDNA synthesis was prepared by using a Reverse Transcription Kit . The primers of i et al. . The priThe relative expression levels of VvOMTs were analyzed and calculated according to our established method . The relp ≤ 0.05 and p ≤ 0.01. Line charts were drawn by using OriginPro 9.1 . The pathway graphs were prepared using Chemdraw 20.0 software .Univariate ANOVA was conducted using SPSS 20.0 software . Significant variations were detected by Fisher’s least significant difference test (LSD) at VvOMT1, VvOMT2, VvOMT3, and VvOMT4 in berry skins  was almost the same as the natural condition (Mock) during the whole experiment period no matter if it was calculated in nanograms per kilogram or picograms per berry in two consecutive years. These results showed that the soaking incorporation treatment did not significantly affect IBMP accumulation in grape berries.−1 (−1 (−1 (−1 (The IBMP concentration in Control and Mock treatments showed a continuous downward trend during the experiment period over the 2 years (2020 and 2021). In 2020, the IBMP content peaked at 142.67 ng∙kg−1 a and 101−1  in two consecutive years. And at harvest, the IBMP concentration of grapes was all below the quantitation limit in the two consecutive years .The relative expression levels of VvOMTs were monitored in two consecutive years (2020 and 2021) to evaluate the impact of DCIP incorporation on IBMP biosynthesis.VvOMT1 in grape berry skins after DCIP incorporation showed a different trend during the experiment period for the two different years. In 2020, the relative expression level of VvOMT1 after DCIP incorporation was obviously increased at 121 and 128 DAA over the Control and Mock  for 2 consecutive years.It is noteworthy in this study that, regardless of whether the IBMP content was expressed in nanograms per kilogram or picograms per berry, the incorporation of DCIP in On the one hand, these results suggest that DCIP might participate in the biosynthesis of IBMP in grape berry. The reason why the IBMP concentration only increased at maturity compared with the Control and Mock treatments, it might be because there are series of steps between the DCIP and IBMP, and the DCIP converts into the corresponding key intermediate at a slow rate, so the IBMP concentration of grape berry would not increase immediately and showed significant increase until the later experiment period.situ, the IBMP biosynthesis process might be accelerated after veraison due to the exogenous DCIP incorporation. Thus, the IBMP content showed a significant increase compared with the Control and Mock treatments at the same time points.On the other hand, the IBMP concentration after DCIP treatment showed a significant increase at maturity  when it is generally believed that IBMP in grape berry is in a degradation stage ,37,38,39In view of the results above, the effect of DCIP on IBMP biosynthesis in grapes deserves to be further explored.The research on the biosynthesis of MPs in grapes is still insufficient knowledge, but the final step involving the HPs being methylated into the MPs had been confirmed ,25,26; eIn the grape berries, 72% of the IBMP is located in the berry skins , so the VvOMT1 was synchronized with the IBMP accumulation in grape, but in 2020, the maximum level of IBMP accumulation was 8 days earlier than the maximum VvOMT1 relative expression. After DCIP incorporation, the relative expression level of VvOMT1 in berry skin was significantly up-regulated at 121 and 128 DAA in 2020, but showed no significant difference from the Control and Mock treatments in 2021.In 2021, the relative expression level of VvOMT3 in berry skin was synchronized with IBMP accumulation in grape in two consecutive years, and the peak relative expression level of VvOMT3 was almost synchronized with the maximum IBMP level. After DCIP incorporation, the relative expression level of VvOMT3 in berry skins was remarkably increased at 67 DAA in 2020 and was significantly up-regulated from 69 to 85 DAA in 2021, which was 16–49 days earlier than the significant increase of IBMP accumulation in grape berries.The expression pattern of VvOMT1 and VvOMT3 play a significant role in regulating the IBMP accumulation; the IBMP content in grape berries was positively correlated with the up-regulated expression of VvOMT1 [VvOMT3 expression pattern was almost synchronized with the IBMP accumulation, which plays a major role in MPs biosynthesis in grape berries due to the fact that it had a high affinity for the HP precursors of MPs [These results suggested that both the f VvOMT1 ,28, but s of MPs ,41.VvOMT2 and VvOMT4 did not show a positive correlation with IBMP accumulation in grape berries during the experimental period over the two years. This may be because VvOMT2 favors the methylation of 3-isopropyl-2-hydroxypyrazine (IPHP) to 3-isopropyl-2-methoxypyrazine (IPMP) in grape berries [VvOMT4 might have little relation to IBMP accumulation in grape, which might be why the expression level of VvOMT4 always had been ignored in other studies about MPs’ accumulation.The relative expression levels of  berries . The VvOOnly a few studies have focused on the metabolic pathways of MPs’ biosynthesis. Interpreting the mechanism of relevant MPs study findings only from the VvOMTs expression pattern, which is involved in the final step of MPs’ biosynthesis pathway, has certain limitations. More research on MPs’ metabolic pathway is needed to further explore.On the whole, the results drawn from this study will provide reference for further understanding and much deeper research on IBMP biosynthesis.situ, and the findings reveal a significant increase in IBMP content of grape berries in response to DCIP incorporation compared with the Mock and Control at maturation in two consecutive years. The expression level of VvOMT1 and particularly that of VvOMT3 was obviously up-regulated in berry skin after DCIP incorporation and closely mirrored the IBMP accumulation pattern in two consecutive years. Therefore, we speculate that DCIP might participate in IBMP biosynthesis in the grapes and plays an important role in regulating IBMP accumulation. Given the importance of IBMP in determining the sensory characteristics of wine, further studies on the effects of DCIP are warranted.This study highlights the effect of DCIP on IBMP biosynthesis in Cabernet Sauvignon grapes in


The cochlear implant (CI) is a standard treatment for patients with severe to profound hearing loss. In recent years, early device activation of the sound processor after 2–3 days following surgery has been established. The aim of this study is to evaluate the residual hearing of CI patients with substantial preoperative low-frequency hearing after early device activation over a period of 12 months.n = 32), and CG (n = 25). Low-frequency residual hearing and speech recognition in quiet and in noise were compared over an observation period of 12 months.Results were compared between an early fitted group (EF) with device activation to less than 15 days after CI surgery and a control group (CG) with device activation after 3–6 weeks. In total, 57 patients were divided into EF group (p > 0.05) in the residual low-frequency hearing PTAlow between EF and CG was found, neither preoperatively (EF 33.2 dB HL/CG 35.0 dB HL), nor postoperatively (EF 46.8 dB HL/CG 46.2 dB HL). In both groups, postoperative residual hearing decreased compared to preoperative and remained stable within the first year after CI surgery. Furthermore, both groups showed no significant differences (p > 0.05) in speech recognition in quiet and in noise within the first year.No significant difference (Early device activation is feasible in CI patients with preoperative low-frequency residual hearing, without an additional effect on postoperative hearing preservation. The cochlear implant (CI) is a standard treatment for patients with severe to profound hearing loss , 2. The The advantage of using a hearing preservation technique for CI surgery –20 is thThe investigation of hearing preservation associated with the early initial processor activation is of interest because it is not clear whether early electrical stimulation has an effect on residual acoustic hearing function.Intracochlear healing processes are completed after a period of 4–5 weeks . It remaThe aim of this study therefore was to examine whether early fitting of the sound processor has an impact on the postoperative low-frequency residual hearing as a potential negative side effect following the early fitting procedure. Therefore, long term data on residual hearing loss and of speech recognition in quiet and in noise were compared over a period of 12 months between patients who received an early device activation (study group) and patients whose processor was initially activated after the standard healing phase (control group). It is assumed that long term preservation of the residual hearing is possible after early initial processor activation.To our knowledge, there are currently no studies that have examined the postoperative residual hearing preservation after CI surgery in association with the early sound processor activation within a long term follow-up of 12 months.low (pure tone average) was calculated as the average hearing loss at 125 Hz, 250 Hz and 500 Hz. Furthermore, speech recognition in quiet was measured using the Freiburg multisyllabic and monosyllabic word test at 65 dB SPL presentation level in free field condition. In addition, the 50% speech reception threshold (SRT) of the adaptive Oldenburg Sentence Test  [0N0). Both OlSa tests were performed in a closed setup. The study data were collected preoperatively, at the third day of the first fitting interval and at the follow-up intervals after 3 and 12 months following CI surgery. The study procedure is shown schematically in Fig. A total of 57 patients were enrolled in the prospective study. The early fitting group (EF) included 32 patients . In this group, the sound processor was initially activated within a maximum of 15 days after CI surgery. In the control group (CG), which included 25 patients  the processor was first activated after a standard healing phase of 3–6 weeks. The CI surgery was carried out in approximately the same time span in both patient groups. All patients had a minimum age of 18 years and fulfilled the inclusion criteria for an EAS/hybrid CI surgery. Therefore, a sufficient residual low-frequency hearing was necessary. The criterion was set at a maximum hearing loss of 70 dB HL at a test frequency of 500 Hz. The assignment to the respective study group (EF or CG) was non-randomized and based on the willingness of the patients to participate in the early fitting process. Patients who did not want an early processor activation were assigned to the CG. The demographic data of the patients are shown in Table t, OlSa)  in quiett, OlSa)  in free t test for connected samples was used. For data that did not show a normal distribution, statistical comparison of the results between groups were carried out via the Mann–Whitney-U-Test. Within groups, comparison of the results were statistically evaluated using the Wilcoxon signed-rank test. A Bonferroni correction was used to analyze the data within the groups after the respective statistical tests had been carried out. A p value of less than 0.05 was considered statistically significant. Statistical analysis were performed using SPSS Statistics 24.0 . To obtain valid statistical statements about certain processes over time, a complete case approach of the data was performed. This resulted in lower case numbers. However, subsequent analyzes comprising all available data showed comparable results. Therefore, for comparison between both study groups, the use of all data was prioritized to perform an analysis with larger case numbers. For data evaluation within groups, the complete case approach was applied to allow better comparability of results over time.Normal distribution of the data was tested using the Kolmogorov–Smirnov test. In case of a normal distribution, the statistical comparison of the results between the groups was performed using the t-test for unpaired samples. The data was first tested for variance equality via the Levene test. To compare the results within the groups, the n = 32) was first fitted after 3.0 days (median) and thus significantly earlier  than the CG (n = 25) after 28.0 days (median). The study data were collected over a period of 12 months. Since not all patients were able to attend their follow-up appointments after 3 and 12 months following initial device activation for personal or therapeutic reasons (e.g. ongoing stationary rehabilitation), not all data sets were complete.The early fitting procedure allowed a much earlier initial activation of the sound processor than after awaiting the standard healing phase. The EF  and CG (n = 14) had better preoperative residual hearing than postoperative. In addition, no substantial change in residual hearing between the first fitting appointment and the follow-up intervals was observed in either group. The postoperative residual low-frequency hearing thus remained stable. In Fig. low for both groups over an observation period of 12 months is shown. Comparing results between groups, no significant differences  in PTAlow between EF and CG within the observation period was present (no complete case approach). Within the EF, a significant difference in PTAlow was found between preoperative data (33.2 dB HL) and postoperative measurements on first fitting appointment  = − 3.44, p = 0.038, t-test/Bonferroni correction (BC)), 3-month  = − 3.93, p = 0.017, t test/BC) and 12-month interval  = -3.88, p = 0.018, t test/BC). Within the CG there was a significant difference in PTAlow between preoperative data (35.0 dB HL) and the results at 12-month interval  = − 3.41, p = 0.040, t test/BC). Due to the small number of cases after complete case approach (EF/CG n = 11), however, the results of the statistical evaluation within the groups can only be interpreted to a limited extent. Complete postoperative hearing loss was recorded in one patient from the EF  and CG  at the time of the 12-month interval.To assess the preservation of residual low-frequency hearing up to 12 months after CI surgery, the pre- and postoperative results for pure tone audiometry were compared for each group  between the EF and CG, neither preoperatively nor postoperatively  in speech recognition between EF and CG, both preoperatively and postoperatively  = − 3.74, p = 0.012, t test/Bonferroni correction (BC)), 3-month interval  = − 4.47, p = 0.006, t test/BC) and 12-month interval  = − 5.43, p < 0.001, t test/BC). When evaluating the results from the CG, a significant improvement (t(16) = − 4.50, p < 0.001, t test/BC) in preoperative monosyllabic word score (22.3%) was found compared to the 12-month interval (53.8%). The results of the OlSa in quiet  = − 1.36, p = 0.195, t test) between EF and CG. Also for the OlSa SRT in noise  no significant difference (t(25) = − 0.51, p = 0.613) between EF and CG was found. However, the case numbers for OlSa in quiet are low EF, n = 8/CG, n = 8), so that the evaluation of these data is of limited value.When analyzing the results of the Freiburg multisyllabic word test, there was no significant difference . At the 12-month interval, both groups showed a significant decrease in residual low-frequency hearing compared to preoperative measurements. In the EF, the PTAlow decreased by 21.2 dB HL, in the CG the PTAlow decreased by 18.3 dB HL. The study by Helbig et al., 2016 [low decreased compared to the preoperative PTAlow. With a deterioration in the PTAlow of 11.7 dB HL, the value is slightly lower than the data shown in the authors’ study, but in the study of Helbig et al., 2016 [low in both EF and CG remained stable and showed no significant differences over time.There was no significant difference in PTABoth study groups showed comparable preservation of low-frequency residual hearing. A harmful impact of the early acoustic and electrical stimulation on the residual hearing can thus be excluded. Although the insertion of the electrode into the cochlear is rather traumatic  and intrBoth study groups showed comparable development of multisyllabic and monosyllabic word score within the first year after CI surgery with a significant improvement in postoperative speech recognition at the 12-month interval  compared to preoperative results . The postoperative results of the Freiburg monosyllabic word score of both study groups are comparable to the results of other studies with similar age groups (50–60 years), in which an average speech recognition rate between 45 and 63% was described , 33–35. In the OlSa in quiet as well as in the OlSa in noise comparable results in speech recognition were found in both study groups. At the 12-month interval there was no significant SRT difference between EF and CG. However, only a limited number of data sets were available for the OlSa both in quiet and in noise.n = 6, 20%/CG n = 5, 21.7%). If sufficient residual low-frequency hearing was present, an EAS/hybrid map with corresponding cut-off frequency was usually created during initial device activation, but the acoustic part remained deactivated. In this time period, the patient used electrical stimulation only. The acoustic component of the EAS/hybrid system was activated at an additional appointment about four weeks after completion the initial fitting interval.A precondition for EAS/hybrid use of the CI system is a successful postoperative preservation of residual low-frequency hearing , 19. In A potential limitation of the study is that the results reported here likely depended on patient selection, which was not randomized. The patients chose freely between early fitting and standard fitting procedure. Subjects in the EF group might have had higher self-motivation toward CI than subjects in the CG. This notion seems to be supported by shorter duration of hearing loss and hearing aid experience. Therefore, a possible self-selected bias of the results cannot be excluded. Furthermore, due to complete case approach of the data, the consecutive limitation of the number of data sets is detrimental for the detection of significant effects.Early fitting of the sound processor had no negative effect on the preservation of the postoperative low-frequency residual hearing. In both EF and CG, long term preservation of the low-frequency hearing was possible. Both groups showed a comparable development of EAS/hybrid CI aided speech recognition within the first year after CI surgery. Due to the sufficient long term preservation of the residual low-frequency hearing, EF patients could benefit to the same extent from EAS/hybrid usage as patients who were fitted after the standard healing phase. The results of this study showed that early device activation is possible in EAS/hybrid CI patients with sufficient residual low-frequency hearing.