Cyanobacterial Biocrust on Biomineralized Soil Mitigates Freeze-Thaw Effects and Preserves Structure and Ecological Functions.

Biocrust inoculation and microbially induced carbonate precipitation (MICP) are tools used in restoring degraded arid lands. It remains unclear whether the ecological functions of the two tools persist when these methods are combined and subjected to freeze-thaw (FT) cycles. We hypothesized a synergetic interaction between MICP treatment and biocrust under FT cycles, which would allow both components to retain their ecological functions. We grew cyanobacterial (Nostoc commune) biocrusts on bare soil and on MICP (Sporosarcina pasteurii)-treated soil, subjecting them to repeated FT cycles simulating the Mongolian climate. Generalized linear modeling revealed that FT cycling did not affect physical structure or related functions but could increase the productivity and reduce the nutrient condition of the crust. The results confirm the high tolerance of MICP-treated soil and biocrust to FT cycling. MICP treatment + biocrust maintained higher total carbohydrate content under FT stress. Our study indicates that biocrust on biomineralized soil has a robust enough structure to endure FT cycling during spring and autumn and to promote restoration of degraded lands.

[Right Atrial Blood Cyst with Patent Foramen Ovale:Report of a Case].

A woman in 70s was diagnosed with lung cancer, and a right atrial mass was discovered incidentally during preoperative examination by contrast-enhanced computed tomography (CT). Transesophageal echocardiography revealed a 20-mm, stemmed, spherical mass with low internal echogenicity and partially high echogenicity extending from the junction of the inferior vena cava to the posterior wall of the right atrium. Patent foramen ovale( PFO) was also confirmed. To avoid embolization and obtain diagnosis, the patient was referred for right atrial tumor resection. Cardiopulmonary bypass was established; the right atrial tumor was removed while the patient was in cardiac arrest. The tumor membrane was thin and easily ruptured, revealing jelly-like blood content and calcified mass. The patient recovered well after surgery and was discharged on day 15. According to the pathological examination, the tumor was a blood cyst. This is an extremely rare case of a blood cyst with PFO.

A novel role for Helicobacter pylori cytotoxin-associated gene A in negative regulation of autophagy in human gastric cells.

BACKGROUND: Autophagy plays an important role in carcinogenesis and tumor progression in many cancers, including gastric cancer. Cytotoxin-associated gene A (CagA) is a well-known virulent factor in Helicobacter pylori (H. pylori) infection that plays a critical role in gastric inflammation and gastric cancer development. However, its role in autophagy during these processes remains unclear. Therefore, we aimed to clarify the role of CagA in autophagy in CagA-related inflammation. METHODS: We evaluated the autophagic index of AGS cells infected with wild-type cagA-positive H. pylori (Hp-WT) and cagA-knockout H. pylori (Hp-ΔcagA) and rat gastric mucosal (RGM1) cells transfected with CagA genes. To identify the mechanisms underlying the down regulation of autophagy in AGS cells infected with H. pylori, we evaluated protein and mRNA expression levels of autophagy core proteins using western blotting and quantitative reverse transcription-polymerase chain reaction (RT-PCR). To determine whether autophagy induced the expression of the pro-inflammatory mediator, cyclooxygenase-2 (COX-2), we evaluated COX-2 expression in AGS cells treated with an autophagy inducer and inhibitor and infected with H. pylori. In addition, we evaluated whether COX-2 protein expression in AGS cells influenced beclin-1 (BECN1) expression with si-RNA transfection when infected with H. pylori. RESULTS: Autophagic flux assay using chloroquine showed that autophagy in AGS cells was significantly suppressed after H. pylori infection. The autophagic index of AGS cells infected with Hp-WT was decreased significantly when compared with that in AGS cells infected with Hp-ΔcagA. The autophagic index of RGM1 cells transfected with CagA was lower, suggesting that CagA inhibits autophagy. In addition, BECN1 expression levels in AGS cells infected with Hp-WT were reduced compared to those in AGS cells infected with Hp-ΔcagA. Furthermore, COX-2 expression in AGS cells infected with H. pylori was controlled in an autophagy-dependent manner. When AGS cells were transfected with small interfering RNA specific for BECN1 and infected with Hp-WT and Hp-ΔcagA, COX-2 was upregulated significantly in cells infected with Hp-ΔcagA. CONCLUSIONS: In conclusion, the H. pylori CagA protein negatively regulated autophagy by downregulating BECN1. CagA-induced autophagy inhibition may be a causative factor in promoting pro-inflammatory mediator production in human gastric epithelial cells.

A novel agonist for the HGF receptor MET promotes differentiation of human pluripotent stem cells into hepatocyte-like cells.

Hepatocyte growth factor (HGF) is the natural ligand of the MET receptor tyrosine kinase. This ligand-receptor couple is essential for the maturation process of hepatocytes. Previously, the rational design of a synthetic protein based on the assembly of two K1 domains from HGF led to the production of a potent and stable MET receptor agonist. In this study, we compared the effects of K1K1 with HGF during the differentiation of hepatocyte progenitors derived from human induced pluripotent stem cells (hiPSCs). In vitro, K1K1, in the range of 20 to 200 nM, successfully substituted for HGF and efficiently activated ERK downstream signaling. Analysis of the levels of hepatocyte markers showed typical liver mRNA and protein expression (HNF4α, albumin, alpha-fetoprotein, CYP3A4) and phenotypes. Although full maturation was not achieved, the results suggest that K1K1 is an attractive candidate MET agonist suitable for replacing complex and expensive HGF treatments to induce hepatic differentiation of hiPSCs.

Clinical and Histopathological Features of the Differentiated Type of Early Gastric Cancer with an Unclear Lateral Demarcation.

INTRODUCTION: Magnifying endoscopy with narrow-band imaging (M-NBI) is useful for determining lateral demarcation of early gastric cancers; however, this is sometimes difficult. Features related to an unclear lateral demarcation remain unknown. We evaluated the clinical and histopathological features of early gastric cancers with unclear lateral demarcation on M-NBI. METHODS: This single-center, retrospective, cohort study analyzed early gastric cancer treated by endoscopic submucosal dissection between January 2013 and August 2015. We evaluated the clinicopathological and immunohistochemical features using anti-p53, anti-Ki-67, anti-MUC5AC, anti-MUC6, anti-MUC2, and anti-CD10 antibody staining. We compared the lateral demarcation between the demarcation clear (DC) and the demarcation unclear (DU) lesions by using M-NBI. RESULTS: A total of 224 differentiated adenocarcinomas (DU group: 18 lesions; DC group: 206 lesions) were analyzed. A history of successful Helicobacter pylori eradication was significantly more frequent in the DU group (p = 0.001). We examined the tissues of 72 lesions (DU group: 18 lesions, DC group: 54 lesions [randomly selected]) immunohistochemically. The nonneoplastic superficial epithelium was observed more frequently in the DU group as compared to in the DC group (p = 0.006). Additionally, compared to the DC group, the DU group showed a significantly higher expression of the gastric phenotype markers (p = 0.023) and had lower p53 scores (p < 0.001) and Ki-67 labeling indexes (p = 0.029). Multivariate analysis revealed the nonneoplastic superficial epithelium and a low p53 score as the significant independent variables associated with an unclear lateral demarcation on M-NBI. CONCLUSIONS: The nonneoplastic superficial epithelium and a low p53 score were associated with difficulties in determining the lateral demarcation in early gastric cancers on M-NBI.

Total arch replacement and frozen elephant trunk for acute type A aortic dissection.

OBJECTIVE: The present study aimed to evaluate the outcomes of total aortic arch replacement with proximalization of distal anastomosis using the frozen elephant trunk technique with the J Graft FROZENIX (Japan Lifeline, Tokyo, Japan) and Gelweave Lupiae (Vascutek Terumo Inc, Scotland, United Kingdom) graft (distal anastomosis performed in zones 1 and 2) in patients with acute Stanford type A acute aortic dissection. METHODS: A total of 50 patients underwent total aortic arch replacement using the frozen elephant trunk technique, deploying the J Graft FROZENIX into zone 1 or 2 (zone 1: n = 17, zone 2: n = 33) in combination with the Gelweave Lupiae graft for acute Stanford type A acute aortic dissection. Patient characteristics, intraoperative data, and early and midterm outcomes were analyzed. RESULTS: The overall in-hospital mortality rate was 4% (2 patients). The in-hospital mortality rate in patients with visceral malperfusion was 11% (1/9). There were no patients with paraplegia and stent graft-induced new entry. Resection or closure of the most proximal entry tear was achieved in 100% of 42 patients who had postoperative computed tomography. The overall survival was 87.9%, 84.1%, and 84.1% at 1, 2, and 3 years, respectively. However, 1 patient required endovascular extension for the dilatation of the descending thoracic aorta 4 months after the initial surgery. CONCLUSIONS: Total aortic arch replacement with the frozen elephant trunk technique (zone 1-2) and Gelweave Lupiae graft was safe and effective in simplifying surgery for acute Stanford type A acute aortic dissection.

Suppression of autophagy promotes fibroblast activation in p53-deficient colorectal cancer cells.

Deficiency of p53 in cancer cells activates the transformation of normal tissue fibroblasts into carcinoma-associated fibroblasts; this promotes tumor progression through a variety of mechanisms in the tumor microenvironment. The role of autophagy in carcinoma-associated fibroblasts in tumor progression has not been elucidated. We aimed to clarify the significance of autophagy in fibroblasts, focusing on the TP53 status in co-cultured human colorectal cancer cell lines (TP53-wild-type colon cancer, HCT116; TP53-mutant colon cancer, HT29; fibroblast, CCD-18Co) in vitro. Autophagy in fibroblasts was significantly suppressed in association with ACTA2, CXCL12, TGFß1, VEGFA, FGF2, and PDGFRA mRNA levels, when co-cultured with p53-deficient HCT116sh p53 cells. Exosomes isolated from the culture media of HCT116sh p53 cells significantly suppressed autophagy in fibroblasts via inhibition of ATG2B. Exosomes derived from TP53-mutant HT29 cells also suppressed autophagy in fibroblasts. miR-4534, extracted from the exosomes of HCT116sh p53 cells, suppressed ATG2B in fibroblasts. In conclusion, a loss of p53 function in colon cancer cells promotes the activation of surrounding fibroblasts through the suppression of autophagy. Exosomal miRNAs derived from cancer cells may play a pivotal role in the suppression of autophagy.

Use of a convolutional neural network for classifying microvessels of superficial esophageal squamous cell carcinomas.

BACKGROUND AND AIM: The morphological diagnosis of microvessels on the surface of superficial esophageal squamous cell carcinomas using magnifying endoscopy with narrow-band imaging is widely used in clinical practice. Nevertheless, inconsistency, even among experts, remains a problem. We constructed a convolutional neural network-based computer-aided diagnosis system to classify the microvessels of superficial esophageal squamous cell carcinomas and evaluated its diagnostic performance. METHODS: In this retrospective study, a cropped magnifying endoscopy with narrow-band images from superficial esophageal squamous cell carcinoma lesions was used as the dataset. All images were assessed by three experts, and classified into three classes, Type B1, B2, and B3, based on the Japan Esophagus Society classification. The dataset was divided into training and validation datasets. A convolutional neural network model (ResNeXt-101) was trained and tuned with the training dataset. To evaluate diagnostic accuracy, the validation dataset was assessed by the computer-aided diagnosis system and eight endoscopists. RESULTS: In total, 1777 and 747 cropped images (total, 393 lesions) were included in the training and validation datasets, respectively. The diagnosis system took 20.3 s to evaluate the 747 images in the validation dataset. The microvessel classification accuracy of the computer-aided diagnosis system was 84.2%, which was higher than the average of the eight endoscopists (77.8%, P < 0.001). The area under the receiver operating characteristic curves for diagnosing Type B1, B2, and B3 vessels were 0.969, 0.948, and 0.973, respectively. CONCLUSIONS: The computer-aided diagnosis system showed remarkable performance in the classification of microvessels on superficial esophageal squamous cell carcinomas.

Aortoduodenal Fistula After Endovascular Aortic Repair for an Inflammatory Abdominal Aortic Aneurysm: A Case Report.

Aortoenteric fistula after endovascular aortic repair for an abdominal aortic aneurysm is a rare but severe complication. Particularly, a case of inflammatory abdominal aortic aneurysm is extremely rare and there are only 3 reported cases. A 70-year-old man underwent endovascular aortic repair for impending rupture of an inflammatory abdominal aortic aneurysm and was medicated steroids for approximately 2 years. Four years after endovascular aortic repair, he developed endograft infection with an aortoduodenal fistula and a left psoas abscess. He underwent total endograft excision, debridement, in situ reconstruction of the aorta using prosthetic grafts with omental coverage, and digestive tract reconstruction to prevent leakage. Pseudomonas aeruginosa was detected in the infected aortic sac. The patient has not experienced recurrence of infection in the 35 months since his operation.

Partial aortic root remodeling for chronic aortic dissection with coronary intimal tear.

While there are many reports on partial aortic root remodeling, it is rarely performed for chronic aortic dissection of the coronary artery. This report presents a case of a 69-year-old man incidentally diagnosed with aortic dissection during routine checkup. He had a history of percutaneous coronary intervention from the left main trunk to the left anterior descending artery and left circumflex artery. Computed tomography revealed a chronic type A aortic dissection with an aneurysmal aortic root. The false lumen of the Valsalva sinus originated from the left anterior descending artery and expanded largely to the non-coronary Valsalva sinus. We performed partial aortic root remodeling, resecting the dissected non-coronary Valsalva sinus. The postoperative course was uneventful. Partial aortic root remodeling was effective, but its use might be controversial for chronic aortic dissection without resection of the primary entry of the left anterior descending artery. Moreover, close follow-up is required.

Thoracic endovascular aortic repair for an ascending aortic pseudoaneurysm.

An 80-year-old woman underwent aortic valve replacement and ascending aortic replacement. Two years later, computed tomography revealed a pseudoaneurysm of the ascending aorta replaced with a prosthesis. The pseudoaneurysm arose from the stump of a side branch of the prosthesis. Endovascular treatment for the pseudoaneurysm was carried out using the aortic extension cuff of an infrarenal endovascular system. The postoperative course was uneventful.

Additional effect of magnifying narrow-band imaging on estimating the invasion depth of superficial esophageal cancer.

BACKGROUND AND AIM: To investigate whether assessment by magnifying narrow-band imaging (M-NBI) based on the classification of the Japan Esophageal Society provides additional value to the estimation of the invasion depth of superficial esophageal squamous cell carcinoma (SCC) compared with assessment by white light endoscopy (WLE) alone. METHODS: Endoscopic images of 211 consecutive superficial esophageal SCCs resected by endoscopic submucosal dissection were separated into WLE and M-NBI images. Depth estimation was performed independently by five evaluators using the numerical depth estimation scale (0 = epithelium (EP)/lamina propria (LPM), 1 = EP/LPM > muscularis mucosa (MM)/shallow submucosa (SM1), 2 = MM/SM1 > EP/LPM, 3 = MM/SM1, 4 = MM/SM1 > deep submucosa (SM2), 5 = SM2 > MM/SM1, 6 = SM2), using primarily WLE images (step 1), and subsequently both WLE and M-NBI images (step 2). The discordance scores, determined by the average of the five evaluators' difference between the estimated score (from 0 to 6) and pathological score (0 for histologically proven EP/LPM, 3 for MM/SM1, and 6 for SM2), were analyzed in steps 1 and 2. RESULTS: The discordance scores significantly decreased in step 2 (0.53 ± 0.06) compared with those in step 1 (0.79 ± 0.07) (P < 0.001). When the discordance score < 1.5 was regarded as a clinically correct diagnosis, the rate of the clinically correct diagnosis significantly increased in step 2 compared with that in step 1 (81% to 91%, P < 0.001). CONCLUSION: M-NBI has an additive value for estimating the invasion depth of superficial esophageal SCCs.

Usefulness of a dedicated mouthpiece for the Valsalva maneuver to visualize the hypopharynx during transoral endoscopy.

Background and study aims Patients with esophageal squamous cell carcinoma (SCC) are at high risk of developing second primary SCCs in the hypopharynx. However, such second primary tumors are difficult to observe because of lumen closure. The Valsalva maneuver using a dedicated mouthpiece is a promising technique to visualize the hypopharynx during transoral endoscopy. In the current study, we investigated the utility of this method. Patients and methods The current study was a randomized, controlled, crossover trial. Patients with esophageal SCC were randomly assigned first to undergo pharyngeal observation using the dedicated mouthpiece followed by observation using a conventional mouthpiece, or vice versa. The primary endpoint was complete visualization of the hypopharynx, which was assessed blindly by three external evaluators. Results A total of 68 pharyngeal examinations were analyzed - 34 with the dedicated mouthpiece and 34 with a conventional mouthpiece. Complete visualization was achieved in 68 % of the examinations (23/34) using the dedicated mouthpiece, whereas none of the examinations using the conventional mouthpiece achieved complete visualization of the hypopharynx. Observation scores of the oropharynx were not significantly different between both types of examination ( P  = 0.50). No serious adverse events (AEs) occurred. Conclusions Endoscopic view of the hypopharynx was markedly improved by the Valsalva maneuver using the dedicated mouthpiece, with no serious AEs. This procedure should be included in the endoscopic examinations for the patients with esophageal SCCs.

[Coil Embolization for an Aberrant Left Subclavian Artery Aneurysm with Kommerell's Diverticulum;Report of a Case].

A 67-year-old woman encountered a traffic accident and had chest computed tomography(CT) examination. It revealed a 24 mm Kommerell's diverticulum associated with a right-sided aortic arch and a 15 mm saccular aneurysm of an aberrant left subclavian artery. We performed intra-aneurysmal coil embolization for the left subclavian artery aneurysm after a balloon occlusion test of the left subclavian artery. The postoperative course was uneventful.

Metabolome Analysis Reveals Dermal Histamine Accumulation in Murine Dermatitis Provoked by Genetic Deletion of P-Glycoprotein and Breast Cancer Resistance Protein.

PURPOSE: P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) are xenobiotic transporters which pump out variety types of compounds, but information on their interaction with endogenous substrates in the skin is limited. The purpose of the present study was to clarify possible association of these transporters in dermal accumulation of inflammatory mediators. METHODS: Dermatitis model was constructed by repeated topical application of oxazolone in wild-type, and P-gp and BCRP gene triple knockout (Mdr1a/1b/Bcrp-/-) mice to observe difference in phenotype. Target metabolome analysis of 583 metabolites was performed using skin and plasma. RESULTS: Dermatitis and scratching behavior in dermatitis model of Mdr1a/1b/Bcrp-/- mice were more severe than wild-type mice, suggesting protective roles of these transporters. This hypothesis was supported by the metabolome analysis which revealed that concentration of histamine and other dermatitis-associated metabolites like urate and serotonin in the dermatitis skin, but not normal skin, of Mdr1a/1b/Bcrp-/- mice was higher than that of wild-type mice. Gene expression of P-gp and BCRP was reduced in oxazolone-treated skin and the skin of patients with atopic dermatitis or psoriasis. CONCLUSIONS: These results suggest possible association of these efflux transporters with dermal inflammatory mediators, and such association could be observed in the dermatitis skin.

Aneurysm resection interposed with a spiral saphenous vein graft in a patient with a popliteal venous aneurysm with thrombosis.

We describe the case of a 46-year-old man with pulmonary thromboembolism caused by a popliteal venous aneurysm with thrombosis. The aneurysm was fusiform and partially saccular with a thrombus, and the caliber of the native popliteal vein was large. Tangential aneurysmectomy with lateral venorrhaphy was difficult because of the aneurysm type, and graft interposition was required because of the large venous diameter of the anastomosis site. The patient underwent aneurysm resection interposed with a spiral saphenous vein graft. The postoperative course was uneventful, and the graft was patent at 1 year after surgery.

Exosomal microRNAs derived from colon cancer cells promote tumor progression by suppressing fibroblast TP53 expression.

The tumor microenvironment offers favorable conditions for tumor progression, and activated fibroblasts, known as cancer-associated fibroblasts, play a pivotal role. TP53-deficient cancer cells are known to induce strong fibroblast activation. We aimed to elucidate the oncogenic role of exosomes derived from TP53-deficient colon cancer cells in fibroblast proliferation and tumor growth. Cancer cell-derived exosomes (CDEs) were isolated from the conditioned media of cancer cells using a sequential ultracentrifugation method. The effects of exosomes on tumor growth were evaluated using human cell lines (TP53-WT colon cancer, HCT116; TP53-mutant colon cancer, HT29; and fibroblasts, CCD-18Co and WI-38) and an immune-deficient nude mouse xenograft model. HCT116 (HCT116sh p53 ) cells deficient in TP53 accelerated cocultured fibroblast proliferation compared to TP53-WT HCT116 (HCT116sh control ) cells in vitro. Exosomes from HCT116sh p53 cells suppressed TP53 expression of fibroblasts and promoted their proliferation. Xenografts of HCT116sh p53 cells grew significantly faster than those of HCT116sh control cells in the presence of co-injected fibroblasts, but this difference was diminished by CDE inhibition. Microarray analysis identified upregulation of several microRNAs (miR-1249-5p, miR-6737-5p, and miR-6819-5p) in TP53-deficient CDEs, which were functionally proven to suppress TP53 expression in fibroblasts. Exosomes derived from TP53-mutant HT29 cells also suppressed TP53 expression in fibroblasts and accelerated their growth. The proliferative effect of HT29 on cocultured fibroblasts was diminished by inhibition of these miRNAs in fibroblasts. Our results suggest that CDEs play a pivotal role in tumor progression by fibroblast modification. Cancer cell-derived exosomes might, therefore, represent a novel therapeutic target in colon cancer.

Optimized protocol for the hepatic differentiation of induced pluripotent stem cells in a fluidic microenvironment.

In the present study, we evaluated the performance of different protocols for the hepatic differentiation of human-induced pluripotent stem cells (hiPSCs) in microfluidic biochips. Strategies for complete and partial on-chip differentiation were tested. Unlike full on-chip differentiation, the transfer of iPSCs from Petri dishes to biochips during the differentiation process produced a heterogeneous tissue with enhanced hepatic features compared with control cultures in Petri dishes. The tissue in biochips was constituted of cells expressing either stabilin-1 or albumin, while no stabilin-1 was detected in controls. Functional analysis also revealed double the production rate for albumin in biochips (about 2,000 ng per day per 106 cells). Besides this, tissues obtained in biochips and controls exhibited the metabolism of a specific bile acid. Whole transcriptome analysis with nanoCAGE exhibited a differential expression of 302 genes between control and biochip cultures and a higher degree of hepatic differentiation in biochips, together with increased promoter motif activity for typical liver transcription factors such as estrogen related receptor alpha ( ESRRA), hepatic nuclear factor 1 ( HNF1A), hepatic nuclear factor 4 ( HNF4A), transcription factor 4 ( TCF4), and CCAAT enhancer binding protein alpha ( CEBPA). Gene set enrichment analysis identified several pathways related to the extracellular matrix, tissue reorganization, hypoxia-inducible transcription factor, and glycolysis that were differentially modulated in biochip cultures. However, the presence of CK19/ALB-positive cells and the ɑ-fetoprotein levels measured in the cultures still reflect primitive differentiation patterns. Overall, we identified key parameters for improved hepatic differentiation on-chip, including the maturation stage of hepatic progenitors, inoculation density, adhesion time, and perfusion flow rate. Optimization of these parameters further led to establish a protocol for reproducible differentiation of hiPSCs into hepatocyte-like cells in microfluidic biochips with significant improvements over Petri dish cultures.

[Thoracic Endovascular Aortic Repair for Traumatic Aortic Injury with Internal Iliac Approach;Report of a Case].

A 40-year-old female was injured in a car accident while driving. Computed tomography confirmed a dissection localized to the distal aortic arch and hematoma around the area. Severe liver damage with surrounding extravasation was also confirmed. Furthermore, multiple rib fractures and lumbar spine fracture were apparent. Thoracic endovascular aortic repair( TEVAR) was performed after transcatheter arterial embolization to treat the liver injury. As both the iliac and femoral arteries were narrow, the internal iliac artery was transected peripherally and used as an access route. This method does not require complex procedures such as artificial vascular anastomosis for vascular repair in the event of injury. Therefore, it may be considered as an option when access from the femoral artery is difficult.