Targeting Pro-Survival Autophagy Enhanced GSK-3ß Inhibition-Induced Apoptosis and Retarded Proliferation in Bladder Cancer Cells.

Advanced bladder cancer (BC) (local invasive and/or metastatic) is not curable even with cytotoxic chemotherapy, immune checkpoint inhibitors, and targeted treatment. Targeting GSK-3ß is a promising novel approach in advanced BC. The induction of autophagy is a mechanism of secondary resistance to various anticancer treatments. Our objectives are to investigate the synergistic effects of GSK-3ß in combination with autophagy inhibitors to evade GSK-3ß drug resistance. Small molecule GSK-3ß inhibitors and GSK-3ß knockdown using siRNA promote the expression of autophagy-related proteins. We further investigated that GSK-3ß inhibition induced the nucleus translocation of transcription factor EB (TFEB). Compared to the GSK-3ß inhibition alone, its combination with chloroquine (an autophagy inhibitor) significantly reduced BC cell growth. These results suggest that targeting autophagy potentiates GSK-3ß inhibition-induced apoptosis and retarded proliferation in BC cells.

Tumor-infiltrating immune cell status predicts successful response to immune checkpoint inhibitors in renal cell carcinoma.

Immune checkpoint inhibitors (ICI) have dramatically changed the treatment of metastatic renal cell carcinoma (mRCC). Although many studies have reported biomarkers as predicting the efficacy of ICI in mRCC, they remain controversial and have challenges to apply in real-world practice. We evaluated prognostic significance of multiple molecules associated with tumor immunity in patients treated with ICI. The molecules were detected in tumor tissues by immunohistochemical staining. We identified CD8-positive T cells and CD68-positive macrophages infiltrating into the tumor tissue as significant favorable prognostic factors for ICI treatment. Conversely, high expression of CD4-positive T cells was associated with poor response to ICI. Furthermore, we demonstrated that scoring for the expression status of these three molecules provides a remarkably accurate biomarker in patients with mRCC. Even the classical approach of immunohistochemistry could predict the outcome of ICI treatment by assessing the combined status of tumor-infiltrating immune cells.

Advanced Bladder Cancer: Changing the Treatment Landscape.

Bladder cancer is the 10th most common cancer type in the world. There were more than 573,000 new cases of bladder cancer in 2020. It is the 13th most common cause of cancer death with an estimated more than 212,000 deaths worldwide. Low-grade non-muscle-invasive bladder cancer (NMIBC) is usually successfully managed with transurethral resection (TUR) and overall survival for NMIBC reaches 90% according to some reports. However, long-term survival for muscle-invasive bladder cancer (MIBC) and metastatic bladder cancer remains low. Treatment options for bladder cancer have undergone a rapid change in recent years. Immune checkpoint inhibitors (ICI), targeted therapies, and antibody-drug conjugates are available now. As bladder cancer is genetically heterogeneous, the optimization of patient selection to identify those most likely to benefit from a specific therapy is an urgent issue in the treatment of patients with bladder cancer.

Development of patient­derived tumor organoids and a drug testing model for renal cell carcinoma.

The selection of effective therapeutic agents is critical for improving the survival of patients with renal cell carcinoma (RCC). The aim of the present study was to develop an ex vivo drug testing assay using patient­derived tumor organoid (TO) cultures. For this purpose, surgical tumor specimens were obtained from 20 patients with RCC. TOs were developed ex vivo from freshly resected RCC tumors, and their histopathological and molecular characteristics were evaluated using histological staining and whole­exome sequencing (WES). Using a cell viability assay, the therapeutic efficacy of standard of care tyrosine kinase inhibitors in RCC TOs was determined. It was found that TOs recapitulated the histological features of primary RCC tumors. Using WES, a strong concordance was identified at the genetic level between the primary tumors and their corresponding TOs. Using patient­derived TO models, a prototype of an ex vivo drug testing assay was developed, and it was found that RCC TOs exhibited differential responses to sunitinib, pazopanib, cabozantinib, axitinib and sorafenib treatment. On the whole, although the predictive value of the current assay has to be tested and validated in future clinical studies, the findings of the present study demonstrate a novel approach for ex vivo drug testing in patient­derived TO models, which may have potential for use in the personalized treatment of cancer patients.

Histone deacetylase 6 inhibition in urothelial cancer as a potential new strategy for cancer treatment.

Histone deacetylases (HDACs) are enzymes that remove acetyl groups from histones and have attracted attention as potential targets for cancer therapy. Several small molecule inhibitors have been developed to target HDACs; however, clinical trials of pan-HDAC inhibitors have found these types of inhibitors to be inefficient and to be relatively highly toxic. In the present study, the role of one HDAC isozyme, HDAC6, in urothelial cancer was investigated. Protein expression levels and subcellular localization of HDAC6 was identified in surgically resected bladder tumors using immunohistochemistry. The antitumor effects of 12 small molecule HDAC6 inhibitors were also examined in vitro using cultured urothelial cancer cells. The HDAC6 inhibitors decreased cell viability, with IC50 values in the low µM range, as low as 2.20 µM. HDACi D, E and F had the lowest IC50 values. HDAC6 has been previously reported to regulate programmed death-ligand 1 (PD-L1) and PD-L1 expression was found to be a predictor of decreased overall survival time. There was no association between the protein expression level of HDAC6 and PD-L1 in tumor tissues; however, HDAC6 inhibition by specific small molecule inhibitors resulted in decreased expression levels of membranous PD-L1 in cultured urothelial cancer cell lines. The results suggested that inhibition of HDAC6 could be a promising novel approach for the treatment of urothelial cancer.

Virilization of a female infant genitalia caused by a maternal androgen-producing adrenocortical tumor: A case report.

A 4-month-old girl presented us with genital ambiguity. The patient had a persistent urogenital sinus, posterior labial fusion with clitoromegaly. MRI reveals ovary-like mass in left inguinal region and right abdominal cavity. Uterus and vagina was also identified. Her mother was diagnosed with a right androgen-producing adrenocortical tumor 14 months after the birth of a virilized infant.

Clinically relevant GSK­3ß inhibitor 9­ING­41 is active as a single agent and in combination with other antitumor therapies in human renal cancer.

Glycogen synthase kinase­3 (GSK­3), a serine/threonine kinase, is involved in a broad range of pathological processes including cancer. GSK­3 has two isoforms, GSK­3α and GSK­3ß, and GSK­3ß has been recognized as a therapeutic target for the development of new anticancer drugs. The present study aimed to investigate the antitumor effects of 9­ING­41, which is a maleimide­based ATP­competitive small molecule GSK­3ß inhibitor active in patients with advanced cancer. In renal cancer cell lines, treatment with 9­ING­41 alone induced cell cycle arrest and apoptosis, and autophagy inhibitors increased the antitumor effects of 9­ING­41 when used in combination. Treatment with 9­ING­41 potentiated the antitumor effects of targeted therapeutics and increased the cytotoxic effects of cytokine­activated immune cells on renal cancer cell lines. These results provided a compelling rationale for the inclusion of patients with renal cancer in studies of 9­ING­41, both as a single agent and in combination with current standard therapies.

Free Tube Graft Urethroplasty for Repair of Hypospadias.

INTRODUCTION: We aimed to assess the outcome of free tube graft urethroplasty for single-stage repair of hypospadias with chordee in children. MATERIALS AND METHODS: We retrospectively evaluated a series of 56 patients (16 months to 9 years old, median 24 months) who underwent free graft urethroplasty for repair of hypospadias with chordee between May 2005 and November 2017. The median follow-up was 7 years (range 1-11). RESULTS: After releasing the chordee, the hypospadiac orifice was retracted to become penile in 32 patients (57%), penoscrotal in 18 patients (32%), and scrotal in 6 patients (11%). Single-stage repair was achieved without complications in 42 patients (75%). Of the remaining 14 patients with postoperative complications requiring surgical intervention, 2 had meatal stenosis, 9 had urethrocutaneous fistula, 1 had urethral diverticulum without meatal stenosis, and 1 had meatal regression. One patient who complained the urine stream went upwards in an arc underwent cutback meatoplasty to correct the stream. In all patients, a neomeatus with a vertically oriented slit-like appearance was eventually achieved at the tip of the glans. CONCLUSION: A free graft is an appropriate choice for repairing hypospadias with chordee. Our procedure achieved favorable functional and cosmetic outcomes with a low postoperative morbidity rate.

9-ING-41, a small molecule inhibitor of GSK-3beta, potentiates the effects of anticancer therapeutics in bladder cancer.

Glycogen synthase kinase-3 beta (GSK-3ß), a serine/threonine kinase, has been identified as a potential therapeutic target in human bladder cancer. In the present study, we investigated the antitumor effect of a small molecule GSK-3ß inhibitor, 9-ING-41, currently in clinical studies in patients with advanced cancer, in bladder cancer cell lines. We found that treatment with 9-ING-41 leads to cell cycle arrest, autophagy and apoptosis in bladder cancer cells. The autophagy inhibitor chloroquine potentiated the antitumor effects of 9-ING-41 when tested in combination studies. Our findings also demonstrate that 9-ING-41 enhanced the growth inhibitory effects of gemcitabine or cisplatin when used in combination in bladder cancer cells. Finally, we found that 9-ING-41 sensitized bladder cancer cells to the cytotoxic effects of human immune effector cells. Our results provide a rationale for the inclusion of patients with advanced bladder cancer in clinical studies of 9-ING-41.

Ulcerative interstitial cystitis in an adolescent successfully treated with complete transurethral ulcer resection: A case report.

INTRODUCTION: Interstitial cystitis is difficult to treat and may affect adolescents. CASE PRESENTATION: A 15-year-old girl presented with severe pain upon terminal micturition that persisted for approximately 2 hours. The pain had been present for more than 1 month. Cystoscopy revealed severe erosion throughout the trigone. Transurethral fulguration did not improve her symptoms. However, complete electric resection of the ulcer markedly reduced the symptom. After complete resection, pain on urination disappeared and she has had no pain without medication for 15 months. CONCLUSION: Complete resection not fulguration of the ulcer is effective for interstitial cystitis in adolescent female patients.

[LONG-TERM OUTCOME OF PEDIATRIC KIDNEY TRANSPLANTATION: A SINGLE-CENTER EXPERIENCES].

(Background) Long-term care is necessary for normal growth and development of pediatric recipients of kidney transplants. We report on our experience with pediatric kidney transplantation (KTx) during the past 19 years. (Methods) We retrospectively analyzed the data from 26 recipients who received KTx between 1996 and 2014 at Niigata University Hospital (one patient underwent two consecutive KTx during the designated period). All recipients were 16 years old or younger at the time of KTx. (Results) The graft survival rates at 1, 5, and 10 years after transplantation were 96%, 96%, and 88%, respectively. Three recipients lost the renal graft function due to graft thrombosis, antibody mediated rejection and steroid resistant rejection. Drug non-adherence was associated with rejection episodes, which led to the increasing of estimated glomerular filtration rate (eGFR) level. In addition, renal graft function was related to the growth after KTx. Eighteen recipients graduated from high school during follow-up periods and 17 recipients obtained employment. (Conclusion) Interventions promoting adherence should be implemented among pediatric recipients and parents to optimize graft survival and growth after KTx. Successful KTx contributed the high rate of social participation and employment after pediatric KTx.

[THE USE OF BORTEZOMIB FOR THE TREATMENT OF CHRONIC ANTIBODY MEDIATED REJECTION AFTER KIDNEY TRANSPLANTATION].

(Backgrounds) The efficacy of bortezomib for chronic antibody mediated rejection (CAMR) after kidney transplantation is still obscure. (Materials and methods) CAMR were persisted in 5 recipients who were treated with plasma exchange, low dose of IVIG, steroid pulse therapy, and rituximab. 1.3 mg/m2 of bortezomib was administered on days 1, 4, 8, 11. Serum creatinine (sCr) levels, anti-HLA antibodies, and histology were analyzed. (Results) Stable sCr levels were obtained in 3 out of 5 recipients. No one lost renal graft function during follow-up periods. Anti-HLA class I antibodies were significantly decreased after bortezomib treatment, however anti-HLA class II antibodies were not changed. Histology showed no improvement at 6 months after bortezomib administration. Two recipients whose sCr levels increased during follow-up had already had interstitial fibrosis and tubular atrophy (IF/TA) in histology before bortezomib treatment. (Conclusions) The use of bortezomib after IF/TA could be detected in histology may not contribute to stabilize renal graft function in CAMR.

Congenital Scaphoid Megalourethra: A Case Report.

A congenital megalourethra is an enlargement of the pendulous urethra without evidence of distal obstruction. A 1-month-old boy presented to us with complaint of weak stream, ballooning of the penis before and during voiding and post voiding dribbling, since birth. Physical examination and cystourethroscope confirmed the diagnosis of congenital scaphoid megalourethra. He underwent reduction urethroplasty. During postoperative follow up, he had normal looking penis with good urinary stream.

[A Case of an Orbital Metastasectomy in a Renal Cell Carcinoma after Sunitinib Treatment : A Case Report].

An 81-year-old man had undergone radical nephrectomy for a renal cell carcinoma (RCC) in October 1998. Twelve years postoperatively, he developed diplopia and photophobia. Orbital metastasis of RCC was suspected and sunitinib treatment was given for 16 months. Computer tomography revealed a partial response of metastatic tumor for the first 2 months and stable disease for the next 12 months. However, it enlarged in the last 2 months. Total resection of right metastatic orbital tumor with deep lateral orbital decompression was done in January 2012. He has been well and without any local recurrence, distant metastasis or vision loss for 24 months.

Identification of marker proteins by orthodontic treatment: relationship of RANKL in the gingival crevicular fluid and of amylase in whole saliva with orthodontic treatment.

Orthodontic medical treatment is performed to move a tooth to the optimal position to obtain optimal occlusion. Orthodontic treatment is accompanied by mechanical stress due to orthodontic force and by psychological stress that is experienced as pain or displeasure. The purpose of this study was to identify stress marker proteins during orthodontic treatment. Levels of receptor activator of NFκB (RANKL) and heat shock protein 70 (HSP70) in the gingival crevicular fluid (GCF) were analyzed as markers of mechanical stress, and levels of chromogranin A (CgA) and amylase in whole saliva were analyzed as markers of psychological stress. GCF was collected from control and experimental teeth at initiation of treatment and 24 h after treatment. Whole saliva was collected before treatment, at initiation of treatment and 24 h after treatment. RANKL was expressed at 24 h after treatment in the experimental GCF, but not in the control GCF. HSP70 appeared to be constitutively expressed in GCF, and its levels showed no major change between the control and experimental groups from initiation of treatment to 24 h after treatment. Amylase activity in whole saliva was enhanced at 24 h after treatment compared to control, but CgA levels showed little change between the groups. These results indicated that RANKL and amylase may be the candidate markers for mechanical and psychological stress, respectively, during orthodontic treatment, even though the total protein concentration and amylase activity displayed a large standard deviation among subjects. Further studies are therefore required to establish these markers for clinical use.

Cephalometric evaluation of maxillary sinus sizes in different malocclusion classes.

We found few studies on the association between maxillary sinus size and malocclusion in an electronic search using PubMed. The purpose of this study was to investigate maxillary sinus size in different malocclusion groups and the association between maxillary sinus size and dentofacial morphology by the use of lateral cephalometric radiographs. A total of 120 lateral cephalograms were used. These radiographs were derived from subjects with skeletal Class I, Class II, and Class III malocclusions, classified on the basis of the A-N-B angle. Each malocclusion group consisted of 20 boys and 20 girls ranging in age from 12 to 16 years. Two linear measurements and three area measurements were made to evaluate maxillary sinus size, and four angular and eight linear measurements were made to evaluate dentofacial morphology. Analysis of variance and Pearson's correlation analysis were performed for statistical comparison. The maxillary sinuses showed no significant differences in size between the different classes of skeletal malocclusion or between sexes. However, the maxillary sinus measurements were significantly correlated with several dentofacial morphological measurements. When formulating an orthodontic treatment plan, orthodontists should take into consideration the fact that the patients 12 to 16 years old with large cranial bases and nasomaxillary complexes tend to have larger maxillary sinuses, but there is no significant association between maxillary sinus size and the A-N-B angle denoting the sagittal skeletal jaw relationship.

Tooth size discrepancies among different malocclusions in a Japanese orthodontic population.

OBJECTIVE: To identify the possible sex differences in anterior and overall tooth size ratios and to evaluate whether any differences exist in tooth size ratios and distributions of subjects with clinically significant tooth size discrepancies among Angle Class I, Class II, and Class III malocclusion groups with the corresponding skeletal characteristics in a Japanese population. MATERIALS AND METHODS: Each malocclusion group comprised 60 subjects (30 males and 30 females). The mesiodistal width from first molar to first molar was measured on each pretreatment cast to the nearest 0.01 mm using digital calipers, and the anterior and overall ratios were calculated. Student's t-test, Welch t-test, analysis of variance, and chi2-test were performed for statistical analysis. RESULTS: No statistically significant sex differences were found in anterior or overall ratio in any group. No significant differences in anterior or overall ratios were found among the malocclusion groups. No significant differences were found between the distributions of subjects with clinically significant tooth size discrepancies, categorized by the Bolton standard deviation definition and by the actual amount of change calculated for tooth size correction in millimeters, among the malocclusion groups except for the mandibular correction for the overall ratio between Class I and Class III subjects. CONCLUSION: Bolton's values can be used with confidence for the typical Japanese orthodontic population. The use of the actual millimeters of correction for the tooth size ratios could help orthodontists avoid underestimating the prevalence of clinically significant tooth size discrepancies.

Evidence for amylase release by cGMP via cAMP-dependent protein kinase in rat parotid acinar cells.

Amylase release from the rat parotid gland is primarily mediated by a cAMP-dependent protein kinase (PKA). We previously reported that cGMP/cGMP-dependent protein kinase (PKG) signaling evokes amylase release. In the present study, we investigated whether cGMP-mediated amylase release might be due to cGMP/PKA signaling, as well as cGMP/PKG pathway. Activation of PKA by cGMP was required 100-1000-fold greater concentration than activation by cAMP in a parotid cytosol fraction. Synergistic activation of PKA by the combination of physiological cAMP and low concentration of cGMP was observed. Amylase release from intact acinar cells was synergistically stimulated by the combination of diBu-cAMP and 8-pCPT-cGMP. cGMP dose-dependently stimulated amylase release from saponin-permeabilized parotid acinar cells. Phosphorylation by cGMP produced phosphorylated proteins of the same size as those produced by cAMP. Phosphorylation by cGMP was inhibited by the addition of PKA inhibitor, H-89. These results suggest that cGMP activates both PKG and PKA. Thus, it appears that both cGMP/PKG and cGMP/PKA pathways mediate amylase release from rat parotid acinar cells.