The study was conducted to evaluate the toxicological effects, functional observation battery tests, and sexual maturity of semicarbazide oral gavage administration to juvenile Sprague-Dawley rats for 70 days at 0, 15, 30, and 60 mg/kg/day weaning to sexual maturity. At 60 mg/kg/day, there was a delay in mean age at acquisition of balano-preputial and vaginal patency and a decrease in body weight and food consumption in males. Treatment increased reticulocyte count, aspartate aminotransferase, and alanine aminotransferase levels in both sexes and decreased hematocrit and protein in males. Increased absolute and relative liver and spleen weight in both sexes were observed. Male rats had lower thymus and testes weights, whereas female rats had lower uterine weights. Semicarbazide caused significant changes in sperm motility, sperm count, and sperm abnormality. Histopathologically, semicarbazide caused cortical hypertrophy in adrenals and increased extramedullary hematopoiesis in the spleen; hepatocellular hypertrophy, follicular epithelial hypertrophy in the thyroid, and degeneration of seminiferous tubules in the testis were observed at 60 mg/kg/day when compared to control. Results suggest that 60 mg/kg/day of semicarbazide can exert systemic toxicity in juvenile rats. The no observed adverse effect level (NOAEL) of semicarbazide for juvenile Sprague-Dawley rats was estimated to be 30 mg/kg/day. The semicarbazide (SEM) is the raw material of semicarbazone and has been shown to possess antifungal and antibacterial activities , 2 and rIn vitro, SEM has been shown to have poor mutagenic activity, but in vivo experimental evidence is still insufficient to make a conclusion [SEM showed toxicity in the bone, cartilage, and the aorta in Wistar Hannover GALAS rats . In vitrnclusion . In suscnclusion . SEM is nclusion . Recent nclusion . Ramos enclusion  showed Snclusion .µg/kg (=25 ppb) but found in higher concentrations in baby foods, possibly because of the higher ratio of the gasket area of food mass for these small pack sizes [µg/kg and 53.9 ± 3.1 µg/kg, depending on the type of food within the glass jars [Semicarbazide (SEM) is a by-product of azodicarbonamide (ADC), which is found in foods sold in glass jars and bottles with metal lids sealed with plastic gaskets, according to the European Food Safety Authority . SEM is ck sizes . Differeass jars . The hazass jars . Howeverass jars . AccordiTaking into account that children can represent a subpopulation with particularly high exposure to SEM, the present study is aimed to evaluate the detailed toxicological effects, Functional Observation Battery (FOB) tests, estrous cyclicity, and sexual maturity of SEM upon oral gavage administration during the most appropriate window of exposure—the juvenile period—namely, from weaning to sexual maturity, in male and female SD rats. Potential target organs and/or tissues have been selected on the basis of the juvenile test on rodents and the literature data on SEM toxicity , 22, 23.SEM was purchased from Sigma–Aldrich .In-house breed offspring along with dams were acclimatized on PND 18–21 for 5 days before the start of the treatment. The offspring were weaned on PND 21 from dams. A total of 40 male and 40 female juvenile Sprague-Dawley (SD) rats weighing 40–55 g males and 36–53 g females (23–26 days old) were divided into four groups (10 rats in each group) by body weight stratification. Twelve dams were used to get 40 male and 40 female juvenile rats. The weight variation did not exceed ±20% of the mean weight for each sex. The animal experiment was performed in accordance with the guideline for the care and use of laboratory animals . The Insad libitum.Two juvenile rats of the same sex per cage were housed in sterilized Polysulfone rat cages with paddy husk as bedding material. Rats were housed in a controlled environment room . Pelleted rodent feed  and purified water in polycarbonate bottles were provided SEM is a crystalline white powder stored under ambient conditions. Daily fresh SEM formulation is prepared by dissolving in Milli-Q water for dose administration.The treatment started from between PND 23 and 26 years, which covers an equivalent human age of approximately 2 years of age . SEM forThe selected male and female juvenile rats were assigned to control and treatment groups as given in Three dose levels of 15, 30, and 60 mg/kg/day were selected based on the results of the in-house 14-day oral dose range finding toxicity study with SEM in juvenile SD rats and available literature , 27. In All rats were observed for clinical signs and mortalities twice a day. Detailed clinical examination was done prior to the test item administration on Day 1 for all animals and at weekly intervals. During the comprehensive clinical examination, all rats were observed for changes in the skin, fur, eyes, mucous membranes, the occurrence of secretions and excretions and autonomic activity , posture, changes in gait and response to handling, the presence of clonic or tonic movements, and stereotypies ).Ophthalmological examination was performed with an ophthalmoscope once before the start of the treatment period and at the end of treatment. Mydriasis was induced before examination using a solution of 1% Tropicamide solution.The following neurological examination was conducted during the last week (10th week) of the treatment period for all rats.All animals were observed in the home cage for posture and for the presence or absence of unusual vocalizations and convulsions.Rats were subjected to an open-field test. The rats were placed in an open-field arena with a clean absorbent paper and observed for 2 min. The following behavioral parameters were scored: gait, posture, mobility score, arousal level, clonic or tonic movements, stereotypic behaviors, bizarre behavior, urination, defecation, rearing, and vocalizations .During sensory reactivity measurements, rats were observed and recorded for the following observations: approach response, touch response, click response, tail-pinch response, pupil response, and aerial righting reflex –31.An electronic animal activity measuring device (Columbus Instruments) was used to monitor the motor activity of rats. Each rat was placed individually within the activity cages of the instrument. The rats were monitored for 60 min. During this motor activity following parameters were measured: the stereotypic time  in seconds, the ambulatory time (large ambulatory movement) in seconds, horizontal counts, and ambulatory counts .The landing foot splay was assessed using a similar procedure as published by Edwards and Parker . The heeThe grip performance of the hind and forelimbs was tested using a computerized dual gripping force measuring device (make: Orchid Scientific). Three trials were performed for each rat, i.e., three trials each for forelimbs and hindlimbs. The average of three trials for both forelimbs and hindlimbs was calculated .Individual body weights were recorded on Day 1 of treatment (prior to treatment) and approximately weekly thereafter . The food consumption was measured at weekly intervals, and the individual food consumption was calculated by using the food consumed at each measuring interval and the number of days in the intervening period to determine the food intake/day.The estrous cycle length and pattern were evaluated by vaginal smears, and the stage of the estrous cycle was recorded daily for two weeks prior to necropsy. The length of the estrous cycle was computed for all females as the period between two successive diestrus stages.The age and body weight at the vaginal opening and preputial separation was recorded .Clinical pathology was performed on all animals (Groups 1–4) for blood chemistry and hematology (Day 71). The animals were fasting the entire night before the blood sample. Blood samples were collected by retro-orbital sinus puncture under isoflurane anesthesia prior to terminal sacrifice. Blood was collected into K2EDTA and heparinized tubes for analysis of hematology and clinical chemistry parameters, respectively. The following hematological parameters were determined using ADVIA 2120 hematology system: Hb, RBC, WBC, Plat, Hct, MCV, MCH, MCHC, DLC, Retic, and MPV. The following clinical parameters were determined using Dimension RxL MaX Clinical Chemistry System: ALT, Alb, ALP, AST, BUN, Creat, GGT, Glu, Pi, K, Na, LDH, T. Chol, T. Pro, T. Bil, Trig, Glob, A/G, Ca, Cl.TSH and T4 were estimated by the Enzyme-Linked Immunosorbent Assay (ELISA) method  for the samples.At necropsy, the right epididymis was collected and frozen for sperm count and the right vas deferens was collected for evaluation of sperm motility and sperm morphology. Sperm motility was evaluated using SCA®  CASA  system .At terminal sacrifice, all animals (Groups 1–4) were euthanized by exsanguination under isoflurane anesthesia after blood collection for clinical pathology. The brain, coagulating glands with seminal vesicles, epididymides, heart, kidneys, liver, ovaries, pituitary gland, prostate, spleen, testes, thymus, thyroid and parathyroid, and uterus with the cervix were weighed. The following organs and tissues from all animals were preserved in 10% neutral buffered formalin for histopathological examination: adrenals, bone marrow smear (from femur), brain , bulbourethral gland, cecum, coagulating glands with seminal vesicles, colon, diaphragm, duodenum, femur with joint, heart, harderian glands, ileum (with Peyer's patches), jejunum, kidneys, liver, lungs, mammary glands, mandibular lymph nodes, mesenteric lymph nodes, ovaries, pancreas, pituitary gland, preputial gland, prostate, rectum, salivary glands , sciatic nerves, spleen, spinal cord , sternum with marrow, stomach, thymus, trachea, thyroid and parathyroid, uterus with the cervix, and urinary bladder. The testes and epididymides were collected in modified Davidson's fluid. The eyes were collected in Davidson's fluid.The tissues were processed for routine paraffin embedding and 4-5 micron sections using a Leica® microtome and mounted on slides, which were subsequently stained with Mayer's Hematoxylin and Eosin.p < 0.05. All results were expressed as the mean ± standard deviation of the mean.Differences between groups were determined by one-way analysis of variance (ANOVA) using Statistical Package for Social Sciences  software for windows and post hoc test for intergroup using the least significant difference, followed by Dunnett's test. Significance was considered at No SEM-related clinical signs and mortality were observed in the animals throughout the experiment. However, a clinical sign of sparse hair loss at the right and left flank was observed in a female of the control group and one male rat of the high dose group was considered a spontaneous finding and not related to treatment. However, incidental slight alopecia of the head and trunk in 2/10 in 60 mg/kg/day males and slight to moderate alopecia of the head and hindlimb in 1/10 in 30 mg/kg/day females were considered spontaneous finding and not related to treatment.An ophthalmological examination did not reveal any eye abnormalities.No treatment-related abnormalities were observed in the home cage and open-field observations at all the treated dose levels in either sex .Automated motor activity  assessmeThere were no treatment-related significant variations observed in hindlimb foot splay and grip performance in either sex when compared to concurrent controls .A statistically significant decreased body weight was observed in male rats at 60 mg/kg during treatment days 50–70 . HoweverFood consumption was significantly decreased  in malesThe calculated mean estrous cycle length was 4.76, 4.43, 4.58, and 4.32 days in vehicle control, 15, 30, and 60 mg/kg/day doses, respectively. The mean estrous cycle length in the treated groups was not significantly different from the vehicle control group .When soft pressure is applied to the animal's prepuce, the prepuce fully retracts from the penis. The average age at acquisition of balano-preputial separation in the control, 15, 30, and 60 mg/kg/day exposure groups were 45.98 ± 1.07, 46.88 ± 2.11, 47.63 ± 1.63, and 48.28 ± 1.94 days, respectively. The mean age at acquisition of balano-preputial was statistically higher in the 30 and 60 mg/kg/day dose group .The noticeable split in the membranous sheath covering the vaginal orifice that causes the vaginal edges to separate is referred to as a vaginal opening. The average duration post weaning of vaginal opening (patency) in the control, 15, 30, and 60 mg/kg/day exposure groups were 36.21 ± 1.26, 36.89 ± 1.13, 37.27 ± 1.2, and 39.11 ± 1.13 days, respectively. The mean age at acquisition of vaginal patency was significantly delayed by 3 days at 60 mg/kg/day when compared to the vehicle control group .Body weights at the age of preputial separation and vaginal opening are shown in An increase in reticulocyte count was observed in both sexes at 60 mg/kg/day.This finding was associated with increased hematopoiesis and the presence of megakaryocytes in spleen, microscopically and was not accompanied by any change in other RBC-related parameters.In males, Hct decreased at 60 mg/kg/day group compared with the vehicle control group. No significant differences in other hematological parameters of the 15, 30, and 60 mg/kg/day treated rats as compared with those of rats in the normal control group .Total protein level decreased in males in the 60 mg/kg/day groups compared with in the control groups. AST and ALT levels were significantly increased in both the sexes at the 60 mg/kg/day group .There were no test item-related changes observed in thyroid stimulating hormone (TSH) and thyroxin hormone (T4) levels in adult males and females at termination .The majority of the tissues examined showed no changes in their absolute and relative weights against body weight when compared to concurrent controls. The absolute and relative weights of the liver and spleen were significantly increased in both the sexes at the 60 mg/kg/day group. However, the absolute and relative weight of the testes and thymus were significantly decreased in males, and a marginal decrease in the absolute and relative weight of uterus with cervix was decreased in females at 60 mg/kg/day group Tables  and 10.Significant decreases in the values for percent progressive motile sperms and percent motile sperms were recorded in the 60 mg/kg/day group. Significant decrease in the percent of normal sperms and an increased percentage of abnormal sperms such as giant heads, round heads, tailless heads, and headless tails were recorded at 60 mg/kg/day dose group, compared to the vehicle control. The commonly observed abnormalities across groups were headless or tailless sperms. The values for cauda epididymis weight, number of sperms per cauda epididymis, and number of sperms per gram of cauda epididymis did not differ significantly among the control and high dose group rats .The histopathological evaluations of the selected organs did not reveal any morphological abnormalities that could be attributed to the administration of SEM to the rats. However, following microscopic changes were observed in 60 mg/kg/day group animals.Increased extramedullary hematopoiesis in the spleen, cortical hypertrophy in adrenals involving zona fasciculata of adrenals in males, and both zona fasciculata and zona glomerulosa in females were observed. The lymphoid depletion in the thymus was observed. A minimal degree of centrilobular hepatocyte hypertrophy in both males and females was observed .The main objective of the present study was to comprehensively evaluate the toxicity of SEM in juvenile Sprague-Dawley rats exposed orally for 70 days from weaning to sexual maturation. Few reports have been published on the subchronic oral toxicity of SEM. However, in the present study, mild hepatotoxicity and sperm parameters were affected, which was not reported previously. The rate of mortality, general behavioral changes, and body weight are preliminary indicators of early signs of toxicity caused by various chemicals and drugs , 37. DurIn humans and animals, the hematopoietic pathway is one of the most critical targets for toxic substances and a significant indicator of physiological and pathological states . An incrSince both the liver and the kidney are needed for an organism's survival, biochemical parameters are considered an appropriate predictor for toxicity evaluation . In addiIn the developing male reproductive tract, SEM seems to exert subtle adverse effects by altering the percentage of testicular tissue programmed for spermatogenesis without affecting spermatogenesis itself . In the Increased extramedullary hematopoiesis in the spleen was correlated with increased reticulocytes count, cortical hypertrophy in adrenals this involves zona fasciculata of adrenals in males and both zona fasciculata and zona glomerulosa in females was observed. Hepatocellular hypertrophy in both sexes was observed.After neonatal age, the thymus is required for proper ovarian growth and function, and abnormal thymus function may result in delayed puberty and altered uterus weight , 45. TheTherefore, overall the results of the present study indicate that the NOAEL in juvenile rats is 30 mg/kg/day for SEM oral gavage administration.


Bisphenol F is a substitute material for bisphenol A and is widely used in household products as a raw material for polycarbonate resin, epoxy resin, and plastic reinforcement. It is known to be mainly used in food containers, thermal paper for receipts, and coatings for water pipes. In some countries, bisphenol F has been detected in drinking water and human urine samples. However, due to the lack of safety evaluation data on bisphenol F, it is difficult to establish appropriate guidelines for the proper use of the substance, and social anxiety is increasing accordingly. This study investigated the use, exposure route, and distribution flow of bisphenol F, a household chemical. To determine the no-observed-adverse-effect level (NOAEL) and target organ of bisphenol F after exposure, a single-dose oral toxicity, dose-range finding , repeated dose toxicity , and genotoxicity  tests were performed. The pharmacokinetic profile was also obtained. The test results are as follows: in the pharmacokinetic study, it was confirmed that single oral exposure to BPF resulted in systemic exposure; in single oral dose toxicity test, the approximate lethal dose was found to be 4000 mg/kg and confusion and convulsion was shown in the test animals; NOAEL was determined to be 2 mg/kg/day for male and 5 mg/kg/day for female, and the no-observed-effect level (NOEL) was determined to be 2 mg/kg/day for males and 1 mg/kg/day for females, and the target organ was the small intestine; genotoxicity tests confirmed that BPF does not induce genotoxicity. Bisphenol A (BPA) is a chemical substance with high strength, heat resistance, and transparency  is mainly used as an epoxy resin, and is used for food packaging, water pipes, inner coatings of cans, thermal paper receipts, inter alia, and is mainly used as a coating for water supply pipes. In Korea, BPF was detected in higher concentrations than bisphenol A in the water intake of the Han River. In Seoul, the replacement of water pipes using BPF coatings was promoted. Since then, higher BPF levels were detected in the Han River samples than in the Yeongsan River and Nakdong River samples . Therefore, our research team conducted a long-term toxicity study (90-day repeated administration toxicity test), genotoxicity, and pharmacokinetic studies on BPF to resolve concerns and avoidance of chemical products that are prevalent in society.The BPF (Lot no.: ZS20190510) was purchased from Orient Chemical Enterprise, China. The test substance displayed properties of crystal foam, with low solubility and dispersibility in polar solvents. After grinding the test material, it was sieved through a 100-mesh screen and processed into powder form. BPF in powder form is uniformly dispersed in corn oil at a 20% concentration; this was therefore, selected as the form and vehicle for the test substance. The test solution showed a change in concentration within ±20%, which is the acceptable range for stability when refrigerated for 7 days, and the coefficient of variation between the upper, middle, and lower layers of the low and high doses was confirmed to be within the homogeneity acceptance range of 5%.®, version 6.3, Pharsight Corporation Mountain View, CA, USA).The test was carried out after approval  of the Institutional Animal Care and Use Committee (IACUC) of Korean Conformity Laboratories (KCL). BPF was orally administered at a dose of 200 mg/kg body weight (BW), to Sprague–Dawley (SD) rats. To measure the concentration of BPF in the blood, blood was collected at a certain time  after administration, and the concentration of BPF was analyzed using liquid chromatography with tandem mass spectrometry (LC/MS/MS) . Based on the analysis, changes in blood BPF concentration and pharmacokinetic parameters were calculated via noncompartmental analysis (NCA)  of the IACUC of KCL under good laboratory practice (GLP) regulation. BPF was orally administered to SD rats to investigate the toxicity symptoms and the approximate lethal dose. Test solutions were administered once each to male and female rats at doses of 640, 1600, and 4000 mg/kg, and compared with the control group treated with a vehicle control of distilled water. During the experiment, the occurrence of dead animals, general symptoms, and changes in body weight were noted; gross findings of surviving animals were observed at the end of the experiment.The test was carried out after approval  of the IACUC of KCL and following internationally recognized test guidelines  and GLP regulation. To determine the dose of the 90-day repeated oral administration, a toxicity study was performed by repeated oral administration of BPF for 4 weeks. The test group comprised SD male and female rats administered doses of 25.6, 64, 160, 400, and 1000 mg/kg/day, and their results were compared with those of the control group. Body weight changes, feed intake, eye test, urinalysis, clinical pathology , organ weight measurement, and macroscopic findings were observed at the time of postmortem.The test was carried out after approval  of the IACUC of KCL. In accordance with the OECD testing guidelines  and GLP regulation, sub-chronic oral dose toxicity of BPF was studied in male and female SD rats at doses of 2, 10, 50, and 100 mg/kg/day in male and 1, 5, 15, and 30 mg/kg/day in female rats, followed by a 28-day recovery period. All dosing groups were compared with the control group. During the study, the animals were observed for clinical signs , functional observations , weekly body weights, food/water consumption, and then subjected to an ophthalmological examination. At the end of treatment period, the blood and urine were collected for urinalysis, urine sediment analysis, hematology, blood coagulation tests, serum biochemistry, and hormone analysis. Subsequently, the animals were killed and subjected to gross pathological examination, uterus cycle analysis , and organ weight measurement. The organs were preserved for histopathological examination.Salmonella typhimurium and WP2uvrA, a tryptophan-requiring strain of Escherichia coli, were used. The test substance of the highest concentration was prepared by dissolving the test substance in dimethyl sulfoxide (DMSO), and the test substance of a low concentration was prepared by diluting it step by step in DMSO. Both the direct method and the metabolic activity method were used.To obtain basic data for confirming whether the test substance (BPF) causes carcinogenicity, a microbial reverse mutation test was conducted following internationally recognized test guidelines  and GLP regulation. In the test, the histidine-requiring strains  of To determine the concentration in the main test, a concentration determination test was conducted in a concentration group of five steps with a mixture of three with 5000 µg/plate as the highest concentration. As a result, growth inhibition was observed in all the strains using the direct method and the metabolic activation method. Compared to the negative control group, no increase in the number of reverse mutated colonies that could be judged as positive was observed. Based on the concentration determination test, the main test was conducted at the following concentrations: TA98 strain of S9 mix (−)/TA1537 of S9 mix (+): 0, 7, 21, 62, 185, and 556 µg/plate.The OECD guidelines 473  informed this test. To evaluate the genotoxicity of BPF, ovarian embryonic cells (CHO-k1 cells) derived from Chinese hamsters were used. The metabolic activation method (+S9 mix) with metabolic activating enzyme system (S9) and the direct method without application (− S9 mix) was used to conduct the chromosomal abnormality test under GLP regulation.The test substance was dissolved in DMSO to prepare the test substance at the highest concentration and subsequently diluted sequentially for lower concentrations of the test substance. To determine the treatment concentration of the test substance in the main test, a cell proliferation inhibition test at concentrations of 2.29, 6.86, 20.58, 61.73, 185.19, 555.56, 1666.67, and 5000 µg/ml was performed. Three mixtures and concentrations were chosen. The concentrations in the main test were as follows: direct method : 2.29, 6.86, and 20.58 µg/ml; direct method : 20.58, 61.73, and 185.19 µg/ml; metabolism activity method : 20.58, 61.73, and 185.19 µg/ml.To evaluate the genotoxicity of BPF, a micronucleus test was performed using bone marrow cells from Institute of Cancer Research mice following internationally recognized test guidelines  and GLP regulation. The test was carried out after approval  of the IACUC of KCL. The concentration of the test solution was as follows: preliminary test: 2000 mg/kg BW/day, 1000 mg/kg BW/day, 500 mg/kg BW/day, main test: 2000 mg/kg BW/day, 1000 mg/kg BW/day, and 500 mg/kg BW/day. In the preliminary test, three males and three females per group were used. As a result of the preliminary test, symptoms such as abnormal walking were observed in the group of females and males administered the highest concentration of test substance (2000 mg/kg BW/day), but no dead animals were observed. Based on the results of the preliminary test, the main test was conducted with the highest dose concentration of 2000 mg/kg BW/day. In this test, males were used because it was determined that there was no difference in sensitivity to toxicity between males and females.p < 0.05. SPSS for Windows version 12.0 software  was used for analysis. Asterisks (*) indicate statistically significant differences compared with the control groups. (1) Analysis of continuous data : the statistical treatment was conducted assuming normality. The differences among the groups were examined and assumed equal variance using a standard one-way analysis of variance (ANOVA). If the test showed statistical significance, the data were analyzed by parametric multiple comparisons to compare the control group with the experimental groups. If equal variance was obtained, Duncan’s test was used, and if the equal variance was not obtained, Dunnett’s t-test was applied. (2) Analysis of non-continuous data : the data were converted by scale conversion and then analyzed using Chi-squared analysis.The differences among the control group and all the dosing groups were analyzed through parametric or non-parametric multiple comparison procedures, as appropriate. Differences were determined to be statistically significant at The concentration of the BPF increased rapidly 0.25 h after administration, and then decreased rapidly for 3 h thereafter. In males, BPF in blood serum vanished after 72 h, and in females, the drug remained even after 72 h. The concentration of BPF in blood over time is shown in Fig. 0-72 h, Cmax, and Vd value of females were ~ 1.5, 1.3, 1.1 times higher than those of males, respectively. Cl of males was ~ 1.5 times higher than that of females. The elimination half-life was increased in females because females have a higher absorption and lower clearance rate. However, there was no difference in the volume of distribution between both sexes. Unlike other chemicals, BPF has a high vanishing half-life, which is believed to be delayed in excretion from the body. In addition, it is judged that drug loss in females is delayed compared to males because females have a higher absorption rate and a lower excretion rate due to increased elimination half-life. As a result of analyzing the difference between males and females, AUCTwo males in the 4000 mg/kg group died on the first day of administration. After treatment, symptoms such as salivation and inanimation were observed in the male and female 640 mg/kg group; salivation, inanimation, stupor , and convulsions  were observed in the male and female 640 mg/kg group; and salivation, inanimation, confusion, and convulsions were observed in the male and female 4000 mg/kg group. On the first day of treatment, contamination around the anus was observed in the male 640, 1600, and 4000 mg/kg groups and the female 4000 mg/kg group, and no specific general symptoms were observed thereafter.p < 0.01) on the 1st day after treatment and on the 7th day in the 4000 mg/kg group (p < 0.05). There was no difference in body weight on the third day after treatment, but a tendency to lose weight was observed. All animals had recovered to their normal weight by the 14th day. In females, on the 1st and 3rd days after treatment, the body weight of the 1600 and 4000 mg/kg groups decreased (p < 0.01), and then recovered normally  to confirm the recovery of the substance.No dead animals were observed during the test. During the administration period, salivation was observed immediately after administration of the test substance in the males dosed > 10 mg/kg/day and females dosed > 15 mg/kg/day. Considering the duration, frequency, and pattern of salivation, it tended to be a dose-dependent effect of the test substance, but it only appeared immediately after administration and not during the recovery period. This is believed to have no toxicological significance as it appears to be due to the temporary promotion of salivation.No significant differences were noted across different groups in both sexes in the main group and the recovery group.During the administration period, significant weight loss  was observed in the males receiving > 200 mg/kg/day and in the females receiving > 30 mg/kg/day. During the recovery period, the degree of weight loss in the male 200 mg/kg/day and female 60 mg/kg/day groups slowed . Although there was some recovery of weight lost, it is considered a toxicological effect because the weight loss was > 10% of the control group, and a dose correlation was observed  level of the male 200 mg/kg/day and the females dosed > 15 mg/kg/day was higher than that of the control group. The response showed a dose dependency and, in particular, both sexes showed the same tendency in the highest dose group. The increase in PT values at the highest dosages exceeded the range of biological fluctuations  at this laboratory. The changes in PT were considered to be an effect of the test substance for two reasons: (1) fat-soluble vitamins are required for the production of coagulation factors related to PT levels. Among the histopathological findings of this study, lymphatic dilatation seems to have inhibited the absorption of fat-soluble nutrients, and (2) the PT level recovered to some extent during the recovery period without administration of the substance. The increase in APTT levels in the recovery group of females receiving 60 mg/kg/day group was not observed in the main group, so it seems that this is of no toxicological significance , high-density lipoproteins (HDL), and low-density lipoproteins levels (LDL) in the males dosed > 50 mg/kg/day and the CHO level in the females dosed > 5 mg/kg/day groups decreased. In the case of HDL, a decreasing trend was observed. It was judged a secondary effect of absorption inhibition of the test substance .The cholinesterase (ChE) levels decreased in the male 200 mg/kg/day group in the main group and the recovery group, and there was no statistical difference, but a decreasing trend was observed even at high dosages in the female group, which is considered to be a change by the test substance. However, both male and female levels were within the biological rage of the test animals , therefore, its toxicological significance is unclear  level increased, therefore, an effect of the test substance cannot be excluded. The relative weight of the right thyroid gland of the male 200 mg/kg/day group and the left thyroid gland of the female 30 mg/kg/day group was increased compared to that of the control. The incidence was unilateral, there was no difference in absolute weight, and the dose dependency was not clear, so it was considered to be a relative change due to weight loss of the test animal (about 8–17% decrease in fasting weight before postmortem compared to the control group). In the case of histopathological examination, no functional or morphological abnormalities of the thyroid gland were identified and recovery was demonstrated during the recovery period. Therefore, the change in the T4 level was not found to have any supporting results, therefore, it was judged that there was no toxicological significance in the hormone analysis . In the recovery group, a minimal level of lymphatic dilatation was observed in the male 200 mg/kg/day and female 60 mg/kg/day groups, and symptoms recovered Fig. .Fig. 5LaUnder the test conditions, weight loss was observed in the males dosed > 200 mg/kg/day and females dosed > 30 mg/kg/day. Lacteal dilatation in the small intestine was observed in the males dosed > 10 mg/kg/day and females dosed > 15 mg/kg/day. Secondary to lacteal dilatation, the CHO level decreased in the males dosed > 50 mg/kg/day and the females dosed over > 5 mg/kg/day. The PT value was prolonged in the male 200 mg/kg/day and female > 15 mg/kg/day groups. All the findings mentioned above showed reversibility during the recovery period in the absence of BPF Fig. . ConsideTherefore, the NOAEL of BPF was determined as 2 mg/kg/day in males and 1 mg/kg/day in females, and the target organ was identified as the small intestine.As a result of the main test, growth inhibition of bacteria was observed in all strains in the direct and metabolic activity methods. Compared to the negative control group, no increase in the number of reverse mutated colonies (which could be judged as a positive result) was observed. As a result of the sterility confirmation test of the test substance and S9 mix, no contamination by microorganisms was observed in the positive and negative controls. The number of colonies induced was within the expected range, indicating that each test was performed properly. From these results, it was determined that BPF did not induce a reverse mutation under these test conditions.In the case of the 24-h continuous treatment group and the 18-h recovery after 6-h treatment group without metabolic activity, the frequency of abnormal intermediate phases increased significantly compared to the control group at all dosages. In the case of the metabolic activity method (6-h treatment and 18-h recovery), no statistically significant increase was observed in the frequency of abnormal metaphase in all treatment groups compared to the negative control group. In the direct method and the metabolic activity method, the frequency of ploidy and intranuclear embedding also failed to increase compared to the negative control. Consequently, BPF did not induce chromosomal abnormalities in CHO-k1 cells under these test conditions.p < 0.01). After administration of the test substance, no statistically significant body weight change was observed in all administration groups compared to the control. The micronucleus incidence in the control and the positive control and the proportion of polyinflammatory red blood cells among the total red blood cells were within the range of historical control data. From the above results, it is believed that BPF does not induce micronuclei in the bone marrow cells of mice under the conditions of this test.As a result of the main test, no dead animals occurred in the test substance administration groups, and abnormal walking was seen in animals receiving 2000 mg/kg/day group. The number of polyinflammatory red blood cells among the total red blood cells did not show a distinct difference, and no inhibition of the proliferation of myeloid cells was observed compared to the control group. The micronucleus induction frequency observed in 4000 polyinflammatory red blood cells per animal did not show statistically significant results compared to the control at all dosages. Meanwhile, a difference was observed in the micronucleus induction frequency of the positive control group compared to the excipient control group  were performed, BPF did not induce reversion mutation or chromosomal aberration. In the in vivo micronucleus test, it was confirmed that BPF did not induce micronuclei in the bone marrow cells of mice. The above three genotoxicity tests confirmed that BPF does not induce genotoxicity.As a result of the single oral dose toxicity test for BPF, symptoms such as confusion and convulsions were observed when administered orally in a single-dose toxicity test in rodents, and the approximate lethal dose was found to be 4000 mg/kg. As a result of a repeated oral administration toxicity test over 90-day, weight loss in the male 200 mg/kg/day and female > 30 mg/kg/day groups, lacteal dilatation in small intestine in the male > 10 mg/kg/day and female > 15 mg/kg/day groups, reduction of total cholesterol in males > 50 mg/kg/day and females > 5 mg/kg/day, and prolonged prothrombin time in males 200 mg/kg/day and females > 15 mg/kg/day were observed. All the findings mentioned above showed reversibility during the recovery period in the absence of BPF. Through the above test results, the NOAEL of BPF was determined as 2 mg/kg/day in males and 5 mg/kg/day in females, and the NOEL of BPF was determined as 2 mg/kg/day in males and 1 mg/kg/day in females. The target organ was identified as the small intestine.Lacteal dilatation in the small intestine may be associated with intestinal lymphangiectasia in humans. Intestinal lymphangiectasis is classified into idiopathic intestinal lymphangiectasia due to congenital malformation and secondary intestinal lymphangiectasia caused by lymphatic obstruction or an underlying disease that increases intra-lymphatic pressure (Vignes and Bellanger Several studies have reported estimated daily intakes through exposure scenarios of BPA including its analogs (Liao and Kannan In the recent research, BPF was detected in several mustard products containing S. alba seeds and in various plants of the orchid family (Zoller et al. Currently, BPF is being used without a suitable substitute, but a risk issue has been raised which mandates national policy intervention for its use (Eladak et al.


In this talk I will describe a series of studies conducted at the Centre for the Developing Brain, King’s College London, that seek to increase our understanding of why infants who are born very early (before 32 weeks’ gestation) are more likely to develop socio-emotional problems when they grow up compared to infants who are born at term. As part of the Evaluation of Preterm Imaging study we carried out multimodal MRI at term in over 200 newborns and studied whether we could identify specific patterns of brain development in those infants who might develop problems with emotion regulation and general mental health as they grow-up. At the behavioural level, we found that very preterm children compared to term-born controls had more mental health problems, including anxiety and autism-spectrum behaviours. Preterm children had lower IQ, were less able to regulate their emotions and inhibit unwanted behaviours. Children’s tendency to attribute negative emotions to daily events, which could lead to increased anxiety, was associated with two main neonatal brain features. These were: 1) weaker structural connectivity in a long-range white matter projection tract called the uncinate fasciculus which connects the frontal lobe with the anterior temporal lobe and 2) altered fronto-limbic functional connectivity, both of which play a critical role in several aspects of social and emotional development. These findings show that early brain changes can be used to predict children’s social and emotional outcomes, hence could be used to inform preventative interventions aimed at averting and targeting emerging emotional disorders.No significant relationships.


Sanghuangporus baumii is a traditional medicinal fungus that produces pharmacological terpenoids, but natural resources are insufficient for applications, and its growth and development mechanisms are poorly understood. Combining metabolomic and transcriptomic analyses, we found four terpenoid hormones and a central gene, isopentenyl diphosphate isomerase (IDI), involved in growth and development. Additionally, an exogenous hormone test was used to further confirm the importance of the four terpenoid hormones. Finally, hormone content determination and qRT−PCR were performed to explore the growth and development mechanism; we found thatcis-zeatin (CZ) plays a major role in the mycelia stage, trans-zeatin (TZ) and gibberellin A4 (GA4) are important in the primordia stage, GA4 is crucial for the fruiting bodies stage, and abscisic acid (ABA) may be a marker of maturity. The IDI gene was also found to affectterpenoid hormone content by regulating the relative gene transcript levels, thereby controlling morphological changes in S. baumii. Our results revealthe growth and development mechanisms of S. baumii and may promote the breeding and utilisation of high-quality varieties. Sanghuangporus baumii (Pilát) L.W. Zhou & Y.C. Dai is a precious fungus with medicinal effects, including improving immunity, a hypoglycemic effect and anticancer and antitumour activities . Th. ThCYP53IDI gene expression level were observed in Myc, followed by Pri and Fru. Furthermore, we revealed a correlation and linkage between TZ and CYP5340A71 and CYP5340A72 gene expression levels in different developmental stages. Similarly, the highest ABA product abundance and the highest ABA4 gene expression levels were observed in Fru, followed by Priand Myc. Our results are in agreement with those of previous studies [Metabolome and transcriptome analyses have become common technical means to explore the underlying molecular mechanisms of fungal metabolic biosynthesis ,30, and  studies ,30, showS. baumii. We compared the differences in terpenoid hormone contents and related gene transcript levels between IT and WT S. baumii and found that overexpression ofthe IDI gene does not increase the contents of all terpenoid hormones, mainly due to competition among hormones and tissue-specific expression [S. baumii CZ, TZ and GA4 levels and transcription of related genes were decreased in Myc and Pri stages, presumably due to increased triterpenoids accumulation caused by high-level expression of triterpenoid biosynthetic genes (Analysis of the underlying mechanism revealed the regulation of growth and development of pression ,35. IT Sic genes . When thic genes . This alic genes B. This pS. baumii are affected by terpenoid hormones, althoughit is unknown whether expressing all terpenoid hormone synthesis genes at the same time can accelerate the whole growth and development stage. Therefore, in future studies, we plan to use the gene coexpression method to study the growth and development of S. baumii [S. baumii.Studies have shown that the growth and development of . baumii  in an at. baumii ,35. The IDI. Subsequently, we measured the transcript levels of terpenoid genes and the content of terpenoid hormones. Analysis of the underlying mechanism revealed that the IDI gene repressed the transcription of TRIT1, CYP5340A71, CYP5340A72 and GGPS genes by increasing the transcript levels of SQS and LS genes, and this consequently decreased the accumulation of CZ, TZ and GA4, which slowed growth in the Myc and Pri stages. The IDI gene increased transcription of the GGPS gene by attenuating the transcriptional activity of SQS and LS genes, resulting in increased GA4 content and accelerated growth in the Fru stage. Specifically, CZ played a major role in the Myc stage, TZ and GA4 functioned in the Pri stage and GA4 influenced the Fru stage. ABA may be a marker of maturity in S. baumii. These findings provide important guidance for molecular breeding of S. baumii.Metabolomics and transcriptomics approaches were applied to identify four terpenoid hormones and the associated key biosynthesis gene,


The RMS roughness is about 0.5 nm. The FWHM of (002) XRD rocking curve is 230 arcsec and the FWHM of (102) XRD rocking curve is 260 arcsec. As result, a hole concentration of 5 × 1018 cm−3 and a resistivity of 1.5 Ω·cm have been obtained. The hole concentration increases due to the incorporation of surface accumulated Mg dopants into suitable Ga substitutional sites with minimal formation of compensatory defects.Metal modulation epitaxy (MME) is a technique in which metal beams  are switched on and off in short periods in an RF MBE system while a continuous nitrogen plasma beam is kept on. We systematically studied the effect of periodic duty cycling on the morphology, crystalline quality, Mg doping concentration, and electrical properties of GaN:Mg films grown by MME. When the metal shutter duty cycling is 20 s open/10 s close, the sample has smooth surface with clear steps even with Mg doping concentration higher than 1 × 10 In fi20 cm−3 ,10. As a20 cm−3 , co-dopi20 cm−3 , superla20 cm−3  and dopi20 cm−3 . HoweverMetal-modulated epitaxy (MME) technology has been reported to greatly increase hole concentration in GaN. MME is a technique in which metal beams  are switched on and off in short periods in an RF MBE system, while a continuous nitrogen plasma beam is kept on . We firsIn metal modulation epitaxy, there are three parameters that can be changed to affect the crystal quality of the sample, namely, the fluxes of the metal source, the opening time of the metal source shutter and the closing time of the metal source shutter. According to my literature survey, the previous studies changed one or two of these parameters. In this article, I have studied the influence of three parameters on GaN:Mg films. The effect of periodic duty cyclings on GaN:Mg films  is systematically investigated in this work, and GaN:Mg films with smooth morphology, high crystalline quality and high hole concentration are obtained.−10 Torr. The templates were cleaned using a dilute hydrochloric acid solution in a ratio of 1:3, and then using ultrasonically cleaned with acetone, isopropanol, deionized water in sequence for 10 min each time, and dried with nitrogen before being loaded into a preconditioning chamber. Templates were first degassed in the preconditioning chamber at 200 °C for 60 min, followed by degassing in the growth chamber at 650 °C for 30 min. For all films, nitrogen was supplied by a RF plasma source with the power of 330 W and N2 flow of 1.5 sccm. Ga was supplied by a standard effusion cell, and Mg was supplied by the Veeco corrosive valve cracker. During growth, the BEP of Ga was 6.8 × 10−8 Torr. An 80 nm thick GaN buffer layer was first grown on templates by regular growth. Then GaN:Mg films were grown at 760 ℃ with the same Ga and Mg flux, but the periodic duty cyclings (open/close) was varied. The total opening time of metal source shutter was 7200 s.All GaN:Mg films were grown by MME in a GEN 20A MBE system on n-type MOCVD-grown GaN templates, with a chamber pressure of approximately 1 × 10A Veeco Dimension 3100 Atomic Force Microscope (AFM) was used to observe the surface morphology of GaN: Mg samples in tapping mode. The electrical characteristics of the films at room temperature, such as hole concentration and mobility, were determined by a Hall effect measurement device. The samples were cut into small pieces of 1 cm×1 cm, using indium dots as contact electrodes, and the current-voltage curve of each sample was linear to ensure that the contact was ohmic. Mg concentration was detected by secondary ion mass spectrometry (SIMS) measurements. With a high-resolution XRD instrument, the crystalline quality of the films was evaluated by the full width at half maximum (FWHM) of the symmetric (002) and asymmetric (102) diffraction patterns of the rocking curves.5 cm−2, which is determined by optical microscopy. Longer metal source shutter opening times means higher III/V ratio on average over one cycle, resulting in smoother surfaces, which is consistent with the theoretical considerations reported by Zywietz et al. [The metal source shutter closing time was kept constant of 5 s, and the metal source shutter opening time was 5 s, 10 s, 15 s and 20 s, respectively, which are recorded as A1, A2, A3 and A4. z et al. . The incThe metal source shutter opening time was kept constant of 10 s, the metal source shutter closing time was 2.5 s, 5 s, 10 s and 20 s, and the samples are marked as B1, B2, B3 and B4, respectively. As can be seen in We chose four different shutter duty cycles, with duty cycling (open/close) of 6s/3 s, 10 s/5 s, 20 s/10 s, 40 s/20 s, but the total metal source shutter opening time was kept at 7200 s. The samples were denoted as C1, C2, C3, and C4 respectively. As shown in 20 cm−3, 1 × 1020 cm−3, 6 × 1019 cm−3, and 1 × 1018 cm−3, respectively, which can be seen from 18 cm−3 and 1.4 × 1018 cm−3, respectively. As there are too many compensation defects in A1, although the Mg doping concentration of sample A1 is the highest, it appears as n-type probably due to the compensatory defects. Due to the excessive metal droplets on the A4 surface, the incorporation of Mg into the lattice is inhibited under too Ga-rich conditions. The doping concentration of Mg on the surface is only 1 × 1018 cm−3, and the generated holes are too few to offset the influence of intrinsic defects, so it appears n-type in the Hall test.According to the SIMS measurement, the Mg doping concentrations of the four samples A1, A2, A3, and A4 are 2.5 × 1017 cm−3, 1 × 1020 cm−3, 2 × 1020 cm−3, and 1.8 × 1020 cm−3, respectively. However, only sample B2 appears as p-type, and the hole concentration is 2 × 1018 cm−3. The other three samples appear as n-type GaN. As the metal source shutter closing time becomes longer, the overall average III/V ratio in the period decreases, and the Mg doping concentration generally increases. The excessively long shutter closing time for sample B4 results into slightly reduced Mg concentration compared to sample B3 probably due to aggravated Mg desorption. Thus, B1 appears as n-type mainly because Mg concentration is too low, and B3 and B4 are attributed to high density compensatory defects.According to the SIMS measurement, as shown in 19 cm−3, 1 × 1020 cm−3, 1.2 × 1020 cm−3, and 1.2 × 1020 cm−3, respectively. Except for C1, the Mg doping concentration is about the same. This is because the III/V ratios of the latter three samples remain the same. C1 may be due to the fact that the metal source shutter closing time is too short. Therefore, Ga and Mg atoms do not have enough time to diffuse and migrate on the growth surface, and find a suitable lattice position for incorporation, so the Mg doping concentrations is reduced.The results of SIMS measurements shown in 18 cm−3 is obtained; for comparison, the highest hole concentration of typical p-GaN in the world is 1–2 × 1018 cm−3, which is a great progress. However, hole mobility is relatively low, about 2 cm2/V·S. This is probably due to the high doping concentration of Mg, which forms a lot of point defects and extended defects, causing serious ionized impurity scattering. A low resistivity of 1.5 Ω·cm is obtained by optimizing the metal source shutter opening time. Since Mg doping concentrations are almost the same for the other three samples, the increase in hole concentration is mainly due to the incorporation of surface accumulated Mg dopants into suitable Ga substitutional sites, while the formation of compensatory defects (N vacancies and extended defects) is minimal.8 cm−2 and 3.86 × 108 cm−2, respectively. The sample C3 shows the lowest edge defect density and the least defect compensation, which is consistent with the highest hole concentration obtained by the Hall test.We measured the X-ray diffraction rocking curve of these three groups of samples and obtained full width at half maximum (FWHM) of (002) plane and (102) plane, as shown in −8 Torr, nitrogen was supplied by a RF plasma source with the power of 330 W and N2 flow of 1.5 sccm, the substrate temperature is 760 ℃, the metal shutter duty cycling is 20 s open/10 s close, and the sample has a high Mg doping concentration of over than 1 × 1020 cm−3 with a smooth surface. The RMS roughness is about 0.5 nm. The FWHM of (002) XRD rocking curve is 230 arcsec and the FWHM of (102) XRD rocking curve is 260 arcsec. As a result, a hole concentration of 5 × 1018 cm−3, corresponding Mg activation rate of 4.2%, and a resistivity of 1.5 Ω·cm have been obtained, which is important to develop optoelectronic device such as laser. The hole concentration increases due to the incorporation of surface accumulated Mg dopants into suitable Ga substitutional sites with minimal formation of compensatory defects.In summary, we systematically studied the effect of the periodic duty cycling on the surface morphology, crystal quality, Mg doping concentration, and electrical properties of Mg-doped GaN films grown by MME technique. It was found that when the BEP of Ga was 6.8 × 10


The literature was searched on the PubMed and Web of Science databases. The search terms used were “OTOPLAN”, “cochlear planning software” “three-dimensional imaging”, “3D segmentation”, and “cochlear implant” combined into different queries. This strategy yielded 52 publications, and a total of 31 studies were included. The review of the literature revealed that OTOPLAN is a useful tool for otologists and audiologists as it improves preoperative surgical planning both in adults and in children, guides the intraoperative procedure and allows postoperative evaluation of the CI.The cochlear implant (CI) is a widely accepted option in patients with severe to profound hearing loss receiving limited benefit from traditional hearing aids. CI surgery uses a default setting for frequency allocation aiming to reproduce tonotopicity, thus mimicking the normal cochlea. One emerging instrument that may substantially help the surgeon before, during, and after the surgery is a surgical planning software product developed in collaboration by CASCINATION AG  and MED-EL (Innsbruck Austria). The aim of this narrative review is to present an overview of the main features of this otological planning software, called OTOPLAN Conventional acoustic hearing aids become less effective for individuals affected by severe to profound sensorineural hearing loss. For a subset of this population, the emergence of cochlear implants (CIs) has offered an alternative therapeutic option .A CI translates acoustic signals into electrical impulses that are then conveyed to the organ of Corti in the cochlea. The regular cochlea has a tonotopic map that conducts characteristic frequencies (CFs) to precise locations along the basilar membrane (BM).CFs correspond to the frequency at which the cochlear BM expresses the highest sensitivity with the lowest sensation sound level .The arrangement of frequencies across space is known as cochlear tonotopy. Nerve fibers extend from the BM to the spiral ganglion, which is situated nearer to the central axis of the cochlear spiral and houses the cell bodies of neurons .To achieve the optimal functional outcome for CI patients, it is expected that tonotopic stimulation should be as precise as possible, meaning that the CI electrodes stimulate in accordance with their postoperative tonotopic positions .The variation in post-surgical speech perception can be affected by the patient’s own physiologic features such as severity of deafness, age at CI surgery, etiology of hearing loss, degree of residual hearing, and several other preimplantation cognitive factors ,6. WhileCurrently, standard CI surgery uses a default setting for frequency allocation that aims to replicate tonotopicity, thus mimicking the normal cochlea. In the CI, electrode arrays are surgically placed within the cochlea to directly activate nerve fibers through electrical signals, circumventing the impaired sensory cells and mechanoelectrical transduction process . This prThe frequency-to-place matching of the CI electrode connection is usually based on the Greenwood function . This teElectrode arrays come in several different designs that influence the allocation of the CI electrodes within the cochlea ; when usWhen pre-set frequency programming is applied, differences in the type of implant, CDL, and degree of insertion may cause discrepancies between the CF of the cochlea and the frequency that the electrode presents to that part of the cochlea, leading to an allocation mismatch.  To solveAt this stage, it became necessary to implement a system that could facilitate a personalized examination of the patient’s anatomy before the scheduled surgery, rather than relying on a standard approach to guide the surgical preparations for CI.®, allows three-dimensional handling that can be used preoperatively by the otologist to study the cochlear anatomy and visualize the electrode insertion depth prior to surgery, and to choose the best electrode array for that patient; postoperatively, it can be applied to check frequency allocation, and the proper CI position.One emerging instrument that may help the surgeon before, during, and after the surgery is an otological planning software product created in cooperation by CASCINATION AG  and MED-EL (Innsbruck Austria) . This sy® otological planning software and its different uses and to discuss the advantages and disadvantages of its use in CI surgery.The aim of this narrative review is to describe the main features of the OTOPLANThe literature was searched on the PubMed and Web of Science databases in May 2023. No time limitations were applied. The search terms were “OTOPLAN”, “cochlear planning software” “three-dimensional imaging”, “3D segmentation”, and “cochlear implant” combined into the following query: ‘cochlear planning software’ * AND [(‘cochlear implant*’ OR ‘cochlear implant surgery*’ OR ‘otosurgery’) AND ]. Papers published until the date of the review that contained this query in the title or the abstract were selected. We limited our search to publications indexed as articles, proceedings or reviews available in the English language.® system but used alternative software; and 8 articles that focused on robotic surgery for CI with or without OTOPLAN®. Finally, a total of 31 studies were included, among which 15 focused on adults only and 13 focused on children or mixed populations of adults and children, whereas 3 did not provide information on the age of the patients. This strategy yielded 52 publications. We excluded 4 articles that were focused not on CI but on other types of middle ear implants or bone-conduction implants; 5 articles that did not use the OTOPLAN® as a tool for preoperative surgical planning and postoperative electrode placement analysis and anatomy-based fitting in CI surgery.This review included several studies addressing the use of OTOPLANOTOPLAN, a standalone software tool, enables the generation of 3D reconstructions from digital imaging and communications in medicine (DICOM) images, offering a multifaceted approach to cochlear parameter calculation and visualization. This software not only facilitates the visualization of postoperative electrode positions but also ensures individualized matching and provides a comprehensive patient report, alongside a detailed postoperative analysis .When exploring cochlear metrics with OTOPLAN, several values are assessed, each providing unique insights into cochlear dimensions. The A-value represents the diameter, defined as the maximum distance from the round window to the lateral wall of the basal turn, passing through the modiolus. Meanwhile, the B-value illustrates the width of the cochlea, measured perpendicularly to the A-value line at the modiolus. Additionally, the H-value indicates the height of the cochlea, quantifying the space between the center of the basal turn and the apex point. These metrics are visually represented in OTOPLAN adeptly localizes the modiolus, round window, and cochlear boundaries to calculate the CDL at the organ of Corti level, utilizing the elliptic–circular approximation (ECA) method . HistoriGiven that cochlear dimensions can fluctuate significantly, ranging from 19.71 to 45.6 mm and potentially varying by 30% to 40% ,28, it iThe capability of OTOPLAN in determining both the angular insertion depth (AID) of the CI  and the Accurate cochlear dimension estimates assist surgeons in selecting the most apt electrode array, ensuring an optimal length for each specific patient. This precision proves particularly insightful in instances of inner ear malformations and during the postoperative activation phase, where the frequency map can be utilized to enhance frequency reallocation.Designed to be compatible with both CT and MRI scans, OTOPLAN has demonstrated a minimal impact of CT slice thickness on CDL measurement . FurtherThe dimensions and form of the cochlea influence CI electrode placement and cochlear coverage, affecting final pitch discrimination ,24. The Utilizing CT imaging and 3D reconstruction, OTOPLAN has been applied in various research contexts to explore ear anatomy . The sofThe research indicates no difference in the A-value measured via DICOM and OTOPLAN, affirming its applicability for pediatric patients. Furthermore, a notable correlation exists between cochlear parameters  and the CDL, with a combination of the A-value and B-value providing a precise CDL estimation. Additionally, men generally present higher cochlear parameters than women, with CDL measurements being comparable to those obtained through the use of alternative techniques ,31,38.While OTOPLAN’s reliability in estimating cochlear parameters has been acknowledged, concerns about its accuracy in malformed ears have been raised . The sofOTOPLAN’s measurements of cochlear parameters in children were used in 3D segmentation software to measure the vestibular aqueduct (VAD) volume. A strong correlation was found between OTOPLAN measurements and VAD volumes, enabling the development of a system to calculate both CT inner ear volume and VAD volumes, particularly in subjects with an enlarged vestibular aqueduct requiring CI surgery. Gender-based differences in inner ear anatomy were also confirmed, and gender and VAD width at the midpoint were identified as significant predictors of the risk of gushing .Measuring the height of the scala tympani is challenging due to the limited resolution of clinical CT scans. OTOPLAN facilitates the generation of personalized, oblique cochlear views using available CT images .Research using OTOPLAN to explore the relationship between mastoid thickness and growth from 6 months to 20 years revealed a logarithmic growth in mastoid thickness that plateaus at 20 years. This information can assist otosurgeons in determining the required drilling depth in the mastoid and potentially inform improvements in CI design to minimize excess electrode lead length .® software. The evaluation noted excellent inter-rater reliability, a high agreement of outcomes, and a reduction in execution time using the automatic method [A retrospective evaluation compared cochlear parameters measured manually by two expert otosurgeons with those automatically obtained using a development version of the OTOPLANc method .The pivotal role of OTOPLAN in CI surgery is illuminated through its utility in assisting surgeons in devising a strategic surgical approach, foreseeing potential intraoperative complications, and choosing an electrode array that harmonizes with the specific cochlear anatomy. A variety of research has explored these aspects, investigating diverse methodologies for segmenting temporal bone surgical anatomy for patient-specific virtual reality simulation. Automated segmentation algorithms have emerged as a flexible and viable approach, particularly useful in managing cases with abnormal anatomy .A notable concern in CI surgery is intraoperative complications, which can lead to endoluminal fibrosis and round window ossification. Ensuring complete array insertion is crucial, not only to secure an optimal tonotopic match but also to stimulate the apex of the cochlea, which is vital for speech understanding and music perception, and where mismatch is notably pronounced .Moreover, OTOPLAN has proven effective in instances of severely advanced otosclerosis, including those with cochlear ossification and anatomical abnormalities, as well as in cases with a normally shaped cochlea, where conventional DICOM viewers were insufficient in providing necessary information. Particularly, OTOPLAN has demonstrated its worth in challenging cases of severely advanced otosclerosis, with comparative studies revealing that patients who underwent CI surgery with OTOPLAN evaluation exhibited slightly better outcomes and encountered no instances of incomplete array insertion, as opposed to a historical group where surgical planning was conducted using CT images ,46,47.Effective surgical planning, which enables the surgeon to sidestep critical issues during CI surgery, should be accomplished using the least invasive approach. Utilizing OTOPLAN-reconstructed imaging provides insightful analysis of the optimal surgical trajectory line through the retro-facial recess towards the round window and also enables the measurement and classification of the size of the facial recess, comparing the retro-facial approach to the standard facial recess approach .Accuracy in estimating the CDL is augmented by three-dimensional reconstructions, which scrutinize the entire 3D structure of the cochlea and are not subject to viewing angle effects, thereby representing the most precise technique for CDL estimation .Choosing the correct electrode array length is another crucial aspect in CI surgery, given the array of electrodes available, which vary in length and structure. OTOPLAN has been utilized to measure the CDL and estimate the frequency allocation map, assisting in selecting the most fitting electrode array . In pediOTOPLAN has also been validated in determining the postoperative AID, CDL, and cochlear place frequency with a lateral-wall array ,52. AID Furthermore, it has been suggested that the customized placement of slim modiolar electrodes, based on CDL measurements using OTOPLAN, can result in improved modiolar proximity . A shortEnsuring a complication-free cochlear implant procedure involves a meticulous preoperative assessment, which aims to facilitate atraumatic insertion and address individual anatomical variations. Nonetheless, achieving optimal audiological outcomes can be intricate due to patient-specific factors, unpredictable comorbidities, and potential surgical complications .The complexity of cochlear differences is well-established. Even when the cochlear morphology seems normal on CT scans, surgical challenges, such as partial insertion due to a smaller cochlea or injury to the lamina spiralis, can arise because traditional CT scans may not be sufficiently sensitive to identify certain issues .In the postoperative phase, the use of OTOPLAN allows surgeons to visualize the electrode insertion status from postoperative CT scans, enabling the application of an anatomy-based fitting map. This map, utilizing patient-specific data, aims to provide a fitting that closely aligns with natural tonotopic perception, considering critical parameters like the calculated tonotopic frequency of each electrode contact, determined by its angular location .In a comparative analysis of two insertion techniques using OTOPLAN, no differences were found in electrode array position or position in the scala tympani, regardless of whether insertion was through the round window or through anteroinferior cochleostomy in the transcanal Veria technique . AnotherInvestigations into the impact of existing frequency-to-place mismatch on speech perception in noise have indicated that a smaller mismatch correlates with improved speech perception in noise after 6 months. However, it has been acknowledged that additional research is needed to explore tonotopic fitting strategies based on postoperative CT images, revealing precise electrode contact positions ,61.Analyzing a postoperative CT using otological planning software allows for creating an anatomy-based frequency map, aiming to reallocate electrode array frequency and minimize mismatch. A comparison between anatomy-based and default frequency allocations revealed significant differences, seemingly related to cochlear coverage .A recent study investigated the impact of frequency reallocation based on an anatomical frequency map on speech perception and observed an improvement in speech perception, particularly in the parameters of Speech Recognition Threshold and Speech Awareness Threshold, following the reallocation .OTOPLAN software serves as a preoperative tool for surgeons, assisting in the selection of a more specific electrode array to optimize speech perception and facilitate postoperative anatomy-based fitting. A consensus regarding its utility in automatic cochlear measurements (CDL and frequency mapping) of A-, B-, and H-values has been noted, enabling a more personalized approach to cochlear implant (CI) surgery ,38.Leveraging automated tools and 3D visualization, the system enables quicker and more precise measurements compared to manual methods using the conventional DICOM viewer. This enhancement in measuring cochlear parameters and automatically calculating the CDL could potentially streamline clinical workflows and provide otosurgeons with greater autonomy. The precision of software-based cochlear reconstructions, even for malformed inner ears, has been substantiated, with no device-related complications reported in the existing literature .However, a few aspects of OTOPLAN deserve comment. Firstly, it necessitates that surgeons, audiologists, and radiologists become acquainted with the procedure, implicating a learning curve and potential variability in measurements due to differing training experiences . The latThe latest OTOPLAN version also introduces the capability to merge CT and MRI images, enhancing preoperative evaluation. Furthermore, integrating OTOPLAN with intraoperative electrophysiologic testing could potentially mitigate misplacement and establish correlations with postoperative acoustic hearing, presenting an additional benefit.A review of the existing literature underscores the utility of OTOPLAN for otologists and audiologists, enhancing preoperative surgical planning for both adults and children by providing detailed anatomical information, guiding intraoperative procedures, and facilitating postoperative evaluation of cochlear implantation. Notably, the system enables automatic CDL calculation, frequency mapping using A and B diameter measurements, and assistance in electrode selection. In postoperative stages, OTOPLAN provides a visualization of the electrode insertion status through postoperative CT scans, allowing audiologists to utilize an anatomy-based fitting map. This map uses patient-specific data to closely approximate natural tonotopic perception. However, additional research is warranted to refine the accuracy of CDL estimation, which is crucial for a thorough evaluation of pitch match and cochlear coverage in residual hearing.


Evidence for use of second-line immunosuppressants for immune-related adverse events (irAEs) is inadequate. Therefore, a multicenter analysis should assess the efficacy of second-line immunosuppressants for severe irAEs associated with different malignant diseases.This descriptive study aims to investigate the effects of second-line immunosuppressants on corticosteroid-refractory irAEs in patients with lung cancer. We analyzed the effects of second-line immunosuppressants on underlying lung cancer and associated adverse effects.p < 0.005). The median progression-free survival and duration of response of underlying lung cancer from second-line immunosuppressant administration were 2.1 and 3.0 months, respectively. Of the patients with irAE, 27.4% developed infections and 5.5% might die due to infection.Our study included 4589 patients who had received immune checkpoint inhibitor treatment, with 73 patients (1.6%) developing irAEs requiring second-line immunosuppressants. The most commonly observed irAE was pneumonitis (26 patients), followed by hepatobiliary disorders (15 patients) and enteritis (14 patients). We found a confirmed response rate of 42.3% for pneumonitis, which was lower than the response rates of 86.7% for hepatobiliary disorders and 92.9% for enteritis. The time from the start of corticosteroid therapy to the addition of a second-line immunosuppressant correlated significantly with the resolution of irAE to Grade 1 (correlation coefficients of r = 0.701, Second-line immunosuppressant response was confirmed in 72.2% of irAEs in patients with lung cancer, with lower response rates observed in irAE pneumonitis compared to other irAEs.The online version contains supplementary material available at 10.1007/s00262-023-03528-x. Immune checkpoint inhibitors (ICIs), including anti-PD-1/PD-L1 and anti-CTLA-4 antibodies, have become the standard of care for lung cancer. ICIs are used for systemic therapy in advanced disease , consoliFor moderate to severe irAEs, corticosteroids are typically used as a first-line treatment. In cases of severe irAEs not controlled by corticosteroids alone, the addition of second-line immunosuppressants, such as infliximab or mycophenolate mofetil, may be considered. These second-line immunosuppressants are expected to increase immunosuppression and resolve severe irAEs. However, evidence supporting the use of second-line immunosuppressants for irAE management has been insufficient despite clinical guideline recommendations , 9. MostIn this study, we conducted a multicenter retrospective analysis of second-line immunosuppressants for irAEs in patients with lung cancer. We analyzed the effect of second-line immunosuppressants and their adverse effect on the underlying disease.We conducted a multicenter retrospective analysis in the TOPGAN group, a Japanese lung cancer research group (TOPGAN 2022-01), with the primary objective of clarifying the effect of second-line immunosuppressants for corticosteroid-refractory irAEs. We defined steroid refractoriness as irAEs that were not controlled by steroids alone. In such cases, second-line immunosuppressants were also administered. Second-line immunosuppressants were also used to taper steroid after irAEs were under control. We collected data on patients with lung cancer who had received ICI treatment until July 2022 and had developed irAEs and received second-line immunosuppressants in addition to corticosteroids. Patient data collected included patient characteristics , type of irAE, effect of second-line immunosuppressants, and clinical course after second-line immunosuppressant administration. Second-line immunosuppressants analyzed in this study were azathioprine, cyclophosphamide, cyclosporine, infliximab, intravenous immunoglobulin, mycophenolate mofetil, tacrolimus, and tocilizumab. This study was approved by the institutional review board of the Cancer Institute Hospital of the Japanese Foundation for Cancer Research (IRB no. 2022-GB-065), and informed consent was waived because of the retrospective nature of the study, with an opt-out option included.Two outcomes were evaluated in this study: (1) Responses to second-line immunosuppressants determined by each investigator using patient medical records and (2) The resolution of irAEs to CTCAE grade 1 (G1) within 90 days of second-line immunosuppressant initiation. The clinical course of the underlying lung cancer was evaluated by measuring progression-free survival (PFS) and duration of response (DOR) from the initiation of second-line immunosuppressant administration. Tumor response was evaluated based on the RECIST v1.1 criteria. Incidence of infections following second-line immunosuppressant administration was also analyzed.p < 0.05. Data analysis was performed using R and SPSS Statistics for Windows version 24.Categorical variables were compared using Fisher’s exact test, and continuous data were compared using the Mann–Whitney U test. The strength of the relationship between two sets of data was determined using Spearman’s rank correlation coefficient. The Kaplan–Meier method was used to estimate the median survival time. Statistical significance was set at Overall, 73 of 4589 patients who received ICI treatment 1.6%) developed irAEs requiring second-line immunosuppressant administration. Of the 358 patients who received combination immunotherapy with nivolumab and ipilimumab, 17 (4.7%) developed irAEs requiring second-line immunosuppressants. The frequency of patients requiring second-line immunosuppressants varied by institution, ranging from 0 to 3.2% . The most common irAE was pneumonitis (n = 26), followed by hepatobiliary disorders (n = 15) and enteritis (n = 14). Infliximab was the most frequently used immunosuppressant (n = 19), followed by cyclophosphamide (n = 15) and mycophenolate mofetil (n = 14). Of the 72 cases evaluated, 52 patients (72.2%) responded to the second-line immunosuppressants. Response rates of second-line immunosuppressants in each irAE are shown in Fig. Among the patients with pneumonitis, 34.6% (9/26) had a history of thoracic irradiation . The median time between radiation completion and pneumonitis onset was 95  days. Responsiveness to second-line immunosuppressants was confirmed in 44.4% (4/9) of cases. This was not substantially different from the 41.2% (7/17) response rate seen in those who had not undergone irradiation. The types of pneumonitis and number of cases among our patients were organizing pneumonia (OP) in 11 cases, nonspecific interstitial pneumonia (NSIP) in five cases, hypersensitivity pneumonia (HP) in one case, diffuse alveolar damage (DAD) in eight cases, and OP plus DAD in one case. The response rates to second-line immunosuppressants for each type were OP, 63.6% (7/11); NSIP, 60.0% (3/5); HP, 100% (1/1); DAD, 0% (0/8); and OP + DAD, 0% (0/1).p = 0.148). In the 34 patients whose irAE resolved to G1 after second-line immunosuppressant addition, there was a significant correlation between the time from the start of corticosteroid administration to immunosuppressant addition and irAE resolution to G1 .  of second-line immunosuppressants in irAE pneumonitis in patients with lung cancer . Other rGuidelines for managing irAEs recommend considering the addition of a second-line immunosuppressant 3–4 days after starting corticosteroid therapy . A retroThe previous clinical trials have reported that patients with lung cancer who discontinued treatment due to severe irAEs tend to have a longer DOR than others . HoweverThe evidence supporting the recommendation of second-line immunosuppressants has been relatively insufficient, relying mainly on retrospective studies from single centers and expert opinions. As shown in this multicenter study, irAE management with second-line immunosuppressants is not always sufficient to resolve all irAEs, particularly pneumonitis, and adverse effects on underlying lung cancer are a concern. Therefore, it is crucial to accumulate further evidence on the use of second-line immunosuppressants in each malignant disease, including this study.There are some limitations to this study. First, as this is a case series, we were unable to compare second-line immunosuppressants with other treatments. Second, due to the retrospective nature of this study, some biases in treatment selection and effects could not be avoided despite conducting multicenter trials to minimize these biases.The response rate of second-line immunosuppressants was confirmed in 72.2% of irAEs in patients with lung cancer. However, the response rate was lower in irAE pneumonitis compared to other irAEs.Supplementary file1 (DOCX 62 KB)Below is the link to the electronic supplementary material.


TM; (i) the Modified Bouchard Activity Record; (j) the Rett Syndrome Behavioral Questionnaire; and (k) the Rett Syndrome Fear of Movement Scale. The authors recommend that service providers consider evaluation tools validated for RTT for evaluation and monitoring to guide their clinical recommendations and management. In this article, the authors suggest factors that should be considered when using these evaluation tools to assist in interpreting scores.Rett syndrome (RTT) is a complex neurodevelopmental X-linked disorder associated with severe functional impairments and multiple comorbidities. There is wide variation in the clinical presentation, and because of its unique characteristics, several evaluation tools of clinical severity, behavior, and functional motor abilities have been proposed specifically for it. This opinion paper aims to present up-to date evaluation tools which have specifically been adapted for individuals with RTT often used by the authors in their clinical and research practice and to provide the reader with essential considerations and suggestions regarding their use. Due to the rarity of Rett syndrome, we found it important to present these scales in order to improve and professionalize their clinical work. The current article will review the following evaluation tools: (a) the Rett Assessment Rating Scale; (b) the Rett Syndrome Gross Motor Scale; (c) the Rett Syndrome Functional Scale; (d) the Functional Mobility Scale—Rett Syndrome; (e) the Two-Minute Walking Test modified for Rett syndrome; (f) the Rett Syndrome Hand Function Scale; (g) the StepWatch Activity Monitor; (h) the activPAL MECP2 coding for methyl CpG-binding protein 2 [Rett syndrome (RTT) is a neurodevelopmental disorder occurring in one of every 10,000 live female births in the US [rotein 2 .Many aspects of the clinical condition are linked to a specific genetic alteration. For instance, mutations such as p.Arg270*, p.Arg255*, and p.Arg168* tend to be associated with more severe symptoms, while mutations such as p.Arg133Cys, p.Arg294*, and C-terminal deletions tend to be associated with milder symptoms [It is a necessity that formal and informal assessments are conducted regularly by caregivers, parents, teachers, and healthcare professionals to suggest and assess clinical care and day-to-day management. Both assessment types inform the design of intervention strategies that are most suited to the goals set for each individual. Approaches that are suitable for RTT need to be considered. For example, the movement skills of many individuals with RTT are affected by apraxia. Despite the existence of many evaluation tools for apraxia conditions , the uniIdentify the evaluation goal—The assessor should have a clear idea regarding the objective of the evaluation and should choose the most appropriate and related evaluation tool. For instance, an evaluation tool that provides a picture of the individual’s skills at a specific time could be adequate for monitoring a person’s improvement (or worsening) but could be insufficient for identifying emergent skills. When the goal of the evaluation is to recognize the impact of an intervention, the assessor should decide which tool to use after identifying the intervention program and goals.Familiar vs. unfamiliar situations—Individuals with RTT may not respond well to new situations and new people . TherefoTiming of assessment—Individuals with RTT have times during the day when they function better ,11. If pEvaluation organization—When the evaluation begins, it should proceed from the tasks that are easier for the girl to those more challenging that require more active effort. In this way, the girl will experience success and will then be more able to tackle complex tasks.Communication—The assessor should remember to talk to the girl throughout the evaluation meeting, modulating their voice to strengthen the relationship, observing carefully, and providing instructions and feedback. Communication should be supported by sufficient non-verbal elements  and augmentative communication tools usually used by the girl, is so far as possible, and caregivers.Ambiance—The assessor should make sure that there is a generally relaxed atmosphere before and during the assessment.Use natural situations—The person may show better abilities in certain natural situations compared to their abilities during an official examination. Therefore, whenever possible, the assessor should observe the person performing a task in situations in which they are used to doing it .Adjust the assessment according to the person’s need—Individuals with RTT may have variable functioning at different times of the day or on different days . TherefoAssess the musculoskeletal condition—The degree of muscular shortening, articular contractures, and muscular tone abnormalities may negatively affect the individual’s functional ability .Before evaluating a person with RTT, it is crucial to consider the following points:Clinical severity assessment: the Rett Assessment Rating Scale (RARS);Assessment of gross motor abilities: the Rett Syndrome Gross Motor Scale (RSGMS), the Rett Syndrome Functional Scale (ReFuS), the Functional Mobility Scale—Rett Syndrome (FMS-RS), and the Two-Minute Walking Test (2MWT) modified for RTT;Manual function assessment: the Rett Syndrome Hand Function Scale (RSHFS);Assessment of physical activity level: the Stepwatch Activity Monitor™ (SAM), the activPAL™, and the Modified Bouchard Activity Record (M-BAR);Behavioral assessment: Rett Syndrome Behavioral Questionnaire (RSBQ);Fear of movement assessment: the Rett Syndrome Fear of Movement Scale (RSFMS).With these considerations in mind, the current opinion paper presents the evaluation tools mostly used by the authors in their clinical and research practice with individuals with RTT and provides essential considerations and suggestions regarding their best use. The current paper’s authors believe that the information provided here could help to construct an evidence-informed short- and long-term evaluation and intervention plan when working with this population. All presented scales were developed or validated to be used with people with RTT and include tools for:Information related to each evaluation tool’s structure, function, and certain psychometric properties are reported in This questionnaire was developed to assess the severity of the disorder in individuals with RTT by parents or caregivers. The tool has been used in previous studies ,35,36,37The tool was adapted from the Gross Motor Function Measure ,48 to beThis scale was developed to detect the functional ability of girls with RTT before, during, or after an intensive motor intervention. The tool is based on independent performance only. The scale is extremely sensitive to small changes, so it is suitable when the intervention period is short. In carrying out an evaluation with this tool, the task execution times without any assistance are measured. Verbal encouragement is possible, and the presentation of favorable individuals (parents/care providers) and motivational factors are recommended (it is important to list and maintain the same conditions in each test) but avoiding physical contact is required. Each item’s score ranges from 0–12 points, so the score in the tool can range from 0 to 372 points. A higher score corresponds to better functional ability. Typical scores of young individuals with RTT who can walk ranged from 130–220 (before intensive intervention) and 200–320 (after intensive intervention) were found . The firThe Functional Mobility Scale (FMS) assesses the functional mobility of children with cerebral palsy and movement disorders ,53. It eThe 2MWT is a functional walking test developed as an alternative to the Six-Minute Walk Test. This test has traditionally been used for individuals who have experienced a stroke or have motor and neurological impairments ,56,57. RThe loss of manual function is typical for individuals with RTT during the initial period of the disorder, and the RSHFS is a valuable tool to score the person’s level of hand functioning. The RSHFS protocol was developed based on the Hand Apraxia Scale, a 10-item measure of manual functioning . The hanStep counters and pedometers are objective, inexpensive, valid, reliable, and relatively easy-to-use measures of physical activity commonly used within the normative population . MoreoveThe activPAL™ device uses a uniaxial accelerometer and inclinometer to determine step count and the time spent in lying down/sitting as well as standing and walking (uptime) positions. The device is attached to the person’s thigh using adhesive pads and provides data measuring the thigh inclination and acceleration . In a rep < 0.001) and less time standing (1 h vs. 2 h 15 min—p = 0.04) than those who walked independently. The M-BAR was validated with 43 individuals with RTT  to evaluate the “uptime” in this population [The BAR is a whole-day diary card in which every 15 min of the day a number can be assigned between one and nine to represent either sedentary behavior or a particular level of physical activity intensity . When vapulation . Moreovepulation  demonstrpulation .Some behavioral features have been associated with RTT ,76. AlthOne of the behavioral characteristics reported in girls with RTT is fear of movement (FOM) . The conThe current opinion paper is focused on evaluation tools dedicated to motor function due to the authors’ experience in this field. Although the existing evaluation tools allow for the assessment of several aspects of RTT, a few of these remain unexplored to their full extent. For instance, objectively scoring this population’s reaching ability is needed. As discussed above, every evaluation tool presents advantages and disadvantages and could be adequate for one specific aim but not another. Therefore, future research projects should focus on completing validation processes for each scale mentioned above. Furthermore, there are physiological measures that have been used in the general population, such as heart rate and respiration measurements, using technological gadgets. The possibility of using such devices for individuals with RTT has been questioned in previous research projects . The valAs individuals with RTT present functional diversity in different areas, validated tools are recommended to assess their abilities as a group as well as before, during, and after treatments. The current article has presented evaluation tools built, modified, and validated specifically for individuals with RTT. These evaluation tools stand alone or can be used together to enable a comprehensive and appropriate assessment of an individual’s function according to a specific research protocol or an intervention program. The proposed evaluation tools cover the evaluation of RTT severity level (RARS), level of gross motor skills (RSGMS), independently performable motor skills (ReFuS), global mobility (FMS and 2MWT), purposeful manual functioning (RSHFS), physical activity , and fear of movement (RSFMS). The authors hope that by presenting the evaluation tools described above, better therapeutic assessments will be implemented by clinicians working with individuals with RTT, enabling finer therapeutic goal setting and better outcome evaluation during clinical interventions and research procedures. Moreover, the authors suggested options for the future development of evaluation tools for RTT and some futuristic research directions.