Optical coherence tomography findings of balloon angioplasty/stenting for in-stent restenosis after carotid artery stenting.

OBJECTIVES: The optimal therapeutic methods for in-stent restenosis (ISR) after carotid artery stenting (CAS) remains controversial. This study aimed to use optical coherence tomography (OCT) to evaluate the in-stent architectures during endovascular angioplasty/stenting for ISR. MATERIALS AND METHODS: Six lesions of ISR after CAS were evaluated by OCT during endovascular angioplasty/stenting. RESULTS: In one lesion, the OCT system could not be crossed because of elongation distal to the ISR lesion. In five lesions, pre-procedural OCT clearly revealed neointimal hyperplasia or neoatherosclerosis. The mean in-stent area stenosis was 84%. After regular balloon angioplasty, tissue compression and dissection of various sizes and layers were detected. After balloon angioplasty (with a mean balloon size of 5.4 mm), the minimum lumen area (from 1.7 ± 0.6 to 11.4 ± 5.3 mm2, p < 0.01) and the minimum in-stent area (12.7 ± 2.6 to 21.8 ± 5.0 mm2, p < 0.01) showed a significant increase. Additional stent was placed in one lesion that developed into a flap by dissection after balloon angioplasty. In another lesion in which sufficient dilatation was not achieved by balloon angioplasty, a major stroke occurred by acute occlusion of the ISR lesion 10 months later. CONCLUSIONS: OCT can detect the in-stent architecture of ISR lesions after balloon angioplasty and additional stent placement. However, which dissection should be treated by additional stent remain problematic.

Arid5a/IL-6/PAI-1 Signaling Is Involved in the Pathogenesis of Lipopolysaccharide-Induced Kidney Injury.

A systemic inflammatory response leads to widespread organ dysfunction, such as kidney dysfunction. Plasminogen activator inhibitor-1 (PAI-1) is involved in the pathogenesis of inflammatory kidney injury; however, the regulatory mechanism of PAI-1 in injured kidneys remains unclear. PAI-1 is induced by interleukin (IL)-6 in patients with sepsis. In addition, the stabilization of IL-6 is regulated by the adenine-thymine-rich interactive domain-containing protein 5a (Arid5a). Therefore, the aim of the present study was to examine the involvement of Arid5a/IL-6/PAI-1 signaling in lipopolysaccharide (LPS)-induced inflammatory kidney injury. LPS treatment to C57BL/6J mice upregulated Pai-1 mRNA in the kidneys. Enzyme-linked immunosorbent assay (ELISA) revealed that PAI-1 expression was induced in the culture supernatants of LPS-treated human umbilical vein endothelial cells, but not in those of LPS-treated human kidney 2 (HK-2) cells, a tubular cell line. Combined with single-cell analysis, endothelial cells were found to be responsible for PAI-1 elevation in LPS-treated kidneys. Administration of TM5441, a PAI-1 inhibitor, reduced the urinary albumin/creatinine ratio, concomitant with downregulation of Il-6 and Arid5a mRNA expressions. IL-6 treatment in LPS model mice further upregulated Pai-1 mRNA expression compared with LPS alone, accompanied by renal impairment. Furthermore, the expression of Il-6 and Pai-1 mRNA was lower in Arid5a knockout mice than in wild-type mice after LPS treatment. Taken together, the vicious cycle of Arid5a/IL-6/PAI-1 signaling is involved in LPS-induced kidney injury.

Genetically proxied HTRA1 protease activity and circulating levels independently predict risk of ischemic stroke and coronary artery disease.

HTRA1 has emerged as a major risk gene for stroke and cerebral small vessel disease with both rare and common variants contributing to disease risk. However, the precise mechanisms mediating this risk remain largely unknown as does the full spectrum of phenotypes associated with genetic variation in HTRA1 in the general population. Using a family-history informed approach, we first show that rare variants in HTRA1 are linked to ischemic stroke in 425,338 European individuals from the UK Biobank with replication in 143,149 individuals from the Biobank Japan. Integrating data from biochemical experiments on 76 mutations occurring in the UK Biobank, we next show that rare variants causing loss of protease function in vitro associate with ischemic stroke, coronary artery disease, and skeletal traits. In addition, a common causal variant (rs2672592) modulating circulating HTRA1 mRNA and protein levels enhances the risk of ischemic stroke, small vessel stroke, and coronary artery disease while lowering the risk of migraine and age-related macular dystrophy in GWAS and UK Biobank data from > 2,000,000 individuals. There was no evidence of an interaction between genetically proxied HTRA1 activity and levels. Our findings demonstrate a central role of HTRA1 for human disease including stroke and coronary artery disease and identify two independent mechanisms that might qualify as targets for future therapeutic interventions.

Photoclick reaction for rapid and simple fluorescence detection of itaconic acid and its derivatives in fungal cultures.

Itaconic acid (IA) and its derivatives produced by fungi have significant potential as industrial feedstocks. We recently developed a method for the detection of these compounds based on their terminal C-C double bonds. However, the presence of reducing agents, such as glucose and other fungal metabolites, leads to undesirable side reactions, and consequently, deteriorates the detection specificity. Therefore, we developed a fluorescence detection method for IA and its derivatives underpinned by a photoclick reaction. The photoclick reaction between conjugated IA and 5-(4-methoxyphenyl)-2-phenyl-2H-tetrazole under UV irradiation affords a fluorescent product. No fluorescence was detected when succinic acid was subjected to the reaction, indicating that a terminal C-C double bond is required to induce fluorescence. Optimal reaction conditions were determined to be a combination of 80% final dimethyl sulfoxide concentration, 30-s UV irradiation, and a pH of 2. Two weeks after the reaction at 4 °C, 89.0% of the initial intensity was retained, indicating that the reaction product was relatively stable. Glucose and kojic acid did not induce fluorescence after the reaction, indicating that these reducing agents did not affect fluorescence. IA was detected in a culture of Aspergillus terreus, and its quantification using the photoclick reaction was in agreement with the results obtained using high-performance liquid chromatography analysis. Interestingly, the IA derivative avenaciolide present in submillimolar quantities was also detectable in a culture of Aspergillus avenaceus using this method. The established method will enable the development of high-throughput screening methods to identify fungi that produce IA and its derivatives.

Upregulation of OASIS/CREB3L1 in podocytes contributes to the disturbance of kidney homeostasis.

Podocyte injury is involved in the onset and progression of various kidney diseases. We previously demonstrated that the transcription factor, old astrocyte specifically induced substance (OASIS) in myofibroblasts, contributes to kidney fibrosis, as a novel role of OASIS in the kidneys. Importantly, we found that OASIS is also expressed in podocytes; however, the pathophysiological significance of OASIS in podocytes remains unknown. Upon lipopolysaccharide (LPS) treatment, there is an increase in OASIS in murine podocytes. Enhanced serum creatinine levels and tubular injury, but not albuminuria and podocyte injury, are attenuated upon podocyte-restricted OASIS knockout in LPS-treated mice, as well as diabetic mice. The protective effects of podocyte-specific OASIS deficiency on tubular injury are mediated by protein kinase C iota (PRKCI/PKCι), which is negatively regulated by OASIS in podocytes. Furthermore, podocyte-restricted OASIS transgenic mice show tubular injury and tubulointerstitial fibrosis, with severe albuminuria and podocyte degeneration. Finally, there is an increase in OASIS-positive podocytes in the glomeruli of patients with minimal change nephrotic syndrome and diabetic nephropathy. Taken together, OASIS in podocytes contributes to podocyte and/or tubular injury, in part through decreased PRKCI. The induction of OASIS in podocytes is a critical event for the disturbance of kidney homeostasis.

Disruption of piRNA machinery by deletion of ASZ1/GASZ results in the expression of aberrant chimeric transcripts in gonocytes.

In the male germline, the machinery to repress retrotransposons that threaten genomic integrity via the piRNA pathway is established in gonocytes. It has been reported that disruption of the piRNA pathway leads to activation of retrotransposons and arrests spermatogenesis before it enters the second meiosis; however, its effects on gonocytes have not been fully elucidated. In this study, we analyzed the effects of Asz1 deletion, which is a crucial component of the piRNA pathway, on the gonocyte transcriptome. In Asz1-null gonocytes, MIWI2, which is responsible for introducing DNA methylation to retrotransposons in a piRNA-dependent manner, disappeared from the nuclei of fetal gonocytes. Transcriptome analysis revealed that retrotransposons targeted by the piRNA pathway and non-annotated transcript variants were upregulated in gonocytes from neonatal Asz1-/- mice. These non-annotated transcript variants were chimeras generated by joining exons transcribed from retrotransposons and canonical genes. DNA methylation analysis showed that retrotransposons that induce the expression of aberrant chimeric transcripts are not fully methylated. This was consistent with the impaired nuclear localization of MIWI2 in Asz1-null gonocytes. Furthermore, heterogeneity of DNA methylation status in retrotransposons was observed in both gonocytes and their descendants. This suggests that the piRNA system in gonocytes can potentially prevent spermatogenic cell populations bearing aberrant chimeric transcripts from propagating later in spermatogenesis. In conclusion, Asz1 is required to repress retrotransposons and retrotransposon-driven aberrant chimeric transcripts in gonocytes through the piRNA pathway.

Thermal desorption gas chromatography-mass spectrometric analysis of polycyclic aromatic hydrocarbons in atmospheric fine particulate matter.

Thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS) is used to analyze polycyclic aromatic hydrocarbons (PAHs) in atmospheric fine particulate matter. However, despite the high sensitivity of TD-GC-MS, the recovery rate of PAHs is greatly influenced by active sites in the equipment. PAHs are decomposed or adsorbed at active sites, decreasing quantitative accuracy. Also, the thermal extraction of PAHs is easily affected by the matrix in PM2.5 samples, decreasing the thermal extraction efficiency. Herein, the analytical sensitivities of PAHs were improved by adding analyte protectant (AP) and thermal desorption aid (TDA) as an auxiliary agent. The combination of 2 µL of 0.5 w/v% D-sorbitol (as AP) and 2 µL of 10 w/v% Tween®20 (as TDA) was found to be most effective in improving the analytical sensitivity of PAHs. The sensitivities of 5-6-ring PAHs with high boiling points increased most when analyzing blank filter papers added with PAHs standard sample or real samples of PM2.5 compared with the samples without the auxiliary agent. When analyzing real samples of PM2.5, the peak areas of 5-ring and 6-ring PAHs in the PM2.5 sample added with the optimized auxiliary agent were 1.40 and 1.96 times that without the auxiliary agent. It is considered that AP in the auxiliary agent covered active sites and protected PAHs undergoing decomposition or adsorption. TDA improved the thermal extraction rate of high boiling point PAHs. When using alternative heat sampling equipment to analyze low concentrations of high boiling point components, the auxiliary agent proposed herein can increase the analytical sensitivity toward the target compounds.

Thermal Extraction of Polycyclic Aromatic Hydrocarbons from Atmospheric 2.5 µm Particulate Matter Collected on a Filter Paper Using a High-Temperature Headspace Method.

Recently, owing to the performance improvement of the headspace (HS)-sampling devices and its consumables, HS vial samples can be analyzed at temperatures up to 300°C. Some studies have attempted to analyze polycyclic aromatic hydrocarbons (PAHs) in atmospheric 2.5 µm particulate matter (PM 2.5) collected on a filter paper by gas chromatography/mass spectrometry (GC/MS) coupled with thermal desorption device. However, no studies have reported the use of an HS-sampling device to quantify PAHs in PM 2.5 filter paper. In this study, we found that the quantification of PAH analysis using HS-GC/MS can be improved by the following steps, so that the accuracy becomes almost the same as that of a solvent-extraction method: 1) replacement of the air in the HS vial with nitrogen, 2) limiting the solvent to toluene, 3) using the hydrolyzed polyimide-covered septum, and 4) optimization of the heating temperature and heating time of the HS vial. As a result, we succeeded in protecting PAHs in an HS vial at a high temperature and in creating an analysis method with a high recovery rate and high repeatability; the limit of quantitation of each PAH in this method was 5.4 pg m-3 in the case of a volume of 10080 m3 of air being collected on the filter paper.

Optimization of computed tomography contrast studies with a new, simple dosing regimen incorporating body size: examination of contrast effects in the thoracoabdominal aorta.

The dosage of contrast agents for computed tomography contrast studies is calculated based on the parameter of actual body weight (ABW) to ensure reproducibility. The use of lean body weight (LBW) and adjustment for physique (lean or obese) improves accuracy. However, this method is complex, because LBW is not a general body parameter and requires a special device to measure. To solve this problem, contrast body weight (CBW), has been proposed as a new and simple parameter that considers physique. CBW is calculated by determining the blood volume ratio based on body height, ABW, and sex and can potentially correct for body size. It can be calculated by entering a formula in a Microsoft Excel sheet. Since CBW can be easily obtained using this general tool, we decided to compare the two body parameters of ABW and CBW. We compared ABW and CBW and demonstrated a higher correlation between CBW-based dosing and the amount of iodine used per body weight than with ABW-based dosing. CBW-based dosing allows correction for body size. This indicates that contrast enhancement over a spectrum of lean or obese examinees can be linearly evaluated. To date, this method has shown good results.

Bonding-Based Wafer-Level Vacuum Packaging Using Atomic Hydrogen Pre-Treated Cu Bonding Frames.

A novel surface activation technology for Cu-Cu bonding-based wafer-level vacuum packaging using hot-wire-generated atomic hydrogen treatment was developed. Vacuum sealing temperature at 300 °C was achieved by atomic hydrogen pre-treatment for Cu native oxide reduction, while 350 °C was needed by the conventional wet chemical oxide reduction procedure. A remote-type hot-wire tool was employed to minimize substrate overheating by thermal emission from the hot-wire. The maximum substrate temperature during the pre-treatment is lower than the temperature of Cu nano-grain re-crystallization, which enhances Cu atomic diffusion during the bonding process. Even after 24 h wafer storage in atmospheric conditions after atomic hydrogen irradiation, low-temperature vacuum sealing was achieved because surface hydrogen species grown by the atomic hydrogen treatment suppressed re-oxidation. Vacuum sealing yield, pressure in the sealed cavity and bonding shear strength by atomic hydrogen pre-treated Cu-Cu bonding are 90%, 5 kPa and 100 MPa, respectively, which are equivalent to conventional Cu-Cu bonding at higher temperature. Leak rate of the bonded device is less than 10-14 Pa m³ s-1 order, which is applicable for practical use. The developed technology can contribute to low-temperature hermetic packaging.

Preoperative assessment of pleural adhesion by Four-Dimensional Ultra-Low-Dose Computed Tomography (4D-ULDCT) with Adaptive Iterative Dose Reduction using Three-Dimensional processing (AIDR-3D).

PURPOSE: To assess the feasibility of Four-Dimensional Ultra-Low-Dose Computed Tomography (4D-ULDCT) for distinguishing pleural aspects with localized pleural adhesion (LPA) from those without. METHODS: Twenty-seven patients underwent 4D-ULDCT during a single respiration with a 16cm-coverage of the body axis. The presence and severity of LPA was confirmed by their intraoperative thoracoscopic findings. A point on the pleura and a corresponding point on the outer edge of the costal bone were placed in identical axial planes at end-inspiration. The distance of the two points (PCD), traced by automatic tracking functions respectively, was calculated at each respiratory phase. The maximal and average change amounts in PCD (PCDMCA and PCDACA) were compared among 110 measurement points (MPs) without LPA, 16MPs with mild LPA and 10MPs with severe LPA in upper lung field cranial to the bronchial bifurcation (ULF), and 150MPs without LPA, 17MPs with mild LPA and 9MPs with severe LPA in lower lung field caudal to the bronchial bifurcation (LLF) using the Mann-Whitney U test. RESULTS: In the LLF, PCDACA as well as PCDMCA demonstrated a significant difference among non-LPA, mild LPA and severe LPA (18.1±9.2, 12.3±6.2 and 5.0±3.3mm) (p<0.05). Also in the ULF, PCDACA showed a significant difference among three conditions (9.2±5.5, 5.7±2.8 and 2.2±0.4mm, respectively) (p<0.05), whereas PCDMCA for mild LPA was similar to that for non-LPA (12.3±5.9 and 17.5±11.0mm). CONCLUSIONS: Four D-ULDCT could be a useful non-invasive preoperative assessment modality for the detection of the presence or severity of LPA.

Investigation of Surface Pre-Treatment Methods for Wafer-Level Cu-Cu Thermo-Compression Bonding.

To increase the yield of the wafer-level Cu-Cu thermo-compression bonding method, certain surface pre-treatment methods for Cu are studied which can be exposed to the atmosphere before bonding. To inhibit re-oxidation under atmospheric conditions, the reduced pure Cu surface is treated by H2/Ar plasma, NH3 plasma and thiol solution, respectively, and is covered by Cu hydride, Cu nitride and a self-assembled monolayer (SAM) accordingly. A pair of the treated wafers is then bonded by the thermo-compression bonding method, and evaluated by the tensile test. Results show that the bond strengths of the wafers treated by NH3 plasma and SAM are not sufficient due to the remaining surface protection layers such as Cu nitride and SAMs resulting from the pre-treatment. In contrast, the H2/Ar plasma⁻treated wafer showed the same strength as the one with formic acid vapor treatment, even when exposed to the atmosphere for 30 min. In the thermal desorption spectroscopy (TDS) measurement of the H2/Ar plasma⁻treated Cu sample, the total number of the detected H2 was 3.1 times more than the citric acid⁻treated one. Results of the TDS measurement indicate that the modified Cu surface is terminated by chemisorbed hydrogen atoms, which leads to high bonding strength.

Adaptive Advantage of Myrmecochory in the Ant-Dispersed Herb Lamium amplexicaule (Lamiaceae): Predation Avoidance through the Deterrence of Post-Dispersal Seed Predators.

Seed dispersal by ants (myrmecochory) is found worldwide, but the benefits that plants obtain from this mutualism remain uncertain. In the present study, we conducted laboratory experiments to demonstrate seed predator avoidance as a benefit of myrmecochory using the annual ant-dispersed herb Lamium amplexicaule, the disperser ant Tetramorium tsushimae, and the seed predatory burrower bug Adomerus rotundus. We compared the predation intensity of Lamium amplexicaule seeds by Adomerus rotundus under the presence or absence of Tetramorium tsushimae. Both the number of seeds sucked by Adomerus rotundus adults and the feeding duration of sucked seeds by nymphs were significantly reduced in the presence of ants. This effect was most likely due to the behavioral alteration of Adomerus rotundus in response to the ant presence, because ants seldom predated Adomerus rotundus during the experiment. Our results demonstrated that the presence of ants decreases post-dispersal seed predation, even when the ants do not bury the seeds. The present study thus suggests that the non-consumptive effects of ants on seed predators benefit myrmecochorous plants.

Primitive neuroectodermal tumor of the transverse colonic mesentery defined by the presence of EWS-FLI1 chimeric mRNA in a Japanese woman.

We report a case of primitive neuroectodermal tumor (PNET) arising in the transverse colonic mesentery. A 24-year-old Japanese woman was admitted to Kagoshima City Hospital with complaints of abdominal pain and sensations of abdominal fullness of 5 months' duration. On palpation, a mass the size of an infant's head was noted in the right flank. Abdominal computed tomography (CT) and ultrasonography showed a huge mass that consisted of multiple cystic components. On arteriography, a slight tumor stain appeared, with stretched and displaced tributaries of the right colic and middle colic arteries. Barium swallow examination demonstrated that the ascending colon was shifted to the right and small intestine to the left. We performed an en-bloc resection of the tumor in the transverse colonic mesentery, including the ascending colon, proximal jejunum (20 cm in length), and greater omentum. The resected tumor was 12 x 10 x 7 cm in size, 590g in weight, elastic soft in consistency, and multicystic. Histologically, the specimens showed a sheet-like proliferation of spindle-to-polygonal cells, and focally, the tumor formed rosette structures. Immunohistochemically, the tumor cells were positive for neuron-specific enolase (NSE) and mic-2. EWS-FLI1 chimeric mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Based on the above findings, we finally diagnosed the tumor as PNET of the colonic mesentery. There has been no recurrence for 20 months after operation. PNET arising in the mesentery is very rare, and we distinguished PNET from other tumors by immunohistochemical examination and by demonstration of the presence of EWS-FLI1 chimeric mRNA in the tumor.

Recurrent nonocclusive mesenteric ischemia after resection of iliac artery aneurysm.

A case of recurrent nonocclusive mesenteric ischemia in a patient with isolated internal iliac artery aneurysm penetrating the sigmoid colon is described. On the day after the aneurysm and the sigmoid colon had been resected, the patient developed necrosis of the left hemicolon. Fourteen and nineteen days after left hemicolectomy, massive intestinal bleedings occurred, requiring ileectomy. On the basis of operative findings of good pulsation of visceral arterial branches; angiography showing patent mesenteric vessels with some spasms; and pathological findings suggesting mesenteric ischemia, these ischemic events were diagnosed as nonocclusive mesenteric ischemia. Low-output syndrome induced by massive intestinal bleeding and atrial fibrillation and sepsis were responsible for the establishment of the nonocclusive mesenteric ischemia. Development of disseminated intravascular coagulation and continuous administration of diuretics for acute renal failure seemed to have further perturbed the mesenteric circulation. The patient died of subsequent multiple organ failure 4 months after the first operation. We should pay more attention to nonocclusive mesenteric ischemia in patients with mesenteric ischemia, and strict circulatory management during the perioperative period is essential in these patients.