Protective role of pyrroloquinoline quinone against gentamicin induced cochlear hair cell ototoxicity.

Gentamicin (GM) is one of the commonly used antibiotics in the aminoglycoside class but is ototoxic, which constantly impacts the quality of human life. Pyrroloquinoline quinone (PQQ) as a redox cofactor produced by bacteria was found in soil and foods that exert an antioxidant and redox modulator. It is well documented that the PQQ can alleviate inflammatory responses and cytotoxicity. However, our understanding of PQQ in ototoxicity remains unclear. We reported that PQQ could protect against GM-induced ototoxicity in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells in vitro. To evaluate reactive oxygen species (ROS) production and mitochondrial function, ROS and JC-1 staining, oxygen consumption rate (OCR), and extracellular acidification rate (ECAR) measurements in living cells, mitochondrial dynamics analysis was performed. GM-mediated damage was performed by reducing the production of ROS and inhibiting mitochondria biogenesis and dynamics. PQQ ameliorated the cellular oxidative stress and recovered mitochondrial membrane potential, facilitating the recovery of mitochondrial biogenesis and dynamics. Our in vitro findings improve our understanding of the GM-induced ototoxicity with therapeutic implications for PQQ.

Pan-cancer analysis and experimental validation of DTL as a potential diagnosis, prognosis and immunotherapy biomarker.

BACKGROUND: DTL has been found to be related with multiple cancers. However, comprehensive analyses, which identify the prediction value of DTL in diagnosis, prognosis, immune infiltration and treatment, have rarely been reported so far. METHODS: Combined with the data online databases, the gene expression, gene mutation, function enrichment and the correlations with the immunity status and clinical indexes of DTL were analyzed. Expression of DTL and the degree of immune cell infiltration were examined by immunofluorescence (IF) and immunohistochemistry (IHC) and analyzed by statistical analysis. Furthermore, the influences of DTL on the cell cycle, cell proliferation and apoptosis were detected by live cell imaging, IF and flow cytometric (FC) analysis. Genomic stability assays were conducted by chromosome slide preparation. RESULTS: DTL was widely expressed in various cells and tissues, while it was overexpressed in tumor tissues except acute myeloid leukemia (LAML). Pan-cancer bioinformatics analysis showed that the expression of DTL was correlated with the prognosis, immunotherapy, and clinical indexes in various cancers. In addition, gene set enrichment analysis (GSEA) uncovered that DTL was enriched in oocyte meiosis, pyrimidine metabolism, the cell cycle, the G2M checkpoint, mTORC1 signaling and E2F targets. Furthermore, the overexpression of DTL, and its association with immune cell infiltration and clinical indexes in liver hepatocellular carcinoma (LIHC), bladder urothelial carcinoma (BLCA) and stomach adenocarcinoma (STAD) were verified in our study. It was also verified that overexpression of DTL could regulate the cell cycle, promote cell proliferation and cause genomic instability in cultured cells, which may be the reason why DTL plays a role in the occurrence, progression and treatment of cancer. CONCLUSIONS: Collectively, this study suggested that DTL is of clinical value in the diagnosis, prognosis and treatment of various cancers, and may be a potential biomarker in certain cancers.

Gene association detection via local linear regression method.

The development of next-generation sequencing technology has provided us with great convenience in genetic association studies and many effective analysis methods were proposed continuously. However, population stratification is still a major issue in current genetic association studies. Many existing methods have been developed to remove the bias due to population stratification for common variant association studies, but such methods may be not effective for rare variant, which will lead to power reduction. Therefore, in this paper, we develop a principal component analysis strategy (called PC-LLR) based on local linear regression method to eliminate population stratification effect in both rare variant and common variant association studies. Simulation results indicate that the new PC-LLR method can eliminate population stratification effect well. It has correct type I error rates in all cases and higher powers in most cases, while most existing methods have inflated type I error rates at least in some cases. We also demonstrate that the PC-LLR is more effective to eliminate population stratification effect through applying the PC-LLR to the whole-exome sequencing data set from genetic analysis workshop 19 (GAW19).

Application of flexible fiberoptic bronchoscopy in the removal of adult airway foreign bodies.

BACKGROUND: Flexible fiberoptic bronchoscopy is a rapid, cost effective and safe procedure. AIM: To analyze demographic information and endoscopic findings in adult patients with airway foreign body aspiration and its removal. METHODS: Fifty-seven adults (40 males, 17 females; average age 40 years old) with airway foreign body aspiration were analyzed. Cough (37, 65%) was the most common clinical presentation. The most common foreign body was bone followed by dental prosthesis and food debris. RESULTS: In the current study, 42 out of the 57 (74%) airway foreign bodies were successfully removed under flexible fiberoptic bronchoscopy. However, it was failed in 15 patients and thus, rigid bronchoscopy was used to remove foreign bodies successfully in 13 of the 15 patients. Thoracotomy was performed for the 2 patients whose foreign body removal was unsuccessful even with rigid bronchoscopy. CONCLUSION: The findings of the current study revealed that flexible fiberoptic bronchoscopy is a safe and effective procedure for the removal of adult airway foreign bodies in the majority of cases. Rigid bronchoscopy can be a backup procedure in case flexible bronchoscopy is failed.

A permutation method for detecting trend correlations in rare variant association studies.

In recent years, there has been an increasing interest in detecting disease-related rare variants in sequencing studies. Numerous studies have shown that common variants can only explain a small proportion of the phenotypic variance for complex diseases. More and more evidence suggests that some of this missing heritability can be explained by rare variants. Considering the importance of rare variants, researchers have proposed a considerable number of methods for identifying the rare variants associated with complex diseases. Extensive research has been carried out on testing the association between rare variants and dichotomous, continuous or ordinal traits. So far, however, there has been little discussion about the case in which both genotypes and phenotypes are ordinal variables. This paper introduces a method based on the γ-statistic, called OV-RV, for examining disease-related rare variants when both genotypes and phenotypes are ordinal. At present, little is known about the asymptotic distribution of the γ-statistic when conducting association analyses for rare variants. One advantage of OV-RV is that it provides a robust estimation of the distribution of the γ-statistic by employing the permutation approach proposed by Fisher. We also perform extensive simulations to investigate the numerical performance of OV-RV under various model settings. The simulation results reveal that OV-RV is valid and efficient; namely, it controls the type I error approximately at the pre-specified significance level and achieves greater power at the same significance level. We also apply OV-RV for rare variant association studies of diastolic blood pressure.

Association detection between ordinal trait and rare variants based on adaptive combination of P values.

Next-generation sequencing technology not only presents a new method for the detection of human genomic structural variation, but also provides a large number of genetic data of rare variants for us. Currently, how to detect association between human complex diseases and rare variants using genetical data has attracted extensive attention. In the field of medicine, many people's health and disease conditions are measured by ordinal response variables, namely, the trait value reflects the development stage or severity of a certain condition. However, most existing methods to test for association between rare variants and complex diseases are designed to deal with dichotomous or quantitative traits. Association analysis methods of ordinal traits are relatively fewer, and considering ordinal traits as dichotomous and quantitative traits will inevitably lose some valuable information in the original data. Therefore, in this paper, we extend an existing method of adaptive combination of P values (ADA) and propose a new method of association analysis for ordinal trait based on it (called OR-ADA) to test for possible association between ordinal trait and rare variants. In our method, we establish a cumulative logistic regression model, in which the regression coefficients are estimated by the Newton-Raphson algorithm and the likelihood ratio test is used to test the association. Through a large number of simulation studies and an example, we demonstrate the performance of the new method and compare it with several methods. The analysis results show that the OR-ADA strategy is robust to the signs of effects of causal variants and more powerful under many scenarios.

Protective Effects of Acupuncture Against Gentamicin-Induced Ototoxicity in Rats: Possible Role of Neurotrophin-3.

BACKGROUND The aim of this study was to investigate the protective effects of acupuncture against gentamicin-induced ototoxicity and explore the possible protective role of neurotrophin-3 (NT-3). MATERIAL AND METHODS Twenty-four rats were divided randomly into 4 groups: control group, gentamicin group, neitinggong group, and tinggong group. Rats in the gentamicin, neitinggong, and tinggong groups received intraperitoneal injection of gentamicin (100 mg/kg) for 14 consecutive days. Rats in the neitinggong and tinggong groups further received acupuncture at neitinggong or tinggong acupoints once every 2 days for 20 days. Rats in the control group received intraperitoneal injection of saline. Auditory brainstem response (ABR) was tested in all rats on the day before treatment (day 0), and again on day 14 and day 20 to determine the average threshold value of ABR for each treatment group. The expression of NT-3 in the cochlear nucleus and the inferior colliculus nucleus were detected by immunohistochemical staining. RESULTS The average threshold value of ABR was significantly higher in the gentamicin group as compared with that of the control group on day 14 (P<0.05). On day 20, the average threshold values of ABR in the neitinggong and tinggong groups were significantly lower than that of the gentamicin group (P<0.05). No statistically significant differences in NT-3 expression in the cochlear nucleus were observed among the groups (P>0.05). However, the expression of NT-3 in the inferior colliculus nucleus in both the neitinggong and tinggong groups was significantly higher than that of the gentamicin group (P<0.01). CONCLUSIONS A decrease in NT-3 expression in the inferior colliculus nucleus may contribute to gentamicin-induced ototoxicity in rats. Acupuncture at neitinggong or tinggong acupoints effectively improved hearing, which was attributed partially to the rescue of NT-3 expression in the inferior colliculus nucleus. Therefore, preserving NT-3 expression in the auditory system may be a viable strategy to counteract gentamicin-induced ototoxicity.

Genome-wide interaction analysis of quantitative traits in outbred mice.

With a large number of quantitative trait loci being identified in genome-wide association studies, researchers have become more interested in detecting interactions among genes or single nucleotide polymorphisms (SNPs). In this research, we carried out a two-stage model selection procedure to detect interacting gene pairs or SNP pairs associated with four important traits of outbred mice, including glucose, high-density lipoprotein cholesterol, diastolic blood pressure and triglyceride. In the first stage, a variance heterogeneity test was used to screen for candidate SNPs. In the second stage, the Lasso method and single pair analysis were used to select two-way interactions. Moreover, the shared Gene Ontology information about the selected interacting gene pairs was considered to study the interactions auxiliarily. Based on this method, we not only replicated the identification of important SNPs associated with each trait of outbred mice, but also found some SNP pairs and gene pairs with significant interaction effects on each trait. Simulation studies were also conducted to evaluate the performance of the two-stage method in different situations.

Nonconvex Robust High-Order Tensor Completion Using Randomized Low-Rank Approximation.

Within the tensor singular value decomposition (T-SVD) framework, existing robust low-rank tensor completion approaches have made great achievements in various areas of science and engineering. Nevertheless, these methods involve the T-SVD based low-rank approximation, which suffers from high computational costs when dealing with large-scale tensor data. Moreover, most of them are only applicable to third-order tensors. Against these issues, in this article, two efficient low-rank tensor approximation approaches fusing random projection techniques are first devised under the order-d ( d ≥ 3 ) T-SVD framework. Theoretical results on error bounds for the proposed randomized algorithms are provided. On this basis, we then further investigate the robust high-order tensor completion problem, in which a double nonconvex model along with its corresponding fast optimization algorithms with convergence guarantees are developed. Experimental results on large-scale synthetic and real tensor data illustrate that the proposed method outperforms other state-of-the-art approaches in terms of both computational efficiency and estimated precision.

Investigating the Effects of Perampanel on Autophagy-mediated Regulation of GluA2 and PSD95 in Epilepsy.

Epilepsy is a chronic neurological disorder characterized by recurrent seizures. Despite various treatment approaches, a significant number of patients continue to experience uncontrolled seizures, leading to refractory epilepsy. The emergence of novel anti-epileptic drugs, such as perampanel (PER), has provided promising options for effective epilepsy treatment. However, the specific mechanisms underlying the therapeutic effects of PER remain unclear. This study aimed to investigate the intrinsic molecular regulatory mechanisms involved in the downregulation of GluA2, a key subunit of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors, following epileptic seizures. Primary mouse hippocampal neurons were cultured and subjected to an epilepsy cell model. The expression levels of GluA2 and autophagy-related proteins were assessed using Western blotting and real-time fluorescent quantitative PCR. Immunofluorescence and immunohistochemistry techniques were employed to investigate the nuclear translocation of CREB-regulated transcriptional coactivator 1 (CRTC1). Additionally, status epilepticus animal models were established to further validate the findings. The epilepsy cell model exhibited a significant decrease in GluA2 expression, accompanied by elevated levels of autophagy-related proteins. Immunofluorescence analysis revealed the nuclear translocation of CRTC1, which correlated with the expression of autophagy-related genes. Treatment with an autophagy inhibitor reversed the decreased expression of GluA2 in the epilepsy cell model. Furthermore, the calcium/calmodulin-dependent protein phosphatase inhibitor FK506 and CaN overexpression affected the dephosphorylation and nuclear translocation of CRTC1, consequently influencing GluA2 expression. Animal model results further supported the involvement of these molecular mechanisms in epilepsy. Our findings suggest that the downregulation of GluA2 following epileptic seizures involves the activation of autophagy and the regulation of CRTC1 nuclear translocation. These intrinsic molecular regulatory mechanisms provide potential targets for developing novel therapeutic strategies to alleviate refractory epilepsy and preserve cognitive functions in patients.

Corrigendum: Role of a novel mouse mutant of the Galnt2tm1Lat/tm1Lat gene in otitis media.

[This corrects the article DOI: 10.3389/fneur.2022.1054704.].

Effectiveness of galvanic vestibular evoked myogenic potential for evaluation of Meniere's disease.

OBJECTIVES: Although the cause of Meniere's disease (MD) is not fully understood, endolymphatic hydrops is widely believed to be responsible for MD. Previous studies have used Air-Conducted Sound (ACS)-induced Vestibular Evoked Myogenic Potentials (VEMPs) to evaluate otolithic function in patients with MD. However, the use of Galvanic Vestibular Stimulation-VEMPs (GVS-VEMPs) with other vestibular tests in MD has been rare. This study aimed to explore the application of galvanic VEMPs in assessing MD. METHODS: Normal individuals and patients with unilateral definite MD were included in this retrospective study. All participants underwent pure tone audiometry. Ocular and cervical VEMPs induced by GVS, and ACS were recorded. The characteristic parameters of VEMPs (n1 latency, p1 latency, amplitude, and AR) were analyzed. RESULTS: The provocation rates of GVS-VEMPs did not differ between MD patients and control individuals. Compared with ACS, GVS could evoke potentials with longer latencies. MD patients presented GVS-VEMPs with lower amplitudes and ACS-cVEMP with shorter latencies and had a higher response rate in GVS-oVEMP. However, no differences or correlations were found in the characteristic parameters of GVS-VEMPs among the different stages of MD. CONCLUSIONS: GVS is as effective as ACS for inducing VEMP, and GVS-VEMP recording can detect retrolabyrinthine degeneration in MD. Further research is needed to assess the utility of GVS-VEMP in the evaluation of MD severity. LEVEL OF EVIDENCE: Level 4.

The advantages of vestibular-evoked myogenic potentials induced by bone-conducted vibration in patients with otitis media.

BACKGROUND: At present, there is no consensus on the optimal methods for the diagnosis of vestibular dysfunction. OBJECTIVE: To explore the advantages of bone-conducted vibration (BCV) related vestibular-evoked myogenic potentials (VEMPs). MATERIAL AND METHODS: Fifty patients with otitis media volunteered for VEMP examination. They were randomly selected with air-conducted sound and BCV stimulation VEMPs assessed in both ears. RESULTS: The provocation rate of BCV-VEMPs was significantly higher than that of air-conducted sound VEMPs. Among 50 affected ears, there was no significant difference in the provocation rate of BCV-VEMPs between patients with air-bone conduction gaps lower or higher than 20 dB. There was no significant difference in the provocation rate of BCV-VEMPs between both ears in 30 patients with unilateral otitis media and the comparison of BCV-oVEMP parameters made no significant difference in amplitude, N1 latency, P1 latency, or N1-P1 duration, except for the threshold. A comparison of BCV-cVEMP parameters between affected and healthy ears revealed no significant difference between groups in terms of threshold, amplitude, N1 latency, P1 latency, or n1-p1 duration. CONCLUSIONS AND SIGNIFICANCE: BCV-VEMPs may be stably induced in patients with conductive hearing loss.

Effect of age on virtual reality-assisted subjective visual vertical and subjective visual horizontal at different head-tilt angles.

OBJECTIVE: Subjective Visual Vertical (SVV) and Subjective Visual Horizontal (SVH) values may reflect bilateral utricle asymmetry. Bilateral utricle static tension balance can be used to evaluate bilateral otolith lesions and otolith-related central neuropathy. Few studies have examined Virtual Reality (VR)-assisted SVV and SVH values at various head-tilt angles across age groups. The present study aimed to determine the effects of age on VR-assisted SVV and SVH values at different head-tilt angles. METHODS: We divided 180 healthy subjects into 6 age groups (n = 30 in each group). VR-assisted SVV and SVH measurements were performed at 9 head-tilt angles (head held vertically, 0°; head tilted 30°, 45°, 60°, and 90° to the left/right) in the roll plane. RESULTS: SVV and SVH values significantly differed with head-tilt angle (p < 0.05). No significant difference was detected in the SVH and SVV values between different age groups (p = 0.632 and p = 0.810, respectively), and no interaction between the age group and the head-tilt angle was found for the SVH and SVV values (p = 0.670 and p = 0.084, respectively). CONCLUSION: These results suggest that age may have little effect on VR-assisted SVV and SVH at different head-tilt angles. Therefore, VR-assisted SVV and SVH can be evaluated as an effective, fast, and simple way to evaluate utricle function. LEVEL OF EVIDENCE: Level 4.

Outcome analysis of immediate and delayed laparoscopic pyeloplasty in infants with severe ureteropelvic junction obstruction.

Objective: The treatment timing of ureteropelvic junction obstruction (UPJO) in infants remains controversial. This study aimed to compare the recovery effect of renal morphology of immediate and delayed laparoscopic pyeloplasty in infants with severe UPJO. Methods: The infants with severe UPJO-induced hydronephrosis who underwent laparoscopic pyeloplasty according to their age at the time of surgery [the immediate treatment (IT) group: ≤1 month of birth, the delayed treatment (LT) group: 3-6 months of birth] in our center between 2010 and 2019 were enrolled in this study. Ultrasonography was used to assess renal morphology, including anteroposterior diameter (APD) of a pelvic, parenchymal thickness (PT), polar length (PL), and Society of Fetal Urology (SFU) grade. Preoperative and postoperative renal morphological outcomes at 6, 12, and 24 months were measured and compared. Results: During this period, a total of 135 patients were assigned to receive either IT (n = 73) or LT (n = 62) and were included for analysis. There were no significant differences in renal morphology indices at baseline between groups of IT and LT. The APD, PT, and PL in both groups all recovered to certain degrees compared with those at baseline, however, the IT group recovered more significantly than the LT group. Despite there being no significant difference in SFU grade between the two groups before and after surgery, the reduction of SFU grade in the IT group was more significant than that in the LT group during the 6-, 12- and 24-month follow-up periods. The PL, SFU, and APD were greater in the IT group than in the LT group at 6, 12, and 24 months of follow-up. At 6 months PL was not significantly higher between the two groups, while the outcome was significantly different at 12 months and 24 months. Conclusion: Immediate laparoscopic pyeloplasty for the infant with severe ureteropelvic junction obstruction is effective, and it can accelerate the recovery of renal morphological indices in infants with severe UPJO-induced hydronephrosis.

Role of a novel mouse mutant of the Galnt2 tm1Lat/tm1Lat gene in otitis media.

Genetic susceptibility is one of the most important causes of otitis media (OM). Mutant Galnt2 homozygote (Galnt2 tm1Lat/tm1Lat) mimics human otitis media in comparable pathology and causes hearing loss. Otitis media is characterized by effusion and dysregulated mucosa proliferation and capillary expansion in the middle ear cavity, which is associated with hearing loss. The mucociliary dysfunction could be seen in the middle ear cavity (MEC) in a patient harboring the disease that develops in severity with age by a scanning electron microscope. Tumor necrosis factor alpha (TNF-α), transforming growth factor-beta 1 (TGF-ß1), Muc5ac, and Muc5b upregulate the expression in the middle ear, which correlates with inflammation, craniofacial development, and mucin secretion. The mouse model with a mutation in the Galnt2 (Galnt2 tm1Lat/tm1Lat) was explored in this study as a novel model of human otitis media.

Large-Scale Transient Production in ExpiCHO-S™ with Enhanced N-Galactosylation-Sialylation and PEI-Based Transfection.

Large-scale transient expression in Chinese Hamster Ovary (CHO) cells provides a rapid protein production method with a potential start-to-end alignment advantage for biotherapeutics drug discovery. In this chapter, experimental protocols are illustrated for transient expression of therapeutic glycoproteins with improved galactosylation and sialylation in ExpiCHO-S™ system. To reduce the production cost, we also describe a novel procedure for PEI-mediated transfection in ExpiCHO-S™ cells that supports therapeutic protein expression comparable to the level with ExpiFectamine™-based transfection.

A two-step method for estimating QTL effects and positions in multi-marker analysis.

Quantitative trait is always controlled by multiple latent genetic loci, and genetic markers have been used to map quantitative trait loci (QTLs) auxiliarily. The method of multiple interval mapping (MIM) provides an appropriate way for mapping QTL using genetic makers. However, the computation in the MIM seems infeasible for a large number of marker intervals. Nowadays, the Dantzig selector (DS) method proves to be a more efficient method to estimate model effects in a linear model when the number of parameters is (much) larger than the sample size, which has not been applied to genetic mapping for QTL. In this paper, we developed a two-step method for mapping QTL based on the MIM, and we also illustrate the feasibility of adopting the DS to estimate marker or QTL effects. Simulation results showed that the proposed method performed satisfactorily well by comparisons with the existing MIM method, and the analysis to real data set also tested the practicability and efficiency of the DS method in genetic mapping.

Differential regulation of mRNA stability controls the transient expression of genes encoding Drosophila antimicrobial peptide with distinct immune response characteristics.

The tight regulation of transiently expressed antimicrobial peptides (AMPs) with a distinct antimicrobial spectrum and different expression kinetics contributes greatly to the properly regulated immune response for resistance to pathogens and for the maintenance of mutualistic microbiota in Drosophila. The important role of differential regulation of AMP expression at the posttranscriptional level needs to be elucidated. It was observed that the highly expressed Cecropin A1 (CecA1) mRNA encoding a broad antimicrobial spectrum AMP against both bacteria and fungi decayed more quickly than did the moderately expressed Diptericin mRNA encoding AMP against Gram negative bacteria. The mRNA stability of AMPs is differentially regulated and is attributed to the specific interaction between cis-acting ARE in 3'-UTR of AMP mRNA and the RNA destabilizing protein transactor Tis11 as shown in co-immunoprecipitation of the Tis11 RNP complex with CecA1 mRNA but not other AMP mRNA. The p38MAPK was further demonstrated to play a crucial role in stabilizing ARE-bearing mRNAs by inhibiting Tis11-mediated degradation in LPS induced AMP expression. This evidence suggests an evolutionarily conserved and functionally important molecular basis for and effective approach to exact control of AMP gene expression. These mechanisms thereby orchestrate a well balanced and dynamic antimicrobial spectrum of innate immunity to resist infection and maintain resident microbiota properly.

A convention-radiomics CT nomogram for differentiating fat-poor angiomyolipoma from clear cell renal cell carcinoma.

This study aimed to construct convention-radiomics CT nomogram containing conventional CT characteristics and radiomics signature for distinguishing fat-poor angiomyolipoma (fp-AML) from clear-cell renal cell carcinoma (ccRCC). 29 fp-AML and 110 ccRCC patients were enrolled and underwent CT examinations in this study. The radiomics-only logistic model was constructed with selected radiomics features by the analysis of variance (ANOVA)/Mann-Whitney (MW), correlation analysis, and Least Absolute Shrinkage and Selection Operator (LASSO), and the radiomics score (rad-score) was computed. The convention-radiomics logistic model based on independent conventional CT risk factors and rad-score was constructed for differentiating. Then the relevant nomogram was developed. Receiver operation characteristic (ROC) curves were calculated to quantify the accuracy for distinguishing. The rad-score of ccRCC was smaller than that of fp-AML. The convention-radioimics logistic model was constructed containing variables of enhancement pattern, VUP, and rad-score. To the entire cohort, the area under the curve (AUC) of convention-radiomics model (0.968 [95% CI 0.923-0.990]) was higher than that of radiomics-only model (0.958 [95% CI 0.910-0.985]). Our study indicated that convention-radiomics CT nomogram including conventional CT risk factors and radiomics signature exhibited better performance in distinguishing fp-AML from ccRCC.