GEP-1 was isolated and purified from Gastrodia elata (G. elata) by hot-water extraction, ethanol precipitation, and membrane separator. GEP-1, which has a molecular weight of 20.1 kDa, contains a polysaccharide framework comprised of only glucose. Methylation and NMR analysis showed that GEP-1 contained 1,3,6-linked-α-Glcp, 1,4-linked-α-Glcp, 1,4-linked-β-Glcp and 1,4,6-linked-α-Glcp. Interestingly, GEP-1 contained citric acid and repeating p-hydroxybenzyl alcohol as one branch. Furthermore, a bioactivity test showed that GEP-1 could significantly promote the growth of Akkermansia muciniphila (A. muciniphila) and Lacticaseibacillus paracasei (L.paracasei) strains. These results implied that GEP-1 might be useful for human by modulating gut microbiota.A novel homogeneous polysaccharide named  Gastrodia elata (G. elata), from the family Ochidaceae, is a traditional Chinese medicine, which is widely used in the clinic. Modern pharmacological research shows that G. elata has a variety of biological activities, such as anti-convulsion 2 [4)-α-Glcp-(1→]10 [6)-β-Glcp-(1]5→4)-α-Glcp-(6→, with three branches of β-Glcp and CA-repeating p-HA attached to the backbone chain at O-3 position of 1,3,6-linked α-Glcp and O-1 position of 1,4,6-linked α-Glcp. Bioactivity tests showed that GEP-1 could promote the growth of A. muciniphila and L. paracasei strains. These results imply that GEP-1 could be employed to modulate human gut microbiota for the treatment of diseases.A homogeneous polysaccharide with molecular weight 20.15 kDa and named GEP-1 was isolated and purified from


Regardless of the unadjusted model or adjusted model, patients who received more frequent IT steroid injections seemed more likely to recover hearing . Six IT injections had the highest rate of hearing recovery (79.1%). In conclusion, IT injection number was an independent factor that was positively associated with hearing recovery, and the optimal number of IT steroid injections was 6. Batroxobin plus higher number of IT steroid injections showed more effective for treating SSNHL.The frequency of intratympanic (IT) steroid injection varies from once daily to once weekly or less among studies and does not reach a uniform standard. This study investigated the potential association between the number of IT steroid injections and hearing recovery to determine the optimal number in sudden sensorineural hearing loss (SSNHL) patients. A retrospective study involving 233 SSNHL patients receiving IT steroids plus batroxobin within 7 days of onset was performed. Patients were followed up for 3 months. More than 15 dB of HL improvement in the pretreatment pure tone average (PTA) was defined as effective. The effective group had a higher IT injection numbers than the ineffective group (≥ 6 times: 84.6 vs. 61.1,  Sudden sensorineural hearing loss (SSNHL) is characterized by hearing loss >30 dB occurring at least 3 consecutive frequencies within 3 days . SSNHL aCurrently, the frequency of IT steroid injection varies from once daily to once weekly or less among studies and does not reach a uniform standard , 11, 12.This retrospective study was approved by the regional ethical standards committee of the First Affiliated Hospital of Zhengzhou University. According to Siegel's criteria and guidelines for sudden hearing loss issued by the Chinese Society of Otorhinolaryngology Head and Neck Surgery in 2015, more than 15 dB of HL improvement in the pretreatment PTA was defined as effective; lower levels of improvement were defined as ineffective , 15. AccWe reviewed the clinical data of new, consecutive SSNHL patients visiting the otolaryngology head-neck surgery department inpatient at the First Affiliated Hospital of Zhengzhou University between January 2017 and December 2020. All patients had an extensive history taken and underwent a series of examinations, including physical examination, neurotological examination, imaging, audiology, and vestibular testing. The studied patients received comprehensive treatment, including treatment with IT steroids and intravenous batroxobin , based on the Chinese guidelines for SSNHL . BatroxoHearing tests were performed by air- and bone-conducted pure tone audiometry and speech audiometry at screening, after 1 and 2 weeks of treatment, and at 3 months of follow-up. According to the guideline for SSNHL ruled by Chinese Society of Otorhinolaryngology Head and Neck Surgery in 2015  was used for statistical analyses. For categorical data, frequencies and percentages were calculated to describe the distributions of subgroups among patients according to sex, tinnitus, aural fullness, vertigo, location of damaged ear, history of diabetes or hypertension, audiogram shapes, number of IT injections and prognosis effect. Categorical data were compared with the chi-squared test or Fisher's exact test if appropriate. Means (±SD) were used to summarize the average levels of quantitative data, such as age, initial treatment time, PTA and number of batroxobin injections. The Shapiro–Wilk test was used to examine whether the measured values could be approximated by a normal distribution. Quantitative data that were distributed normally were compared by a two-sample During the study period, 763 SSNHL patients were included. Among these patients, 233 patients met the inclusion criteria, and 530 were excluded according to the exclusion criteria. The mean (SD) age of the patients was 44.15 ± 11.71 years, and 48.9% were women. Vertigo, tinnitus and aural fullness were present in 48.1%, 84.5% of patients and 65.7% of patients, respectively. The mean hearing threshold of the affected ear was 72.30 ± 31.85 dB. After analyzing the audiograms, 14.2% indicated low-frequency hearing loss; 16.7%, high-frequency hearing loss; 32.2%, flat-type hearing loss; and 36.9%, profound hearing loss. The mean number of IT steroid injections was 6.69 ± 1.64 injections. After treatment, 61.4% of treatments were found to be effective .P < 0.001). The symptom of vertigo differed significantly between the effective and ineffective groups . The effective group had a shorter initial treatment time than the ineffective group . Audiogram shapes also differed significantly between the effective and ineffective groups (P = 0.001). Compared to the ineffective group, the effective group had higher IT injection numbers . However, sex, location of affected ear, PTA before treatment, number of batroxobin injection, symptoms of aural fullness and tinnitus, hypertension and diabetes did not differ significantly between the effective and ineffective groups .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.


Background/Objectives: Low-uptake of hearing aids among older adults has long dogged the hearing care system in the U.S. and other countries. The introduction of over-the-counter hearing aids is set to disrupt the predominantly high-cost, specialty clinic-based delivery model of hearing care with the hope of increasing accessibility and affordability of hearing care. However, the current model of hearing care delivery may not be reaching everyone with hearing loss who have yet to use hearing aids. In this study, we examine the group of people who do not use hearing aids and describe their characteristics and health care utilization patterns. We also consider what other healthcare pathways may be utilized to increase access to hearing treatment.Design: Cross-sectional, the 2017 Medicare Current Beneficiary Survey.Setting: Non-institutionalized adults enrolled in Medicare, the U.S. public health insurance program for older adults (65 years and older) and those with qualifying medical conditions and disabilities.Participants: A nationally representative sample of 7,361 Medicare beneficiaries with self-reported trouble hearing and/or hearing aid use.Measurements: Survey-weighted proportions described the population characteristics and health care utilization of those with hearing loss by hearing aid use, and the characteristics of those with untreated hearing loss by health care service type utilized.Results: Women, racial/ethnic minorities, and low-income Medicare beneficiaries with self-reported hearing trouble were less likely to report using hearing aids than their peers. Among those who do not use hearing aids, the most commonly used health care services were obtaining prescription drugs (64%) and seeing a medical provider (50%). Only 20% did not access either service in the past year. These individuals were more likely to be young and to have higher educational attainment and income.Conclusion: Alternative models of care delivered through pharmacies and general medical practices may facilitate access to currently underserved populations as they are particularly high touch-points for Medicare beneficiaries with untreated hearing trouble. As care needs will vary across a spectrum of hearing loss, alternative models of hearing care should look to complement not substitute for existing access pathways to hearing care. We certify that this work is novel and provides important contributions to the literature by highlighting the disparities in access to hearing care and alternative pathways of providing hearing care that could address existing disparities among Medicare beneficiaries.- Women, racial/ethnic minorities, and low-income Medicare beneficiaries are less likely to use hearing aids.- They more commonly visit pharmacies and general medical practices for health care.Why does this matter?Less that one in five Medicare beneficiaries with hearing loss use hearing aids. Delivering hearing care through alternative pathways may improve hearing care access.In 2020, 44.11 million adults in the U.S. were estimated to have hearing loss; with the aging of the population, the number is expected to increase to 73.50 millions in the next 40 years , and hasHearing aids have been shown to improve hearing-related quality of life, listening ability, communication, and social and emotional function , 9. EarlBarriers to hearing aid use include high cost and lack of, or inadequate, insurance coverage, perceived stigma by others, significant resources required to navigate current processes, including transportation, mobility, and know-how as well as a lack of clear recommendations or guidance by primary care providers , 14.Current hearing care delivery in the U.S. follows a medical model of specialty clinic-based care, primarily through audiologists and otolaryngologists, which may be costly and time-consuming; even those who have the time and money needed may find it frustrating . The curAlternative approaches to the specialty clinic-based hearing care model tackle some of the shortcomings of the typical clinic-based model, and include community-delivered hearing care, mobile health applications, tele-audiology, pharmacies, retail clinics such as Costco and Walgreens, and involve primary care providers in hearing care . CommuniRecognizing the importance of treating hearing loss and the presence of barriers to hearing care access at the national level, the President's Council of Advisors on Science and Technology and the National Academies of Sciences, Engineering, and Medicine released reports with recommendations to improve the accessibility of hearing care for older adults , 20, andThe OTC Hearing Aid Act required the Food and Drug Administration (FDA) to develop regulations by August 2020 for the sale of over-the-counter hearing aids to treat mild to moderate hearing loss. The FDA has missed this statutory deadline but are scheduled to release these recommendations by the end of the year .By allowing the sale of OTC hearing aids that would be regulated to ensure the safety and efficacy of these devices, people with self-perceived hearing loss will be able to purchase hearing aids without assessment or counseling from a hearing care professional. However, ensuring proper access to hearing care services is essential to promote optimal hearing loss management and maximal benefit from hearing aids for those who need them. In a randomized-controlled trial, people who self-selected their own pre-programmed hearing aids via an OTC service-delivery model, compared to those who received hearing aids via an audiologist-based service-delivery model, were less likely to be satisfied with their hearing aid or purchase one after the study .From a public health perspective, it is unclear to what extent alternative models currently available reach people with hearing loss who have yet to use hearing aids. People with untreated hearing loss have different health care utilization patterns and costs compared to those without hearing loss,  and compIn this study, we identify who the current model of hearing care is serving and how those individuals differ from Medicare beneficiaries with untreated hearing trouble. We then explore the other health care service patterns of the population with untreated hearing trouble and describe the populations accessing the most common health care services by sociodemographic characteristics and health status, to demonstrate the population that could be potentially reached via alternative delivery models of hearing care. We focus in on medical providers and pharmacies as those services have the highest utilization across the Medicare population.We used the 2017 Medicare Current Beneficiary Survey and Cost Supplement file, a nationally representative survey of Medicare beneficiaries linked to administrative claims data. Medicare is the publicly-funded health insurance program in the United States for adults aged 65 years and older and those under 65 years who qualify based on medical condition or disability. For this study, 7,361 Medicare beneficiaries who self-reported a little or a lot of trouble hearing or reported using hearing aids were included in the analytic sample.The analysis was separated into two sub-populations: untreated functional hearing impairment (those who self-reported a little or a lot of trouble hearing but no hearing aid use), and those who did report using a hearing aid.via the internet, they were counted as not having a prescription drug touch point as we are trying to identify in-person visits that might lend themselves to receiving other healthcare services.The primary outcome of interest was healthcare utilization by service type, including inpatient, outpatient , medical provider , prescription drugs, home health, and skilled nursing facilities services. These utilization variables were derived from both survey report and administrative claims data and through an adjudication process developed by the MCBS administrators . As the Medicare beneficiaries who reported untreated functional hearing loss were described according to the health care service types utilized. Population characteristics used to describe the groups included sociodemographic characteristics, health and functional status, and access to online information. Sociodemographic characteristics included age, gender, race/ethnicity, educational attainment, income relative to federal poverty level, living arrangement, urban/rural status, and supplemental insurance coverage. Supplemental insurance coverage includes Medicaid for low-income adults, Medicare Advantage (the private arm of Medicare), and Medigap the supplemental plans that correspond with the public program. Medicare Advantage plans may include coverage of hearing aids. Health-related variables included severity of hearing trouble, functional vision impairment, number of chronic conditions, cognitive impairment, number of activities of daily living limitations (ADLS), and having a helper to carry out ADLs. Access to online information included having a personal computer at home and using the Internet to get information.This study is a descriptive analysis of socio-demographic characteristics among Medicare beneficiaries with untreated hearing loss across different groups based on health care utilization. All analyses used the survey weights provided by the Centers for Medicare and Medicaid to account for the complex survey design of the MCBS including the over-sampling of black and Hispanic populations, and survey non-response. Weighted proportions were used to describe the characteristics and health care service utilization of Medicare beneficiaries with functional hearing loss by hearing aid use, and the characteristics of people with untreated functional hearing loss by health care service type utilized. Group comparisons were made using Pearson's chi-squared test of independence. StataSE 14 was used to conduct all analyses.Overall, 7,361 Medicare beneficiaries reported hearing trouble or hearing aid use, representing a weighted sample of 28,195,657 Medicare beneficiaries. Among them, 87% reported not using hearing aids . A greatThe health care services most accessed by Medicare beneficiaries with hearing trouble in 2017 were obtaining prescription drugs, seeing a medical provider, and utilizing outpatient services . Among tThe characteristics of Medicare beneficiaries with untreated functional hearing loss are presented by service type utilized . Fifty pSixty-four percent of Medicare beneficiaries with untreated functional hearing loss obtained prescription drugs in the past year. Compared to those who did not, a greater proportion of those who obtained prescription drugs were non-White, a woman, and younger than 65 years old and older than 75 years. A larger proportion of individuals had low educational attainment and income, had supplemental insurance coverage, especially Medicaid, Medicare Advantage, or Medigap, were living alone or with children and other family members, rather than living with a spouse. A larger proportion of people who obtained prescription drugs in the past year relative to those who did not obtain prescription drugs had chronic conditions, cognitive impairment, a lot of trouble (rather than a little trouble) hearing, trouble seeing, limitations in activities of daily living, and had a helper. Compared to those who did not obtain prescription drugs, fewer Medicare beneficiaries who did obtain prescription drugs reported having a computer or using the internet for information.Among Medicare beneficiaries with untreated functional hearing loss, 20% did not see a medical provider or obtain prescription medicines in the past year. Compared to those who utilized any of these services, those who did not were younger (<65 years old), White, men and had higher educational attainment and income. Among those who did not obtain prescription drugs or see a medical provider in the past year, 69% did not have supplemental insurance coverage, compared to 89% of those who utilized at least one of the services, and 57% lived with a spouse, compared to 49%. A smaller proportion of those who did not access any of the services compared to those who did had chronic conditions (87 vs. 95%), cognitive impairment (6 vs. 11%), limitations in activities of daily living, a lot of trouble hearing (rather than a little trouble hearing), trouble with vision, and had a helper. Of those who did not obtain prescription drugs or see a medical provider in the past year, 76% had a personal computer at home and 70% used the Internet to get information, compared to 67 and 61%, respectively, among those who accessed any of the services.This study reinforces the low uptake of hearing aids among Medicare beneficiaries who report having hearing trouble and the differences in socio-demographic characteristics and health services utilization between those who use hearing aids and those who do not. Almost nine in ten (87%) beneficiaries who identify hearing problems are not serviced by the current model of hearing care, which involves high-cost devices and a predominantly specialty clinic-based approach to hearing care. Those with untreated functional hearing loss are more often women and racial/ethnic minorities. They have lower incomes, and more functional limitations than those who use hearing aids. This corroborates the financial and physical barriers to accessing hearing aids as found in previous analyses , 28.Many studies highlight the importance of hearing to one's health and well-being. From a health system perspective, untreated hearing loss is associated with higher health care costs and poor health outcomes . The curWhile these alternative delivery models are being trialed in specific locations, this analysis provides a nationally representative picture of health care access patterns among Medicare beneficiaries with self-reported hearing trouble to provide insights into which alternative models of hearing care may be the most accessible to these individuals. Sixty-four percent of beneficiaries with untreated functional hearing trouble visited a pharmacy and 50% visited a medical provider. Our analysis suggests that a hearing care program run through a pharmacy would reach a population with greater financial and physical barriers, than a program run through a medical provider clinical only. Those who visited the pharmacy were more likely to be lower income, in a minority racial or ethnic group, with greater comorbidities, and functional limitations, than those who did not go to the pharmacy. Pharmacies have taken on a greater role in delivering health care to communities over time across other aspects of health including administering vaccinations, medication reconciliation, and patient education .Interestingly, 20% of the untreated functional hearing trouble group did not visit a pharmacy or medical provider in the previous 12 months, suggesting that these approaches would not capture all beneficiaries with self-reported hearing trouble. These non-users tended to be aged 65-74, men, white, with higher income and living with a spouse. They were more likely to report a little rather than a lot of hearing trouble. They were also more likely to have a personal computer at home and use the internet, suggesting they might be good candidates for an online model of hearing care, such as through mobile health applications and tele-audiology. The coronavirus pandemic, which has significantly disrupted access to clinic-based health care, has increased the need for alternative models of hearing care, particularly those that incorporate an online or tele-audiology component. As evidenced in the UK during the pandemic, there remain both provider- and patient-side barriers to delivering tele-audiology services .The purpose of this analysis is to better inform the planning for increasing access to hearing care at a time when the existing model is already undergoing substantial change. The introduction of the Over-the-Counter Hearing Aid Act (2017) which will regulate the sale of hearing aids to treat mild- to moderate-hearing loss over the counter and is expected to spawn a broader array of more affordable, quality-controlled devices available direct to the consumer. If these low-cost devices are placed in pharmacies, or doctors' offices, it may result in greater uptake of hearing care among older adults who have not previously engaged in the existing model of care.With many alternative delivery models focused on increasing access to affordable devices without hearing care services come questions of quality of care. While early analysis of self-fitting devices compared to audiologist supported fitting suggests that comparable outcomes can be achieved , the litUltimately, there is unlikely to be a one-size-fits-all approach to hearing care. Even across two common delivery models such as medical provider and prescription drug there was 34% overlap among Medicare beneficiaries with untreated functional hearing trouble. Alternative delivery methods should consider how to complement the existing model rather than substitute for it. Defining a pathway for hearing care in the U.S. that covers the spectrum from prevention of hearing loss to treatment, and addresses the physical, financial, and emotional barriers to seeking care is a crucial goal of the system. This will require engagement from older adults and current hearing care providers, as well as other stakeholders both inside and outside the health care system.Many alternative or complementary models of care will require changes to existing workforce and reimbursement structures, including forward thinking on scope-of-practice legislation, investments in training and certification of paraprofessionals, such as community health workers, and the reimbursement of education and counseling separate from the cost of the device , 43.The limitations of the study reflect the challenge of refining the measures for hearing loss and utilization based on the survey and administrative data available. Firstly, hearing loss is measured by self-reported trouble hearing, not by a professional examination. The population captured in this analysis is therefore those who recognize they have some degree of hearing trouble and does not reflect the entire population who could potentially benefit from some degree of hearing care. Previous studies have shown that individuals often underestimate their hearing loss and that underestimation can vary by sociodemographic characteristics . For theSecondly, the categories of utilization provided by the MCBS contain broader categories of medical provider and prescription drugs than primary care and pharmacy visits, respectively. Our methods detail the ways that we attempted to refine these variables; however, it is possible that our estimates of potential reach of alternative delivery models may over-estimate those attending primary care clinics or pharmacies. The prescription drug measure does not account for visits to the pharmacy for over-the-counter purchases. It is therefore also possible that this pathway is under-estimated. Finally, we do not assume that attendance at these alternative pathways equates to receiving hearing care treatment.This analysis highlights the ongoing inequities in the existing hearing care system which may be attributable to the financial and physical barriers associated with high-cost devices and clinic-based hearing care. Disruption to the clinic-based delivery model brought on by the introduction of FDA-regulated, direct-to-consumer hearing aids, has resulted in piloting of models in the community, pharmacies, and medical systems. This analysis suggests that these new delivery models may create greater equity in hearing care access by reaching currently unserved populations.cms.gov.The data that supports the findings of this study are available from the Centers for Medicare and Medicaid Services (CMS) but restrictions apply to the availability of these data, which were used under license for the current study, and are not publicly available. Data can be requested from AW, LA, and NR designed the study and drafted the manuscript. AW conducted the data analysis. AW, LA, CN, CM, FL, and NR interpreted the data. CN, CM, and FL critically revised the manuscript for important intellectual content. All authors contributed to the article and approved the submitted version.This work was supported by the Commonwealth Fund (grant number 20192345). This manuscript was also supported in part by funding from the Cochlear Center for Hearing and Public Health at the Johns Hopkins Bloomberg School of Public Health. CN was funded by NIA/NIH grant K23AG059900. NR was funded by NIA/NIH grant 1K23AG065443. Funders and sponsors had no role in the design, methods, analysis, or preparation of the paper.AW reports receiving a speaker honorarium from the American Speech-Language-Hearing Association and consulting honorarium from BioMedical Insights. CN serves on the non-profit board of directors for Access HEARS and the Hearing Loss Association of America. NR is a scientific advisory board member of Shoebox, Inc and Good Machine Studio but does not receive financial compensation. FL reports being a consultant to Frequency Therapeutics, receiving speaker honoraria from Caption Call, and being the director of a public health research center funded in part by a philanthropic gift from Cochlear Ltd to the Johns Hopkins Bloomberg School of Public Health. CM is on the scientific advisory board for Good Machine Inc but does not receive financial compensation. The remaining author declares 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.


Background: The purpose of the present study was to explore the association between resting metabolic rate (RMR) and protein-energy wasting (PEW) risk in Chinese hemodialysis patients by age and gender subgroup.Methods: RMR and body composition (body cell mass (BCM) and fat mass) of 774 patients undergoing hemodialysis were estimated by bio-electrical impedance analysis (BIA). Anthropometric data were collected by a standard measurement protocol, and the upper arm muscle circumference (AMC) was calculated. Biochemical nutritional and dialysis parameters were obtained. Linear regression analysis was used to analyze the relationship among RMR, body composition and nutritional factors.Results: The mean age was 54.96 ± 15.78 years. RMR level in patients was 1463.0  kcal/d. In multiple linear regression models, BCM, left calf circumference (LCC), fat mass were the determinants association with RMR (P<0.001). Among the patients in the sample, 133 (17.2%) had been diagnosed with PEW per International Society of Renal Nutrition and Metabolism (ISRNM) criteria and 363 (46.9%) were being at risk PEW. The area under the receiver-operating characteristic curve (AUC) of RMR for predicting risk PEW was greater than RMR/BCM and RMR/body surface area (BSA). When the cutoff of RMR was 1481 kcal/d it had the higher sensitivity and specificity (82 and 42%), and the AUC was 0.68 in elderly maintenance hemodialysis (MHD) patients (P<0.001). After adjustment for potential confounders, lowest RMR quartile level (<1239) increased the risk of PEW  in all patients.Conclusions: Older patients with PEW have a lower RMR reduction. RMR and RMR/BCM may play the role in objective screening to detect risk PEW in MHD patients, especially in males. The kidney is not only an excretory organ, but also an important energy metabolism organ . MalnutrBesides physical activity, basal energy expenditure (BEE) accounts for 60–75% of total daily energy expenditure, and basal metabolic rate (BMR) is a classical index for daily energy expenditure. Currently, resting metabolic rate (RMR) is used to evaluate energy expenditure in clinical practice instead of BMR due to being easily detected and only 10% higher . Bio-elePatients aged 18–90 years underwent routine MHD at 11 dialysis centers in Guizhou, Southwest China, were enrolled in this multicenter, prospective study. Inclusion criteria: (i) dialysis facilities chosen from Grade III general hospitals and voluntary participation in the study, (ii) patients were assessed by BIA and regular hemodialysis three times a week for 4 h each time. We excluded patients who did not sign the informed consent, had acute illness in the last 3 months, severe sepsis, shock, multiple organ dysfunction syndrome (MODS) and recent change in dialysis modality.2 surface area high-flux polysulfone dialyzers with bicarbonate-based dialysate . The prescribed duration time was 4 h with a blood flow rate of 250 ml/min and a dialysate flow rate of 500 ml/min. The adequacy of dialysis was assessed by using Kt/V . According to Kidney Disease Outcomes Quality Initiative (KDOQI) 2015 update, the target Kt/V should be 1.2 per session.All patients were dialyzed using 1.6 m2 (kg/m2). Upper arm circumference (AC) and left calf circumference (LCC) was measured with soft feet. The measuring point of AC was the midpoint from shoulder to olecranon connection with upper limbs naturally drooping, and that of LCC was the strongest level of left calf. Triceps skinfold thickness (TSF) was performed with calliper and read the needle to the nearest 0.1 mm approximately. To minimize intra-operator variability, the averages of three consecutive measurements were recorded. Upper arm muscle circumference (AMC) was calculated as AC − 3.14 × TSF. Grip strength (GS) was obtained using a hand-held dynamometer. The patient squeezed the dynamometer with all of their strength, typically three times with left hand. Body surface area  was calculated as 0.0061 × height + 0.128 × weight − 0.1529.To evaluate nourishment status in the subjects, we assessed their height (cm) and weight (kg) with light clothes. Body mass index (BMI) was calculated as weight/heightBody capacitance, lean body mass (LBM), fat mass, body cell mass (BCM), intracellular water (ICW) and RMR were assessed using a five-compartment model of bio-electrical impedance analyzer  with an electrical current of 50 kHz. Measurement was performed before a mid-week dialysis session on the non-access site of the body. Patients were laid in the quiet room with the temperature 22–24°C. Two pairs of sensor electrodes were placed on the patient’s right hand and wrist, and right foot and ankle.Blood samples were also obtained in the morning after an 8-h overnight fast. Laboratory parameters included: pre-dialysis serum creatinine (Scr), blood urea nitrogen (BUN), parathyroid hormone (PTH). Biochemical indicators were tested by automatic biochemical detector .2, AMC < 19.62 cm  were classified for the presence of risk PEW. According to the number of items that meet the diagnostic criteria, we defined the mild risk of PEW (one out of three categories), moderate risk of PEW (two categories) and diagnosed PEW  [Diagnostic criteria provided by the International Society of Renal Nutrition and Metabolism (ISRNM) were used to identify risk PEW in our patients. In the present study, serum albumin <38 g/l, BMI < 23 kg/megories) .. Categorical variables were estimated for association of RMR and PEW by the chi-square test. In multivariate linear regression analysis, RMR was modeled as the independent variable and other variables with P≤0.10 were inserted. The receiver-operating characteristic (ROC) curve was carried out for the assessment of accuracy of diagnostic tests. Associations of RMR with presence of PEW risk was performed by logical regression. Throughout the study, P<0.05 was taken as the minimum level of statistical significance.SPSS software  were used for statistical analyses. Means ± SDs or median with interquartile range were used for normally and non-normally descriptive statistics of continuous variables, respectively. Two-way Multiple Analysis of Variance (two-way MANOVA) was used to find the difference of RMR in the groups cross-classified by age and gender. Skewed data among the groups were analyzed using Kruskal–Wallis test2 = 8.228, P=0.042), and there was no statistical significance between gender groups .Only 133 cases (17.2%) were diagnosed with PEW per ISRNM criteria; 33.1% of cases were classified as having moderate risk of PEW and 13.8% of cases were classified as being at mild risk of PEW (met one of four PEW criteria) and 35.9% of subjects fulfilled no PEW criteria . CompareP<0.05). However, statistical differences in RMR levels between age and gender were eliminated after adjustment for LBM. Compared with the younger group, elderly patients had a lower serum creatinine, body capacitance, BCM, LBM, ICW, LCC and GS, there were statistically significant interaction between age and gender groups (P<0.05). On the other hand, female in elderly group had a greater fat mass than men and both in younger group (P<0.05). In additional, age and BMI were higher, and PTH was lower in elderly group compared with younger group, but there were no statistical significance between gender groups. Dialysis duration, Kt/V, BUN, AMC and TSF no statistically significant interaction between age and gender groups (P>0.05).Data are shown in β = 0.646, 0.198, 0.120 and −0.122, respectively, P<0.001) values maintained independent factors association with RMR in all patients. Same factors were also the independent determinants of RMR in male, age < 65, as well as age ≥ 65 patients, data are shown in Supplementary Table S1. However, in female patients, besides BCM, LCC and fat mass, serum creatinine  instead of capacitance was the different determinant association with RMR (Supplementary Table S1).To further investigate the independent correlation of RMR in different individuals, multiple linear regression models were constructed using a model adjusted for LBM, BMI, PTH, GS and ICW. It revealed that BCM, LCC, capacitance and fat mass (Standardized P<0.01). As shown in For predicting PEW, the area under the receiver-operating characteristic curve (AUC) and cut-off value of RMR for males were 0.73 and 1628 kcal/d , for females 0.65 and 1278 kcal/d , respectively. In age subgroups, 0.68 and 1481 kcal/d  for elderly patients and 0.68 and 1609 kcal/d  for age < 65 patients, respectively  increased the risk of PEW in this group , reduced risk with higher RMR level  was also statistically significant .After adjusting for potential confounders , data in Energy monitoring for disease state has become an important indicator of guiding the nutritional support treatment . In gene2 consumption, CO2 production and energy expenditure in the human body [Our previous studies have found that RMR levels in patients with MHD were associated with anthropometry level, such as calf circumference and AMC, which may become a new index to evaluate energy consumption and malnutrition ,24. Bodyman body . The maiman body . Anotherman body . Our stuAdditionally, there is no consensus about whether gender affects RMR, which is especially scare in elderly patients with ESRD . It was A large number of studies have shown that the high prevalence of PEW among HD patients ranged from 31 to 75% when separately assessed with different nutritional methods, including the malnutrition inflammation score and SGA questionnaire . When usSome weaknesses and limitations in the present research should be noted. Firstly, indirect calorimetry is recognized as ‘the gold standard’ which can determine energy wasting. The RMR in the present study is obtained based on the analysis of BIA, which eliminates the effect of fluid loading on RMR. However, whether this method is specific to hemodialysis patients remains to be further studied. Finally it is important to point out that a gender difference in the prediction of PEW in RMR assessed by BIA after adjustment of BCM and BSA, and the sensitivity of male is higher than that of female. Therefore, it is necessary to further explore and improve the indicators for predicting PEW in female patients.In the present study, we have found that older patients with PEW have a lower RMR reduction. RMR and RMR/BCM may play the role in objective screening to detect PEW in MHD patients, especially in males. The current study is valuable because it is one of the largest scale studies to explore the relationship of RMR and PEW in MHD patients.We have evaluated the cut-off value for screening PEW in MHD, which will be a noteworthy tool in clinical practice.Click here for additional data file.


Older adults with hearing loss have difficulties during conversation with others because an elevated auditory threshold reduces speech intelligibility, especially in noisy environments. Listening and comprehension often become exhausting tasks for hearing-impaired elders, resulting in social isolation and depression. The aim of the present study was to investigate the advantages of hearing aid use in relation to relief from listening-related fatigue, which is still controversial. Participants included a sample of 49 hearing-impaired elders affected by presbycusis for whom hearing aids were prescribed. The Modified Fatigue Impact Scale was used to assess cognitive, physical and psychosocial fatigue. The vitality subscale of the Short Form Health Survey 36 and a single item of the multi-dimensional Speech, Spatial and Quality Hearing Scale (“Do you have to put a lot of effort to hear what is being said in conversation with others?”) were also used. The Cognitive Failures Questionnaire was used to investigate daily errors related to lack of memory and reduced mindedness. Hearing aids rehabilitation resulted in improved speech intelligibility in competing noise, and a significant reduction in cognitive and psychosocial fatigue and listening effort in conversation. Vitality was also improved and a significant reduction in the Cognitive Failures Questionnaire scores was observed. Findings from the study indicate that the use of hearing aids in older impaired-listeners provide them not only with an increased auditory function but also with a reduction in listening-related fatigue and mindedness. Fatigue is very common in older people and interferes with daily activities, worsening their quality of life. Its prevalence increases with age  and alsoFor subjects with hearing impairment, the sensory loss results in a degradation of the quality of peripheral signals, which results in the requirement of increased cognitive resources to recognize relevant information from the speech stream, analyze context and recall prior phonological knowledge ,7,8,9. TMental fatigue, in turn, is proved to be more detrimental to goal-directed (top-down) rather than stimulus-driven (bottom-up) attentive efforts, which potentially creates a self-reinforcing loop in hearing-impaired subjects ,13. UnfoOn the contrary, mental fatigue as a chronic condition in hearing-impaired people should be more appropriately considered as a result of increased listening effort and cognitive load over a long period of time . As a maBesides this, mental fatigue in hearing-impaired older adults is also due to both depression and cognitive decline, which are frequently associated ,23 and cThe study cohort was composed of 46 older hearing-impaired subjects, receiving a first prescription for hearing aids at the Tertiary Audiological Centre of the University Hospital of Modena . Epidemiological data are presented in The study was approved by the regional (AVEN) Ethical Committee (n° 260/2009). Informed consent was obtained from all participants.All subjects were aged ≥65 years and presented bilateral, symmetrical sensorineural hearing loss  and a downward sloping configuration of the audiogram for which no cause could be found other than presbycusis . The useSubjects underwent a first evaluation session including a collection of sociodemographic data and medical history. The otologic examination included otoscopy, pure tone audiometry, tympanometry and an acoustic reflex threshold test. Sensorineural hearing loss was confirmed by a normal tympanogram and by a speech-frequency pure tone average (PTA) of thresholds at 0.5, 1, 2, and 4 kHz in the better ear of above 15 dB .Subjects selected to be included in the baseline assessment of pre-fitting functioning were then examined by two senior psychiatrists: the presence of a diagnosable mood, anxiety or psychotic disorder, which were ruled out by the administration of the Structural Clinical Interview (SCID-I) screening tool , and preThe single item of the multi-dimensional Speech, Spatial and Quality Hearing Scale (SSQ) by Gatehouse and Noble : “Do youModified Fatigue Impact Scale (MFIS) , ItalianThe vitality subscale of the 36-item Medical Outcomes Study Short Form Health Survey (SF-36), Italian version . It consThe Cognitive Failure Questionnaire (CFQ) , ItalianSubjects admitted to the study after psychiatric evaluation were asked to fill in the following questionnaires:Binaural, behind-the-ear (BTE) hearing aids were prescribed. All hearing aids were at the minimum feature settings with digital feedback reduction, digital noise reduction and automatically activated fixed directional systems. These systems automatically engage the directional microphone instead of the omnidirectional one when background noise is present, via signal processing decisions made for a given listening environment, and they employ a stationary polar pattern.About six months later, after optimization of amplification and acclimatization to hearing aids, the same subjects were asked to fill in the aforementioned questionnaires again and submitted to a simplified procedure of speech audiometry in aided and unaided conditions.Speech audiometry material contained 5 lists of 20 common two-syllable recorded words, the so-called spondees . The patThe number of words correctly repeated was computed as a percentage. An average use per day (expressed in hours) was obtained by data-logging. A single patient’s amplification and other technical parameters of the hearing aids were not assessed as variables in this study.Statistical analysis was performed using SPSS 20. Descriptive statistics are reported as mean, standard deviation of the means and minimum/maximum values. The Wilcoxon signed-rank test was the non-parametric statistical tool used to compare the repeated measurements on a single sample and a 0.05 level of statistical significance was adopted. Data are reported separately for patients with mild and moderate hearing loss. Since only three patients with severe hearing loss were recruited, they were excluded from the statistical analyses. Effect sizes for non-parametric variables were computed by dividing the test statistic z by the square root of the number of observations (n° of cases × 2) over the two time points according to the formula: r = Z/√n . AccordiThe degree of hearing loss expressed in dB HL was mild (26 to 40) in 10 (21.7%) cases and moderate (41 to 55) in 36 (78.3%). The mean duration of daily use of hearing devices was 10.1 h , slightly less in comparison with previous reports on retired, over 65-year-old hearing-impaired individuals .Z-value and asymptotic, two-tailed significance are reported. The PTA in the aided condition was significantly improved compared to the unaided one and had a large effect size (r = 0.65).All variables are presented in p < 0.005) and had a large effect size (r = 0.65). The CFQ score showed a significant improvement (p < 0.05) and the effect size was medium (r = 0.49). The single item of the SSQ “Do you have to make a lot of effort to hear what is being said in conversation with others?” showed a significant reduction in perceived listening effort after acclimatization to hearing aids (p < 0.05) and the effect size was large (r = 0.62)The improvement in the intelligibility of words was also significant (p < 0.05) and the effect size was large (r = 0.54). A significant improvement was also observed in the psychosocial fatigue subscale (p < 0.05) with large effect size (r = 0.59). The physical subscale scores did not show any significant change at the final assessment (p > 0.05). Finally, the score of the vitality subscale (SF-36) significantly increased in patients with hearing aids (p < 0.05) and the effect size was medium (r = 0.49).The MFIS cognitive subscale scores indicated a remarkable improvement in perceived mental fatigue in hearing-impaired elders after acclimatization and use of hearing aids (p < 0.005) and the effect size was medium (r = 0.38). The MFIS cognitive subscale scores also suggested significant improvement of perceived cognitive fatigue and the effect size was medium (r = 0.32). However, a remarkable improvement was observed in the psychosocial fatigue subscale (p < 0.001), but the effect size was medium (r = 0.45). The score of the vitality subscale (SF-36) significantly improved at final assessment and the effect size was medium (r = 0.39).All variables are presented in Hearing loss is a chronic and often worsening type of sensory deprivation, with related neuropsychological consequences when left untreated . It has This study confirmed that the use of auditory devices allows for better word comprehension in competing noise in elders affected by both mild and moderate hearing loss. It should be noted that signal and noise in the experimental setting were spatially separated so as to involve a lower cognitive load on perceptual and cognitive resources for speech understanding .Moreover, the signal-to-babble ratio of 10 dB is probably more favorable for aided elders than those encountered in realistic sound scenarios .Listening-related fatigue is a common complaint among elders with presbycusis and is also thought to depend on cognitive overload, which is required to compensate for hearing impairment. It is known that cognitive load and listening effort are reduced by use of hearing aids in elders, even when they do not significantly improve speech recognition tests in laboratories . The perIn this sample, no amelioration of physical fatigue was reported. A possible explanation is that auditory deprivation generates a high demand on cognitive resources that could be alleviated by hearing aids but is not related to functional limitation, pain, or muscular weakness, causing physical fatigue such as in patients with cancer . PhysicaAdditionally, patients’ perception of vitality was significantly increased after hearing aids use; the magnitude of the effect was considerable and the minimally important difference (5 points) for the scale was reached  in subjeThe fact that the CFQ score was significantly improved by the use of hearing aids can be interpreted by the finding that a higher score is generally associated with a better selective attention, maybe due to a more efficient active inhibition of distractors . Since sOn the other hand, the strength of individuals’ reports of attention failures over a relatively long time interval  provides the examiners with a better understanding of how cognitive resources support hearing-impaired elders in everyday life and improve appreciation of underestimated cognitive issues related to sensory deprivation.Research into the benefits of hearing aid signal processing has focused its effects on laboratory speech perception tests, evaluating changes in intelligibility and listening effort. However, laboratory speech perception tests have generally failed to capture the complexity of the impairment in understanding everyday life conversation, overlooking cognitive and listening-related fatigue. Further analysis and better understanding of the dimension of fatigue seems particularly important in view of recent findings on fatigue and vigor in patients with hearing impairment, which indicates that fatigue does not directly correlate with the degree of hearing loss , but witThis study shows that susceptibility to both cognitive fatigue and cognitive failures in elders affected by both mild and moderate hearing loss are alleviated by the use of hearing aids, in the absence of major psychiatric disorders. These results also suggest that it would be useful to include a multidimensional fatigue scale and a questionnaire to assess everyday life cognitive lapses in the standard assessment of elder subjects. This could be helpful in providing hearing-impaired older subjects with feedback on their sensory frailty and to improve it through hearing rehabilitation.


Co-treatment with CP55,940 and TCS-1102 also led to increased CB1R-OX1R colocalization in the ventral striatum. This was not seen following co-treatment with TCS-OX2-29, nor in CB1R-OX2R colocalization. The magnitude of effects following co-treatment with CP55,940 and either the DORA or OX2R-selective antagonist was greater in males than females. These data show that CB1R-OX1R colocalization in the ventral striatum underlies cataleptic additivity between CP55,940 and the DORA TCS-1102. Moreover, cannabinoid-orexin receptor interactions are sex-specific with regards to brain region and functionality. Physical or molecular interactions between these two systems may provide valuable insight into drug-drug interactions between cannabinoid and orexin drugs for the treatment of insomnia, pain, and other disorders.The endocannabinoid and orexin neuromodulatory systems serve key roles in many of the same biological functions such as sleep, appetite, pain processing, and emotional behaviors related to reward. The type 1 cannabinoid receptor (CB1R) and both subtypes of the orexin receptor, orexin receptor type 1 (OX1R) and orexin receptor type 2 (OX2R) are not only expressed in the same brain regions modulating these functions, but physically interact as heterodimers in recombinant and neuronal cell cultures. In the current study, male and female C57BL/6 mice were co-treated with the cannabinoid receptor agonist CP55,940 and either the OX2R antagonist TCS-OX2-29 or the dual orexin receptor antagonist (DORA) TCS-1102. Mice were then evaluated for catalepsy, body temperature, thermal anti-nociception, and locomotion, after which their brains were collected for receptor colocalization analysis. Combined treatment with the DORA TCS-1102 and CP55,940 potentiated catalepsy more than CP55,940 alone, but this effect was not observed for changes in body temperature, nociception, locomotion, or  Within the endocannabinoid system (ECS), lipid-based agonists, anandamide (AEA) and 2-arachidonoylglycerol (2-AG) primarily bind to 2 subtypes of the cannabinoid G protein-coupled receptors (GPCR), cannabinoid receptor type 1 (CB1R) and cannabinoid receptor type 2 (CB2R). The orexin system shares many physical and functional similarities with the ECS . It conspathways . Moleculin vivo  the receptor subtype targeted, and (2) separate versus combined compound treatments. Activating CBRs while blocking orexin receptors causes sedation or sleep-like effects . In contCP55,940 (Cat # 90084) and TCS-1102 (Cat # 18495) were purchased from Cayman Chemical Company . TCS-OX2-29 was purchased from Abcam . All compounds were stored at −20°C until use. CP55,940 was first dissolved in 100% methanol, then added to a vehicle solution consisting of: 1 part ethanol, 1 part Kolliphor EL , and 18 parts 1 M phosphate-buffered saline (PBS) . The concentration of stock solution used for CP55,940 varied based on animal weight and treatment dose. TCS-OX2-29 and TCS-1102 were first dissolved in a 10% DMSO solution in PBS, then added to a vehicle solution consisting of 1 part ethanol, 1 part Kolliphor EL, and 18 parts PBS. TCS-OX2-29 was prepared as a 5 mg/mL stock solution. TCS-1102 was prepared as a 1.5 mg/mL stock solution. All compounds were prepared at room temperature, after which they were stored at 4°C overnight before use the next morning.ad libitum access to food, water, and environmental enrichment. All mice were maintained on a 12 h light:dark cycle (07:00-19:00/19:00-07:00). Mice were randomly designated to receive 2 intraperitoneal (i.p.) injections (1 on each side) of the following treatment combinations: vehicle and CP55,940 at 5 doses , vehicle and TCS-OX2-29 at 4 doses , vehicle and TCS-1102 at 4 doses , 1 mg/kg CP55,940 and TCS-OX2-29  at 4 doses, or 1 mg/kg CP55,940 and TCS-1102  at 4 doses, totaling 22 treatment groups (n = 6 per group). CP55,940 doses were based on previously published studies from our group in the same battery of in vivo assays  and approved by the Animal Research Ethics Board and the Scientific Merit Review Committee for Animal Behavior at the University of Saskatchewan. In keeping with the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines, power analyses were conducted to determine the minimum number of mice required for the study, and mice were purchased, rather than bred, to limit animal waste  were purchased from Charles River Labs . Mice were group housed  with o assays , 2021. Tal waste .via a rectal thermometer (°C). Thermal anti-nociception was assessed by the tail flick latency test 20 min following the injections. Mice were restrained with their tails placed ∼ 1 cm into 52 ± 2°C water. The time until the tail was removed from the water was recorded as the tail flick latency (s). Tails were removed after 20 s if they had not been removed already . Locomotion was measured in the open field test 25 min following the injections. Mice were placed in the 55 cm × 55 cm square-shaped open field for 10 min, during which they were free to roam. Distance traveled (m) and average velocity (cm/s) were measured with EthoVision XT . Distance and velocity scores were then normalized and expressed as a percentage of vehicle means (%Vehicle). For statistical processing, tetrad scores were averaged between mice in the same drug treatment group . The trial was completed when the mouse turned its head or body, made 3 consecutive escape attempts, or at 60 s of immobility . Internal body temperature was recorded 15 min after the injections nt group , 2021.™. C.T. Compound , then sliced at a thickness of 20 μm using a cryostat held at −20°C. Slices were mounted on Superfrost™ Plus microscope slides  then stored at −20°C until they were used for immunohistochemistry.A separate set of mice were euthanized, and their brains were collected 30 min after i.p. injections. This tissue collection time was based on the tetrad timeline, as treated mice finished the tetrad test 30–35 min post-injection. Mice were placed in a rodent vapor chamber, which delivered a mixture of oxygen and isoflurane for approximately 2 min before the animal was fully anesthetized. Mice were then transcardially perfused with 5 mL of ice cold 0.9% saline solution, followed by 5 mL of ice cold 4% paraformaldehyde solution. Brains were rapidly dissected from the skull then submerged in 4% paraformaldehyde solution on ice. The perfused mouse brains were then stored at 4°C for 1 day before being submerged in 30% sucrose solution and stored at 4°C for another 1–1.5 days. Once the brains sunk to the bottom of the sucrose solution, the sucrose solution was drained, and the brains underwent flash-freezing using liquid nitrogen. Frozen brains were stored at −80°C prior to slicing. For slicing, frozen brains were embedded in Tissue-Plus2O2 at room temperature for 10 min, (2) rinsing in 1 M PBS 3 times for 5 min each time, (3) blocking non-specific binding at room temperature by incubating in 10% fetal bovine serum at room temperature for 2 h, (4) incubating with primary antibodies at 4°C for 24 h, (5) rinsing in 1 M PBS 3 times for 5 min each, (6) incubating in secondary antibodies at room temperature for 1 h, at which point all steps proceeded in the dark due to the secondary antibodies’ light sensitivity, (7) rinsing in 1 M PBS 3 times for 5 min each, and (8) mounting the immuno-stained slices using ProLong™ Gold Antifade Mountant with DAPI . All immune-stained slices were stored at 4°C before imaging. The following primary antibodies were used: cannabinoid receptor CB1R monoclonal antibody  diluted at 1:500, orexin receptor 1 polyclonal antibody  diluted at 1:50, orexin receptor 2 polyclonal antibody  diluted at 1:200. The following secondary antibodies were used: Goat Anti-Mouse IgG H&L (Alexa Fluor® 594)  diluted at 1:500 was used for the mouse anti-CB1R primary antibody, while Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)  diluted at 1:500 was used for the rabbit-anti-OX1R and -OX2R antibodies. Each brain region was triple-labeled with (1) DAPI-CB1R-OX1R or (2) DAPI-CB1R-OX2R.The immunohistochemistry procedure consisted of the following steps: (1) blocking endogenous peroxidase by incubating with 0.3% Hn = 6 cells in the same sex and drug treatment group.A Zeiss LSM700 confocal microscope  equipped with Zeiss ZEN Black (version 2.3 SP1) software  was used to obtain fluorescent 3D images of the immuno-stained brain slices. 10–12 Z-stacks encompassing an average tissue depth of 15 μm were collected at 63X oil immersion from each ventral striatum and primary motor cortex. ImageJ (version 2.1.0)  was used to merge the Z-stacks to form 3D images for analysis. Within each image, DAPI-immunolabeled cells were randomly chosen for colocalization analysis on ImageJ and its Fiji package . Pearson’s correlation coefficients were calculated for (1) CB1R-OX1R, and (2) CB1R-OX2R . For staED50 and Emax values . For data without a clear dose-response , Emax was reported as the maximum response observed. All Emax data are reported as mean ± SEM. ED50 data are reported as the mean with 95% confidence interval (CI). Isobolographic analyses for body temperature were conducted using ED50 data with 95% CI only because ED50 could not be estimated in other data sets. Homogeneity of variance was confirmed using Bartlett’s test. Statistical analyses for tetrad data were conducted by two-way analysis of variance (ANOVA) to account for both sex and drug treatment. Immunohistochemistry colocalization data are presented as a mean ± SEM where “n” represents individual cells counted within the brain region of a single mouse. Colocalization means are denoted as Pearson’s correlation coefficients. Statistical analyses for the colocalization data were conducted by two-way ANOVA to account for both sex and drug treatment. Post hoc analyses were performed using Tukey’s (two-way ANOVA) test. Significance was set as p < 0.05.Tetrad data are presented as mean ± SEM where “n” represents the number of animals per treatment group. Data from the ring holding assay and tail flick assay are reported as percent maximum possible effect (%MPE) for catalepsy and %MPE for anti-nociception, respectively. Results from the open field test are stated as a percentage of vehicle scores (%Vehicle). Dose-response curves were fit using a three parameter non-linear regression to yield the p < 0.001) (p = 0.0159) , TCS-OX2-29 (1–30 mg/kg), TCS-1102 (0.1–10 mg/kg), and co-treatments of 1 mg/kg CP55,940 and either TCS-OX2-29 (1–30 mg/kg) or TCS-1102 (0.1–10 mg/kg). Treatment with CP55,940 alone or 1 mg/kg CP55,940 + TCS-1102 produced a dose-dependent increase in catalepsy in both sexes . Co-trea< 0.001) . Looking 0.0159) . There wp < 0.001) (p = 0.0032)  . Thus, t 0.0032) .p < 0.001). Co-treatment with 1 mg/kg CP55,940 + TCS-1102 was also more efficacious in producing hypothermia compared to TCS-1102 alone in males (p < 0.001) and females (p = 0.0049)  . There wp < 0.001) and females (p = 0.0049) (p < 0.001)  . Co-trea< 0.001) . BecauseED50 values from Isobolograms comparing compound ED50 was estimated to be greater than the maximum dose evaluated (p < 0.001), and TCS-1102 alone , respectively  (p < 0.001) (p = 0.0293) and both co-treatments of CP55,940 + TCS-OX2-29 (p < 0.001), and CP55,940 + TCS-1102 (p < 0.001), had higher analgesic effects in males compared to females  . Moreove< 0.001) . There w< 0.001) . Given t< 0.001) . With re females . Sex difED50 values could not be estimated for 1 mg/kg CP55,940 + TCS-1102 as no plateau was observed   (p = 0.0005) (p < 0.0001)  . Also in 0.0001) . In fema 0.0005) . Co-trea 0.0001) . No othep < 0.001) (p < 0.001)  compared to TCS-OX2-29 alone in both sexes (< 0.001) . Similar< 0.001) . Locomot< 0.001) . BecauseThirty min post-injection, brain tissue was collected from the following drug treatment groups: 1 mg/kg CP55,940, 1 mg/kg TCS-OX2-29, 1 mg/kg TCS-1102, 1 mg/kg CP55,940 + 1 mg/kg TCS-OX2-29, and 1 mg/kg CP55,940 + 1 mg/kg TCS-1102. Immunohistochemical experiments focused on two brain regions: (1) ventral striatum, a brain region thought to facilitate cataleptic effects  and (2) In both sexes, CB1R-OX2R colocalization in the ventral striatum was higher in 1 mg/kg TCS-OX2-29-treated mice compared to mice co-treated with 1 mg/kg CP55,940 + 1 mg/kg TCS-OX2-29 . Comparein vitro, or investigated their physiological interdependence in complex disease models related to sleep, appetite, and reward. One other study has assessed their dual modulation of body temperature, pain, locomotion, anxiety, and memory in healthy and transgenic male mice  the preoptic anterior hypothalamus and control of body temperature , (2) spiUnlike body temperature, nociception, and locomotion, co-treatment with CP55,940 and TCS-1102 produced longer-lasting catalepsy than CP55,940 or TCS-1102 alone, demonstrating additivity where each drug had an equal role in producing catalepsy. This was not observed for the combination of CP55,940 and TCS-OX2-29, suggesting that OX1R antagonism is more critical in potentiating CP55,940-induced catalepsy. Additivity between these two drugs may be better explained by regional differences in CB1R-OX1R and CB1R-OX2R interactions or heterodimerization in the ventral striatum, a sub-cortical region that expresses all three receptor subtypes to modulate catalepsy . Beyond Sex differences were observed in two scenarios. First, male C57BL/6 mice were more sensitive to the cataleptic effects of 1 mg/kg C55,940 + 1 mg/kg TCS-1102 compared to treatment-matched females. Sex differences in catalepsy were not detected in the TCS-OX2-29-administered groups, indicating that males are more sensitive than females to simultaneous CBR and OX1R- but not OX2R-manipulation. Closer examination of the receptor colocalization results revealed that in the absence of cannabinoid and orexin drug administration, males and females had similar levels of CB1R-OX1R and CB1R-OX2R expression in the ventral striatum. Following treatment with TCS-OX2-29 alone, cells from the ventral striatum of males had higher levels of CB1R-OX1R/OX2R colocalization compared to females. In all other drug treatments, no significant difference in cannabinoid and orexin receptor colocalization was observed between sexes. Male rodents have lower hypothalamic mRNA levels of OXA and OXB precursor, prepro-orexin , as well9-THC compared to female rats in late proestrus (The second sex difference observed was within the tail flick test. Males were more sensitive to the analgesic effects of CP55,940 alone, as well as both combination drug treatments. Most behavioral studies have reported that females are more sensitive to the cataleptic and anti-nociceptive effects of phyto- and synthetic cannabinoid agonists . With reroestrus . Estrus Cannabis is one of the most highly consumed psychoactive drugs in the world . Many peThe raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The animal study was reviewed and approved by Animal Research Ethics Board and the Scientific Merit Review Committee for Animal Behaviour at the University of Saskatchewan.in vivo experiments and edited the manuscript. RBL aided in the design of the experiments, analyzed the data, and edited the manuscript.HJK designed and executed the experiments, analyzed the data, as well as wrote and edited the manuscript. AZ, AMS, UCE, MJB, and TH assisted with 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.


Editorial on the Research TopicHearing Loss: Mechanisms and PreventionHearing Loss: Mechanisms and Prevention has encompassed 34 contributions from experts who are dedicated to recent advances in the mechanisms and prevention of hearing loss in ways of hair cell damage and prevention, spiral ganglion cell development and inherited hearing loss in animal models and patients, as well as a range of novel treatment approaches for hearing loss.Hearing loss, which is often referred as an “invisible disability,” is one of the most common sensory impairments worldwide. The prevalence of hearing loss is high and it is estimated that 20 percent of the world’s population is affected by hearing loss to some degree in 2021 according to the WHO report. The resulting consequences are significant burdens to the economy and society. There are numerous contributing factors to hearing loss, such as noise exposure, congenital, infectious, traumatic, and immune-mediated causes and the severity of hearing loss is always related with age. To develop preventative and treatment strategies specific to the underlying causes, it is crucial to understand the pathophysiology of these contributing factors. This Frontiers Research Topic entitled Sun et al. identified that the G protein-coupled receptor 125 is expressed in multiple cell types dynamically in the developing and mature cochlea in mice. Stereocilia play an important role in hearing and balancing sensation. Du et al. investigated the role of the Rho GTPase cell division cycle 42 in mice and reported it as a vital regulator in stereocilia development of cochlear hair cells. In recent decades, new technologies have emerged in inner ear research which help researchers reveal the development of hair cells. As such, single-cell sequencing technology is a powerful tool for analyzing gene expression variations across different cell types, and it has also been proven to be useful in inner ear research. Wu et al. reviewed recent applications of single-cell sequencing in inner ear research, covering from identifying unknown cell subtypes, discovering novel cell markers, to revealing dynamic signaling pathways during development. Meanwhile, by using single-cell RNA sequencing analysis, Chen et al. Identified different cell subtypes in the greater epithelial ridge cells in the cochlear duct related to their degeneration during postnatal development in rats. During P1 and P7 rats, five cell clusters reduced significantly while four clusters, enriched with genes associated with the degeneration of the greater epithelial ridge cells, had high similarity in gene expression patterns and biological properties. Besides utilizing single-cell sequencing technique, Wu et al. from a different group developed a 3D imaging technique for the three-dimensional examination of the microstructure of the full thickness of the tympanic membranes in mice. With this imaging technique, they discovered the 3D form of the elastic and collagen network, as well as the close spatial relationships between the elastic fibers and the elongated fibroblasts in the tympanic membranes, which provides important information for hair cell development.Hair cells are most important part in sound conduction and balancing sensation. Their development is tightly correlated with the function of cochlea. To reveal cochlear development and hearing, Liu et al. reported excessive accumulation of calcium due to acoustic overexpression and slow clearance around the presynaptic ribbon might lead to disruption of calcium homeostasis by compared the consequences of noise-induced cochlea synaptopathy of C57BL/6J and CBA/CaJ mice. The susceptibility of noise-induced cochlear synaptopathy in CBA mice is caused by mitochondrial dysfunction of inner hair cells. Xiao et al. investigated the molecular behavior of high-mobility group box 1 (HMGB1) in the cochlea following noise exposure both in mice and in vitro and reported that HMGB1 has a possible negative effect on cellular lifespan indicated by the higher cell viability observed in the HMGB1 knocked-down mice after stimulation with H2O2. In addition to resolving the intrinsic cause of noise-induced hearing loss, researchers are also working on the approaches to protecting noise-induced damge. The use of FK506 (tacrolimus) to treat noise-induced hair cell loss and noise-induced hearing loss (NIHL) has been applied clinically and He et al. identified the downstream mechanisms of FK506-attenuated NIHL. They found that FK506 treatment not only inhibits calcineurin activity to attenuate moderate-noise-induced outer hair cell loss and hearing loss, but also inhibits reactive oxygen species and activates autophagy. Badash et al. demonstrated that endolympahtic hydrops are correlated with noise-induced cochlear synaptopathy by exposing live CBA/CaJ mice to various noise intensities and using optical coherence tomography to measure endolymph volume. Liang et al. also found a positive role sirtuin-3 in protecting cochlear hair cells against noise-induced damage via the superoxide dismutase 2/reactive oxygen species signaling pathway. Many different factors are also involved in the development of hair cells and contribute to hearing loss. Ding et al. identified that the ototoxicity of 2-hydoxypropyl-beta-cyclodextrin (HPβCD) spread from the high-frequency base towords the low-frequency apex of the cochlea from P3 to P28 and the HPβCD-induced outer hair cell (OHC) death is correlated with the upregulation of prestin in OHCs. In addition, 4–6 weeks post-HPβCD treatment, there is a second, massive wave of degeneration involving inner hair cells, pillar cells, auditory nerve fibers and spiral ganglion neurons. An interesting effect of caffeine in cochlear hair cells was identified by Tang et al. They showed that caffeine induces autophagy and apoptosis in auditory hair cells via the SGK1/HIF-1ɑ pathway, which suggests overdoes of caffeine may lead to hearing impairment. Gong et al. described the importance of claudin h in morphogenesis and auditory function of the hair cells. Zebrafish with deficiency of claudin h have significant reduction of otic vesicle size and loss of utricle otolith and loss of hair cells in neuromasts caused by the deficiency of claudin h can be rescued by claudin h mRNA in zebrafish. Tu et al. demonstrated that the deficiency of small muscle protein, x-linked (SMPX) causes stereocilia degeneration in cochlea and progressive hearing loss using an Smpx null mouse model by CRISPR-Cas9 technique. Kwesi et al. presented a study of effect of high jugular bulb (HJB) on the hearing loss in patients with large vestibular aqueduct syndrome (LVAS). LVAS patients with concurrent HJB show higher air conduction thresholds.A great effort has been made to study the cause of hair cell damage and the approaches to protecting them. Noise can induce cochlear hair cell damage and it is the most common cause of hearing impairment. Noise-induced hearing loss involves different mechanisms and pathways. Li et al. Although hearing loss can be caused by various factors, new approaches to treating hearing loss are emerging. Dong et al. revealed the positive function of optic atrophy1 (OPA1) in hearing by examining the ability of OPA1 to protect against cisplatin-induced cochlear cell death both in vitro and in vivo. They showed overexpression of OPA1 prevented cisplatin-induced ototxicity, which suggests a possible role of OPA in ototoxicity and/or mitochondria-associated cochlear damage. It has been demonstrated before that neither N-acetylcysteine nor dexamethasone can protect hair cells from oxidative stress when at ineffective concentrations, but Bai et al. reported when these two drugs combine together, they show a better therapeutic effect both ex vivo and in clinical patients. Chen et al. developed a stable and effective to deliver dexamethasone (DEX) via an electrode coated with polycaprolactone. This device maintains stabilityof DEX concentration for more than 9 months and shows promising application in cochlear implantation. New technologies are also making contributions to treatment of hearing loss, such as stem cell-based therapies as reviewed in He et al. and nanoparticle treatment as reviewed by Huang et al. In the prior review, they fully described the ways of inducing the differentiation of stem cells, the implantation operation and regulation of exogenous stem cells after implanted into the inner ear, and elaborated the relevant inner ear signal pathways and the clinical applications of new materials. In the latter review, they summarized recent developments challenges of nanoparticles in diagnostics and treatment of hearing loss.Apart from cochlear hair cell damage, hearing loss can also relate several neurological disorders, such as Alzheimer’s disease, Parkinson’s disease, Huntington’s disease and autism spectrum disorder, as thoroughly reviewed in Sun et al. explored the regulatory mechanisms of atrial natriuretic peptide (ANP) underlying functional properties of auditory neurons in vitro and reported that ANP could support and attract neurite outgrowth of sprial ganglion neurons (SGN) and possesses a high capacity to improve neuronal survival of SGNs against glutamate-induced excitotoxicity via triggering the natrieretic peptide receptors-A/cGMP/PKG pathway. Ma et al. indicated that the expression levels of vesicle transporter protein 3, glutamate/aspartate transporter protein, and Na+/K+-ATPase ɑ1 are disrupted in spiral ganglion cells in mice after noise exposure, suggesting that disruption of glutamate release and uptake-related protein expression may exacerbate the occurrence of synaptopathy. Some gene mutations are also participated in the development of spiral ganglion neurons. Qiu et al. investigated the pathological role of mutant ATP6V1B2 in the auditory system with transgenic mice carrying c.1516 C > T (p.Arg506∗) in Atp6v1b2, Atp6v1b2Arg506∗/Arg506∗. They showed the transgenic mice have hidden hearing loss at early stages and developed late-onset hearing loss. The degeneration of spiral ganglion neurons are induced by apoptosis activated by lysosomal dysfunction and the subsequent blockade of autophagic flux, which then further impairs the hearing. Meanwhile, scientists are searching for potential approaches to protecting spiral ganglion neurons. Wang et al. described a transgenic mice with tumor necrosis factor 2/4 double knockout show the attenuation of spiral ganglion neuron degeneration by the differential regulation of some core molecules. Chen et al. explored a way to reprogram cochlear Sox2+ glial cells into functional spiral ganglion neurons by induction of small molecules. In the field of regeneration research of spiral ganglion neurons in the inner ear, utilization of specific genetic tool of animal models is a common research approach. For example, a specific spiral ganglion neuron damage approach is described by Hu et al. They generated a strain of transgenic mice exhibiting inducible SGN-specific Cre activity in the inner ear which may serve as a valuable SGN damage model.Spiral ganglion neurons are bipolar neurons connecting the primary auditory receptor cells, the hair cells, with the auditory brain stem. Their development and protection are highly linked with the auditory system. Xu et al. identified a spontaneous mutation of coiled-coil domain-containing 154 gene as a new osteopetrosis-related gene can induce congenital deafness. In porcrine model, Ren et al. studied the population statistics, hearing phenotype, and pathological changes of congenital single-sided deafness (CSSD) which is highly resembled with human non-syndromic CSSD disease. The deaf cochlear of this strain show cochlear-saccular degeneration. In vitro, Wen et al. investigated the mechanisms of Waardenburg syndrome (WS) by inducing an iPSC line derived from a WS patient with SOX10 mutation. The induced cells differentiated into neural crest cells (NCCs) and SOX10 deficiency had a significant impact on the gene expression patterns throughout NCC development in the iPSC model. In children, Liang et al. identified 18 new potential genes associated with congenital deafness and 87 potential new genes associated with otitis media by using a network-based method incorporating a random walk with restart algorithm, as well as a protein-protein interaction framework. In patients, Wang et al. reported a phenotypic heterogeneity of post-lingual and/or milder hearing loss with the GJB2 c.235delC homozygous mutation. Zhu et al. reported a compound heterozygous variant of the OTOF gene in familial temperature-sensitive auditory neuropathy and the auditory neuropathy can be diagnosed by the presence of cochlear microphonics with absent or markedly abnormal auditory brainstem responses (ABRs). Wang et al. further demonstrated the significance of genetic testing for auditory neuropathy patients with p.E818K in the ATP1A3 gene. All these findings have made contributions to a genetic understanding of inherited deafness and provide novel biomarkers for clinical screening.Although various environmental insults can cause damage to hair cells and spiral ganglion neurons, inherited factors will also lead to hearing loss verified both in animal models and clinical patients. In mice, In conclusion, the collection of research articles and reviews presented in this Research Topic provides a comprehensive set of information on the factors attributing to hair cell development, the mechanisms of hair cell damage and the approaches to protect them, as well as spiral ganglion neuron development and protection and genes involved in the inherited hearing loss. Most importantly, there are various new potential treatment approaches to hearing loss. Together, the achievements included in this Research Topic make huge contributions to further understand the underlying causes of hearing loss and may facilitate the development of novel therapies to treat hearing loss in the near future.


Patient. A 63-year-old patient presented with a recurrent cholesteatoma of the left petrous apex. The patient had a long-term history of cholesteatoma and MRI with diffusion-weighted imaging (DWI) detected a suspicious lesion in the left petrous apex on follow-up. Intervention. The cholesteatoma could be completely removed from the petrous apex with partial superior semicircular canal plugging and removal with hearing preservation. Outcomes. Cholesteatomas of the temporal bone are managed by surgery with complete excision of the lesion.  To introduce a novel surgical approach to petrous apex lesion (PA) with superior semicircular canal plugging for hearing preservation.  The translabyrinthine approach, generally useful in nonhearing ears, could be utilized with the additional technique of superior semicircular canal plugging to preserve hearing in this patient.  This case highlights the possibility of a hearing preservation strategy for PA cholesteatomas using a translabyrithine approach. The petrous apex (PA) is embedded in the medial portion of the temporal bone, bordered by several vital structures . A variePA pathologies are usually found incidentally on imaging for unrelated symptoms. If present, they are heterogeneous. The most common complaint is hearing loss, but aural fullness and vertigo secondary to compression of the 8th cranial nerve are also common. These symptoms are present in the majority of patients with petrous apex pathology. Additional symptoms include headaches due to the involvement of the trigeminal nerve, diplopia from compression of the sixth cranial nerve, and seventh-nerve-related facial spasm or weakness. Extended cholesteatoma may also result in Gradenigo's syndrome including purulent otorrhea, abducens palsy, and otalgia . EvaluatThe basic goal of cholesteatoma surgery is the complete removal of the squamous epithelium to minimize the risk of recurrence. Cholesteatoma is a life-threatening disease due to its intracranial complications . Due to Semicircular canal plugging was originally introduced to treat patients with intractable benign paroxysmal positional vertigo and was subsequently applied to other hydropic ear diseases such as Meniere's disease , 9. SemiThe translabyrinthine approach provides the straightest route to the petrous apex , 6, 13. A limited number of studies appear to demonstrate the possibility of hearing preservation in patients after translabyrinthine cholesteatoma removal. Therefore, this case report further highlights the potential mechanisms for hearing preservation after labyrinthectomy. However, further studies are required to implement this approach to preserve hearing for extended cholesteatomas of the petrous apex region.


Minichromosome maintenance complex component 7 (MCM7) is involved in replicative licensing and the synthesis of DNA, and its overexpression is a fascinating biomarker for various cancer types. There is currently no effective agent that can prevent the development of cancer caused by the MCM7 protein. However, on the molecular level, inhibiting MCM7 lowers cancer-related cellular growth. With this purpose, this study screened 452 biogenic compounds extracted from the UEFS Natural Products dataset against MCM protein by using the in silico art of technique. The hit compounds UEFS99, UEFS137, and UEFS428 showed good binding with the MCM7 protein with binding energy values of −9.95, −8.92, and −8.71 kcal/mol, which was comparatively higher than that of the control compound ciprofloxacin (−6.50). The hit (UEFS99) with the minimum binding energy was picked for molecular dynamics (MD) simulation investigation, and it demonstrated stability at 30 ns. Computational prediction of physicochemical property evaluation revealed that these hits are non-toxic and have good drug-likeness features. It is suggested that hit compounds UEFS99, UEFS137, and UEFS428 pave the way for further bench work validation in novel inhibitor development against MCM7 to fight the cancers. Cancer chemotherapy has historically targeted DNA replication because cancer cells proliferate uncontrollably compared to most non-cancerous cells . The McmThe toroidal Mcm2-7 complex, unlike other replicative helicases, is composed of six distinct and essential subunits (numbered 2–7) . Even thOver the last several decades, improvements in anticancer drug research and development have resulted in the discovery and approval of more than 100 anticancer medicines by the FDA ,15. BaseThere are only limited treatment options for cancer, including chemotherapy, radiation, and surgical removal. However, these treatment methods are less successful because of recurring cancer, chemical resistance, and the effect of non-targeted cells. Furthermore, many antitumor agents are plagued by issues of rather undesirable side effects. This affirms the impediment of cytotoxic chemotherapy. Thus, new agents to extend cancer prevention chemotherapy remain vital to be discovered .Drug development necessitates the selection of a few potential hits from a vast library of chemical components. Although, it can be challenging to screen such an extensive compound library using wet-lab assays . MoleculThe 3D structure of the MCM7 protein retrieved from protein data bank [PDB ID: 6XTX] is a hetero 11-mer. The chain ‘F’ is the MCM7 having 719 amino acids . All theNatural compounds were accessed from the ZINC database under the catalogue “UEFS Natural Products” which is a collection of biogenic compounds with an “In-Vitro” activity level . 452 bioUsing the AutoDock Vina program in PyRx software, the prepared library of biogenic compounds was computationally screened against the catalytic site of MCM7 protein . The SDFThe best-screened hits were further docked to the MCM7 protein active site using the ‘Autodock4.2’. A semi-flexible docking technique was employed. The ligand was flexible while the protein was treated as a rigid molecule. The energy of docked ligands (hit compounds) was minimized using the MMFF94 force field. The Autogrid tool was used to create grid maps with specific coordinates for docking hit compounds into the active site of the MCM7 protein. Grid points were kept as 40 × 40 × 40 Å with 0.375 Å spacing. However, the X, Y, and Z coordinates were set as 212.0711, 231.253, and 164.717, respectively. Docking simulation was accomplished using the Solis & Wets local search and the Lamarckian genetic algorithm method. For each docking, hundreds of different runs were used with a total limit of 2,500,000 energy assessments. The binding pose with the highest negative binding energy (BE) value was deemed the most promising. http://www.swissadme.ch/, accessed on 15 April 2021), a web tool with competent in-house methods like iLOGP and the BOILED-Egg, was utilized to generate robust prediction models for physicochemical characteristics  was used to predict toxicity of the best three compounds. ProTox-II accepts a two-dimensional chemical structure/smile ID as input and produces a toxicity profile for 33 models with confidence scores, an overall toxicity radar graphic, and three most comparable compounds with documented acute toxicity [SwissADME  and Cl− (chloride). After minimization, soft coupling with the modified berendsen thermostat (NVT) was applied to heat the system for 100 ps. Periodic boundary conditions (PBC) were applied with a constant number of particles in the system, constant pressure, and constant temperature simulation criteria (NPT) to carry out 30 ns simulations for MCM7 structure and complex MCM7 with UEFS99.GROMACS 2018.1  was usedComputer-assisted drug design has become one of the essential techniques in modern drug design, as it can minimize the cost, time, and labour involved in the drug development process . Since cIn this study, we performed a computational screening of 452 biogenic compounds extracted from the UEFS Natural Products dataset to target MCM7 protein. Among them, the top three hits UEFS99, UEFS137, and UEFS428 showed good binding with the MCM7 protein. The docking analysis showed that 12 amino acid residues  of the MCM7active site were involved in interactions with UEFS99. Pro383, Val385, Ala386, Ser388, Gln389, Asp445, Glu446, and Lys449 residues of MCM7 were interacted with UEFS99 via van der Waals interaction . UEFS137BE and inhibition constant for ‘UEFS99-MCM7’, ‘UEFS137-MCM7’, and ‘UEFS428-MCM7’ complexes were found to be ‘−9.95 kcal/mol and 2.49 µM’, ‘−8.92 kcal/mol and 5.13 µM’, and ‘−8.71 kcal/mol and 9.53 µM’, respectively . An enzyme’s active site is a region of the enzyme with a certain shape that causes it to associate with a specific substrate, causing it to undergo a chemical reaction . The actHydrogen bonds aid in determining the efficacy of the inhibitor against the target protein and play a key role in its stability with the enzyme/protein . Tyr345 50 of 632 µM. Gly384, Val385, Ala386, and Gln389 were the common interacting residues of MCM7 protein with ciprofloxacin and the selected hit compounds  , which rBiogenic compounds and their derivatives are an excellent source of molecules to be investigated for anticancer activities. In cancer pharmaceutics, 49% of chemotherapy drugs are derived from or inspired by natural sources such as plants, microbes, and marine organisms . Vinca aChemotherapy, radiation therapy, and surgical removal are the only treatment options available for cancer patients; phytochemical compounds are considered an alternative therapeutic option to improve treatment efficiency. Additional issues, such as the side effects of standard treatments like chemotherapy ,44 and r.Several drug development failures can be attributed to inadequate pharmacokinetics and bioavailability, in addition to potency and toxicity. Two pharmacokinetic characteristics that should be estimated at various phases of the drug development process are gastrointestinal absorption and brain access . The phy.MD simulations of the protein and protein-ligand complex were carried out to illustrate the inhibition of MCM7 by the UEFS99. A comprehensive analysis of the dynamic trajectories which includes root mean square deviation (RMSD), RMS fluctuations (RMSFs), the radius of gyration (Rg), solvent accessible surface area (SASA), and H-bond profiling of MCM7 and its docked complex, retrieved through simulation was carried out. The analysis revealed very minutes differences between MCM7 and the complex. The RMSD values of the Ca atoms in MCM7 concerning the complex were reported across a 30 ns time frame, indicating a significant increase up to 1 ns, equilibrium at roughly 2.5 to 10 ns, followed by a small increase up to 30 ns, as shown in Figure 5The RMSD values of MCM7 and the complex are presented to show how the compounds affect structural stability and integrity. MCM7 structure revealed a marked increase in RMSD up to 2 ns in this plot and was subsequently optimized up to 27 ns at 0.7 nm, after which the RMSD of the complex reached 1 nm A. The comprehensive analysis of the RMSF profile of the MCM7 and the complex revealed variations of amino acids in the catalytic and non-catalytic regions. The catalytic site of MCM7 revealed fluctuations in the range of 0.2 to 0.3nm. However, the non-catalytic region in the complex structure has a high level of mobility, with fluctuations ranging from 0.2 to 1nm, as shown in 2. However, the SASA of MCM7 showed an average area of 360 nm2, as shown in The solvent accessible surface area (SASA) of the complex and MCM7 structures were computed for 30 ns simulation. The SASA analysis revealed that the complex was exposed to the solvent region with an average area of 390 nmThe radius of gyration (Rg) was measured for both structures. The radius of gyration computed for 30 ns illustrates the compactness of the protein with protein folding and unfolding by employing thermodynamic principles. Rg values for MCM7 protein and complex describe that both the structures maintain equilibrium with the average Rg value of 3.25 nm, as shown in Hydrogen bond occupancy and their number were calculated during 30 ns simulation. Intra-protein hydrogen bond occupancy within the MCM7 and the MCM7-UEFS99 complex remained stable throughout the study. Since the two graphs are overlapping, indicating high stability and good binding affinity of the ligand.Multiple human cancers are caused by the overexpression of the DNA replication licensing complex component MCM7.The present article describes the biogenic compounds screening against MCM7 protein using the in silico art of technique to find promising MCM7 inhibitors. It is suggested that the hit compounds UEFS99, UEFS137, and UEFS428 show good binding with MCM7 protein, with UEFS99 also demonstrated stability as revealed by MD simulation. This study paves the wave for further wet lab validation (in vitro and in vivo study) of these compounds in the form of novel inhibitor development against MCM7 to fight cancers.