This preliminary study aimed to develop a regression model to estimate the resting metabolic rate (RMR) of young and middle-aged Koreans using various easy-to-measure dependent variables.The RMR and the dependent variables for its estimation  were measured in 53 young  and middle-aged  healthy adults. Statistical analysis was performed to develop an RMR estimation regression model using the stepwise regression method.1 regression models estimated only by FFM was 66.7% (R2) and 66.0% (adjusted R2), while mean standard errors of estimates (SEE) was 219.85 kcal/day. Additionally, mean explanatory power of RMR2 regression models developed by FFM and age were 70.0% (R2) and 68.8% (adjusted R2), while the mean SEE was 210.64 kcal/day. There was no significant difference between the measured RMR by the canopy method using a metabolic gas analyzer and the predicted RMR by RMR1 and RMR2 equations.We confirmed that FFM and age were important variables in both the regression models based on the regression coefficients. Mean explanatory power of RMR1 = 24.383 × FFM + 634.310, RMR2 = 23.691 × FFM - 5.745 × age + 852.341.This preliminary study developed a regression model to estimate the RMR of young and middle-age healthy Koreans. The regression model was as follows: RMR The total energy expenditure in 24 hours consists of RMR, physical activity energy expenditure (PEE), and diet-induced thermogenesis (DIT). The RMR represents approximately 60-75% of daily energy expenditure (DEE) in a 70 kg person3, accounting for the largest contribution to the 24-hour energy expenditure. It consequently has a large impact on the regulation of body composition and energy balance. An abnormally high RMR is associated with pathologic and inflammatory conditions. It also tends to decrease with aging, with a low RMR playing an important role in the pathogenesis of obesity and age related chronic diseases6. Therefore, an accurate measurement of the RMR is very important.Resting metabolic rate (RMR) is the total number of calories burned when the body is completely at rest. It is proportional to the lean body mass and decreases by approximately 0.01 kcal/min for each 1% increase in the body fat9. However, both the methods require tedious procedures and are time- and cost- consuming. Thus, many researchers have developed various regression models for RMR estimation14. Determining the contribution of the RMR to the DEE is an important calculation for understanding, developing, and executing body weight- related interventions4. For example, RMR estimation regression model is applied to determine the target energy intake in weight loss programs, develop dynamic prediction models of weight gain and loss, identify the patients with potential metabolic abnormalities, design public health programs promoting obesity prevention in diverse populations, and assess the potential energy deficits in metabolically stressed patients4.Methods for measuring the RMR include direct and indirect calorimetry, with the latter being used commonly due to a more efficient measurement. It is further divided into two methods, one using doubly labeled water; and the other using a human metabolic chamber, a hood-, or a mask system10 was the most commonly used, but only 50-75% of the RMR variability could be explained by this equation. Its major disadvantage was overestimating RMR by at least 5%. Additionally, RMR was estimated only by age, height, and weight and the regression rate, R2 value, did not exceed 0.7. Previous attempts to overcome these shortcomings failed to show high regression rates14, as they also estimated RMR with only age, height, weight, and lean body mass. Therefore, developing a regression model, with a higher regression rate using various dependent variables, is important for accurate RMR measurements.The previous regression models were developed using only height, age, weight, and fat-free mass (FFM) and hence did not have a high correlation and regression rate. The Harris–Benedict regression modelThe present study was a preliminary study and it aimed to evaluate the Korean adults  to generate regression equations to predict the RMR from age, height, body mass index (BMI), FFM, fat mass, % body fat, systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), pulse pressure (PP) and resting heart rate (HR).7. All subjects were of Korean origin, with a stable weight for at least 3 months prior to the measurements, and without a history of thyroid disease, diabetes mellitus-I or II, cardiovascular disease, or severe hypertension in the past 6 months. There was also no history of orthopedic disease or other medical issues over the past year in the pre-screening surveys. All the subjects received a medical clearance for their participation and were explained about the purpose, procedures, and potential risks of the study. All proceedings of the study were approved by the Institutional Review Board of Konkuk University (7001355-201903-HR-305) in Korea and were conducted according to the Declaration of Helsinki. All subjects arrived at the laboratory early in the morning (8:00 AM) after overnight fasting (≥ 8 hour) and rested for 30 minutes, after which their blood pressure, body composition, and resting HR was measured, followed by the RMR measurement. All the subjects were instructed to sleep for at least 8 hours before the RMR measurement and to stay awake during the process. If they fell asleep, their shoes or toes were squeezed to keep them awake.Fifty three young  and middle-aged  healthy adults were included in the present study <Height, BMI, body weight, FFM, fat mass, and % body fat were measured using the bioelectrical impedance analysis equipment . All subjects fasted overnight prior to body composition measurement. The subjects wore lightweight clothing and were asked to remove any metal items.After all the subjects were sufficiently rested for more than 20 min, their blood pressure was measured twice using an autonomic blood pressure monitor  and the average value was used for analysis. The blood pressure parameters measured were SBP, DBP, MAP, and PP. The resting HR was measured using an autonomic HR monitor .2 and 5% CO2) before the measurement. All the RMR testing procedures were performed in a 9 m (width) × 7 m (length) × 3 m (height) chamber with a temperature of 23 ± 1 °C and a humidity of 50 ± 5%, regulated by the environmental control chamber . Subjects were asked to limit their physical activity and abstain from alcohol intake one day before the measurement. The subjects took a rest for 30 min, prior to the measurement. The RMR was measured in a supine position for 30 min, and the average value of the last 25 min was used for the analysis.All subjects fasted overnight, prior to the measurement of the RMR by indirect calorimetry using a metabolic gas analyzer  with a flow-dilution canopy hood system. Calibration was performed using the calibration gas . Regression analysis using the stepwise method was used to predict the RMR from age, height, BMI, FFM, fat mass, % body fat, SBP, DBP, MAP, PP, and HR. A two-tailed student’s paired t-test was used to detect differences between measured and predicted RMR. Bias was calculated as the difference between measured and predicted RMR. The authors rigorously conformed to the basic assumptions of a regression model . Statistical Package for the Social Sciences (SPSS) version 24.0  was used for the statistical analysis and the level of significance  was set at 0.05.The means and standard deviations were calculated for all the measured parameters. The Shapiro-Wilk test verified the normal distribution of all the outcome variables. To perform the linear regression analysis, we verified the independent variables by checking the regression coefficient  was statistically significant <2), adjusted coefficients of determination (adjusted R2), and standard errors of estimates (SEE) were calculated for the regression model. The mean explanatory power of RMR1 regression models estimated only by FFM was 66.7% (R2) and 66.0% (adjusted R2), while the mean SEE was 219.85 kcal/day <2 regression models developed by FFM and age were 70.0% (R2) and 68.8% (adjusted R2), while the mean SEE was 210.64 kcal/day <The coefficients of determination  <In the present study, there was no significant difference between the measured RMR by canopy method, using a metabolic gas analyzer, and the predicted RMR by RMR1 = 24.383 × FFM + 634.310, RMR2 = 23.691 × FFM - 5.745 × age + 852.341).A preliminary study was conducted to develop a regression model for estimating the RMR of healthy young and middle-aged Korean adults using various easy-to-measure dependent variables. Based on the data obtained, our study developed two regression models  and 70.0% (RMR2 equation) of the variance in the criterion variable of RMR was attributable to the variance of the combined predictor or independent variables.10 developed separate RMR estimation models  for males (n = 136) and females (n = 103). Only 50-75% of the RMR variability could be explained by this equation, with the disadvantage of overestimating the RMR by at least 5%. Further, the RMR was estimated only by age, height, and weight and the regression rate, R2 value, did not exceed 0.7. Schofield12 developed an RMR estimation model for Italian males based on multiple sample sizes . However, the samples had a disproportionate number of Italian military cadets, soldiers, workers, and miners, who did not represent the typical Italian population. These constituted 56% of the 18 - 30 years old male cohort2. Additionally, the regression rate of the RMR estimation model was low . Hayter & Henry16 developed an RMR estimation model using Schofieldʼs database, which predicted the RMR in Northern Europeans and Americans, except Italians. However, this model also had a regression rate similar to that of Schofield Previously, Harris and Benedict11 re-evaluated the Harris-Benedict equation using the age, height, and weight used in the latter’s estimation model using normal subject data (n = 239) from the study and additional data, which were obtained from the subjects spanning a wider age range (n = 98). As a result, a model with a relatively higher regression rate  was developed. Also, Mifflin et al.13 developed a predictive equation for RMR from the data of 498 healthy subjects, including females (n = 247) and males (n = 251), aged 19-78 yrs (45 ± 14 yrs). Normal weight (n = 264) and obese (n = 234) individuals were studied and the RMR was measured by indirect calorimetry. This model showed high regression rates for both males and females .Conversely, Roza and Shizgal11 and Mifflin et al.13 using FFM and age, with a smaller sample size. This was due to the strict adherence to the basic assumptions of a linear regression model. Compared to a study by Piers et al.9, which developed an RMR estimation model using similar sample sizes and independent variables, the present model showed a higher regression rate.The present preliminary study developed an RMR estimation model with a regression rate that was similar to Roza & Shizga1 = 24.383 × FFM + 634.310, RMR2 = 23.691 × FFM - 5.745 × age + 852.341. The bias  and correlation  between the estimated RMR and the measured RMR were reasonable.In conclusion, we developed a regression model using FFM and age to estimate the RMR of young and middle-aged healthy Korean adults through preliminary experiments. The developed model was as follows: RMRHowever, the present study was a preliminary study and had its limitations. The regression model for individual genders could not be developed due to the small sample size, and a validity test could not be performed. Further research to overcome these limitations is thus recommended.


Scientific Reports 10.1038/s41598-019-43669-7, published online 14 May 2019Correction to: This Article contains two errors in the Discussion section under the subheading ‘Effect of material properties on the cell interactions’.74, we observed that the un-normalized adhesion of a polysaccharide with cells was very weak, only about 2 nJ for WA07 and 2.7 nJ for HepG2 after 30 s in contact ”“In accordance with previous findings measured with a different method and other cell typesshould read:74, we observed that the un-normalized adhesion of a polysaccharide with cells was very weak, only about 1.6 fJ for WA07 and 2.7 fJ for HepG2 after 30 s in contact ”.“In accordance with previous findings measured with a different method and other cell typesSecondly,“The corresponding un-normalized adhesion values for LN-521 interactions were 3.8 nJ for WA07 and 33 nJ for HepG2 ”should read:“The corresponding un-normalized adhesion values for LN-521 interactions were 3.8 fJ for WA07 and 33 fJ for HepG2 ”.Additionally, the Supplementary Information that accompanies this Article contains errors in Table S1, S2 and S3.In Table S1, the units of adhesion energy are incorrectly given as nJ instead of fJ. Additionally, three values of adhesion energy for CNF in panels (b), (d) and (f) are incorrect.In Table S2, two values of radius are incorrect for CNF in panel (a) and Col I in panel (b).In Table S3, the units of adhesion energy are incorrectly given as nJ instead of fJ.The correct Tables S1, S2 and S3 appear below as Tables


Human cathepsin L belongs to the cathepsin family of proteolytic enzymes with primarily an endopeptidase activity. Although its primary functions were originally thought to be only of a housekeeping enzyme that degraded intracellular and endocytosed proteins in lysosome, numerous recent studies suggest that it plays many critical and specific roles in diverse cellular settings. Not surprisingly, the dysregulated function of cathepsin L has manifested itself in several human diseases, making it an attractive target for drug development. Unfortunately, several redundant and isoform-specific functions have recently emerged, adding complexities to the drug discovery process. To address this, a series of chemical biology tools have been developed that helped define cathepsin L biology with exquisite precision in specific cellular contexts. This review elaborates on the recently developed small molecule inhibitors and probes of human cathepsin L, outlining their mechanisms of action, and describing their potential utilities in dissecting unknown function. Lysosomes play critical roles in human biology receiving, trafficking, processing, and degrading biological molecules from seminal cellular processes, such as endocytosis, phagocytosis, autophagy and secretion. Discovered by the ground-breaking work of de Duve, these single-membrane enclosed cytosolic organelle maintain an acidic ~4.5–5) pH environment, and house close to sixty proteolytic enzymes ,2. Among.5–5 pH ecis Golgi by UDP-N-acetylglucosamine:N-acetylglucosaminephosphotransferase enzyme . ,44. 5,44survival . In addisurvival .Different forms of cathepsin L enzyme have been isolated in distinct cell types and their intracellular and extracellular environment . At specCtsl−/− mice showed that they developed many key traits of dilated cardiomyopathy, such as interstitial myocardial fibrosis, cardiac chamber dilation, and impaired contraction. In addition, the newborn Ctsl−/− mice acquired increased number of acidic organelles, although with altered morphology and function. These studies indicate that the inhibition of cathepsin L activity in these cells is detrimental to the proper functioning of cardiac cells. This was further corroborated by an overexpression study of cathepsin L in mice cardiomyocytes that exhibited cardioprotective effect by inhibiting the Akt signaling pathway . T. TD)-Az-de chain , exhibitde chain  resembleR,R)-(EtO)-Azi-Leu-Pro-OBzl  analog; however both exhibited only a marginal selectivity over cathepsin B with diminished eudysmic ratio. The authors then superimposed and analyzed the structures of certain epoxide and aziridines and postulated that these inhibitors can assume different orientations in the active site while still binding within the enzyme pockets. This scaffold was further explored by Vicik et al. who extended the previous work and developed a series of compounds in which Boc-(S)-Leu-(S)-Azy--Azi(OBn)2 (The type III inhibitor class  is comprzi(OBn)2 , Type I 1 position. Subsequently, they also varied the R2 position while keeping either morpholine or N-methyl piperazine group intact , and Leishmania major [IC50 = 12.5 µM], and did not exhibit any significant toxicity against human aortic endothelial cells and zebrafish. Although thiocarbazate motif showed promise as an inhibitory scaffold, the lack of reasonable stability (it decomposes even in DMSO) and only modest inhibitory activity in cell-based assays probably ceased any further development of the scaffold methionine-labeled proteins only detected two separate bands at 36 kDa and 39 kDa, which correspond to the intracellular inactive precursors of cathepsin L and B respectively. This indicated that the inactive precursor proteins did not react with P1, demonstrating its unique ability to quantify only active protein. Further, P1 was utilized to probe active cathepsin L and B in different human tissues as well as in lysosomes and whole cells [2]Tyr-Ala-CHN2  isomer of the biotinylated probe was incubated with cathepsin L and subjected to gel electrophoresis, electro-transferred to a membrane, and exposed to streptavidin-alkaline phosphatase conjugate, a strong labeling of the enzyme-inhibitor complex was observed. However, when the enzyme was treated with E-64, an active site-directed competitive and irreversible cathepsin inhibitor, prior to incubation with P3, the labeling was diminished, clearly demonstrating that P3 competes for the active site of cathepsin L. In line with this observation, a desthiobiotinylated analog also exhibited the same trend but with reduced labeling due to its weaker binding affinity to streptavidin. Although P3 has a modest binding affinity (Ki = 1.4 µM) to cathepsin L, it exerted a 36-fold selectivity over cathepsin B. Certainly, the affinity labeling technique not only served as an ABP for cathepsin L but also provides a premise for developing aziridine-based chemical tools for functional proteomics.Gelhaus et al. first developed biotinylated aziridine-2,3-dicarboxylate and demonstrated its anti-plasmodial activity using cell-based studies . Since, 2, respectively, prior to the probe treatment [Although covalent modifiers of proteins have been vastly exploited as chemical tools for target identification and functional proteomics, photoaffinity probes offer unique mode of action. They bind proteins non-covalently  first and form a non-selective covalent bond with the closest amino acid residue only upon irradiation. Torkar et al. took advantage of this technique and developed the first photoaffinity-based probe  a two-photon dye, DL-1 pyrimidine derivative), at P1 position, and (b) Disperse Red 1 dye, a fluorescence quencher, in the place of aldehyde moiety. The resulting imaging probes , to validate their approach and to further evaluate the influence of isotopes on enzyme binding specificity. The newly developed probes exerted promising selectivity toward both recombinant proteases and proteases from cancer cell lines, HCT-116 and MDA-MB-231.The authors were able to simultaneous detect the activities of proteases in HCT-116 cells. Moreover, each of the protease-specific probes exerted a similar labelling efficiency in HCT-116 cells, regardless of their metal counterparts, which further reinforces the compatibility of this class of probes as cytometry-labelling agents. They extended their investigation to THP-1 cells, a non-adherent monocyte-like cell line, that expresses both cathepsin B and L but contains very low levels of legumain enzyme. Probe treated THP-1 showed a clear labeling of both cathepsin B and L with no detectable activity of legumain. This finding was consistent with their transcriptional data. Finally, the developed probes not only allowed to detect the activome of cathepsin L, B, and legumain in peripheral blood mononuclear cells (PBMC) but also enabled to categorize NK cells in two distinct populations based on protease activome levels. This strategy, unlike many existing technologies, thus allowed the simultaneous detection of target proteases, thereby providing a more holistic understanding of the activome. This certainly merits the future development of TOF-based probes for multiplexed enzyme activity detection.Although fluorophores-containing molecules have been appreciated as useful imaging probes, commonly used fluorophores often suffer from spectral overlapping that limits the number of targets that can be analyzed concomitantly . To addrThe inhibition of cathepsin L has continue to emerge at the forefront of drug development for several human diseases. Yet, no inhibitory agents targeting cathepsin L have advanced to clinical trials. While inhibitors of cathepsin B and S are currently being evaluated in clinical trials, recent failure of Odanacatib, a cathepsin K inhibitor for osteoporosis, in late stage clinical trial has made pharmaceutical industries wary of targeting cathepsins. The key challenge remains gaining inhibitor selectivity with respect to the other members of cathepsins and directing them to the targeted cell types for selective functional perturbation. With its ubiquitous expression profile, the function of cathepsin L in individual cell types must be precisely defined first; this is especially important since several isoforms of cathepsin L have been reported in the distinct cellular locations, and their activity and functional profile in individual cell types still remain poorly documented. While recently developed cathepsin inhibitors and probes have significantly advanced our understanding of cathepsin L function in both normal and disease cells, more efforts are needed for the development of isoform-selective reagents for further advancement; perhaps new allosteric modules on cathepsin L enzyme can be explored and exploited for precision targeting. Fortunately, several structural coordinates for cathepsin L enzyme forms, some with a diverse set of complexing ligands and some without, are now available . These c®) for CD30-positive relapsed or refractory Hodgkin′s lymphoma. In addition, protease cleavable prodrug strategy has also inspired the development of cathepsin B selective probe and even enabled real-time monitoring of drug release [In recent years, disease-associated protease activatable prodrugs have gained much recognition and garnered breakthrough therapies in the area of antibody-drug conjugation (ADC) . The maj release ,183. Int release . This st


Escherichia coli results in misfolded and non-active protein aggregates in the cytoplasm, so-called inclusion bodies (IB). In recent years, a change in the mindset regarding IBs could be observed: IBs are no longer considered an unwanted waste product, but a valid alternative to produce a product with high yield, purity, and stability in short process times. However, solubilization of IBs and subsequent refolding is necessary to obtain a correctly folded and active product. This protein refolding process is a crucial downstream unit operation—commonly done as a dilution in batch or fed-batch mode. Drawbacks of the state-of-the-art include the following: the large volume of buffers and capacities of refolding tanks, issues with uniform mixing, challenging analytics at low protein concentrations, reaction kinetics in non-usable aggregates, and generally low re-folding yields. There is no generic platform procedure available and a lack of robust control strategies. The introduction of Quality by Design (QbD) is the method-of-choice to provide a controlled and reproducible refolding environment. However, reliable online monitoring techniques to describe the refolding kinetics in real-time are scarce. In our view, only monitoring and control of re-folding kinetics can ensure a productive, scalable, and versatile platform technology for re-folding processes. For this review, we screened the current literature for a combination of online process analytical technology (PAT) and modeling techniques to ensure a controlled refolding process. Based on our research, we propose an integrated approach based on the idea that all aspects that cannot be monitored directly are estimated via digital twins and used in real-time for process control.Overexpression of recombinant proteins in • Monitoring and a thorough understanding of refolding kinetics are essential for model-based control of refolding processes.• The introduction of Quality by Design combining Process Analytical Technology and modeling ensures a robust platform for inclusion body refolding. Escherichia coli results in inclusion bodies (IBs) which generally are biologically inactive and need to be processed to yield an active, final product. However, this disadvantage has to be considered in view of several benefits of IBs like high purity, simple separation from cells and stability against mechanical, thermal, and proteolytic stress  is used for the analysis of protein folding on the basis that folded and unfolded proteins show different spectra  can be applied to predict the secondary structure of proteins by analyzing which wavelengths of radiation in the infrared region of the spectrum are absorbed by the sample Tatulian . TherebyFTIR can illuminate the structural related changes during the refolding process and can be used to determine when to terminate the process to avoid protein loss due to aggregation. In comparison to CD, salt solutions are not problematic and turbid samples can be analyzed  enable measurements of concentrations as low as 0.08 g/L at feedback times of 5 min  provides information on the molecular and geometric composition of covalent bonds, while additionally enabling assessment of non-covalent bonds between neighboring atoms Wüthrich . In refoNMR is not suitable for routine real-time monitoring of refolding processes, as it requires protein purification and concentration prior to measurement  is a measurement technique applicable in real time during refolding processes to monitor protein refolding and aggregation. Fluctuations in light scattering are detected to determine the hydrodynamic size of the particles  which increases the robustness of the measurement by measuring the scattered light at multiple angles simultaneously and, thus, preventing to omit populations present in the sample  lower than 10 μg/mL. The minimum sample concentration is 0.3 g/L assuming a sample volume of 2 μL. However, RP-HPLC is a time-consuming technique that can take 20–80 min for the high-resolution analysis of a sample. Consequently, a timely depiction of refolding kinetics can be problematic. Furthermore, overloading should be avoided by applying less than 1 mg per 1 mL of the column to have a better resolution of the resulting peaks  even when using a sampling device, such that other measurements are needed as information sources concerning monitoring and control of the process.Further processing of samples using size-exclusion HPLC (SEC-HPLC) facilitates the monitoring of aggregates during refolding. Thereby, a timely depiction of the changes in aggregate kinetics is possible  can be defined. Since different analytical methods differ not only in robustness, sensitivity, and accuracy but also in real-time measurement capabilities, the additional use of modeling is a promising possibility to enhance and align several methods. Some of the monitoring techniques described do not feature sufficiently low feedback times for process control. However, these techniques nevertheless provide process knowledge that can be used in modeling approaches which will be discussed in section “Models of refolding processes.” For instance, solubilized IBs and native protein could be quantified offline through SEC-HPLC and RP-HPLC for model refinement, while the refolding process itself can be monitored in real-time through detecting native protein via FTIR and aggregation through DLS or combinations of the aforementioned methods. Another problem arises for the spectroscopic methods with changing reference spectra due to the feeding of the dilution by fed-batch mode. Therefore, updated measurements of the reference spectrum are necessary.A model describes the change of a dynamical system over time. Therefore, it enables predictions of the system behavior into the future with given initial conditions. In the case of refolding processes, a model can be utilized to estimate the targeted product forms from other measurements; hence, it functions as a state observer providing additional indirect measurements of otherwise unmeasurable states. Mechanistic models describe a dynamical system with one or a set of differential equations. They are derived from physical first principles such as mass, momentum, heat, or energy balances  such as yield and space-time-yield of the refolding reaction can be the system outputs. Both KPIs can be determined online by soft sensors using the total protein concentration in the refolding vessel if the concentration of the native protein can be directly measured or accurately estimated. Simple examples of successful process estimation are based on mass, energy, or elemental balancing. Considering the law of mass conservation, unknown states can be directly calculated from others.For more complex system descriptions, subject to non-linear reaction kinetics and internal dynamics, the most probable internal state can be estimated with non-linear Bayesian filters, such as the extended , redox potential, and feeding rate) and KPIs  need to be monitored through direct measurements or soft sensors to enable their control  with constant feed rate versus varying feed rates with identical final total protein concentration shows that higher yields are reached when the concentration of solubilized protein in the refolding vessel at each time step is smaller. This is achieved through either lower final total protein concentration or lower feeding rates. To further emphasize the relation between yield and space-time-yield, multiple refolding simulations by dilution in fed-batch mode with feeding rates from 0.015 to 10 mL min−1 are used to visualize this behavior in a Pareto plot  and the normalized root mean square error (NRMSE) of model simulations and experimental data (Daume et al. To transform this proposal into a working refolding process with monitoring and control, the defined model needs to be parametrized and validated to ensure its predictive power and applicability. Parametrization is usually performed by applying the weighted residual sum of squares WRSS, and model validation is performed by calculating and evaluating the goodness of fit (Although biopharmaceutical manufacturing has been transforming along the QbD principles in the recent years, industrial refolding is still conducted based on empirically established procedures instead of sound process knowledge. The reason behind this lies within the complex nature of these processes requiring cross-disciplinary methodology and knowledge to identify the key correlations. Until these black box processes are resolved, considerable efforts need to be made in terms of refolding process development that is costly, requires time, and yet results in non-scalable and strictly product-specific techniques with low yields.However, the generation of mechanistic process knowledge on refolding kinetics could establish universally applicable methodologies within product families. To achieve this, real-time monitoring of relevant process variables with online sensors and soft sensors is indispensable. The analytical methods described in this review could provide a good basis for this; however, as previously discussed, the monitoring of refolding kinetics is still very challenging due to low protein concentrations. Additionally, not every process variable is directly measurable and related analytical methods have certain limitations. To meet these challenges, a modeling approach can be applied in addition to analytical techniques to estimate the non-measurable process variables in real time, thereby illuminating the complete picture of the process. Furthermore, when all relevant variables and their correlations are available, more complex model systems can be established to predict the process progression. Based on these, optimal and adaptive control trajectories are possible to achieve more efficient IB recovery processes, better product quality, and higher yields. Furthermore, in accordance with the current trends in the industry aiming at the digital transformation of manufacturing processes, an adequate software environment is needed to support this endeavor. Such a software needs to support the real-time integration and calculation of complex models, based on the acquisition of current process data and subsequently, it should be able to provide the optimized output trajectories for the manufacturing system to realize closed-loop control. Eventually, not only refolding processes can be steered, but as a next step, the complete IB downstream processing chain could be optimized.


Despite the high prevalence of sensory impairment in older adults, there are no standard practices for its consideration in cognitive studies. We conducted a systematic review to identify and survey prospective cohort studies collecting cognitive data in older adults in order to determine whether and how hearing and vision were considered. Among 81 cohorts that responded, 30 (37%) objectively assessed hearing, with audiometry as the most frequently-used method; 61 (75%) used patient-report and 12 (15%) used provider-report to subjectively assess hearing. Forty-one (51%) cohorts objectively assessed vision, half of which measured distance or near visual acuity; 55 (68%) used patient-report and 10 (12%) used provider-report to subjectively assess vision. Nineteen (23%) cohorts offered hearing accommodations and 30 (37%) offered vision accommodations during cognitive testing. Findings indicate variation in methods used to assess hearing and vision as well as in accommodation practices that could impact estimates of cognition among older adults.


The speech reception threshold (SRT) was investigated depending on the separation angle between speech and noise. The lowest (best) SRT was obtained for a separation of target and interfering source from S0N0 at an angle of about S±60°N∓60°. The derived normative curve was comparable to SRTs predicted by the binaural-speech-intelligibility-model. The systematic separation of signal and noise showed a significant improvement in speech intelligibility for normal-hearing people even for small separation angles. This experimental setting was verified. This study aimed to assess the effect of small sound source separation on binaural hearing and speech perception.Speech perception in noise is challenging and is improved by binaural hearing. Since signal processing of assistive hearing devices often modifies or masks the peripheral binaural head-shadow or better-ear effects, central binaural processing should be measured separately. In a prospective study, 10 listeners with normal hearing were tested with the German matrix sentence test in a set-up with two loudspeakers located at opposite angles in the horizontal plane with respect to S Understanding speech in noisy environments is challenging even for listeners with normal hearing and becomes even more difficult if the surrounding noise is competing speech ,2. Heariper se, while the binaural summation and even more the squelch effect might be viewed as higher order influence in the auditory processing pathway [Beside the monaural assessment of the head shadow effect, binaural signal processing may result in binaural summation, which is the difference in SRT between listening with two ears and listening with the better monaural ear, or the binaural squelch . The bin pathway .In people with hearing loss using assistive hearing technologies the ability to process ILD and ITD is limited . Binaura0N0 presentation for binaural summation to multiple loudspeaker settings [0N±90, N0S±90) [For clinical testing, a procedure is needed which reduces the influence of the rather peripheral effects and focus on the central binaural effects. Typical sound source settings for the evaluation of binaural hearing with speech in noise tests range from a simple Ssettings  and multsettings ,17,18. O N0S±90) .Gabriel et al.  found thIn our study on normal-hearing subjects, the head shadow and better-ear effect were minimized by focusing on small symmetrical spatial separation between speech and noise sources, both separated in the frontal position. The SRT improvement as a function of separation angle determines the benefit of spatial separation of signal and noise and might serve as a tool for evaluating hearing aid and speech processor settings which are presumed to provide some improvement to the user .The head shadow effect was assessed separately by measuring the speech and noise separately in a binaural dummy head recording and using a binaural speech intelligibility model (BSIM). To our knowledge, BSIM was used for the first time to predict SRT for symmetrically separated sound sources .This study aimed to assess the effect of small sound source separation on binaural hearing and speech intelligibility. A normative curve of SRT improvement for normal-hearing subjects was recorded as well as the prediction of BSIM for the measured setup verified.We conducted a prospective experimental trial involving ten healthy adults with normal hearing . Their pure-tone hearing level threshold averaged across 0.5–4kHz was between 1.25 and 11.25 dB  with a maximal side difference of 6.25 dB. The maximal threshold was 20 dB for one participant at 6 kHz, any other thresholds did not exceed 15 dB. Participants had no acute or chronic ear diseases or health conditions relevant to the study question . All participants signed a written informed consent. The protocol for this study was according to the Declaration of Helsinki. It was approved by the Ethics Committee of the Martin-Luther-University Halle-Wittenberg .We used the German matrix sentence test  to measThe measurements were done in a sound insulated room with a reverberation time of 0.35 s. All sound stimuli were generated by a PC, converted by Fireface 400 interface  to an analog signal and sent to a POA-800 power amplifier . Two CD 1020 loudspeakers  were mounted in a custom-made semicircle in the horizontal plane . The angTo familiarize the participants with the test and reduce the training effect, two sentence lists with 20 sentences each were presented prior to the actual measurement. This step was followed by tests with systematic, symmetric separations of the loudspeakers with angles of 0°, 1°, 2°, 4°, 6°, 9°, 13°, 20°, 30°, 45°, and 90°. Additional presentations at 60° were applied for five random participants and at 75° for three random participants, respectively. The order of presentation was inverted for five of the ten participants.φ = ±2.8°). Therefore, the influence of the height adjustment for angles larger than 6° was investigated. It was changed for five participants at an angle of ± 9°, and for the other five, it was kept constant for the entire measurement for all angles. For six participants, speech was presented on the left and the noise on the right side, and for four participants, this was reversed.Because of the geometrical dimensions, it was not possible to adjust the loudspeakers at the same height in the frontal position  –SRT(S0N0)) was calculated for each loudspeaker position and averaged over all participants. Since the noise and speech directions were randomized between the subjects SRTs were compared between the sides by an analysis of variances (ANOVA). The averaged SRT over both sides were then compared between the presentation angles by an ANOVA for repeated measures. Bonferroni corrections were used for post hoc comparison.The change in speech reception threshold by sound source separation in reference to the SRT at SThe angle at which the SRT improvement reached significance and the angle with the lowest (best) SRT were identified. Since a SD for the test–retest differences in e.g., cochlear implant users of around 1 dB was reported , we assuFor comparison of the observed results with model predictions, the binaural speech intelligibility model (BSIM) was used . For thi0N0, the mean SRT across all participants was –7.3 dB (SD = 2.2 dB) SNR relatively to a noise level of 60 dB SPL. 0N0 and used as reference (ΔSRT = 0). For all participants, the trend of the curve was the same, and the ΔSRT improved when the sources were separated. At approximately S±60°N∓60°, the SRT reached a plateau or a minimum and in most cases increased slightly until the maximum separation angle of S±90°N∓90° in this experimental series.At Sp<0.001). Post hoc comparisons showed that increased angles led to reduced SRTs. SRT measures with angles from 4° and above were significantly different from the S0N0 condition .The averaged ΔSRT values are shown in ±5.2°N∓5.2°. Thus, a symmetric separation of signal and noise in the front of 10.4° led to an SRT improvement of 2 dB SNR. For small angles up to 13° , the improvement can be described with a linear fit with a slope of –0.4 dB SNR/°. 0N0 was 11.3 dB SNR (SD: 0.58 dB SNR), which was measured at the S±60°N∓60° loudspeaker position .The figure also shows the SNR improvement of 2 dB at a loudspeaker position of SMeasured binaural dummy head recording was used to calculate the BSIM. Up to an azimuth of 40°, the model data is within the range of measured normative data.This study aimed to assess the effect of small sound source separation on binaural hearing and speech perception.0N0 presentation of both, signal and noise, the SRT was –7.3 dB SNR, which is similar to values of –7.1 dB SNR reported for the used speech test by others [Under the frontal Sy others . The resy others  and alsoy others . This imy others . With thWe are aware of the limitations we are faced with because of the small group of participants, fixed speaker positions and a non-random presentation of the stimuli. To get more precise results the number of participants can be increased. Smaller steps in changing the angles would lead to a better resolved curve. Nevertheless the data already fit to the predicted BSIM data. Although the SRT improvement was not unexpected, it could be shown, for the first time, for symmetrical separation of sound sources with high angle resolution. The SRT predicted by the BSIM fit to the measured SRT up to sound source separations of ±40° . Thus, o0N0 presentation, we assume an improvement of 2 dB as being relevant. This improvement was already achieved at a relative small source separation angle of 10.4° (±5.2°). Nevertheless this size of 2 dB improvement has to be further investigated for future setups with hearing impaired patients.The high angle resolution was used to focus on small symmetric angle separations where the head-shadow is very small. Even for small angles, the SRT improved linearly with the separation angle. The improvement was –0.4 dB SNR / degree. A significant improvement compared to the frontal presentation of both sound sources was reached at an angle of 4°. Those improvements are often within the standard deviations for the test–retest differences in e.g., cochlear implant users of around 1 dB . However0N0 or Intelligibility Level Difference tests. Even if only a few angles would be used for sound presentation the long recording time would limit such application in clinical routine and might be decreased by using the Oldenburg sentenced test for children (OLKISA) matrix test, which reduces the presented items to 60% as compared to the OLSA used in this study. Since the use of small presentation angles requires a stable head orientation in the room, a good fixation of the head position in the sagittal plane (0°) as done in this study appears crucial.The results and experiences from the experimental procedure could be translated to patients with hearing aids, cochlear implants or binaural listeners to assess binaural SRT improvements. It could be the basis for evaluation of beamforming algorithms or future developed algorithms. By including small symmetrical source separations more sensitive information about binaural benefits would be recordable than with the commonly used sound presentations at S0N90 settings.In their recent cohort study, Hoppe et al.  showed t0N0 or S0N90 setups show constraints in investigating those effects. Future studies should investigate its feasibility in patients with hearing loss. Following that, the development and evaluation of fitting procedures and other methods with a focus on binaural hearing might be supported.The strength of the proposed setting is the assessment of a binaural SRT as soon as it appears at small angles without being superimposed by the above-mentioned larger “passive” head shadow or better-ear effects. The experimental setting has the potential to support evaluation of fitting procedures and other methods with a focus on binaural hearing. The standardized SS1 Rawdata(PDF)Click here for additional data file.S2 Rawdata(PDF)Click here for additional data file.


The low pathogenic H9N2 AIV caused the serious impact on the poultry industry and public safety. Our purpose was to investigate the molecular evolutionary characteristics of the new isolated H9N2 virus and investigate the intracellular target protein of H9N2 AIV replication in sensitive cells.AIV A/chicken/Shandong/LY1/2017 (H9N2) was isolated from the cloaca of the healthy chicken in Shandong, and the full-length eight gene segments of this isolated H9N2 AIV were amplified by RT-PCR and analyzed. MDCK cells were used as the target cell model, and VOPBA assay and LC-MS/MS were carried out to identify the virus-binding protein of H9N2 AIV. MDCK cells were pre-treated with the special antibody and siRNA, and treated with H9N2 AIV to detect the virus replication. Additionally, Vimentin-pcDNA3.0 was successfully constructed, and transinfected into MDCK cells, and then H9N2 AIV mRNA was detected with RT-PCR.Phylogenetic analysis revealed that HA, NA, PB2, PB1, PA, NP and M seven genes of the isolated H9N2 AIV were derived from A/Chicken/Shanghai/F/98, while NS gene was derived from A/Duck/Hong Kong/Y439/97. The cleavage site sequence of HA gene of the isolated H9N2 AIV was a PARSSR G pattern, and the left side sequence (224 ~ 229) of receptor binding site was NGQQGR pattern, which were similar to that of A/Chicken/Shanghai/F/98. Following VOPBA assay, we found one protein of about 50KDa binding to H9N2 AIV, and the results of LC-MS/MS analysis proved that vimentin was the vital protein binding to H9N2 AIV. The pre-incubation of the specific antibody and siRNA decreased the viral RNA level in MDCK cells treated with H9N2 AIV. Furthermore, we found that over-expressed vimentin increased H9N2 AIV replication in MDCK cells.These findings suggested that the isolated H9N2 AIV might be a recent clinical common H9N2 strain, and vimentin protein might be one vital factor for H9N2 AIV replication in MDCK cells, which might be a novel target for design and development of antiviral drug. H9N2 subtype avian influenza virus (AIV) has become responsible for the increasingly serious influence on poultry production and human health. Since 1994, H9N2 AIV was prevalent rapidly in many chicken farms and waterfowl populations, and became the most popular subtype of AIV in China , 6. AlsoInfluenza viruses internalized and became into the early endosomal Endosomes (EEs) through the binding of HA protein with membrane surface receptor sites N-acetyl neuraminic acid (Neu Ac) and hydroxyacetyl neuraminic acid (Neu Gc), and then developed the late endosomal Endosomes (LEs) . The virIt was important to investigate AIV attachment to trachea in many avian species . AIVs ma2 at 37 °C.H9N2 AIV used in this study was isolated from the cloaca of the healthy chicken in Shandong 2017, which was collected as samples of routinely ongoing surveillance. The hemagglutination inhibition with the special antibody confirmed that the isolate might be H9N2 subtype AIV. The virus was thrice propagated in 9-day-old specific-pathogen-free (SPF) embryonated chicken eggs, and then gene fragments were sequenced and comparatively analyzed. Madin-Darbycanine kidney (MDCK) cells were maintained in Dulbecco modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 5% COAnti-vimentin monoclonal antibody was purchased from Abcam (ab45939) and anti-hemagglutinin polyclonal antibody was purchased from Jianchun Biotechnology (Nanjing). Anti- Glycerophosphate dehydrogenase (GAPDH) antibody (ab8245) was purchased from Abcam. Alkaline phosphatase conjugated goat anti-rabbit IgG and HRP-conjugated goat anti-mouse IgG secondary antibody was purchased from Univ Biotechnology (Shanghai). Vimentin siRNA was purchased from Santa Cruz .Based on the whole genome sequence of H9N2 AIV published in GenBank database of US National Center for Biotechnology Information (NCBI), showed in Table To eliminate the nucleotide acids error of eight genes in cDNA clones obtained using RT-PCR, five samples for each gene were sequenced by Nanjing Qing Ke biological company , and eight genes fragments were amplified. Also, MEGA5.3 was used to diagram the phylogenetic tree of each gene fragment , and to The samples of MDCK cells were obtained to extract all the intracellular proteins after ultrasonication and proteolysis. The collected proteins samples were analyzed with 12% SDS-PAGE, and transferred onto polyvinylidene fluoride (PVDF) membranes. After blocked, the transferred membranes were incubated with 1 multiplicity of infection (MOI) H9N2 subtype AIVs, and then incubated with rabbit polyclonal antibody special to HA protein of H9N2 AIV, and then incubated with goat anti-rabbit IgG secondary antibody. After screened, the results were developed with high resolution image acquisition system.Simultaneously, the proteins samples were analyzed and stained with Coomassie staining, and the bands equivalent to the above western blot of the major virus binding band were recovered and analyzed for liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis by Shanghai Zhongke Xinsheng Life Biotechnology Co., Ltd. . Simply, after reduction and alkylation, Trypsin was added into the examined samples to enzymatic hydrolysis for 20 h at 37 °C. After desalination and freeze-drying, the samples were dissolved in 0.1% FA solution, and then performed on Trap column for mass spectrometry. After MS2 scan, the raw files were obtained and searched in the related databases through Mascot 2.2 software. The detailed search parameters were showed in Table MDCK cells were incubated with 100 μg/mL anti-Vimentin antibody (ab45939) or rabbit IgG control at 37 °C for 2 h. After washing twice with phosphate buffer saline (PBS), the incubated MDCK cells were infected by 0.1 MOI H9N2 virus at 37 °C for 1.5 h, and cultured with DMEM for 36 h. Following Trizol instruction, the total RNAs of MDCK cells were extracted to be usaed as templates for RT-qPCR. The primers used for RT-qPCR were showed in Table MDCK cells in 12-well plate were transfected with vimentin siRNA and control siRNA according to Lipofectamine-3000 (Invitrogen) protocol. After 24 h, the transfected MDCK cells were infected with 0.1 MOI H9N2 AIV at 37 °C for 1 h, and were cultured with DMEM for 36 h, and then were collected to detect the vimentin mRNA and viral mRNA levels by RT-qPCR. Also, the knock-down expressed vimentin proteins were detected by western blot with vimentin antibody (ab45939) as the previous reported . ControlAccording to the published sequence NM_001287023.1 in Genbank database from NCBI, the primers of Vimentin were designed by Primer Premier 6.0, as showed in Table Vimentin-pcDNA3.0 was transfected by using Lipofectamine-3000 (Invitrogen) in 6-well plate, and vector pcDNA3.0 transfected MDCK cells were used as control. At 60 h post-transfection, vimentin mRNA in the transfected MDCK cells were detected by RT-qPCR, and the expression of vimentin proteins were checked by western blot with vimentin antibody (ab45939). Additionally, at 24 h post-transfection, the Vimentin-pcDNA3.0 transfected cells were incubated 0.1 MOI H9N2 AIVs for 1 h at 37 °C, and cultured in DMEM with 10% FBS for 36 h to detect H9N2 virus units by RT-qPCR.t-test or one-way ANOVA with significantly difference less than 0.05.Results were illustrated in bar graphs as means ± standard deviation (SD) of three independent experiments. The statistical significances were analyzed by To verify the detailed subtype,, the isolate were obtained after thrice propagated in 9-day-old SPF embryonated chicken eggs for comparative analysis of gene fragments. Eight genes fragments of AIV  were amplified, respectively, and the full length genome of LY1 was obtained , A/swine/Jiangsu/C1/2008 (JSC1) and A/chicken/Hubei/C1/2007 (HBC1), in which homologies between eight full-length segments of LY1 and those of three viruses ranged from 99.1 to 100% (Table The cleavage site sequence of LY1 strain was a PARSSR↓G pattern, which was consistent with that of the Y280 subline (Clade h9.4.2, Table The potential glycosylation sites analysis on the HA of LY1 showed that LY1 had seven potential glycosylation sites at the same positions as those of three recently isolates, which were less than that of three typical strains Table . CompareIt was observed that the deletions in the stalk and potential N-glycosylation sites of NA gene in LY1 were same to that of F98, JS7, JSC1 and HBC1 strain Table . CompareThe receptor binding sites of NA protein in LY1 mainly existed at three positions, 367 KVDSRS 372, 400 SDNW 403 and 431 PQE 433. It was found that the KVDSRS pattern in NA protein of LY1, which was same to that of the three recently strains JS7, JSC1 and HBC1, and different to that of three typical strains F98, Y280 and BJ1 strains Table .To identify the protein in MDCK cells involved in H9N2 virus attachment, we employed VOPBA with polyclonal antibody against HA protein of H9N2 virus, and found one strong binding-band with molecular mass about 50KDa, in which the same bands appeared in the both parallel samples Fig. , suggestTo determine the role of membrane vimentin protein in H9N2 virus entry, MDCK cells were preincubated with anti-vimentin antibody, and then infected with H9N2 AIV. The qPCR results showed that the viral RNA level was significant lower in anti-vimentin antibody treated cells than that of IgG control Fig. a. It wasTo further detect the function of vimentin in H9N2 virus reproduction, MDCK cells were pretreated with siRNA specific to vimentin, and then infected with H9N2 AIV for 36 h. The results showed that the vimentin expressions were decreased in siRNA treated cells, compared with that of control (Fig. The qPCR results showed that the viral RNA levels of siRNA-transinfected group cells were 5.55, while control-siRNA cells were 14.45, in which H9N2 virus replication in siRNA-transinfected group were significantly decreased, compared to that of control-siRNA Fig. e. These The 1398 bp vimentin gene was amplified Fig. a, and wa. Epidemiological investigation showed H9N2 subtype avian influenza virus might be the donor for the internal genes of H7N9 , 9 and HThe connecting peptide of HA of LY1 was PARSSR↓G motif, a characteristic of H9N2 viruses of land-based poultry . In thisIt was found that there were same hemadsorbing sites of the NA gene between that of JS7, JSC1, HBC1, and LY1, which were different from that of F98, and Y280. Unexpectedly, one amino acid mutant from NST to NNT were occurred in NA protein of JS7, JSC1, HBC1, but did not present in LY1. This finding suggested that the potential biological significance of this molecular marker in LY1 isolate remained to be elucidated.The mechanism of influenza virus entry and replication in cells needs to be further investigated. In this paper, employing VOPBA and mass spectroscopy analysis, we found the vimentin might be the potential binding proteins to LY1 strain in MDCK cells. It has been reported that vimentin protein was associated with multiple cellular functions, and was required for parvoviral infection . Also, vIn summary, the isolated H9N2 AIV might be a recent clinical common H9N2 strain, and vimentin was identified as the binding protein to H9N2 AIV, and the results of the specific antibody, siRNA and over-expression proved that vimentin protein was one vital factor for H9N2 virus replication in MDCK cells, which might be a novel target for antiviral drug design and development.Additional file 1: Table S1. The reference strains of H9N2 subtype influenza virus and their abbreviations.Additional file 2: Table S2. Specific primers for eight gene segments of LY1 strain.Additional file 3: Table S3. The detailed search parameters of mass analysis.Additional file 4: Table S4. The qPCR primers for HA gene of H9N2 Virus and Vimentin.Additional file 5: Table S5. The PCR primers of Vimentin for construction of vimentin-pcDNA3.0.


Mixing alkali-resistant glass fiber (ARGF) into grouting slurry can prevent the development of cracks; thus, understanding the properties of ARGF grouting material is important for applications in engineering. Two types of ARGFs (Cem-FIL®60 and Anti-Crak®HD) were selected as mixing materials, and their performance was tested in four areas, namely, compressive strength, tensile strength, flexural strength, and impervious performance, under four different mixing amounts of fiber . Results demonstrate that the addition of ARGF increased the compressive strength and tensile strength of the grouting slurry, and the best performance was at 0.5%. The effect on the flexural strength and impervious performance was related to the mixing amount, and the fiber may have induced a counter-effect for certain amounts of added ARGF. Mixing ARGF could increase the early strength ratio of grout; however, a high early strength ratio did not necessarily result in high strength, as the flexural strength did not change synchronously with the early strength ratio; a similar pattern was found for the impermeability. Cem-FIL®60 had a better effect on the properties of grouting materials than Anti-Crak®HD. These results were successfully applied in the water-plugging and reinforcement engineering of a karst tunnel. Grouting is one of the most effective methods used to solve geological problems, and the characteristics of a grouting material can have a direct and critical impact on the effectiveness of grouting. While the development of geological structures is complex, the diversity in the environment contributes to the richness and diversity of grouting materials. Different grouting materials have different advantages with different applications. Therefore, mastering the properties of new grouting materials is the key to ensuring the effectiveness of grouting. In order to improve the effectiveness of grouting materials, many researchers conducted studies on the development and performance of grouting materials with rich and distinctive results.Research and development of chemical grouting materials. He et al.  studied Development of special geological grouting materials. Kim et al.  developeResearch on environmentally friendly grouting materials. Park et al.  studied Perez-Garcia et al.  mixed grResearch on fiber grouting materials. Nishimura et al.  studied Although much research on grouting materials and their properties was conducted, the main areas of focus were on chemical grouting materials, special geological grouting materials, and some environmentally friendly grouting materials. However, there was very little research on fiber grouting materials, whereas the existing research mainly focused on nano fiber and polyurethane fiber as added material in terms of their effect on the properties of slurry injection. Few studies were done on the flow pattern of alkali-resistant glass fiber grout, whereas the compressive strength, tensile strength, flexural strength, and impermeability of alkali-resistant glass fiber grouting materials were not studied. ARGF is an environmentally friendly material with excellent alkali resistance and corrosion resistance. The addition of ARGF can avoid the adverse effect of cement alkaline corrosion on the admixture, improving the performance of grouting materials to a certain extent. ARGF can play a role in preventing crack and impermeability, as well as improve the grouting effect. Therefore, it is necessary to carry out experimental research on the properties of ARGF grouting materials in order to grasp the characteristics of the influence of the type and amount of materials, as well as the effect of other conditions on the grouting materials.In this study, laboratory tests and engineering application methods were used. Two ARGFs, Cem-FIL60 and HD, were selected to study the compressive strength (CS), tensile strength (TS), flexural strength (FS), and impervious performance (IP) of ARGF with four mixing amounts . The performance of the slurry with mixed ARGF in terms of CS, TS, FS, and IP, as well as its early strength ratio of CS (ECSR), early strength ratio of TS (ETSR), early strength ratio of FS (EFSR), and early permeant ratio (EPR), was analyzed. In addition, the nature of the influence of material types and mixing amount on grouting performance was obtained. The material was applied to karst tunnel grouting projects, and it achieved good grouting results, providing an important reference for the application of ARGF grouting slurry.The cement used was PC 32.5R composite Portland cement produced by Sinoma Zhuzhou Cement Co., Ltd., while alkali-resistant chopped strands produced by Taishan Glass Fiber Co., Ltd.  were used for the ARGF. Cem-FIL60 and HD were selected, and their characteristic parameters are shown in C-ARGF was made by mixing water, cement, and ARGF in a certain proportion. To make the study of the strength properties of slurry stones representative, the water–cement ratio used in this experiment design was 1:1 (water–cement ratio refers to the ratio of the weight of water to that of cement). When the water–cement ratio is 1:1, C-ARGF grout is characterized as a Bingham fluid ,31. The In order to facilitate specimen classification, specimens adulterated with Cem-FIL-type of glass fiber grout are represented by CFIL . Specimens adulterated with HD-type ARGF are represented by HD, and the slurry specimens without ARGF are classified as NCH-0. For CS and TS testing, the specimen size was 100 mm (length) × 100 mm (width) × 100 mm (height). For the FS test, the specimen size was 400 mm (length) × 100 mm (width) × 100 mm (height). For the IP test, the specimens were circular-truncated cones, with a size of 175 mm (top diameter) × 185 mm  × 150 mm (height). According to the specification requirements ,34, the The slurry configuration, specimen preparation, and hardening were carried out at a room temperature of 20 ± 5 °C. When the specimen hardening time reached 24 h, the mold was removed; then, the specimens were put into a 20 ± 2 °C environment for curing. The test was conducted after the specimens reached the test time, according to the maintenance requirements. At present, there are many testing methods for glass fiber cement products in the world ,36. ThisIn terms of the impermeability test, the seepage height method was adopted. The Vaseline-coated specimens were pressed into a test mold, which was installed on the HP-4.0 impermeability tester, as shown in By adopting the above experimental methods, the experimental results of CS, TS, FS, and PS for each experimental group are obtained, the results are displayed using intuitive graphics, and a systematic analysis is performed. The values of CS, TS, and FS are the average values of the three specimens in each group, and the value of impermeability height is the average height of the six specimens in each group.The penetration height (PH) is an index used to characterize the IP. A larger PH denotes a worse IP, and, as PH decreases, the IP of ARGF improves.In summary, the analysis of the performance indicators of the slurry showed that, based on the strength analysis, the CS and TS of ARGF improved significantly, and the effect of FS and IP was directly related to its mixing amount. For example, for an HD mixing amount of 1.0%, a negative effect on FS was realized. Based on the analysis of the early strength ratios, adding ARGF increased the ECSR, ETSR, and EFSR of the grouting materials; however, an increase in EFSR did not necessarily correspond to an improvement in FS. Based on the PH ratio analysis, adding ARGF reduced the EPR and improved the IP in the early stage. Based on the analysis of the mixing amount, the CS and TS increased first and then decreased with an increase in the mixing amount. However, how FS and IP vary with mixing amount is uncertain, and some amounts were shown to have the opposite effect. Based on the material type analysis, Cem-FIL60 ARGF is superior to HD materials in terms of CS, TS, and IP, while both materials have their own advantages in terms of FS. However, in general, the effect of adding the Cem-FIL60 ARGF is better than that of the HD material.Qiyueshan Tunnel is a controlled project of Liwan Expressway in Lichuan City, Hubei Province. The area is mainly dominated by limestone development, with shale development in the middle. Both areas are extremely fragmented rock masses and well-developed karsts. Serious karst water gushing can be found at the excavation exposure, as shown in Tunnel water gushing is the result of complex geological conditions such as karst development and geological structure fragmentation; the karst geological condition is hidden and cannot be directly revealed for processing. In this case, grouting is an effective and direct solution.The original grouting scheme adopted pure cement grouting, in which the grout was diluted and washed out by karst water. After the grouting, large-scale water seepage (or even effluent) occurred in the surrounding rocks, and the anti-alkali phenomenon was serious, resulting in poor grouting, as shown in Owing to the complicated development of karst and fissures, the large amounts of gushing water, the richness of the karst water, and a certain dynamic pressure, the grouting pressure was selected to be 3.0–5.0 MPa, and the diffusion radius was 1.1–1.3 m. When the single-hole grouting reached the design pressure and was maintained for 5 min, or when the deformation of the surrounding rock exceeded the allowable value, the grouting was stopped. For this project, the grouting reinforcement process was carried out as shown in After grouting, the phenomenon of infiltration occurred only in local areas without large-scale infiltration or dripping , and theIn order to test the effect of grouting and water plugging and make the inspection hole representative, the inspection hole was set near the first layer of grouting circle formed around the water outlet point, and four detection holes were provided from the periphery to the water outlet point. CFIL60 ARGF grouting inspection holes were numbered T1–T4, and pure cement slurry inspection holes were numbered M1–M4. The specific layout is shown in As can be seen from The use of ARGF as an additive material of grouting slurry was examined. Through the experiment, the influence of the types and mixing amounts of ARGF on the CS, TS, FS, and IP of the slurry was studied. Meanwhile, the influence of the types and mixing amounts of ARGF on the ECSR, ETSR, EFSR, and EPR of the slurry was analyzed. The mixing amounts of the optimal volume and types of ARGF were determined, providing a basis for the engineering application of this grouting material.(1) The addition of ARGF was found to improve the CS and TS of the grouting material, and the CS and TS of Cem-FIL60 and HD ARGF reached a maximum when the mixing amount was 0.5%.(2) Different mixing amounts of ARGF were shown to have different effects on the FS, even resulting in a counteracting effect at certain amounts. For example, for mixing amounts of Cem-FIL60 and HD of 1.0% and 0.5%, respectively, the FS was lower than that of the pure cement slurry for the same curing time. For mixing amounts of Cem-FIL60 and HD of 0.25% and 1.0%, respectively, the FS reached a maximum, along with the IP. For a mixing amount of HD of 1.0, the PH was higher than that of pure cement slurry at 7–28 d; however, this phenomenon only occurred for an HD mixing amount of 1.0%.(3) Mixed ARGF resulted in increased ECSR, ETSR, and EFSR. Meanwhile, the CS and TS increased along with the rise of ECSR and ETSR. However, as for FS, it was lower than that of the pure cement slurry, although the EFSR increased under certain mixing amounts. Mixed ARGF could reduce the EPR of the slurry; however, the reduction in EPR did not represent an improvement in the IP. Under some mixing amounts, the EPR decreased, while the IP was weaker than that of pure cement slurry.(4) In comparing the superiority between the Cem-FIL60 and HD ARGFs, for mixing amounts of 0.25% and 0.5%, Cem-FIL60 was shown to be better than HD in CS, TS, and IP. However, for a mixing amount of 1.0%, HD was shown to be better than Cem-FIL60 in FS. Therefore, Cem-FIL60 ARGF was revealed to be generally superior to HD. Thus, CFIL60 ARGF can be the initial choice for mixing material in grouting engineering applications. Some problems were found in the application of C-ARGF grouting material, and the most common problem was pipe plugging. After field analysis, it was found that the problem is mainly related to the length and mixing amount of ARGF. In order to ensure the application effect of the new grouting material, the best combination of length and dosage of ARGF should be implemented to solve this problem in the future.


In all cases, amphotericin B formulations were administered as first-line therapy; in eight cases, they were also administered in combination with an echinocandin or triazole. Seven patients underwent surgical debridement procedures. The six-week mortality was 58%. Among the patients diagnosed between 2009–2015, one of the six survived, and of those diagnosed between 2016–2020, four of the six were salvaged. Conclusions: Disseminated mucormycosis is a life-threatening and often fatal disease, and improved diagnostic and therapeutic strategies are needed. Nevertheless, in this population-based study, five patients (42%) were salvaged through combined liposomal amphotericin/triazole treatment and extensive surgical interventions. Background: Mucormycosis is a life-threatening infection with a tendency for angioinvasion that may lead to progressive dissemination. Disseminated mucormycosis, defined as the involvement of two or more non-contiguous sites, is rare in children, and data concerning its management and outcome are scarce. The aim of this study was to assess the contemporary management strategies and outcomes of disseminated mucormycosis in the pediatric population. Methods: We conducted a retrospective search in six large tertiary medical centers for all cases of disseminated mucormycosis that occurred between 2009–2020 in patients aged 1–20 years. Results: Twelve cases were identified. Underlying conditions included hematological malignancies ( Mucormycosis is a life-threatening infection in immunocompromised children, caused by fungi of the order Mucorales. It is characterized by a rapid clinical course, with a tendency for angioinvasion that may lead to progressive dissemination, tissue infarction, and necrosis . SeveralRapid diagnosis, prompt initiation of antifungal therapy, reversal of immunosuppression, and surgical debridement are cornerstones of mucormycosis management ,8,9. BecIn this multicenter retrospective study, our aim was to assess the management strategies and outcomes of all children with disseminated mucormycosis diagnosed in six large tertiary medical centers in Israel during the study period.We conducted a retrospective search for cases of disseminated mucormycosis in six large tertiary medical centers. We included all children in these hospitals, which encompassed all six pediatric hematology–oncology departments in Israel. In each center, the medical records, microbiology databases, and pathology information systems were searched for all cases of disseminated mucormycosis that occurred between 1 April 2009 and 1 May 2020 in patients aged 1–20 years. Mucormycosis was categorized as proven, probable, or possible, based on the definitions of the European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) for invasive fungal disease . MucormyFive patients had been included in a previous general study , and oneIdentification of the cultured Mucorales was performed at various institutional microbiology laboratories using standard phenotypic methods. When possible, fresh specimens were sent to a central reference laboratory  for DNA sequencing and analysis. DNA was extracted from clinical samples using the QIAamp DNA Mini Kit , and was amplified using a semi nested polymerase chain reaction (PCR) assay targeting specific zygomycete regions at the 18S rDNA, as well as a fungal-broad range PCR reaction targeting the 28S rDNA. The PCR products were separated by electrophoresis in ethidium bromide stained 2% agarose gels, then sequenced on a 3130 Genetic Analyzer capillary electrophoresis DNA sequencer  and analyzed using the Basic Local Alignment Search Tool (BLAST). Antifungal susceptibility testing was performed at the National Mycology Reference Laboratory . The minimal inhibitory concentrations (MICs) of the antifungal agents for each isolate were determined using broth microdilution.n = 5; acute myeloid leukemia (AML), n = 4; and mature B-cell (Burkitt) leukemia, n = 1), one had been diagnosed with a solid tumor (metastatic medulloblastoma), and one patient was a solid organ (liver) transplant recipient. Two patients underwent hematopoietic stem cell transplantation (HSCT) prior to mucormycosis diagnosis. One patient, with relapsed ALL, was an allogeneic HSCT recipient, and the other, with medulloblastoma, was an autologous HSCT recipient. Nine patients were diagnosed during the course of their chemotherapeutic regimens, as follows: five during the first month of remission induction and four after consolidation chemotherapy courses. None of the patients had a refractory, uncontrolled underlying malignancy. The median duration of neutropenia prior to mucormycosis diagnosis was 12.5 days (range 0–35 days). Twelve pediatric cases of disseminated mucormycosis, occurring between 2009–2020, were identified for inclusion in the study. The median age of the study group was 13.5 years (range 8.9–19.2 years). Ten patients had an underlying hematological malignancy (acute lymphoblastic leukemia (ALL), n = 8), bowel (n = 6), lung (n = 6), brain (n = 4), and skin (n = 2). Three cases were presumably of a sinopulmonary origin, four were from the gastrointestinal tract, and one was from a liver allograft; in the four other cases, the origin was unclear. All patients had been evaluated by cranial, thoracic, and abdominal computed tomography (CT). It is noteworthy that in six patients, bacterial sepsis preceded mucormycosis; four of them had already been admitted to the intensive care unit prior to mucormycosis diagnosis.Ten of the twelve patients presented with prolonged fever. Other presenting symptoms included abdominal pain, chest pain and dyspnea, convulsions, hemiparesis, and obtundation. Anatomical sites involved included the liver (n = 11) and/or culture (n = 7). In addition, in seven cases, the pathogens were molecularly confirmed by PCR. Isolates included Rhizopus (n = 4), Mucor (n = 2), and Lichtheimia species (n = 4). In two cases, the specific pathogen was unknown (n = 1) or aspergillus (n = 3) were diagnosed in addition to mucormycosis. The radiographic and histopathological features are shown in All cases were classified as proven by EORTC/MSG criteria, with evidence by pathology ( unknown . Antifun unknown . One pat unknown . In fourn = 4) or liposomal amphotericin  were administered as first-line therapy. Of the eight cases treated with liposomal amphotericin, three were treated at a dose of 5 mg/kg/day, and five with higher doses of 7.5–10 mg/kg/day. In eight cases, combination antifungal therapy, comprising of AmB/L-AmB along with an echinocandin (caspofungin or anidulafungin) or triazole (posaconazole or isavuconazole), was administered as first-line or salvage therapy. All survivors were treated with prolonged step-down therapy.The treatment modalities are summarized in Seven of the twelve patients underwent surgical debridement procedures, mainly bowel resections and the excision of visceral organs . Pulmonary lobectomy and functional endoscopic sinus surgeries (FESS) were also performed.2 of dexamethasone, with subsequent blood count recovery within several days. Adjunctive therapies included granulocyte colony-stimulating factor (G-CSF) in nine patients and granulocyte infusions in five. Two patients with ALL were rapidly weaned from corticosteroid therapy. In one patient, after liver transplantation, immunosuppression was stopped. A patient with AML (patient no. 11) had a high fever and persistent severe pancytopenia, despite G-CSF therapy. Bone marrow aspiration on day 14 from mucormycosis diagnosis demonstrated prominent hemophagocytosis H. Blood Seven patients died with active mucormycosis, all within six weeks of diagnosis  was diagnosed with mucormycosis following four autologous HSCTs for the treatment of metastatic medulloblastoma. He was treated with an L-AmB/echinocandin combination, followed by L-AmB/intravenous posaconazole, in addition to bowel resections, two partial hepatectomies, and extensive debridement procedures. He received oral posaconazole step-down treatment for 9.5 months from the mucormycosis diagnosis. A 13.5-year old boy (patient no. 12) was diagnosed with disseminated skin mucormycosis after a final course of AML therapy I. He wasIn three of the five survivors, treatment of the underlying conditions was continued as soon as mucormycosis was deemed to be under control. A 10-year old girl, after liver transplantation (patient no. 2), successfully underwent liver re-transplantation, along with the re-institution of immunosuppression. She continued oral posaconazole for 12 months thereafter. A 13.5-year old girl (patient no. 8) was diagnosed with mucormycosis during induction for Philadelphia-positive ALL. She underwent two extensive surgical procedures, with multiple bowel and visceral resections A,B and aIn this multicenter study, we report twelve children diagnosed with disseminated mucormycosis at six tertiary medical centers in Israel over a period of eleven years. Of the twelve patients in our study cohort, seven died with active mucormycosis within six weeks. Among the patients diagnosed between 2009–2015, one of the six patients survived, and of those diagnosed between 2016–2020, four of the six patients were salvaged.All twelve patients in our study were treated with amphotericin B products as first-line therapy, with a gradual transition from AmB to L-AmB over the years, in accordance with the current recommendations ,8,9. TheTreatment of invasive fungal infections has evolved considerably in recent years with the introduction of new azoles, posaconazole and isavuconazole, to the therapeutic armamentarium. Because of concerns regarding the variable bioavailability of posaconazole oral suspension, more stable delayed-release tablets and an intravenous formulation have recently been made available. Isavuconazole, intravenous posaconazole, and delayed release posaconazole tablets are currently recommended for salvage therapy in cases of refractory mucormycosis or intolerance of first-line antifungal agents, and are moderately recommended as first-line therapy ,21. ThesSeven of the twelve patients with disseminated mucormycosis in our study received combination therapy consisting of AmB/L-AmB with an echinocandin (caspofungin) or triazole (posaconazole/isavuconazole). In view of the high mortality rates of mucormycosis, combined antifungal therapy seems an attractive option. A possible benefit has been shown in some murine models and patient series, but its efficacy has not been demonstrated in a clinical study . In thisHowever, it is difficult to attribute improved salvage rates to pharmacological factors alone . NonpharRapid diagnosis is a significant challenge in disseminated mucormycosis, as a result of the nonspecific nature of the clinical presentation. Most of our patients had prolonged neutropenic fever. Several were otherwise asymptomatic and were diagnosed following empirical imaging studies. Gastrointestinal involvement occurred in six of the twelve patients in our study group. Patients with gastrointestinal involvement are characterized by a clinical presentation that typically resembles neutropenic enterocolitis, with a rapid progression and imminent risk of gastrointestinal perforation. Sudden, unexplained bowel necrosis in neutropenic patients with hematological malignancies should arouse suspicion of mucormycosis, and the microbiology laboratory should be urgently alerted. In this study, cultures were positive for mucormycosis in only seven of the twelve cases, and molecular tests performed on fresh clinical specimens in the majority of cases significantly contributed to a swift verification of the diagnosis . FurtherSurgical interventions are considered a cornerstone of mucormycosis management, and current guidelines strongly recommend early complete surgical debridement procedures . In our The role of surgical interventions has been firmly established in mucormycosis management. The benefits of repeated extensive debridement with clean margins have been shown in rhino-orbito-cerebral mucormycosis , as wellIn our study, four of the five survivors had hepatic involvement A–H. One n = 4) did not.The reversal of immunosuppression is of paramount importance in the management of mucormycosis, and underlying conditions significantly impact outcome. In developed countries, the most common underlying conditions in patients with mucormycosis are hematological malignancies and HSCT. Dissemination reportedly occurs more frequently in these severely immunosuppressed patients. In this study, none of the patients had an uncontrolled, refractory hematological malignancy; in such cases, the reversal of immunosuppression is virtually impossible, leading to a poor outcome . The twoThere are a lack of data from controlled clinical trials to guide the management of mucormycosis. Disseminated mucormycosis is a rare entity, with limited data regarding the outcome and management, especially in children. Our population-based study, despite its limited number of patients, highlights several important issues concerning the treatments and outcomes of children with this entity. In our study cohort, we note improved salvage rates in recent years, concurrent with the availability of new antifungal agents. Other factors that may have contributed to an improved outcome are patients’ underlying conditions, specific disease localizations, and a heightened index of suspicion.In this multicenter population-based study, we report twelve children diagnosed with disseminated mucormycosis over a period of eleven years. Disseminated mucormycosis is a life-threatening and often fatal disease, and improved diagnostic and therapeutic strategies are needed. Nevertheless, in our study, five patients were salvaged, most of them in recent years, through combined L-AmB/triazole antifungal therapy and extensive surgical interventions.


GJB2 mutations have shown to be successfully managed by CI. Furthermore, cochlear implantation provides the auditory rehabilitation for the most common etiology of hearing loss in adults and age-related hearing loss (ARHL) or presbycusis. However, in some cases, cochlear implantation have been associated with some challenges. Regarding implantation in children, studies have shown that sometimes parents seem to have unrealistic expectations regarding the ability of CI to provide auditory rehabilitation and speech improvement. Given the evidence revealing the beneficial effects of early intervention via CI in individuals with hearing disorders especially hearing loss due to genetic etiology, early auditory and genetic screening efforts may yield better clinical outcomes. There is a need to better understand genotype-phenotype correlations and CI outcome, so that effective genetic counseling and successful treatment strategies can be developed at the appropriate time for hearing impaired individuals.The use and utility of cochlear implantation has rapidly increased in recent years as technological advances in the field have expanded both the efficacy and eligible patient population for implantation. This review aims to serve as a general overview of the most common hearing disorders that have favorable auditory outcomes with cochlear implants (CI). Hearing loss in children caused by congenital cytomegalovirus infection, syndromic conditions including Pendred Syndrome, and non-syndromic genetic conditions such as hearing impairment associated with  Hearing loss is a common neurosensory disorder affecting humans. Studies have long documented the significant prevalence, financial burden, and societal ramifications of hearing loss . A WorldThere are three main types of hearing loss: sensorineural, conductive, and mixed . Of thesDespite the initial introduction in 1957 and commercialization, CI became widely used in the last 20 years to provide auditory rehabilitation to individuals having severe to profound SNHL . The mosGiven that both children and adults who suffer from severe to profound SNHL are eligible for CI, 324,200 cochlear implants have been implanted worldwide as of December 2012, with 58,000 devices implanted in adults and 38,000 in children in the U.S. alone . Note, hOne of the most significant effects of untreated SNHL in young children is delayed language, reading, and behavioral development . While cSNHL in children can be congenital or acquired. Of children with congenital hearing loss, approximately 50% of cases are due to environmental causes, while another approximately 50% are due to genetic factors . Within COL4A3, COL4A4, and COL4A5.The many genetic causes of congenital hearing loss in children can be further categorized into syndromic and non-syndromic conditions, which, respectively, constitute 30% and 70% of the genetic causes of congenital SNHL . There aAlport syndrome can be inherited in either an X-linked or autosomal recessive fashion and leads to hearing loss, among other symptoms . The heaPendred syndrome, the most common syndromic cause of SNHL, is another syndromic condition that is inherited in an autosomal recessive fashion and usually causes congenital SNHL, often at birth, via enlarged vestibular aqueduct, a common inner ear deformity . StudiesThere are several other genetic causes of congenital hearing loss, and the most common are outlined in In the following section, we will discuss the outcomes of CI in relation to the genetic etiology of hearing loss based on the expression of genes in the cochlea or SGNs.ACTG1 codes for γ−Actin, the main actin isoform found in IHC and OHC and is vital for cytoskeletal maintenance , the speech intelligibility rating (SIR), and speech perception scores compared to matched controls. This study was limited by its small sample size and homogenous, Han Chinese population  after CI , a hearing disorder characterized by a functional cochlea but a non-functional auditory nerve. So while sound transduction may occur, the signals are not sufficiently transmitted to the brain  codes for Pejvakin, a stereociliary rootlet protein expressed in HCs, SGN, and auditory brain stem nuclei , are responsible for approximately half of the cases of hereditary SNHL in developed nations  mutations had excellent CAP scores 3 years after implantation  demonstrated a better CAP score than children with no detected mutation (n = 75) (-implant . Moreove(n = 75) . Age is (n = 75) .TMPRSS3 codes for a type II transmembrane serine protease expressed in hair cells, supporting cells, sulcus cells, interdental cells, cochlear neurons, and the SGN. TMPRSS3 mutations can lead to either a severe, pre-lingual hearing loss phenotype or a milder, later−onset, progressive, post-lingual hearing loss. In both cases, higher frequencies are typically lost first , or presbycusis, is a multifactorial disease with varying etiologies and can be attributed to progressive and irreversible degeneration of the cochlea, inner ear, or the auditory nerve . Given iEYA4, NID2, ARHGEF28, and CTBP2. A few genes have also been associated with other forms of hearing loss such as CDH23, which is associated with early onset congenital deafness  is a rapid onset deafness, usually of one ear, that affects 5–20 out of every 100,000 people . Though With cochlear implantation, as with any surgical procedure, there exists the potential for surgical complications and associated consequences. Despite advances in surgical techniques and the design of less traumatic electrodes, CI may be associated with the loss of residual hearing in the implanted ear . ResearcAdditionally, the individuals having auditory neuropathy spectrum disorder (ANSD) sometimes do not significantly benefit from CI depending on the site of lesion . In ANSDAt a cursory glance, the importance of maintaining residual hearing is not immediately apparent as cochlear implants are placed in individuals who have severe to profound hearing loss, However, as we will discuss, studies have documented numerous benefits that maintenance of residual hearing after CI contributes positively to auditory rehabilitation and patient self-confidence. Furthermore, the preservation of residual hearing will continue to gain relevance as an important outcome of CI surgery as the eligibility criteria for implantation continue to broaden, and as patients who may have unilateral or moderate hearing loss become eligible to receive CIs. The reasoning for the importance of preserving residual hearing stems from the fact that it can augment both the frequency and range of hearing, especially at lower frequencies . This auIncreasing understanding of the importance of residual hearing, in concert with advances in CI technology, has led to the recognition of electrode array design and surgical techniques as the keystone principles that determine success in the preservation of residual hearing. A systematic review further exposited on these principles by identifying five factors that seem to determine hearing preservation during CI: (1) minimally invasive surgery, (2) suitable electrode insertion route, (3) gentle insertion technique, (4) control of inflammatory reaction after CI, and (5) atraumatic electrodes . In summWhile preservation of residual hearing is currently a primary challenge associated with cochlear implantation and a topic of much research, there are other potential complications of CI surgery. Minor complications are mostly temporary and include post-operative dizziness, taste disturbances, transient facial nerve palsy, and the possibility of new-onset of tinnitus. The major complications include facial nerve stimulation, skin flap complications, and rarely infection . InfectiImproved post-operative performance on hearing loss has been well-received, with regards to patient satisfaction, in both adolescent and adult patient populations, with a few specific exceptions. A study used the Satisfaction with Amplification in Daily Life (SADL) questionnaire to evaluate patient satisfaction in children in Brazil who received CI as a means to treat auditory neuropathy-induced hearing loss. It was observed that overall satisfaction had a mean of 8, with a minimum of 7 and a maximum of 9, on a 10-point Likert scale . The samParents of Dutch adolescents with unilateral CI completed prospective surveys prior to surgery for a second implant and 12 and 24 months after the procedure . This stCI is widely used to provide auditory rehabilitation for a variety of hearing disorders in both children and adult populations. CI seems to be highly effective in the majority of cases of hearing loss. As CI technology becomes more refined, the patient population eligible for such implants will likely increase in the future. Furthermore, it is expected that genetic screening will continue to play a significant role for the identification of appropriate children and adults with congenital or inherited hearing disorders. The promising results of early intervention via CI as well as guidelines that formalize appropriate and timely screening and intervention could lead to population-level health improvements. However, some patients with hearing loss due to mutations in specific, genes especially those expressed in SGNs, do not benefit adequately from CI. Our understanding is still very limited regarding phenotype-genotype correlation and CI outcome, especially in minority and Hispanic populations leading to a significant knowledge gap. Given the high surgical implantation and clinical management cost of CI (>$1 million lifetime cost), prospective identification of the worst performers would reduce unnecessary procedures and healthcare costs as well as surgical risks. Since the severity of hearing loss due to genetic etiology varies in different ethnic groups and hence CI outcome, there is a need to establish genotype-phenotype correlations. Establishing a genotype-phenotype association will pave the way to expand the indication of CI to hearing-impaired individuals as determined by their genetic cause of HL and will provide genetic-based CI performance metrics. Better knowledge about genotype-phenotype correlation and CI outcome will help in providing effective auditory rehabilitation to hearing-impaired individuals in pursuit of improving their quality of life.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.AE is a consultant and received research funding from MED-EL Corporation. The remaining 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.