Chemotherapy and radiotherapy in oncology have repercussions in hearing health, and can damage structures of the inner ear. These repercussions usually, result in a bilateral and irreversible hearing loss.To identify sensorineural hearing loss cases with complaints of tinnitus and difficulty in speech understanding and investigate their relationship with the types of chemotherapy and radiotherapy the patients received.Cross-sectional, clinical, observational, analytical, historical cohort study of 58 subjects treated in a public hospital in the state of Sergipe, diagnosed with neoplasia. The subjects were submitted to anamnesis, conventional pure tone audiometry, and speech recognition threshold.p = 0.035; R = 0.196). The auditory complaint analysis shows that most of the subjects had tinnitus and speech understanding difficulty, even with a normal auditory threshold.Of the 116 ears, 25.9% presented sensorioneural hearing loss characterized by changes in high frequencies. There was a positive correlation between hearing loss and the association of chemotherapy and radiotherapy (Cancer treatment causes hearing loss, associated with the administration of chemotherapy and radiotherapy. Cyclophosphamide increased the risk of causing hearing loss. Complaints of tinnitus and speech understanding difficulty were observed. Currently, cancer is considered a public health problem with high prevalence. There is a projection of 27 million new cases for the year 2030 worldwide and 17 million deaths from the disease. In Brazil, estimates for the year 2014/2015 report approximately 580,000 new cases of the disease. Among the most prevalent forms, non-melanoma skin cancer, prostate, breast, colorectal, lung, and stomach cancers prevail.The basic modalities in cancer treatment involve surgery, chemotherapy, and radiotherapy. Currently, the combination of chemotherapy and radiotherapy has improved patient survival. However, none of these treatment modalities is free of side effects. Among their effects, hearing loss caused by ototoxicity, can be documented.4In recent years, the number of studies reporting the influence of chemotherapeutic agents on hearing function has increased. The drugs employed belong to different classes, and some of them – aminoglycosides, antineoplastic agents, antibiotics, nonsteroidal anti-inflammatory drugs, diuretics and antihypertensives – are considered to be ototoxic drugs. Drugs included in the platinum group are the most devastating, generating auditory symptoms such as tinnitus and hearing sensitivity change.Radiotherapy can also damage the auditory organ. This therapeutic modality promotes tumor cell destruction by its ionizing radiation beams. A pre-calculated dose of radiation is applied to tumor cells during a specified time period, in a volume of tissue that includes the tumor. Hearing loss is most commonly found in the treatment of head and neck tumors. Although there are few studies focused on the side effects of radiotherapy on hearing health, the literature shows a wide variation in the incidence of ototoxicity.7In both treatments, the higher concentration of toxic substances in the body is able to reach the organ of Corti and sensory epithelia of the posterior labyrinth, through the labyrinthine fluids. These substances compromise mainly the outer hair cells and can lead to cochlear symptoms; however, vestibular disorders may arise in a slow or insidious way, even after the end of treatment. Usually, hearing loss is bilateral, irreversible, associated with tinnitus and is high-frequency in audiometric configuration.8Studies investigating hearing in oncologic patients using conventional audiometric frequencies, report variable symptoms. It is common to observe tinnitus, difficulty understanding conversation in noisy environments, and changes in speech discrimination.Studies in this area also report that the hearing loss caused by ototoxic substances is often underestimated. Even in the presence of a hearing disorder, patients only report hearing complaints in specific situations, such as in noisy environments. Other patients only exhibit partial understanding of a message, which makes it more difficult for relatives to detect the hearing loss.Studies on oncology and hearing health sought to promote early detection of hearing impairment and to implement preventive measures in order to improve the quality of life of this population. This research was specific; its objectives were to identify cases of sensorineural hearing loss and their relationship with chemotherapy and radiotherapy session means, and with tinnitus and speech understanding difficulty complaints, as well as their relation to the use of various chemotherapy drugs.This was a cross-sectional, clinical, observational, analytical, historical cohort study. All ethical guidelines were followed, and this project was approved by the Research Ethics Committee, under protocol No. 0066.0.107.000-11.Data collection was conducted from March to November of 2013 in the physical space of the cancer ward of a public hospital in Sergipe. The sample comprised 58 subjects aged 25–59 years with pathological diagnosis of cancer who were undergoing chemotherapy and/or radiotherapy. The following drugs were utilized for chemotherapy: carboplatin, cisplatin, doxorubicin, epirubicin, farmorubicin, vincristine, actinomycin, gemcitabine, oxaliplatin, cyclophosphamide and fluorouracil.Subjects were excluded from the study if they: were over 60 years of age (to avoid hearing loss by age); had a history of previous ear surgery; worked in a setting with noise exposure; had previous acoustic trauma; had received ototoxic medication prior to current treatment; had infections or congenital syndromes; exhibited outer ear canal obstruction and/or had previously received chemotherapy and/or radiotherapy. In addition, subjects with a diagnosis of head and neck cancer or conductive hearing loss were also excluded.After signing the informed consent, the patients’ history was obtained and meatoscopy and conventional pure tone audiometry were performed, with a survey for speech reception threshold.The anamnesis scheme was developed by the researchers, and consisted of information concerning the past history of the disease, treatment performed, drugs used, time of use of the drug, initiation of therapy, and overall hearing health.The equipment used to perform pure tone audiometry and a speech reception threshold survey was the Interacoustics Ad Model 229 B audiometer with a TDH-39 headset in a soundproof booth that followed the equipment calibration standards proposed by the Brazilian Federal Council of Phonoaudiology on environmental noise measurement . On examination of pure tone audiometry, hearing thresholds were surveyed by interoctave frequencies of 0.25–8 kHz by air conduction and of 0.5–4 kHz by bone conduction. The speech recognition threshold survey was conducted to confirm the tonal findings.Thus, using the tonal audiometry findings, the sample was organized into two groups: Group 1, consisting of ears with normal hearing thresholds, and Group 2, ears with hearing loss.p ≤ 0.05 and the admitted α value was 0.1.Data were tabulated and processed by the software Statistical Package for Social Sciences (SPSS), version 17.0. For a description of data, tabular presentation of means, medians, standard deviations, maximum and minimum values, and percentages were used. Sample normality was verified using the Shapiro–Wilk test. To evaluate the relationship between variables, the Mann–Whitney and bivariate correlation (Spearman coefficient) tests were utilized. Values were considered significant at The sample consisted of 58 subjects (116 ears) with female predominance (93.1%), and mean age of 47.19 (±9.36) years. For Group 1, the mean age was 46.56 (±9.38), and for Group 2, 51.93 (±9.38) years.The most frequent diagnosis was breast cancer (69%), followed by cervical (8.6%) and uterine (5.2%) cancers. Among the therapeutic procedures recommended by the medical staff, it was noted that 84.5% underwent surgery, and patients were also referred for chemotherapy and/or radiotherapy. Chemotherapy and radiotherapy usually occurred after surgery; 37.9% underwent chemotherapy, 15.5% radiotherapy, and 46.6% both chemotherapy and radiotherapy.The number of chemotherapy sessions ranged from one to 36, with a mean of 7.56 sessions; while the number of radiotherapy sessions ranged from one to 70, with a mean of 18.31. By the time audiological evaluations were carried out, 1.7% of the population were starting treatment; 53.4% were in progress, and 44.8% had completed treatment.n = 2), cisplatin (n = 6), doxorubicin (n = 6), epirubicin (n = 9), farmorubicin (n = 1), vincristine (n = 1) actinomycin (n = 1), gemcitabine (n = 6), and oxaliplatin (n = 3); the most frequently used agents were fluorouracil (n = 17) and cyclophosphamide (n = 18).The studied population used either a single chemotherapeutic agent or a combination of several drugs. The patients used the following drugs: carboplatin  exhibited sensorineural hearing loss, predominantly in the high frequencies. Audiometric analysis of the hearing loss cases revealed that worsening of tonal auditory thresholds began at the frequency of 4 kHz and gradually increased at the higher frequencies of 6 and 8 kHz .Table 1Dp-value = 0.035 and correlation coefficient R = 0.196. No significant relationship was observed for these treatments in isolation, as may be seen in In cancer treatment, a positive bivariate correlation was found between hearing loss and the combination of chemotherapy and radiotherapy, with Regarding the type of chemotherapy drug used in the population presenting hearing loss, only cyclophosphamide showed a significant correlation .Table 3pp = 0.79), and also occurred in conjunction with complaints of difficulty in understanding speech (p = 0.46).With respect to self-reported hearing complaints by Group 1, 62.8% reported having a perception of tinnitus and 32.6% had difficulty listening; while in Group 2, 60.0% complained of tinnitus and 40.0% had difficulty listening. The presence of tinnitus was often observed, even without hearing loss (Of the 15 subjects (30 ears) who experienced hearing loss, 23.3% reported worsening of hearing after chemotherapy and 6.7% after radiotherapy.In this study, most participants were female, with prevalence of breast cancer, which confirm the findings in the literature for the state of Sergipe in 2014/2015.The population studied had a mean age over 40 years, similar to that found in previous studies.Most respondents (84.5%) underwent surgery, and then were treated with chemotherapy and/or radiotherapy. The adopted medical management was also described in the literature, considering that, in the early staging process, the recommended treatment is surgery. This procedure enables a complete removal of the tumor and provides a better chance of survival. Subsequently, radiotherapy and chemotherapy are performed, independently or together.Most often, the medical team opted for a combination of chemotherapy and radiotherapy (46.6%), followed by chemotherapy (37.9%) and radiotherapy (15.5%). It is known that the type of treatment recommended is related to the type of tumor; and in the literature, high values for chemotherapy were found.With respect to the use of chemotherapeutic agents, it was found that cyclophosphamide was more significant in the determination of changes in auditory threshold; in the literature, this agent is referred to as ototoxic, possibly injuring cochlear hair cells.Hearing loss was mainly sensorineural, which is consistent with hearing loss caused by cochlear ototoxic substances, with early involvement of high frequencies.16In the audiometric analysis, hearing threshold worsening occurred at and above the frequency of 4 kHz. Among the most affected frequencies, changes in 6–8 kHz have been diagnosed, which is in line with the literature.It was found that, of the 15 subjects with hearing loss, 23.3% reported worsening of post-chemotherapy hearing thresholds, and 6.7% reported after radiotherapy, with consequent hearing complaints. Studies have indicated that when there is change at the 4 kHz frequency, hearing complaints may occur. Thus, these data explain the hearing worsening in the population studied, since it was possible to detect hearing change from the frequency of 4 kHz onwards.The presence of tinnitus was considered as a relevant complaint. Tinnitus occurred in 62.8% of Group 1 and in 60.0% of Group 2. The literature also indicates a high incidence of tinnitus – a common occurrence in patients undergoing cancer treatment, which can be observed even in the absence of hearing loss.18There was no correlation between “difficulty in listening” and “suffering hearing loss.” This finding can be explained by the fact that the most affected frequencies are the highest ones, and the complaints are more frequent when they reach the speech range.With respect to cancer treatment, the means of radiotherapy and chemotherapy sessions were similar for both groups. The literature reports they need to consider that the greater the time of presence of toxic substances in the body, the greater the adverse effect, although there are individual differences in response to toxic agents, as well as variables that enable ototoxicity, e.g., family antecedents with deafness and individual susceptibility, among others.19Hearing loss in subjects undergoing cancer treatment was observed, and the combination of chemotherapy and radiotherapy was instrumental for the change in hearing thresholds. Cyclophosphamide was the most frequently administered chemotherapeutic agent and was positively correlated with hearing loss. In the population submitted to cancer treatment, a high prevalence of tinnitus and speech understanding difficulty were found, even in the absence of hearing loss.The authors declare no conflicts of interest.


Kyoto Encyclopedia of Genes and Genomes pathway in downregulated DEPs in the central retina. Antithrombin-III, fibrinogen gamma chain, and fibrinogen beta chain were identified as hub proteins for myopia progression, which were upregulated in the far-periphery retina. Proteomic analysis in this study suggested that oxidative stress can be the primary pathogenesis of myopia progression and that the far-periphery retina plays a role as the key responder.Uncovering region-specific changes in the myopic retina can provide clues to the pathogenesis of myopia progression. After imposing form deprivation myopia in the right eye of 6-week-old rabbits, we investigated the proteome profile of each retinal region , using accurate high-resolution mass spectrometry. Protein expression was analyzed using gene ontology and network analysis compared with that of the control, the left eyes. Among 2065 proteins detected from whole retinal samples, 249 differentially expressed proteins (DEPs) were identified: 164 DEPs in the far-periphery, 39 in the mid-periphery, and 83 in the central retina. In network analysis, the far-periphery retina showed the most significant connectivity between DEPs. The regulation of coagulation was the most significant biological process in upregulated DEPs in the far-periphery retina. Proteasome was the most significant  Oxidative stress and chronic inflammation have gained importance in the pathogenesis of myopia. The prevalence of myopia is increased in inflammatory diseases, such as diabetes, systemic lupus erythematous, and allergic disease ,2. The eThe retina is one of the primary oxygen-consuming tissues regulated by vascular supply and its alterations . AdditioIn myopia progression, axial elongation has been related to the thinning of the retina and the reduction in retinal pigment epithelium (RPE) of the peripheral area in emmetropization. In contrast, the thinning of RPE density or retinal thickness has not been observed in the macular area . The undIn this study, we analyzed the region-specific retinal proteome profile after myopia induction in a rabbit model using high-resolution accurate mass (HRAM) mass spectrometry. Additionally, we investigated myopia-specific hub proteins in each retinal region, comparing proteomes between control and myopia-induced eyes. It was expected that a more comprehensive understanding of myopia progression mechanisms could be achieved by clarifying proteomic changes in each retinal region.p < 0.01, control vs. myopia-induced eyes, p < 0.01, control vs. myopia-induced eyes, We compared the axial length and refraction between the myopic and control eyes. After 6 weeks of form deprivation, the axial length of the myopia-induced eye significantly increased more than that of the control eye  test, We segmented each retina in the myopia-induced and control eyes into the central, mid-periphery, and far-periphery retina. Proteins from six groups were analyzed using LC-MS/MS following trypsin digestion followed by processing with MaxQuant. A total of 2065 proteins were identified from the whole retinal sample B.We compared each of the six groups using principal component analysis (PCA) A. PCA dip < 0.01), followed by that of alpha-2-HS-glycoprotein , sodium/potassium-transporting ATPase subunit alpha , histidine-rich glycoprotein , and lambda-crystallin . In the far-periphery retina, DEPs showed, on an average, the most significant fold changes compared with those at other regions (p < 0.01), apolipoprotein A-1 , cochlin , and selenium binding protein 1  increased in the far-periphery. After myopia induction, 249 DEPs were identified in the whole retina A. The ex regions B. The exp-value, which was transformed to –log10 values ranging from 0 (white) to 5.5 (dark red). Closer connectivity between each node was presented by more edges with a thicker width. The connectivity was the most pronounced in the far-periphery retina (We constructed protein–protein interaction (PPI) network models using DEPs from each regional retina to identify the essential proteins involved in critical biological processes for myopia induction A. Using y retina A.Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation was also performed for each retinal region. Our results showed that the far-periphery retina mainly plays a role in changing DEPs after the induction of myopia. For whole DEPs, the most important biological processes were the positive coagulation of vasoconstriction, mitochondrion organization, and cell junction assembly on the far-periphery retina  was searched to investigate the biological processes and cellular components. y retina B. ATPasey retina B. The miy retina C. For DOy retina C. On they retina . Proteasp-value) significance was visualized with color from 0 (white) to 8 (dark red). Detailed GO terms in each group were shown in the same color  algorithm using the CytoHubba plugin. The proteins with the top 10 MCC values were considered hub proteins A. In theme color B. The reme color C. No GO Here, we have reported three main findings that are distinct from previous studies: (1) proteomic changes in retinal protein were the most significant in the far-periphery retina after myopia induction, followed by the central retina; (2) coagulation regulation, complement, and coagulation cascades were the most significant and upregulated biological process for myopia induction in the far-periphery retina; and (3) antithrombin-III (SERPINC1), fibrinogen gamma chain (FGG), and fibrinogen beta chain (FGB) are potential novel biomarkers for myopia induction in the retina.The regulation of coagulation was enriched as the most significant biological process in UP-DEPs of the far-periphery retina. The result was consistent with GO analysis for hub proteins in the far-periphery retina. This consistency can confirm the importance of coagulation regulation as a key biological process of myopia induction. Hub proteins with the top 10 MCC values in this study included SERPINC1, FGB, APOA1, AHSG, and HRG, which had been identified in previous studies using human aqueous humor of pathologic myopia; however, Xu et al. had reported that APOA1 was the most significant hub protein .DEPs associated with the regulation of coagulation included SERPINC1, FGB, FGG, HRG, ANXA1, and MYH9. SERPINC1 inhibits leukocyte and platelet recruitment to the post-ischemic retina, subsequently exerting a neuroprotective effect against retina ischemia–reperfusion injury . FGB andwww.genome.jp/kegg/, 6 December 2022). Proteasome degrades misfolded proteins in response to environmental stress [In the central retina, hub proteins such as PSMC6, PSMA1, PSMA8, PSMC2, and PSMD12 are known subunits of the proteasome endopeptidase complex , which il stress . Under ol stress . Based oNotably, the far-periphery retina showed the most significant change after myopia induction despite form deprivation rather than lens-induced myopia induction. Lens-induced myopia induction develops peripheral hyperopic defocus; however, form deprivation methods provide blur imaging to the whole retina. Both involve a reduction in spatial contrast, whereas lens-induced myopia may also involve a pathway initiated by the sign of defocus signal . Form deThere are a few limitations to this study. First, this study was performed in a rabbit retina. Therefore, the result of this study cannot directly be applied to the pathogenesis of myopia in humans. However, the central fovea in humans has a high concentration of macular pigments, such as lutein and zeaxanthin, acting as powerful antioxidants and anti-inflammatory agents . This prIn conclusion, the results of this study reinforced the importance of the far-periphery retina as the key responder to the reduction in spatial contrast caused by form deprivation-induced myopia based on bioinformatics. In addition, it is suggested that oxidative stress on the retina could be associated with the pathogenesis of myopia. Antithrombin-III (SERPINC1), fibrinogen gamma chain (FGG), and fibrinogen beta chain (FGB) are potential novel biomarkers for myopia. Based on this study, continuous changes in choroid and sclera in myopia should be more region-specifically studied in an animal model and humans, leading to identifying therapeutic clues for the progression of myopia.We obtained 5 6-week-old New Zealand white rabbits  from the Department of Laboratory Animal Resources, Yonsei Biomedical Research Institute, Yonsei University College of Medicine. All the animals were raised under a controlled temperature of 25 °C on 12 h light and dark cycle, with lights on at 9:00. Food and water were freely available. The experimental protocol was approved by the Animal Care and Ethics Committee of Yonsei Biomedical Research institute.The right eyes of all the rabbits were assigned as the myopia group, with the left eyes as the control group. Myopia was induced using the form deprivation method with tarsorrhaphy. Under general anesthesia with zolpidem  and rompun , local anesthesia was administered on the upper and lower eyelid margin using xylocaine. Upper and lower eyelids were split along the gray line. The epithelial layer on the gray line was removed with bipolar cauterization. The inner layer was approached and sutured with 6-0 vicryl . The outer layer was approached and sutured with 5-0 nylon . Eyelids were closed entirely, and the eyeball surface could not be observed grossly. Bleeding was controlled meticulously. Ofloxacin ointment  was applied. To prevent secondary infection and inflammation on the operation site, subcutaneous amoxicillin  and ibuprofen  were injected immediately after the operation. The wound site was dressed with 0.9% normal saline, and ofloxacin ointment was applied once a day for 7 days. Subcutaneous amoxicillin was injected once a day for 7 days. After 7 days, the nylon suture on the outer layer was removed. The closed eyelids were thus maintained for 6 weeks. When the tarsorrhaphy site was ruptured, and the eyeball surface was exposed, re-suture was performed immediately. After 6 weeks, the closed eyelid was split under general anesthesia.All measurements were performed under general anesthesia induced with zolpidem (0.3 mL/kg) and rompun (0.2 mL/kg). Measurements of the ocular biometry were taken with A-scan ultrasonography . After topical anesthetic use of proparacaine hydrochloride 0.5% , applanation of the probe on the cornea was performed. Proper alignment of the probe was defined as strong ultrasonic wave reflection peaks of lens surfaces and retinas, which stood high from the level cursor line position. More than seven consecutive measurements of ocular components, including axial length, depth of the anterior segment, and lens thickness, were recorded and averaged after proper alignment was obtained. If the standard deviation of measurements was more than 0.04 mm, we repeated the measurements.Cycloplegic refractions were performed using a streak retinoscope operated by an experienced ophthalmologist, who was unaware of which eye was induced and the purpose of the study. Three measurements with two axes of each were averaged as the spherical equivalent of all eyes, with readings reported to the nearest 0.25 D. Measurements were performed before the induction of myopia and after 6 weeks of myopia induction just before sacrifice.After measurements of ocular biometrics at 6 weeks of myopia induction, a lethal dose of KCL  was administered. The eyes were enucleated immediately after death, and the globe was separated from residual orbital tissue . Marking18 Harvard macro spin column. The resulting peptides were dried and stored at −80 °C.Proteins from pooled retinal tissues (100 μg) were digested into peptides by in-solution digestion. Briefly, 8 M urea in 100 mM ammonium bicarbonate  was added to each pooled sample, and the mixture was homogenized using a disposable homogenizer  until the tissues were no longer lumped. The samples were then briefly vortexed and sonicated on ice for 10 s. Samples were incubated for 20 min at room temperature and then centrifuged at 14,000 rpm for 15 min. Protein concentration of the supernatant was determined in triplicate using the micro BCA protein assay according to the manufacturer’s protocol ; then, 10 mM dithiothreitol  for reduction and 30 mM iodoacetamide  for alkylation were used to denature the proteins. Trypsin , was added to the samples (1:50 = trypsin:sample) and incubated at 37 °C overnight. The activated trypsin reaction was quenched with 0.4% TFA , and peptides were desalted with a C18 particles at an electric potential of 1.8 kV. Complete MS data were acquired in a scan range of 400–2000 Th at a resolution of 70,000 at m/z 200, with an automated gain control target value of 1.0 × 106 and a maximum ion injection of 100 ms. The maximal ion injection time for MS/MS was set to 100 ms at a resolution of 17,500. The dynamic exclusion time was set to exclude previously sequenced peaks for 30 s within a 10 ppm isolation window, and higher-energy collisional dissociation fragmentation was conducted under 30% of normalized collision energy. The proteomics data have been deposited to the ProteomeXchange consortium  with dataset identifier PXD038552 via the PRIDE partner repository [Peptides were re-suspended in 0.1% formic acid (FA) in water and analyzed using the Q Exactive Orbitrap Hybrid Mass Spectrometer coupled with the EASY-nLC 1000 system . For global proteome profiling analysis, the gradient was as follows: from 5% to 50% of solvent B for 85 min, from 50% to 80% of solvent B for 1 min, holding at 80% of solvent B for 8 min, and equilibrating the column at 1% of B for 30 min . The peptides were eluted through a trap column and ionized through an EASY-spray column (50 cm × 75 μm ID) packed with 2 μm Cpository .Raw files were searched using the MaxLFQ algorithm integrated with MaxQuant software   against p-value of less than 0.05 from the Student’s t-test in LFQ intensity were classified as DEPs.Statistical and bioinformatics analyses for proteomics data were conducted based on the normalized protein abundance using MetaboAnalyst software version 5.0, a web-based tool for quantitative data analysis (downloaded on 1 June 2021). Clustering analysis, including PCA and hierarchical clustering using “Euclidean distances” and “ward” linkages, was presented to show the variance and differences among samples. Proteins with an expression greater than ±2-fold change and a https://apps.cytoscape.org/apps/cluego, accessed on 1 October 2022), a Cytoscape plugin, was used to analyze and visualize the functionally organized GO terms. The analysis was based on Uniprot GO annotation. The gene ontology biological process and cellular component terms associated with total identified protein and DEPs were performed using the Database for Annotation, Visualization and Integrated Discovery and g:Profiler online tool . Functiohttps://apps.cytoscape.org/apps/cytohubba, accessed on 1 October 2022), a plugin for Cytoscape, provides 11 topological algorithms for detecting hub genes. Among all the algorithms, the MCC score performs better in hub gene prediction [CytoHubba . The Kolmogorov–Smirnov test was used to confirm the normality of the data. To statistically compare data between groups, we used the Mann–Whitney U test or Wilcoxon signed-rank test for non-normally distributed data. We used the repeated measures ANOVA test to analyze repeated measured data between groups. With the assumption of sphericity with repeated measures ANOVA, Greenhouse–Geisser correction was used. A


Plants have the ability to regulate their growth and development according to available light . Light pArabidopsis thaliana). In dark conditions, basic helix–loop–helix (bHLH) proteins called PHYTOCHROME INTERACTING FACTORS (PIFs) accumulate to bind and repress transcription from light response elements. In this way, photomorphogenesis is repressed, including the expression of PHYTOENE SYNTHASE (PSY) gene, which encodes the first enzyme in the carotenoid biosynthesis pathway , the molecular scenario must be very different as carrots accumulate carotenoids in the roots during dark conditions. Carrots are one of the vegetables that accumulate the most carotenoids, alongside mint and parsley (PSY and CTRI (CAROTENE DESATURASE)  . GeneticATURASE) .Plant Physiology, PHYA (PHYTOCHROME A), PIF4, and PAR1 , LCYB2 (LYCOPENE β-CYCLASE), CHXB1, and CHXB2 (β-RING CAROTEN HYDROXYLASE), were downregulated. The authors hypothesized that this might occur via an interaction of PAR1 with other transcription factors, such as PIFs. Finally, the authors found increased chlorophyll and lutein content in PAR1-silenced plants. This suggested that the absence of PAR1 might contribute to chloroplast formation. Whether PAR1 could be involved not only in carotenoid biosynthesis but also in chromoplast formation remains to be investigated further.To investigate this scenario further, the authors carried out several experiments. First, the authors designed carrot plants with reduced PAR1 that play a role in fine-tuning carotenoid biosynthesis in carrots requires further investigation.Altogether this study showed that increased gene expression of the bHLH protein PAR1 in carrot plants in dark conditions induces the expression of carotenoid biosynthesis genes, including the phytoene synthase enzyme that triggers carotenoid accumulation in the chromoplast. The interconnection between lutein and chlorophyll biosynthesis and carotenoid biosynthesis and accessory partners of Conflict of interest statement. None declared. F1 Crossref missing


Presbycusis is a common disorder in the elderly, which causes hearing loss and may contribute to the development of some psychiatric disorders, leading to isolation due to communication difficulties in the social environment.To identify through the WHOQOL , the quality of life of hearing impaired individuals before and after hearing aid fittings.We had 30 individuals with hearing loss, all over 60 years of age - patients from a Speech Therapy Clinic. The patients answered the WHOQOL questions without the use of hearing aids; and after the effective use of a sound amplification device for a period of three months they answered it again. The WHOQOL - Bref consists of 26 questions, two general quality-of-life questions and 24 associated with four aspects: physical, psychological, environmental and social relations.There was a significant improvement in quality of life in general, as far as leisure activities were concerned, there were no major changes regarding the frequency of negative feelings; even after the hearing aid fitting, the patients continue to have such feelings.The use of hearing aids favored the overall quality of life of the individuals evaluated. There are changes which make it difficult for the individual to adapt himself to the environment, exactly because of the lack of conditions which favor biopsychosocial aging. The individual's appearance changes, enabling one to give him/her an age estimate with very little margin for error. The skin wrinkles in consequence of dehydration and loss of elasticity of the underlying dermal tissue. Teeth are lost, there are muscle atrophy and joint sclerosis causing movement disorders. Osteoporosis plagues the skeleton and renders it subject to fractures. The heart changes in function, the organs of the senses are impairedThe elderly are more vulnerable to insidious degenerative diseases, such as cardiovascular and cerebrovascular, cancer, mental and other disorders, which affect the locomotion system and the organs of the senses. Undeniably, there is a systematic reduction in the degree of social interaction as one of the most evident signs of agingHearing loss is one of the most devastating sensorial deficiencies, because it compromises communication and causes emotional, social and occupational sequelae. Hearing loss is usually a harbinger of agingAccording to IBGE data (2005), because of demographic transition, Brazil is among the 10 countries with the higher number of elderly, with the forecast that, in 2025, 14% of our population will be over 65 years of ageAge-related hearing loss may start at any time; being more common in individuals above 60 yearsPresbycusis is one of the most common disorders reported by elderly individuals, and for most of them, descending bilateral sensorineural hearing loss. Speech understanding difficultyBesides the hearing limitation stemming from the acquired hearing loss, some problems must be stressed, such as: the hearing loss and the hearing handicapHearing loss in advanced age is a serious limiting factor for the individual; and it may also lead to psychiatric problems, causing those with hearing loss to isolate themselves because of the difficulty in communicating in the social environment where they live. Often times, the family of hearing impaired patients do not have the patience to deal with the hearing problem and, usually, do not keep normal dialogues with the patient, only informing him/her of essential matters. The elderly feel embarrassed because of his/her difficulty in hearing, which may contribute to depression.Besides physical problems, the elderly also have higher prevalence of depressive symptomsAs a result of the forecasted aging of the population, the number of patient candidates to fitting an Individual Sound Amplification Device (ISAD) will greatly increase in the next yearsToday, in the current demographic settings, it is urgent to establish the guidelines for the development of diagnostic programs, acquisition of individual sound amplification device and, especially, of a specific hearing reeducation program for the elderly with hearing loss, so that they can participate and enjoy social relations, having a good quality of lifeQuality of life associated with the health and the subjective health status are similar concepts, centered in the patient's assessment, but necessarily connected to the health status impact on the individual's capacity to live fullyIt is an eminently human notion, which has been brought closer to the degree of satisfaction found in family life, love life, social and environmental lives and their own existential esthetics. The term involves numerous meanings, which reflect knowledge, experience and values from individuals and the collection which report to him at different eras, spaces and histories, being, therefore, a social construction with the brand of cultural relativityAccording to the World Health Organization's Quality of Life concept, it is the individual's perception of his/her own projection in life, within the context of culture and value systems in which he lives and in relation to objectives, expectations, patterns and concerns”Quality of Life may be assessed by means of a standardized questionnaire by the group which studies quality of life at the World Health Organization, the World Health Organization Quality of Life Questionnaire , which is an abridged version of a longer questionnaire, with 100 questionsSome authors have stated that, the better a person's adaptation to life at earlier ages, the better will be such adaptation at old age. The elderly with better fitting qualities  would not manifest mental disorders when facing similar life conditions. One of the most important psychological aspects when one talks about old age concerns exactly the ability of people to adapt; thus, old age means a great challenge to contemporary humansThus, with the increase in the number of elderly citizens with hearing loss, and their consequent isolation, the goal of the present paper is to use WHOQOL  to assess the individual's quality of life before and after fitting a hearing aid (ISAD).This study was carried out with the approval of the Ethics in Research Committee, under protocol #002/2009. All participants in this study consented with the publication of its results.The study was made up by 30 individuals with a mean age of 76.8 years, from both genders, seen at the Department of Selection and Fitting of Hearing Aids in the period between August, 2009; and May of 2010.•Signature in the Free and Informed Consent Form.•Age range between 60 and 90 years.•Diagnosis of bilateral moderate sensorineural hearing loss.•Good general health status, starting from the assumption that if the patient was able to go the clinic for care; he/she would also be able to participate in the study.•Without experience concerning the use of ISAD on the first use of the questionnaire.•Have understanding capacity to answer the WHOQOL-Bref questionnaire.•Effective use of the ISAD (daily for more than 6 hours) for three months for the second questionnaire.•Come to the return visits schedule by the service. Exclusion criteria:•No depression reported by the individual during the psychological interview and at the time of the study according to criteria from the institution's psychologist. The test adopted was the Beck's Depression Inventory - (BDI-II).•Hearing loss preventing the questionnaire's understanding.Inclusion criteria:After proper audiologic diagnosis, the patient answered the questions of the WHOQOL - BrefWe selected and fit the sound amplification devices according to the need of each subject; and each patient was advised and educated concerning the use and cleaning of their devices.The patients returned monthly to check for the need of doing fine adjustments in the programming of their hearing aids, and clarification of doubts, thus guaranteeing proper use of the hearing aids. It is worth stressing that there was no hearing training during this period.The elderly made an effective use of the device for three months and, after this period, answered the questions again (the questionnaire was again applied by the researcher), with the aim of checking whether the use of amplification provided for a quality of life improvement.The effectiveness concerning the ISAD use was considered when the individual reported its daily use for longer than six hours/day during the three months considered for employing the WHOQOL - Bref questionnaire.The WHOQOL - Bref is made up of 26 questions, having two general questions, associated with quality of life and 24 representing each one of the 24 aspects which make up the original instrument. The information from which this abridged version stemmed was taken from field tests done in 20 centers in 18 different countries. This questionnaire cropped up from the need for short questionnaires which require less time to answer; however with reasonable psychometric characteristics1.Physical: corresponds to questions associated with pain, discomfort, energy, fatigue, sleep, rest, mobility, daily life activity, medication or treatment dependence and work capacity.2.Psychological: encompasses issues about positive feelings, thinking, learning, memory and concentration, self-esteem, body image and appearance, negative feelings, spirituality, religion and personal beliefs.3.Environment: involves physical safety and protection, home environment, financial resources, health and social care: availability and quality, opportunities to acquire information and skills; participation in and opportunities for recreation/ leisure, physical environment (pollution/noise/ traffic/climate) and transportation.4.Social relations: questions about personal relations, social support and sexual activity. Individuals answered based on gradations, such as: very bad, bad, neither bad nor good, good and very good or very unhappy, unhappy, neither unhappy nor happy, happy, very happy, depending on the aspect investigated the questionnaire offers these options.The WHOQOL-Bref bears the following demand:After filling out the questionnaire, we calculated the general values and those by aspect , enabling an assessment of the individual's quality of life. This analysis was carried out according to the syntaxes proposed by the translators, with the help of the Statistical Package for Social Science (SPSS) 10.0 for Windows, when counting and manual analysis was advised against because of a greater likelihood of errorsThe results consider the 30 participants who returned in the three-month period and answered all the questions in the questionnaire - 17 men (56.6%) and 13 women (43.4%).After comparing the results from the WHOQOL questionnaire, answered before and after fitting a hearing aid, one can notice a significant improvement in quality of life in general, since, after its fitting, all the patients ranked their quality of life as good or very good. As far as health is concerned, after fitting the hearing aid, all patients seemed happy according to the questionnaire, suggesting that for some patients who were hearing challenged before the hearing aid, it was associated with a health problem.Graph 2RAs to leisure opportunities, there were no major changes, given that the answers to the questionnaire given afterwards were well split among the patients, and very similar to the answers found in the pre-fitting questionnaire .Graph 3RWhen asked about satisfaction with personal relations, between the patient and friends, relatives, colleagues and acquaintances, half of the participants remained very pleased. The other half, were less happy when the pre fitting situation was compared to the post-fitting one; conversely, the mean response increased, and there were no answers of unhappiness..Graph 4RAs to the frequency of negative feelings, we noticed that, even after fitting the hearing aid, the patients continued having such feelings, even when they were not often or frequent. This is due to the fact that loosing hearing acuity is not the only reason why the elderly feel depression, anxiety and other noxious emotions .Chart 5RIn today's world, the elderly are seen by our society as disabled people, without a social role to play, compromising productivity since it stresses youth vitality. Modern society is exclusively founded on productivity and, considering social functions, youth is seen as being better. Thus, the elderly feel disappointed with losing their roles, previously lived in full and successfully, causing social isolation and privation from the sources of communication and information, responsible for keeping the individual active in societySome authors stated that, the better the person's adaptation to life in earlier ages, the better the person will adapt him/herself in older ages. Elderly who are better able to cope , did not have emotional disorders when having to face similar life situations. One of the most important psychological aspects of older age is associated to the person's capacity to cope. Thus, growing old becomes a major challenge for our contemporary societyHaving all these challenges, the elderly still have to face a difficulty to communicate with others, stemming from hearing loss resulting from aging itself, compromising their relationships with family and friends, one more impact to their psychosocial livesThe isolation of the elderly and the consequent drop in the quality of their communication, because of sensorial deficits, have a profound impact, since it is the constant flow of communication and information that keeps the individual active in societyTo be a senior citizen with acquired hearing loss is something which goes beyond the hearing impairment itself, causing severe psychosocial implications for his life and that of those who live with himPoor social relations are a health risk factor, and it has been considered as harmful as smoking, high blood pressure, obesity and the lack of physical activitiesEarly diagnosis and treatment of age-related hearing loss are fundamental for a good quality of life for senior citizens. Studies and research in this field point to the possibility of a functional change based on brain plasticity, even when considering adult individualsResults have shown that, after using ISAD, there is improvement in the individuals quality of life as a whole , stressiIn the physical realm, there was a significant difference, with score improvements, and one can see a global improvement when comparing pre and post-fitting data. Such data differ from those in the study carried out in Porto AlegreThe psychological aspect improved with the ISAD, stressing the importance of hearing for an individual's general quality of life and healthAs far as the environment is concerned, there was no statistically significant difference. The questions associated with it encompass physical safety, home environment, financial resources; healthcare and social ; skills, information opportunities, recreation and leisure; physical environment  and transportationThere was a significant improvement as far as social relations are concerned. It is believed that those individuals who participated more actively in the groups, had a more active participation in society avoided social isolation. More than one month to have the questionnaires answered again may have favored the resultsMoreover, it is necessary to create or implement programs aiming at reintegrating the individual to society, especially the elderly. One alternative would be to refer these individuals to group meetings, in order to improve their social relations.Together with another studyThe primary tool used in the rehabilitation of people with hearing loss is the individual sound amplification device; however, there is also the traditional hearing rehabilitation, which includes hearing training and instructions for speech understandingIn this study, the patients followed the same criteria of the other patients seen in the clinic, with monthly return visits to have questions answered and go through the necessary adjustments in their ISAD. Hearing training was not given. Communication strategies are part of the post-fitting instructions and are given to all patients equally; thus, they were not considered to be discussed in the present study.This study showed that using an ISAD improved the overall quality of life of the individuals assessed.


Agaricus bisporus (button mushroom), Pleurotus ostreatus (oyster mushroom), Lentinus edodes (shiitake) and some closely related species. Large effects of substrate chemistry, strain, developmental stage and ecological interactions result in a wide variation of the concentrations of some metabolites in both mycelial cells and fruiting bodies. This is obviously a result of the high adaptation abilities required to survive in natural habitats. Fungal bioprocesses are decoupled from agricultural production and can be operated anytime, anywhere, and on any scale according to demand. It is concluded that fungal biomass, if produced under food-grade conditions and on an industrial scale, could provide a safe and nutritious meat substitute and protein isolates with a high biological value for future vegan foods.Edible mushrooms are widely appreciated for their appealing flavours, low caloric values and high content of presumably health-protecting metabolites. Their long history of safe use together with the looming worldwide food crisis have revived the idea of generating meat analogues and protein isolates by the controlled fermentation of mycelia of these edible fungi as a dietary option. The occurrence of proteins, polysaccharides, smaller metabolites, metal ions and toxins in mycelia and fruiting bodies is compared among the three most popular species,  Horizon Europe 2021/22 and the Green Deal of the European Commission define the creation and safeguarding of a sufficient and nutritious diet for the growing population of the world as one of the “Grand Challenges” [The millennium goals of the United Nations, the program llenges” . In a tellenges” . ConversLess meat—less heat” [Chlorella and Spirulina), insects  and moulds (Quorn from Fusarium venenatum). However, cultivation of plants requires arable land, a stable environment with sufficient rain, pesticides and harvesting machinery. In addition, harvesting is possible only once per season. Algae turn black when heated or extruded [The shortest remedy was formulated by a famous musician: “ss heat” . Alternaextruded , insectsIn this context, it is surprising that the long-standing idea to propagate the mycelia of edible fungi on a fermenter scale  has not Agaricus bisporus (J. E. Lange) Imbach, Pleurotus ostreatus (Jacq.: Fr.) Kumm., and Lentinus edodes (Berk.) Singer (shiitake) [The central question to be answered by the following comparison of the chemical constituents of fermenter grown mycelia and fruiting bodies of edible fungi is whether such mycelia and the protein-rich fractions derived from them can be regarded as safe and used as food or food ingredients, if produced in a food-grade environment. For the sake of brevity, this text will restrict itself to recent work on the three best investigated and most popular species: hiitake) .The shape of a typical epigeal fruiting body with stipe, pileus and gills a differsL. edodes. An Illumina HiSeq based study counted 2080 differentially expressed genes. 1503 were upregulated in the mycelium, and 577 in a mature fruiting body. The fruiting body-specific transcripts were related to replication, karyogamy, and meiosis during fruiting body maturation [Recent studies using high-throughput techniques have shed light on the differential gene expression during fruiting body formation in turation . Differeturation . Lectinsturation . In addituration .Agaricus blazei (now Agaricus subrufescens Peck), an edible species closely related to A. bisporus, yielded more than 10,000 putative genes, and many of them were differentially expressed in mycelia, primordia and fruiting bodies [μmol/L concentrations of methyl jasmonate and salicylic acid as potent inducers of ergosterol synthesis in liquid cell cultures. Within a short incubation, the concentrations of ergosterol in the mycelium exceeded those in the fruiting body [A. bisporus were preferentially expressed in the peel of the fruiting body, which agrees with their presumed role as a protective hydrophobic layer on the pileus surface [A recent study on g bodies . The latg bodies . A folloing body . In cont surface .P. ostreatus, some were abundant in mono- and dikaryotic mycelia, others were expressed only in mono- or dikaryotic mycelia, and others again were expressed preferably in fruiting bodies. The substrate, natural light and temperature affected hydrophobin gene expression and the subsequent growth rate and primordia formation [P. ostreatus [The bipolar hydrophobin proteins tend to self-assemble in monolayers, which explains both their physiological role as a coating and their possible technological function as an emulsifying agent. They can serve as a guideline for gene expression studies in different stages of the fungal life cycle. Of the 40 hydrophobin genes that were differently expressed temporally and spatially in ormation . The infstreatus .Pleurotus eryngii (kings trumpet), a close relative of P. ostreatus [A recent transcriptome analysis is also available for streatus . A compaAgaricus species ranged from 24 to 48% in dry matter, for Pleurotus species from 15 to 33% and in Lentinula from 10 to 20% [If fermenter produced mycelia were to be introduced as alternative protein sources, a high protein concentration would be desired. Depending on the literature source, cultivation conditions and strain, the total protein concentration of fruiting bodies of the 0 to 20% ,24,25,260 to 20% ,27. Thes0 to 20% . In the 0 to 20% .Quorn, a food preparation containing protein isolated from an ascomycete, was introduced on the UK food market in the early 1980s and no adverse effects have been reported since [Protein isolates can be obtained from disintegrated mycelia by common combinations of extraction, (isoelectric) precipitation and centrifugation. ed since . LiteratA. bisporus, laccase concentration increased during mycelial growth and declined at the onset of fruiting body formation, while cellulase activity increased at fruiting. No such changes were observed for most other enzymes [P. ostreatus was regulated by developmental processes, phytoeffectors and abiotic stress [β-1,3-glucanase of L. edodes occurred after the harvest of fruiting bodies but was not detectable in mycelia and young fruiting bodies [A complex metabolism driven by enzymes lies beneath the protein concentrations summarised above. In  enzymes . The expc stress , whereasc stress , the spec stress . A lentig bodies .A. bisporus [P. sapidus and P. sajor-caju [Lentinula mycelium [L. edodes) to BV 82 (Pleurotus sp.), thus equalling or even exceeding common plant sources, such as bean (BV 58) or even soy, the most valuable plant protein (BV 74) [Pleurotus fruiting bodies cultivated on mixed wheat straw and sugar beet compost, a large inter-species and even inter-strain variation in protein concentration  from 22.4 to 34.7% and glutamate  from 12.8 to 18.7% was observed. L. edodes contained only 15.2% protein, but the highest glutamate concentration with 20.9% [All essential amino acids were present in mycelial proteins, but similar to the protein fraction, absolute numbers deviate from source to source. Glutamic acid was highest in bisporus , P. sapijor-caju , whereasmycelium . When th (BV 74) . Among Pth 20.9% . The datth 20.9% .A. bisporus fruiting bodies, cultured mycelia, and fermentation broth showed that the polysaccharides of the fruiting body were mainly composed of mannose, whereas the mycelia contained mainly glucose and the cultivation medium mainly galactose [L. edodes [P. ostreatus showed very similar properties as “immunoceuticals” [P. eryngii concluded “that the polysaccharides from mycelia can be an effective substitute of polysaccharides from fruiting body” [Since ancient times, the consumption of edible fungi has been linked with health claims, such as preventing Parkinson’s and Alzheimer’s disease, diabetes, hypertension, stroke as well as antitumor, antibacterial, immune boosting and cholesterol lowering properties . Many ofalactose . Dynamic. edodes . A recen. edodes . Focussiuticals” . To prodng body” .A. bisporus and P. ostreatus were cultured in different solid and liquid media [P. ostreatus mycelia accumulated higher concentrations of ergosterol and phenolic compounds than the corresponding fruiting body and showed cytotoxicity toward tumour cell lines. In contrast, fruiting bodies of A. bisporus, when compared to young and old mycelia, contained higher concentrations of antioxidant phenols and ergothioneine  [A comparison of the production of volatile compounds by liquid, surface, and solid support cultures with the fruiting body of  mycelia . This is. 55.5%) . When cuL. edodes contained a low concentration of mannitol (ca. 1% in dry mass) whereas the fruiting body contained 20 to 30% in stripe and pileus, respectively, which was exceeded by the pileus of A. bisporus . The most likely explanation for the high mannitol concentration is the need to maintain the osmotic pressure of the soft fruiting bodies until the onset of sporulation [Mycelia of rulation . Such surulation , which wA. bisporus and L. edodes are a source of lovastatin, a cholesterol-lowering compound (L. edodes (around 1 mg/100 g dry mass). However, the amount of lovastatin released from the extracts of the examined species into digestive juices was highest for the mycelium from in vitro cultures of L. edodes (0.51 mg/100 g dry mass). It was concluded that L. edodes mycelium represented “a product with increased hypolipidemic activity” [Like many other edible fungi, the fruiting bodies of compound b. A compctivity” .A. bisporus, the lowest concentrations of Ni, Pb, Cd, and Cr were found in mycelium samples. As expected, rather high concentrations of Cu, Zn, Fe and Mn were found in fruiting bodies [The accumulation of heavy metals including radionuclides in fruiting bodies is well-known. Comparing young and old mycelia and fruiting bodies of g bodies . Accumulg bodies .A. bisporus. Addition of Zn ions to in vitro cultures increased Mg uptake, while addition of l-tryptophan stimulated the uptake of Zn and Fe [A. bisporus ranged from about 14 to 160 μg/g dry mass, and the l-tryptophan enriched in vitro cultures reached about 119 to 312 μg Fe/g dry mass [P. eryngii [L. edodes, while much lower concentrations were reached in fruiting bodies [Observations of this kind stimulated the idea to supply growing mycelia or fruiting bodies with metal salt solutions during growth to complement possible deficiencies in human nutrition (“biofortification”). An extensive study measured similar concentrations of Mg ions in mycelium and fruiting body of n and Fe . Cd concdry mass . Similar eryngii . Its abig bodies . Thus, iA. bisporus and P. ostreatus were able to accumulate Li and were suggested “as an alternative, more calibrated and bio-accessible source” [P. ostreatus. The authors assumed that the longer period of digestion and uptake would result in more favourable pharmacokinetic properties for the Li ions and thus reduce the side effects. The mycelia of these fungi appear to contain efficient metal ion transporters. Thus, the necessary concentrations of metal ions have to be carefully assessed in order not to produce excessively high concentrations.The concept also worked with supplemented Lithium salts . Lithium source” . EnrichmThe strong dependence of growth and metabolite formation on the chemical surroundings reflects the high potential of fungi to adapt to different and changing environmental conditions. This ability allows the use of agro-industrial side-streams as growth substrates of submerged cultivated mycelia. The side-streams should ideally satisfy certain criteria. They should be of food origin, low priced, high volume, locally produced, rich in nutrients, compatible with the physiology of the fungus, free from growth inhibitors, and easy to mill and sterilise .Agaricus subrufescens , a close relative of A. bisporus, was successfully cultivated on straw from asparagus, maize, bean and cotton, and on cottonseed hulls, corn cob sawdust and woodchips, and significant differences in yield, polysaccharide and antioxidant concentrations were found [The almond mushroom re found ,67.P. ostreatus were cultivated on sunflower husks and barley straw supplemented with corn husks, wheat bran, rye malt, and soy flour [Pleurotus and the emerging fruiting bodies were compared and analysed for presumed health-promoting constituents [Three strains of oy flour . The suptituents . FruitinP. ostreatus and P. sajor-caju [P. ostreatus, in contrast to the low activity in P. sajor-caju. High activities of the horseradish-like peroxidase and laccase were found in the primordia and a high concentration of methoxyphenols in the mycelia of P. ostreatus, which was about 100 times higher compared with P. sajor-caju.Investigating lignolytic enzymes, large differences in the activities of laccase, horseradish-like- and Mn-peroxidase and superoxide dismutase were determined in the mycelia, primordia and fruiting bodies of jor-caju . For exaL. edodes found pronounced effects of the substrate, and in particular of the C/N-ratio, on the formation of soluble sugars, polyols, organic acids, sweet and essential amino acids, umami 5′-nucleotides, trehalose, arabitol, malic acid, succinic acid, mannitol and citric acid [L. edodes and P. eryngii, and two common substrates  a high affinity of L. edodes to Cd was found, whereas other edible species selectively accumulated Al, Ni, Pb or Hg [L. edodes [Not only aroma attributes , but alsric acid . CompariPb or Hg . Mixed a. edodes .Tyromyces floriformis, a sesquiterpene producing basidiomycete with uncertain palatability, showed dramatic responses of submerged cultures to polysaccharides and lipids in the growth medium [w/v) 32-α-l-arabinofuranosyl-xylobiose resulted in a significant suppression. In contrast, the supplementation of the cultivation medium with acetyl donors boosted the sesquiterpene concentration. The effects on sesquiterpene synthesis correlated with the intracellular sesquiterpene cyclase activity. Similar results were obtained using submerged cultures of Cerrena unicolor, Postia placenta, and Coprinopsis cinerea, indicating that the metabolism of mycelia is likewise highly susceptible to the extracellular chemistry.Most of the works cited in this chapter were driven by attempts to improve large-scale production of fruiting bodies and showed the immense effects of the chemical environment on the formation of smaller metabolites, accumulation of minerals and activity of enzymes. Comparisons with mycelia were largely missing. However, a study on h medium . Rye araIn the complex course of the cell cycle of higher fungi, monokaryotic (haploid) cells fuse to dikaryots, and dikaryots fuse in the basidia of mature fruiting bodies. Plasmogamy and karyogamy generate single diploid cells, which enter meiosis, a process eventually resulting in four new monokarytic spores with slightly different genomes. The new cell lines arising from spore germination differ accordingly in growth rate as a global parameter and also in their biochemical properties. The haploid cells may develop into a new mycelium or fuse, if appropriate mating types meet, to a new diploid cell with two individual nuclei. This diploid state may persist until new fruiting bodies are formed. As a result, a large genetic space is formed in which mono- and dikaryotic cells co-exist and grow worse or better depending on their ability to adapt to the actual habitat .P. sapidus, a close relative of P. ostreatus, and grown submerged in standard nutrient liquid (SNL) or in minimal medium supplemented with lemon or orange peel [trans-anethole to p-anisaldehyde was selected as an indicator reaction to evaluate biochemical differences. In contrast to the faster growth of all cultures in the presence of orange and lemon peel, it had a negative effect on the enzymatic activity. Significant differences between monokaryotic strains, and in comparison with the dikaryotic parent strain, were detected for the production of p-anisaldehyde -valencene to the grapefruit flavour compound (+)-nootkatone was measured as an indicator reaction [These findings have inspired the obvious idea to cross the “best” monokarya for a new generation of dikarya with superior properties. However, results were often disappointing, because the selected over-average traits of monokaryotic strains simply did not add up in the new dikarya, a long-standing experience of mushroom growers, but rarely scientifically evaluated and quantified . Our ownreaction . The sloreaction . These rreaction .A. bisporus and related Agaricales and viruses [A. bisporus were observed [Bacillus velezensis is protective against mould contamination [Bacillus megaterium and Pseudomonas putida either alone or in combination are effective in improving growth and yield (“biofertilisers”) [Mycothermus thermophilus were identified [Many land plant species rely on mycorrhizal relationships with fungi to provide them with inorganic compounds and trace elements, a prime example of mutualistic interactions vital for terrestrial ecosystems. Only recently, mutualistic and also antagonistic interactions were described between  viruses , benefic viruses ,85, syne viruses  and adve viruses ,87. Althobserved . Bacillumination , and Bacentified . Certainentified .P. ostreatus compared with a virus-cured strain [Glutamicibacter arilaitensis, a bacterium belonging to the phylum Actinomycetota and found on some French cheese, promoted growth and yield [P. ostreatus against competing fungi [Sauerkraut, where one microorganism paves the way for the next one. It is expected that studies in the near future will shed light on the metabolic changes leading to the improved growth and yield observed for the beneficial “contaminants”.A mycovirus impeded growth and formation of extracellular lignolytic enzymes in d strain , whereasnd yield . Based ong fungi . Such suBundesinstitut für Risikobewertung almost every “edible” fruiting body may cause symptoms of intolerance [A. bisporus may accumulate up to 0.5 g agaritine per kg, a fortunately heat-labile hydrazine and precursor to carcinogenic compounds capable of adding on DNA . Fomes fomentarius (touchwood) is feared as dry rot that destroys entire houses. This leads to the question of whether fermenter grown mycelia of edible fungi, if grown under food-grade conditions, should be legally classified as “novel foods”. Considering both the highly flexible biochemistry of mycelia and fruiting bodies and the lack of a prototypic fruiting body chemistry, it may be derived from the above comparisons that the application of the novel food regulation (EU) 2015/2283 encounters borders marked out by the nature of these peculiar foods. The appendix of (EU) 2015/2283 approved a “chitin-glucan” from the ascomycete Bundesamt für Strahlenschutz has pointed out that increased concentrations of Cs-137 are still present in some regions affected by the Chernobyl fallout of 1986 [Finally, commercially available mushrooms may be contaminated with microorganisms, insect eggs, agro-chemicals and heavy metals, even if properly produced. Wild mushrooms suffer more severely from such problems. The German  of 1986 . All of The quality and safety of foods is subject to legal regulations set by the responsible authorities worldwide. They will question if fermenter grown mycelia of edible fungi are chemically safe if they are worse or better than the fruiting bodies we have long consumed. Questions remain about how to prevent consumers being misled and whether mycelia differ from fruiting bodies to such an extent that their expected consumption would be nutritionally disadvantageous. Answers can only be based on a scientific evaluation of the composition, nutritional value, metabolism, and the level of (un)desirable constituents.A. niger may be cited, which is regarded safe, after it was shown that mycotoxin formation is suppressed under industrial production conditions. This must be proven analogously for any industrially produced mycelia from basidiomycetes  178/2002). Appropriate labelling in the list of ingredients of a food item will be mandatory. The risk of consuming undesired or toxic constituents in mycelia of edible basidiomycetes appears to be much lower than in consuming ascomycete sources such as Quorn.The data discussed above suggest that there is little scientifically founded reason to doubt the safety and nutritional value of mycelia from edible fungi if they are produced under controlled conditions in a defined environment. The production of citric acid by


Pleurotus (Oyster mushroom) is an important cultivated edible mushroom across the world. It has several therapeutic effects as it contains various useful bio-molecules. The cultivation and crop management of these basidiomycete fungi depends on many extrinsic and intrinsic factors such as substrate composition, growing environment, enzymatic properties, and the genetic makeup, etc. Moreover, for efficient crop production, a comprehensive understanding of the fundamental properties viz. intrinsic–extrinsic factors and genotype-environment interaction analysis is required. The present study explores the basidiocarp formation biology in Pleurotus mushroom using an in silico response to the environmental factors and involvement of the major regulatory genes. The predictive model developed in this study indicates involvement of the key regulatory pathways in the pinhead to fruit body development process. Notably, the major regulatory pathways involved in the conversion of mycelium aggregation to pinhead formation and White Collar protein (PoWC1) binding flavin-chromophore (FAD) to activate respiratory enzymes. Overall, cell differentiation and higher expression of respiratory enzymes are the two important steps for basidiocarp formation. PoWC1 and pofst genes were participate in the structural changes process. Besides this, the PoWC1 gene is also involved in the respiratory requirement, while the OLYA6 gene is the triggering point of fruiting. The findings of the present study could be utilized to understand the detailed mechanism associated with the basidiocarp formation and to cultivate mushrooms at a sustainable level. Pleurotus is a widely grown edible mushroom and its rank moved from second to first position. Pleurotus contributes to 26% of global mushroom production [Pleurotus, knowledge of fruiting is essential, but the complete fruiting mechanism has not yet been fully understood. The biological phenomenon of transition from vegetative phase to reproductive phase in mushrooms is always a topic of curiosity amongst researchers. Since mushrooms contribute to a healthy diet vis à vis being storehouses of bioactive compounds, it is important to understand the mechanisms of fruiting to cultivate mushrooms at a sustainable level. Pleurotus production is largely concentrated in Asian countries such as China, Japan, South Korea, Taiwan, Thailand, Vietnam and India. Besides the Asian countries, it is significantly grown in more than 25 countries. The Pleurotus species is the largest among cultivated edible mushrooms at global level. There has been a rapid increase in Pleurotus cultivation in the last three decades. The total production registered a many-fold increase from 1987 to 2019. In 2019, the total global production of Pleurotus was recorded at around 11.18 million tonnes [Pleurotus begins as spores, which germinate and form mycelial threads that colonize on lignocellulose wastes. The mycelia digest the substrate via its active enzyme system and convert it into colonized mycelial mass. The colonized substrate produces the fruit bodies under favorable conditions. The fruit body produces a white-colored spore mass from the gills. Compatible spores with different mating types fuse to produce a dikaryotic mycelium which continues the life cycle in nature. This life cycle in Pleurotus is known as the heterothallic life cycle [Pleurotus as a genus is well known for its species diversity and easy cultivation practices. However, its full productivity is yet to be realized due to the limited availability of potential germplasm with variable response to the growing environment and the agronomic practices. The genotype × environment interaction is a critical factor to enhance the Pleurotus productivity, as environmental factors are directly linked to its fruiting initiation, development and maturity. The fruiting of oyster mushrooms is affected by light, humidity, pH of the substrate, oxygen concentration, and temperature, and thus plays a crucial role in the fruiting mechanism [Pleurotus species [Pleurotus [Pleurotus requires different stages viz. hyphal knot formation, primordial initiation, primordial formation and fruit body formation [Schizophyllum commune [Coprinopsis cinerea [Agrocybe aegerita [Pleurotus mushroom. The studies were also undertaken on simulation, rate kinetics of constructed model, network analysis and topological studies of network of fruiting in Pleurotus spp.oduction ,2. The hoduction . Moreoveoduction , dye makoduction , cosmetileurotus . In respormation ,12. Vari commune , Coprino cinerea  and Agroaegerita . In mostaegerita . Theoretaegerita . In manyaegerita , oxidatiaegerita . The comaegerita . TherefoPleurotus mushroom, the literature was accessed by various sources, such as Pubmed, Google Scholar, and Research Gate, etc. The role of the genes responsible for fruiting and their probable mechanisms were noted and the system biology was studied. Using systems biology graphical notation (SBGN), the system biology studies were conducted in Cell Designer4.1. The Cell Designer software enables the biologist to depict biological process  and molecules  in cells as a diagram/model to show the biological network pathways using graphical notation symbols. In the present study, a model was constructed consisting of a biological network (Pleurotus spp. Complex Pathway Simulator (COPASI), Systems Biology Markup Language (SBML) and ordinary differential equation (ODE) Solver were used for channels simulations in Cell Designer software for identification of principal parameters, variable quantity and simulation. Cell Designer required the Java programming language for operations on operating system (OS) platforms of Windows [To find out the molecular mechanism of fruit body development with response to external stimulus in the  network  and path Windows . The simmax  calculations for each reaction can be performed for a complicated model. The SBML squeezer plugin has a unique feature which harvests data from the model of all biological components in SBGN representation and then chooses to withdraw the necessary data from the Systems Biology Ontology (SBO) annotations [In the constructed model, each reaction was calculated for kinetic rate equations using SBML (Systems Biology Markup Language) squeezer version plugin 1.3 of Cell Designer. SBML is a machine-readable format that helps in describing biological processes such as metabolic pathways, cell signaling pathways, etc. . The ratotations ,26. The otations . QuantitPleurotus spp. fruiting mechanism model was visualized for molecular interaction. The data of the model developed by Cell Designer was exported and used in Cytoscape. BiNoM is also used for analyzing complex biological networks using SBML, SBGN, and BioPAX formats for complicated network structures. In the present study, we used Cytoscape for investigating and visualizing the crucial genes, proteins and paths responsible for Pleurotus fruiting.Network Analyzer is an effective and efficient tool present in Cytoscape that enables us to compute topological network parameters in both undirected and directed networks . This opPleurotus spp., previous studies were conducted on various extrinsic and intrinsic factors that affect fruiting but no clear mechanisms or pathways have been suggested to understand the biology of fruit body development. Using system biology graphical notation (SBGN), we tried to establish a fruit body development model accommodating some of the most important environmental parameters responsible for fruiting. Here, the models were created to attempt to find out the responsible key genes, enzymes and pathways to obtain insight into the fruit body development as well as the effect of the environment on the development and biology. The models showed similarities to earlier studies found in the existing literature and identify some key components responsible for fruiting in Pleurotus [For leurotus . The modPleurotus White Collar complex protein were between 0.0 and 2.0 to study the effect on pinhead formation and fruit body devolvement. All the dynamics are provided in The prediction and behavior by simulation are used to understand the series of events taking place during the fruit development process in mushrooms. The predicted dynamic behavior ascertains the effect of the expression of genes in different environments. Simulation studies are helpful in the interpretation of the fate of various molecular species and metabolites with the degree of importance in the model. In simulation, although it is difficult to estimate the amount or concentration of molecular species/metabolites, an approximation was set up using previous wet laboratory-based studies. Knowing the amount of molecular species in quantitative terms will help us to find links and association amongst species ,32. In oThe model analyzed for the network analysis showed the series of events predicted to occur during fruiting, and a total of 89 nodes and 90 edges were developed. The simple topology parameters are number of nodes, number of edges, connected components, clustering coefficient, network diameter, network radius, shortest paths, average neighbors numbers, density, radius, heterogeneity, centralization, self-loops and characteristic path length . The netk, which means that in the network, the edges are between low-connected and highly connected nodes [Pleurotus fruiting. The other nodes in yellow color had the least regulatory effect on the fruiting mechanism pathways. The green-color nodes show the moderate effect on the model pathway  PoWC1-binding flavin-chromophore FAD to activate respiratory enzymes re-37. The other important pathways according to importance were re-20, re-24, re-27, re-12, re-14, re-22, re-3, re-36, re-25, re-39, re-4 and re-9  protein, are important in fruiting and mostly in the fruit body development. The Pleurotus mushroom is considered as one of the most important mushrooms not only for its edible use but also for its myco-remediation potential [Pleurotus genetic improvement this research may be helpful in the development of early varieties using the important fruiting genes  and a pore-forming protein has been found to have a role in fruiting [OLYA6 gene, also known as Ostreolysin A6 (Aa-pri1 gene) transcript, is found in primordial stages and in immature fruit bodies. These proteins are associated with the membrane-attack complex/perforin (MACPF) domains and have various biological roles, such as defense and attack, organism development, cell adhesion and signaling [To find more detailed fruiting mechanism pathways and their response to different environmental factors, an in silico experiment was designed. Under the specific environmental conditions, development of the fruiting body starts with mycelium/aerial hyphal aggregation. Nutrient, light, humidity and temperature, etc., are some of the factors responsible for fruit body development ,50,51. Tleurotus . The prefruiting . The OLYignaling . Proteinignaling . It is oignaling . Moreoveignaling . It is aignaling , but theOLYA6 may be the triggering point of fruiting. Phenotypically hyphae aggregation may be governed by this protein. In our simulation study, it was found that the Pleurotolysin is an important protein for mycelium aggregation   had someregation .Schizophyllum commune [Fbh1 gene is transcribed and translated in high humidity and produces hydrophobin proteins that interfere in aerial hyphae, spores and fruiting body development. Structurally, the Fbh1 and POH1 hydrophobin proteins contain two cysteine and 113 amino acids clusters [Pleurotus ostreatus, the POH1 (P. ostreatus hydrophobin 1) gene showed a regulating effect for limiting pinhead development while POH2 and POH3 expressed specific levels of hydrophobins at the vegetative phase [vmh3 and Fbh1 from P. florida that were expressed in vegetative mycelium and fruiting body development, respectively [Another important step after mycelium aggregation is pinhead formation. In a humid environment, the hydrophobin gene is the most highly expressed gene of all during fruit body development. Hydrophobin is cysteine-rich and abundantly expresses fungal proteins ,58. Hydr commune ,41. In tclusters . During ve phase . Penase ectively ,43. Variectively , while iectively .Pofst3 gene is responsible for controlling the number and shape of sporocarps. Pofst3 is involved in the regulation by inhibiting the cluster formation in P. ostreatus [Pleurotus, as reported in studies, blue light significantly contributes to the growth of pileus [Pleurotus White Collar 1 combines with FAD (flavin adenine dinucleotide) to form the PoWC1–FAD complex that induces the transcription and expression of respiratory genes by binding to their promoter site and inducing gene expression that leads to fruiting body development. In the model, it can be seen that both Glucose-6 phosphate dehydrogenase (G6PD) and phosphofructokinase (PFK) enzymes are involved in phosphoenol pyruvate (PEP) formation  have been overexpressed in the formation of the prophase of mycelium vegetative growth in P. ostreatus and are so closely related to lignin degradation [PoWC1 protein  in streatus . Hyphae f pileus . Transcrf pileus . A photoormation . The G6Pmushroom . A similelopment . After Pelopment . PEP entelopment ,67. Durielopment . Moreoveing body ,69. PoLaormation . The PoLstreatus . Other Lradation . In netw protein . In simuPAL), are protein genes, which were highly expressed in the phenylpropanoid pathway [PAL genes expression are found under abiotic stresses, mainly UV light, temperature, salts, etc., that lead to the collection of phenolic compounds [PAL genes, i.e., PAL1 and PAL2 are identified in P. ostreatus genome during the developmental stage, the expression of both genes was shown in all developmental stages but the highest expression was observed in fruit gills [PAL2 gene significantly up-regulated the process during primordia, fruiting body and spore development in P. ostreatus [PALs, along with tyrosinases, are involved in pileus pigment formation [Some other genes shown in the model, such as Phenylalanine ammonium-lysae , the Fbh1 gene (hydrophobin) and the Pofst gene. PoWC1 is involved in fulfilling both structural changes and helps to fulfill respiratory requirements, while the Fbh1 gene (hydrophobin) and Pofst gene are mainly responsible for structural change. Moreover, OLYA6 may be the triggering point of fruiting but does not act as the governor of key regulatory pathways. Several other biosynthesis genes  and respiratory enzymes  are regulating the entire fruiting body development process during the primordial, fruiting body and spore development process in the Pleurotus mushroom. To understand the specific role of differentially expressed genes might help determine good candidates for in-depth experimental studies. Additional comprehension of different environmental factors and genes involved enhancing or supporting fruiting body induction and development is censorious from the trade point of view and will be helpful in the future for commercial Pleurotus mushroom cultivation.This study widens our knowledge of the advancements in the omics analysis technology by using different bioinformatics tools, demonstrating how the developmental and regulated genes/enzymes in the model are responsible for the fruiting body development of the edible