Long-term outcomes of choledochoduodenostomy for choledocholithiasis: increased incidence of postoperative cholangitis after total or distal gastrectomy.

PURPOSE: Choledochoduodenostomy (CDD) is performed to treat choledocholithiasis (CDL) cases where endoscopic stone removal is difficult. Recognizing CDD characteristics is important for CDL treatment planning. METHODS: A total of 116 patients, including 33 patients ≥ 80 years old (29 with previous total gastrectomy, 19 with previous distal gastrectomy, 20 with built-up stones, 19 with periampullary diverticulum, 10 with confluence stones, 8 with repetitive recurrent stones, 4 with hard stones, 3 with endoscopic retrograde cholangiography [ERC] not available due to lack of cooperation, 2 with a history of pancreatitis post-ERC, and 2 in whom ERC could not be performed due to a disturbed anatomy) underwent CDD for CDL. Postoperative complications and long-term outcomes were evaluated. RESULTS: The in-hospital mortality rate was 0%. The morbidity (grade ≥ IIIA according to the Clavien-Dindo classification) rates in the elderly (≥ 80 years old) and non-elderly (51-79 years old) patients were 3.0% (1/33) and 2.4% (2/83), respectively (p = 0.85). Long-term complications included cholangitis in eight (7%) patients, of which three cases were repetitive and seven had an operative history of total or distal gastrectomy. The incidence of postoperative cholangitis after total or distal gastrectomy was 15% (7/48), which was significantly higher than that involving other causes (1.5%, 1/68; p < 0.01). Two patients with cholangitis after total gastrectomy experienced early recurrence of lithiasis at 2 and 9 months after surgery. CONCLUSIONS: CDD is safe, even in elderly patients. However, a history of total gastrectomy or distal gastrectomy may increase the incidence of postoperative cholangitis.

Gelatin Sponge as an Anchorage for Three-dimensional Culture of Colorectal Cancer Cells.

BACKGROUND: Compared to two-dimensional cultures, three-dimensional (3D) cultures have many advantages in cancer studies. Nevertheless, their implementation is unsatisfactory. This study aimed to develop an anchorage-dependent 3D culture model for colorectal cancer research. MATERIALS AND METHODS: Human HCT116, DLD-1 and SW620 colorectal cell lines were cultured in a gelatin sponge, and its applicability for morphological examination was studied. RESULTS: The resulting specimens were suitable for scanning electron microscopy, transmission electron microscopy, and immunohistochemical examination. HCT116 formed smaller structures and migrated through the pores of the sponge. DLD-1 formed larger structures with tight cell-to-cell adhesion. SW620 also formed large structures but small clustered cells tended to attach to the anchorage more favorably. Immunohistochemical staining demonstrated phosphorylated yes-associated protein (YAP) localized near the attachment site in HCT116 cells. CONCLUSION: Because the gelatin sponge provided suitable anchorage and the cultured cells formed distinguishable 3D structures, this method may be useful for further colorectal cancer research.

Blood group O is a risk factor for delayed post-polypectomy bleeding.

BACKGROUND: Blood group O of ABO blood group system is considered as a risk factor for various bleeding events, but the relationship with endoscopic treatment-associated bleeding has yet to be investigated. This study aimed to evaluate whether blood group O is associated with delayed bleeding after colorectal endoscopic resection. METHODS: This was a retrospective observational study based on medical records at four university hospitals in Japan. We reviewed the records for consecutive patients who underwent colorectal endoscopic resection from January 2014 through December 2017. The primary outcome was the incidence of delayed bleeding, defined as hematochezia or melena, requiring endoscopy, transfusion, or any hemostatic intervention up to 28 days after endoscopic resection. Multivariate logistic regression analysis was performed to adjust the impact of blood group O on the delayed bleeding. RESULTS: Among 10,253 consecutive patients who underwent colorectal endoscopic resection during the study period, 8625 patients met the criteria. In total, delayed bleeding occurred in 255 patients (2.96%). The O group had significantly more bleeding events compared with the non-O group (A, B, and AB) (relative risk, 1.62 [95% confidence interval, 1.24-2.10]; P < 0.001). In multivariate logistic regression analysis, blood group O remained an independent risk factor for the bleeding (adjusted odds ratio, 1.60 [95% confidence interval, 1.18-2.17]; P = 0.002). CONCLUSIONS: Blood group O was associated with an increased risk of delayed bleeding in patients undergoing colorectal endoscopic resection. Preoperative screening for ABO blood group could improve risk assessments.

[A difficult case of a stent-stone complex removal with endoscopic lithotomy].

An 88-year-old patient had undergone endoscopic stenting for the treatment of acute cholangitis caused by choledocholithiasis. After a year and two months, he presented with cholangitis caused by common bile duct stones that formed a stent-stone complex. Another stent was observed adjacent to the old stent;however, the cholangitis relapsed in a short term. Thus, we planned to remove as many stones as possible. These stones were not free-floating and had affected the bile duct. Endoscopic mechanical lithotripsy was attempted;however, it failed. He was successfully treated using peroral cholangioscopy and electrohydraulic lithotripsy. After three months, he developed cholangitis because of the recurrence of choledocholithiasis. After removing as many stones as possible and performing endoscopic stenting, he was followed up as an outpatient. He had no symptoms for seven months after the procedure.

Effect of Phosphoribosyltransferase Down-regulation on Malignant Glioma Cell Characteristics.

BACKGROUND/AIM: Nicotinamide phosphoribosyl-transferase (NAMPT) is a rate-limiting enzyme in the pathway synthesizing nicotinamide adenine dinucleotide (NAD (+)) from nicotinamide (NAM). Glioma tissues exhibit up-regulated NAMPT expression associated with a poor prognosis of patients. To determine if NAMPT can be a molecular therapeutic target, we investigated the effects of short hairpin RNA (shRNA)-mediated NAMPT down-regulation. MATERIALS AND METHODS: We designed shRNA to NAMPT and transfected to T98G cells. The characteristics of these cells were analyzed. RESULTS: The NAMPT shRNA-transfected cells exhibited delayed cell growth. However, there was no difference in the increase of sensitivity to temozolomide (TMZ) or X-ray irradiation between the NAMPT and scramble shRNA-transfected cells. The expression of NAMPT in the NAMPT shRNA-transfected cells increased with cell passage. Additionally, the shRNA-mediated transfection was associated with enhanced expression of quinolinic acid phosphoribo-syltransferase (QPRT). CONCLUSION: shRNA-mediated NAMPT down-regulation may not decrease the NADt to a sufficient level to increase TMZ/radiation sensitivity.

Best period to replace or change plastic stents with self-expandable metallic stents using multivariate competing risk regression analysis.

In endoscopic biliary drainage (EBD) for various benign and malignant biliary disorders, the appropriate timing to replace or change a plastic stent (PS) with a self-expandable metallic stent (SEMS) remains unclear. This study aimed to define the best period to replace or change a PS with a SEMS. Between January 1, 2012, and December 31, 2018, 1,887 consecutive EBD procedures, including 170 SEMS placements, were retrospectively identified. The period to recurrent biliary obstruction (PRBO) was estimated and compared between the malignant and benign groups and according to each disease using time to event analysis and competing risk analysis. Compared with the benign group, the malignant group had significantly shorter median PRBO with interquartile range (IQR) after PS placement [108 (39 - 270) vs. 613 (191 - 1,329) days, P < 0.001], even on multivariate analysis, with a subdistribution hazard ratio (SHR) of 3.58 (P < 0.001). The shortest PRBO distribution from the first quartile of the non-RBO period was seen in Mirizzi syndrome cases (25 days, P = 0.030, SHR = 3.32) in the benign group and in cases of pancreatic cancer (32 days, P = 0.041, SHR = 2.06); perihilar bile duct cancer (27 days, P = 0.006, SHR = 2.69); and ampullary cancer (22 days, P = 0.001, SHR = 3.78) in the malignant group. Our study supports that stent replacement for the benign group is feasible after 6 months, and the best period to replace or change a PS with a SEMS should be decided on the basis of the underlying disease to prevent RBO.

Involvement of Corticotropin-releasing Hormone-related Peptides in Cellular Stress Caused by Anticancer Drugs in Colorectal Cancer.

BACKGROUND/AIM: The expression of corticotropin-releasing hormone (CRH)-related peptides involved in stress response in colorectal cancer has been reported. We examined the involvement of CRH-related peptides in cellular stress caused by anticancer drugs in colorectal cancer. MATERIALS AND METHODS: Changes in the expression levels of CRH-related peptides and their receptors in HCT116, DLD-1, and SW480 cell lines after fluorouracil (5-FU) loading were evaluated. Effects of the receptor antagonist against DNA synthesis disorder caused by 5-FU and SN-38 were evaluated using the 3H-labeled deoxyribonucleoside incorporation assay. RESULTS: No changes in the mRNA expression levels of CRH-related peptides (UCN and UCN2) -and their receptors (CRHR1 and CRHR2) were observed. Addition of antagonists to cells with DNA synthesis disorder caused by 5-FU and SN-38 showed no differences in the incorporation of 3H-labeled deoxyribonucleoside. CONCLUSION: CRH-related peptides showed no effect on the stress response to anticancer drugs nor on DNA synthesis disorder in colorectal cancer.

Endoscopic submucosal dissection in a patient with esophageal adenoid cystic carcinoma.

We report the first use of endoscopic submucosal dissection (ESD) for the treatment of a patient with adenoid cystic carcinoma of the esophagus (EACC). An 82-year-old woman visited our hospital for evaluation of an esophageal submucosal tumor. Endoscopic examination showed a submucosal tumor in the middle third of the esophagus. The lesion partially stained with Lugol's solution, and narrow band imaging with magnification showed intrapapillary capillary loops with mild dilatation and a divergence of caliber in the center of the lesion. Endoscopic ultrasound imaging revealed a solid 8 mm × 4.2 mm tumor, primarily involving the second and third layers of the esophagus. A preoperative biopsy was non-diagnostic. ESD was performed to resect the lesion, an 8 mm submucosal tumor. Immunohistologically, tumor cells differentiating into ductal epithelium and myoepithelium were observed, and the tissue type was adenoid cystic carcinoma. There was no evidence of esophageal wall, vertical stump or horizontal margin invasion with pT1b-SM2 staining (1800 µm from the muscularis mucosa). Further studies are needed to assess the use of ESD for the treatment of patients with EACC.

Association between Yogurt Consumption and Intestinal Microbiota in Healthy Young Adults Differs by Host Gender.

Human intestinal microbiota are influenced by various factors viz. diet, environment, age, gender, geographical, and socioeconomic situation, etc. among which diet has the most profound impact. However, studies investigating this impact have mostly included subjects from diverse geographic/socioeconomic backgrounds and hence the precise effects of dietary factors on gut microbiota composition remain largely confounded. Herein, with an aim to evaluate the association between dietary habits, specifically yogurt consumption, and the gut microbiota in healthy young adults sharing similar age, lifestyle routine, geographical setting, etc., we conducted a cross-sectional study wherein 293 collegiate freshmen answered a questionnaire about their frequency of yogurt consumption over the last 2 months and provided stool specimens for microbiota analysis. Fecal microbiota were analyzed by highly sensitive reverse-transcription-quantitative-PCR assays targeting bacterial 16S rRNA molecules. Fecal organic acids were measured by HPLC. Overall, the gut microbiota were predominated (97.1 ± 8.6%) by Clostridium coccoides group, Clostridium leptum subgroup, Bacteroides fragilis group, Bifidobacterium and Atopobium cluster. Interestingly, after adjusting the data for yogurt consumption, females were found to have higher total bacterial (P = 0.013) and Bifidobacterium (P = 0.046) count and fecal pH (P = 0.007) and lower fecal concentration of total organic acids (P = 0.030), succinic acid (P = 0.007) and formic acid (P = 0.046) as compared to males. Altogether, yogurt consumption showed positive linear association with Lactobacillus and Lactobacillus gasseri subgroup in both male and female subjects; however, several gender-specific disparities were also detected in this yogurt-microbiota association. Yogurt consumption demonstrated a negative association with L. sakei subgroup, Enterobacteriaceae and Staphylococcus in males but shared a positive association with L. casei subgroup and succinic acid in female subjects. The study manifests the association between yogurt consumption and gut microbiota in a healthy homogeneous cohort and show how this association can differ by host gender. The findings should be helpful for prospective studies investigating the diet-microbiome interaction in human health and disease.

Temozolomide does not influence the transcription or activity of matrix metalloproteinases 9 and 2 in glioma cell lines.

Glioblastoma multiforme (GBM) is a treatment-resistant malignancy with poor prognosis. Temozolomide (TMZ) is widely used as a first-line drug for GBM. Although this improves patient prognosis, it does not completely eradicate the tumour. Even after total surgical resection, GBM can exhibit uncontrollable invasiveness at the tumour margins owing to activation of matrix metalloproteinases (MMPs) such as MMP-2 and -9; these degrade collagen IV in the basement membrane, which normally prevents cancer invasion. TMZ induces DNA damage and activates transcription factors including c-jun, c-fos, nuclear factor-κß, and early growth response protein-1, which have putative binding sites on the MMP-9 promoter. TMZ may therefore enhance tumour invasion by stimulating MMP-9 transcription and enzymatic activity. To test this hypothesis, we investigated MMP-2 and -9 mRNA transcription and activity in GBM cell lines treated with TMZ. Human A172 GBM cells were exposed to TMZ (25% and 50% inhibitory concentrations) for 24 or 48h; cell cycle distribution and mRNA levels of MMP-2 and -9 were evaluated using flow cytometry and semi-quantitative reverse transcription PCR, respectively. MMP-2 and -9 enzymatic activities were assessed using gelatin zymography in human A172 and U373 MG GBM cells exposed to TMZ under the same conditions. TMZ altered A172 cell cycle distribution, but not MMP-2 or -9 mRNA levels. TMZ did not affect MMP-2 or -9 enzymatic activities in A172 or U373 MG cells. These findings indicated that TMZ is therefore unlikely to promote GBM invasiveness.

Novel quantitative analysis of the S100P protein combined with endoscopic ultrasound-guided fine needle aspiration cytology in the diagnosis of pancreatic adenocarcinoma.

Specimens obtained with endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) are often tiny and fragmented leading to an inconclusive and doubtful diagnosis. To overcome the limitations of EUS-FNA in the cytological diagnosis of pancreatic adenocarcinoma (PCA), we evaluated whether quantification of the S100P protein combined with EUS-FNA reliably discriminated between PCA and benign pancreatic lesions (BPL). A high sensitivity sandwich ELISA for S100P protein was developed to aid in the detection of PCA in small samples obtained using EUS-FNA. After experimental verification of the sandwich ELISA with cell lines and mouse xenograft tumors, 27 consecutive patients with suspicious PCA who underwent EUS-FNA were enrolled in the present study examining the combination of S100P protein assessment and EUS-FNA cytology. The concentration of the S100P protein in EUS-FNA samples from the PCA group was significantly higher than that in the BPL group (P=0.04). Using receiver operating characteristic curve analysis, we determined the S100P protein cut-off value for PCA diagnosis to be 99.8 ng/ml. The S100P protein levels combined with EUS-FNA cytology to detect PCA showed the following diagnostic values: sensitivity, 94.4% [95% confidence interval (CI), 75.7-99.1%]; specificity, 88.9% (95% CI, 51.8-99.7%); positive predictive value, 94.4% (95% CI, 72.7-99.9%); negative predictive value, 88.9% (95% CI, 51.8-99.7%); accuracy, 92.6% (95% CI, 75.7­99.1%); and area under the curve, 0.92 (95% CI, 0.79-1.00). We established a novel quantitative analysis for the S100P protein in EUS-FNA samples which, when combined with EUS-FNA cytology, could provide promising results for the reliable diagnosis of PCA.

Evaluation of performance of the Omni mode for detecting video capsule endoscopy images: A multicenter randomized controlled trial.

BACKGROUND AND STUDY AIMS: Olympus recently developed a new algorithm called Omni mode that discards redundant video capsule endoscopy (VCE) images. The current study aimed to demonstrate the non-inferiority of the Omni mode in terms of true positives (TPs) and the superiority of the Omni mode with regard to reading time against a control (ordinary ES-10 system). PATIENTS AND METHODS: This multicenter prospective study included 40 patients with various small bowel diseases. VCE images were evaluated by 7 readers and 3 judging committee members. Two randomly allocated readers assessed the VCE images obtained using the 2 modalities for each patient. The order of the modalities was switched between the 2 readers and the interval between readings by the same reader was 2 weeks. The judging committee predefined clinically relevant lesions as major lesions and irrelevant lesions as minor lesions. The number of TPs for major and minor lesions and the reading times were compared between the modalities. The predefined non-inferiority margin for the TP ratio of the Omni mode compared with the control was 0.9. RESULTS: The estimated TP ratios and 95 % confidence intervals for total, major, and minor lesions were 0.87 (0.80 - 0.95), 0.93 (0.83 - 1.04), and 0.83 (0.74 - 0.94), respectively. Although non-inferiority was not demonstrated, the rate of detection of major lesions was not significantly different between the modalities. The reading time was significantly lower when using the Omni mode than when using the control. CONCLUSIONS: The Omni mode may be only appropriate for the assessment of major lesions.

Cell Membrane Perforation with Photosensitizer and a Brush-shaped Soft-polymer Sheet Using a Malignant Glioma Cell Line.

BACKGROUND: Transduction of foreign molecules into cells is an important technique to investigate the functions of corresponding molecules and/or targets. Recently, a mass-producible nanoprinting perforator was devised enabling for large-scale, high-performance drug or nucleic-acid transfer into cells without cell damage. Since little is known on the performance of the system, we investigated its effects on a malignant glioma cell line. MATERIALS AND METHODS: Photosensitization was performed by the Cell Stamper CP-01. The malignant U373MG glioma cell line was used for transduction. RESULTS: Photosensitization transduced FITC-conjugated albumin into cells. Trypan blue inclusion test demonstrated membrane disintegration by the procedure and scanning electron microscopy disclosed perforation of the cell membrane. CONCLUSION: Local oxidation reaction during the nanoprinting caused reversible membrane perforation. Morphological findings from the current study support the above mechanism, therefore the specific printing system might be convenient for transduction of foreign molecules into malignant glioma cells.

Improving the performance of an electronic nose by wine aroma training to distinguish between drip coffee and canned coffee.

Coffee aroma, with more than 600 components, is considered as one of the most complex food aromas. Although electronic noses have been successfully used for objective analysis and differentiation of total coffee aromas, it is difficult to use them to describe the specific features of coffee aroma (i.e., the type of smell). This is because data obtained by electronic noses are generally based on electrical resistance/current and samples are distinguished by principal component analysis. In this paper, we present an electronic nose that is capable of learning the wine related aromas using the aroma kit "Le Nez du Vin," and the potential to describe coffee aroma in a similar manner comparable to how wine experts describe wine aroma. The results of our investigation showed that the aromas of three drip coffees were more similar to those of pine and honey in the aroma kit than to the aromas of three canned coffees. Conversely, the aromas of canned coffees were more similar to the kit coffee aroma. In addition, the aromatic patterns of coffees were different from those of green tea and red wine. Although further study is required to fit the data to human olfaction, the presented method and the use of vocabularies in aroma kits promise to enhance objective discrimination and description of aromas by electronic noses.

Secretion of urocortin I by human glioblastoma cell lines, possibly via the constitutive pathway.

Corticotropin-releasing factor (CRF) and its family of peptides, i.e., urocortins (UCNs), play a critical role in systemic and peripheral stress-response systems and are widely expressed not only in normal tissues but also in various types of cancer cells. Given limited understanding of the mechanism of UCN I secretion, we investigated the UCN I secretory pathway in human neural stem cells (HNSCs) and in two glioblastoma cell lines, e.g., A172 and U-138 MG. Immunoreactivities for CRF receptors were detected in A172 glioblastoma cells, but not in HNSCs or U-138 glioblastoma cells, while UCN I immunoreactivity was detected in A172 and U-138 MG glioblastoma cell lines by both light field and electron microscopy. Interestingly, electron microscopy revealed UCN I immunoreactivtiy in vesicle-like structures in the plasma membrane of the glioblastoma cells. Tracking of a hybrid fluorescent protein containing a UCN I signal peptide expressed in A172 human glioblastoma cells revealed that fluorescence in secretory granules could be decreased by cycloheximide (100µg/ml), indicating that the forward transport of secretory granules containing fluorescent protein was not altered by the inhibition of protein synthesis by cycloheximide. Retrograde transport and the fusion of fluorescent granules in A172 human glioblastoma cells was induced by brefeldin A (10µg/ml), indicating that UCN I secretory granules may be transported via the constitutive pathway. Based on these results, it appears that UCN I is secreted from human glioblastoma cells by exocytosis through constitutive secretory granules, indicating that transcription of UCN I mRNA may be correlated to secretion of UCN I protein.

A novel second-generation multibending backward-oblique viewing duodenoscope in ERCP.

INTRODUCTION: Successful access to the papilla and cannulation of the desired duct can be technically challenging. A novel second-generation multi-bending backward-oblique viewing duodenoscope (2nd M-D scope) was developed to overcome this difficulty. The aim of the present study was to evaluate the utility of 2nd M-D scope during biliary ERCP. MATERIAL AND METHODS: This was a retrospective review of 53 patients with native papilla who underwent biliary ERCP with the 2nd M-D scope. RESULTS: Biliary cannulation and interventional procedures were successfully completed in all patients. In two of these patients, ERCP was initially attempted with a conventional single bending duodenoscope, but biliary cannulation was unsuccessful; one had Billroth-I gastrectomy, and the other had a Type III choledochal cyst. However, with the 2nd M-D scope, biliary cannulation and interventional procedures were successfully achieved. Finally, in nine patients, upward or downward angulation of the proximal bending portion in addition to the distal bending portion was used during ERCP, and the operator's comments demonstrated that it was helpful for obtaining an appropriate en face view to look up to the papilla and facilitate successful biliary cannulation. CONCLUSION: The newly developed 2nd M-D scope could safely facilitate biliary cannulation in selected patients during ERCP.

Abnormal number cell division of human thyroid anaplastic carcinoma cell line, SW 1736.

Cell division, during which a mother cell usually divides into two daughter cells during one cell cycle, is the most important physiological event of cell biology. We observed one-to-four cell division during imaging of live SW1736 human thyroid anaplastic carcinoma cells transfected with a plasmid expressing the hybrid protein of green fluorescent protein and histone 2B (plasmid eGFP-H2B). Analysis of the images revealed a mother cell divided into four daughter cells. And one of the abnormally divided daughter cells subsequently formed a dinucleate cell.

Magnetically guided capsule versus conventional gastroscopy for upper abdominal complaints: a prospective blinded study.

OBJECTIVES: Upper gastrointestinal endoscopy is mostly performed under sedation and has a low yield of relevant gastric lesions in patients without alarm symptoms. Simpler screening tests such as capsule endoscopy could be helpful, but gastric visualization is insufficient with the current passive capsules. A magnetically guided gastric capsule was prospectively evaluated in patients with routine indications for gastroscopy. METHODS: A total of 189 symptomatic patients (105 male; mean age 53 y) from 2 French centers subsequently and blindly underwent capsule and conventional gastroscopy by 9 and 6 examiners, respectively. The final gold standard was unblinded conventional gastroscopy with biopsy under propofol sedation. Main outcome was accuracy (sensitivity/specificity) of capsule gastroscopy for diagnosis of major gastric lesions, defined as those lesions requiring conventional gastroscopy for biopsy or removal. RESULTS: Twenty-three major lesions were found in 21 patients. Capsule accuracy was 90.5% [95% confidence interval (CI), 85.4%-94.3%] with a specificity of 94.1% (95% CI, 89.3%-97.1%) and a sensitivity of 61.9% (95% CI, 38%-82%). Accuracy did not correlate with lesion location, gastric luminal visibility, examiner case volume, or examination time. Of the remaining 168 patients, 94% had minor and mostly multiple lesions; the capsule made a correct diagnosis in 88.1% (95% CI, 82.2%-92.6%), with gastric visibility and lesion location in the proximal stomach having significant influence. All patients preferred capsule gastroscopy. CONCLUSIONS: In a prospective and strictly blinded study, magnetically guided capsule gastroscopy was shown to be feasible in clinical practice and was clearly preferred by patients. Improvements in capsule technology may render this technique a future alternative to gastroscopy.

Voltage-gated calcium channels in the human adrenal and primary aldosteronism.

Calcium channel blockers can efficiently be used in the treatment of primary aldosteronism (PA) related hypertension, but details on the localization of calcium channel (CC) in the human adrenal and its disorders, including PA, have remained unclear. Therefore, in this study we analyzed the known α subunits of L-, N- and T-type CCs in 74 adrenocortical aldosterone-producing adenomas (APA) and 16 cortisol-producing adenomas (CPA) using quantitative RT-PCR (qPCR). We also examined the status of L-(CaV1.2, CaV1.3), N-(CaV2.2) and T-(CaV3.2) CC subunits in five non-pathological adrenals (NA), five idiopathic hyperaldosteronism (IHA) cases, and 50 APA using immunohistochemistry. After qPCR evaluation, only CaV1.2, CaV1.3, CaV2.2, and CaV3.2 mRNA levels could be detected in APA and CPA. Among those, only CaV3.2 mRNA levels were significantly correlated with plasma aldosterone levels (P=0.0031), CYP11B2 expression levels (P<0.0001) and the presence of KCNJ5 mutations (P=0.0019) in APA. The immunolocalization of CCs in NA and IHA was detected in the zona glomerulosa (ZG), with a predominance of CaV3.2 in APA. These findings suggest that different types of CC can be involved in calcium-related aldosterone biosynthesis.

Effects of silica and titanium oxide particles on a human neural stem cell line: morphology, mitochondrial activity, and gene expression of differentiation markers.

Several in vivo studies suggest that nanoparticles (smaller than 100 nm) have the ability to reach the brain tissue. Moreover, some nanoparticles can penetrate into the brains of murine fetuses through the placenta by intravenous administration to pregnant mice. However, it is not clear whether the penetrated nanoparticles affect neurogenesis or brain function. To evaluate its effects on neural stem cells, we assayed a human neural stem cell (hNSCs) line exposed in vitro to three types of silica particles (30 nm, 70 nm, and <44 µm) and two types of titanium oxide particles (80 nm and < 44 µm). Our results show that hNSCs aggregated and exhibited abnormal morphology when exposed to the particles at concentrations = 0.1 mg/mL for 7 days. Moreover, all the particles affected the gene expression of Nestin (stem cell marker) and neurofilament heavy polypeptide (NF-H, neuron marker) at 0.1 mg/mL. In contrast, only 30-nm silica particles at 1.0 mg/mL significantly reduced mitochondrial activity. Notably, 30-nm silica particles exhibited acute membrane permeability at concentrations =62.5 µg/mL in 24 h. Although these concentrations are higher than the expected concentrations of nanoparticles in the brain from in vivo experiments in a short period, these thresholds may indicate the potential toxicity of accumulated particles for long-term usage or continuous exposure.