Modified Intravascular Stent for Microvascular Suture in a Rat Superficial Femoral Artery.

BACKGROUND: Intravascular stent (IVaS) is sometimes used for suturing small vessels, but removing the stent after suturing is difficult. To overcome this problem, we developed an IVaS that integrates a stent and a manipulating string. This study aimed to investigate the usefulness of the modified IVaS (M-IVaS) by comparing it with conventional sutures (CS) and conventional IVaS (C-IVaS). METHODS: Forty-five superficial femoral arteries from rats were resected and sutured. The rats were randomly divided into the following 3 groups: CS, M-IVaS, and C-IVaS, with 15 rats per group. Patency rate, operating time, and ultrasonographic blood flow dynamics were examined immediately after suturing. Patency tests, ultrasonographic evaluations, and histological investigations were performed 1 week (n = 5), 2 weeks (n = 5), and 6 weeks (n = 5) after surgery. RESULTS: The 3 groups showed vessel patency in all cases immediately after suturing and at 1 week, 2 weeks, and 6 weeks after surgery. The mean operative time was 22.6 minutes for the CS group, 21.5 minutes for the M-IVaS group, and 25.9 minutes for the C-IVaS group. There were no significant differences in peak flow velocity and stenosis rate among the 3 groups as evaluated by ultrasonography. Histopathological evaluation revealed a similar recovery process of endothelial cells and no damage to the vascular wall. CONCLUSION: The surgical time using M-IVaS was significantly shorter compared to that using C-IVaS. The M-IVaS reduced the inconvenience of C-IVaS removal. M-IVaS showed the same effectiveness as did the CS in terms of patency rate, operating time, ultrasonographic blood flow dynamics, and histological evaluation. M-IVaS can be used in the field of microsurgery.

Longitudinal Analysis of Mitochondrial Function in a Choline-Deficient L-Amino Acid-Defined High-Fat Diet-Induced Metabolic Dysfunction-Associated Steatohepatitis Mouse Model.

Metabolic dysfunction-associated fatty liver disease (MAFLD) is one of the most common chronic liver diseases worldwide. Some patients with MAFLD develop metabolic dysfunction-associated steatohepatitis (MASH), which can lead to severe liver fibrosis. However, the molecular mechanisms underlying this progression remain unknown, and no effective treatment for MASH has been developed so far. In this study, we performed a longitudinal detailed analysis of mitochondria in the livers of choline-deficient, methionine-defined, high-fat-diet (CDAHFD)-fed mice, which exhibited a MASH-like pathology. We found that FoF1-ATPase activity began to decrease in the mitochondria of CDAHFD-fed mice prior to alterations in the activity of mitochondrial respiratory chain complex, almost at the time of onset of liver fibrosis. In addition, the decrease in FoF1-ATPase activity coincided with the accelerated opening of the mitochondrial permeability transition pore (PTP), for which FoF1-ATPase might be a major component or regulator. As fibrosis progressed, mitochondrial permeability transition (PT) induced in CDAHFD-fed mice became less sensitive to cyclosporine A, a specific PT inhibitor. These results suggest that episodes of fibrosis might be related to the disruption of mitochondrial function via PTP opening, which is triggered by functional changes in FoF1-ATPase. These novel findings could help elucidate the pathogenesis of MASH and lead to the development of new therapeutic strategies.

The piggyBac-Based Gene Delivery System Can Confer Successful Production of Cloned Porcine Blastocysts with Multigene Constructs.

The introduction of multigene constructs into single cells is important for improving the performance of domestic animals, as well as understanding basic biological processes. In particular, multigene constructs allow the engineering and integration of multiple genes related to xenotransplantation into the porcine genome. The piggyBac (PB) transposon system allows multiple genes to be stably integrated into target genomes through a single transfection event. However, to our knowledge, no attempt to introduce multiple genes into a porcine genome has been made using this system. In this study, we simultaneously introduced seven transposons into a single porcine embryonic fibroblast (PEF). PEFs were transfected with seven transposons containing genes for five drug resistance proteins and two (red and green) fluorescent proteins, together with a PB transposase expression vector, pTrans (experimental group). The above seven transposons (without pTrans) were transfected concomitantly (control group). Selection of these transfected cells in the presence of multiple selection drugs resulted in the survival of several clones derived from the experimental group, but not from the control. PCR analysis demonstrated that approximately 90% (12/13 tested) of the surviving clones possessed all of the introduced transposons. Splinkerette PCR demonstrated that the transposons were inserted through the TTAA target sites of PB. Somatic cell nuclear transfer (SCNT) using a PEF clone with multigene constructs demonstrated successful production of cloned blastocysts expressing both red and green fluorescence. These results indicate the feasibility of this PB-mediated method for simultaneous transfer of multigene constructs into the porcine cell genome, which is useful for production of cloned transgenic pigs expressing multiple transgenes.

Extraction spectrophotometry using a lithium-ion selective metallacrown: temperature effect on extraction reaction and application to determination of lithium in serum and seawater.

A metallacrown-type ionophore, 2,3-pyridinediolate-bridged (3,5-dimethylanisole)ruthenium trinuclear complex, has a high extraction selectivity for Li+, but the extraction reaction is very slow. To solve this problem, the effect of temperature on the rapidity and equilibrium of the extraction of Li+ and Na+ as picrates from water to toluene with the metallacrown was investigated in this study. While the extraction of Li+ requires 6 h of shaking for equilibration at 25 °C, the distribution ratio becomes nearly constant after 4 h and 2 h of shaking at 37 °C and 50 °C, respectively. The extraction equilibrium constants (Kex) and associated thermodynamic parameters determined for Li+ and Na+ indicate that the extraction reactions are exothermic and enthalpy-driven: ΔH° = - 53 kJ/mol, ΔS° = - 0.03 kJ/(mol K) for Li+; ΔH° = - 28 kJ/mol, ΔS° = - 0.03 kJ/(mol K) for Na+. Although the extraction ability for Li+ and selectivity for Li+/Na+ decrease with increasing temperature, the values of Kex and Kex(Li+)/Kex(Na+) are 1.0 × 107 and 1.3 × 104, respectively, even at 50 °C, indicating that both are sufficiently high. In the determination of Li+ by extraction spectrophotometry using this metallacrown, extraction at 50 °C for 2 h was employed to speed up the analysis. The method was applied to seawater and serum samples containing a large amount of coexisting ions such as Na+ and Mg2+, and trace amounts (10-6-10-5 mol/L order) of Li+ in microvolume samples (sub-mL order) could be successfully determined.

Alaska Pollock-derived Gelatin Sealant has Higher Sealing Strength than, and Comparable Biocompatibility with, Fibrin Sealant in Porcine and Rat Dural Injury Models.

STUDY DESIGN: Burst strength study in porcine dural models and functional and histological study in rat dural models. OBJECTIVE: This study aimed to investigate the sealing strength and biocompatibility of Alaska pollock-derived gelatin (ApGltn) and fibrin sealants in disrupted dural injuries. SUMMARY OF BACKGROUND DATA: Disruption of the dura mater occurs during spine surgery, leading to cerebrospinal fluid leakage. Fibrin sealant is usually applied to ruptured sites; however, it lacks sealing strength. A novel biocompatible sealant composed of ApGltn was recently demonstrated to have good burst strength and biocompatibility in the porcine aorta. METHODS: Ten porcine dura maters with central holes were covered with ApGltn and fibrin sealants (five samples per group). The maximum burst strength of each sealant was measured, and histological examination was performed after burst testing. Twenty-seven dura maters of male Wistar rats were used for functional and histopathological evaluations. The rats were treated with three surgical interventions: defect + ApGltn sealant; defect + fibrin sealant; defect alone (nine rats per group). Macroscopic confirmation of the sealant, hindlimb motor function analysis, and histopathological examination were performed at two, four, and eight weeks after the procedure. RESULTS: The maximum burst strength of the ApGltn sealant was ~4.4 times higher than that of the fibrin sealant (68.1±12.1 vs . 15.6±8.7 mmHg; P <0.001). Histological examination confirmed that the ApGltn sealant showed tight adhesion to the dural surface, whereas a gap was observed between the fibrin sealant and the dura mater. In the rat model, the ApGltn sealant resulted in spinal function and dural histological findings similar to those of the fibrin sealant. CONCLUSION: The ApGltn sealant had a higher sealing strength than, and comparable effect on dura regeneration with, the fibrin sealant.

Percutaneous Coronary Intervention Strategy for Spontaneous Coronary Artery Dissection of Left Main Coronary Artery with Extensive Intramural Hematoma in the Main Side Branch.

A 46-year-old pregnant woman, presented with worsening episodes of intermittent chest pain. The patient was diagnosed with a non-ST-elevation myocardial infarction. On arrival, she had a stable hemodynamic status without chest pain. She was initially treated with conservative medical therapy. One day later, she complained of severe chest pain, and an electrocardiogram showed ST elevation in leads I, aVL, and V2-5. Emergency coronary angiography showed total occlusion of the left anterior descending artery (LAD) and intermediate stenosis of the left main coronary artery (LMCA). The intravascular ultrasound (IVUS) revealed an intramural hematoma (IMH) from the LMCA to the LAD, extending to the left circumflex artery (LCX) ostium. This finding was consistent with spontaneous coronary artery dissection (SCAD). After stent implantation from the LMCA to the LAD, severe stenosis was noted at the proximal site of the LCX. IVUS showed that the IMH extended to the LCX. The provisional crush stent technique was performed, and the final angiography revealed satisfactory results with thrombolysis in myocardial infarction flow grade 3 in the LAD and LCX. This case report highlighted that stent implantation in the SCAD lesions facilitated the extension of the IMH longitudinally and laterally into the side branch, resulting in stenosis or occlusion. Therefore, the side branch should be evaluated using IVUS before stent implantation. In cases where the IMH extends to the ostium of the side branch, two-stent techniques that do not require guidewire recrossing, such as crush stents, should be considered to avoid side branch occlusion.

Distinction of surgically resected gastrointestinal stromal tumor by near-infrared hyperspectral imaging.

The diagnosis of gastrointestinal stromal tumor (GIST) using conventional endoscopy is difficult because submucosal tumor (SMT) lesions like GIST are covered by a mucosal layer. Near-infrared hyperspectral imaging (NIR-HSI) can obtain optical information from deep inside tissues. However, far less progress has been made in the development of techniques for distinguishing deep lesions like GIST. This study aimed to investigate whether NIR-HSI is suitable for distinguishing deep SMT lesions. In this study, 12 gastric GIST lesions were surgically resected and imaged with an NIR hyperspectral camera from the aspect of the mucosal surface. Thus, the images were obtained ex-vivo. The site of the GIST was defined by a pathologist using the NIR image to prepare training data for normal and GIST regions. A machine learning algorithm, support vector machine, was then used to predict normal and GIST regions. Results were displayed using color-coded regions. Although 7 specimens had a mucosal layer (thickness 0.4-2.5 mm) covering the GIST lesion, NIR-HSI analysis by machine learning showed normal and GIST regions as color-coded areas. The specificity, sensitivity, and accuracy of the results were 73.0%, 91.3%, and 86.1%, respectively. The study suggests that NIR-HSI analysis may potentially help distinguish deep lesions.

Functional analysis of coiled-coil domains of MCU in mitochondrial calcium uptake.

The mitochondrial calcium uniporter (MCU) complex is a highly-selective calcium channel. This complex consists of MCU, mitochondrial calcium uptake proteins (MICUs), MCU regulator 1 (MCUR1), essential MCU regulator element (EMRE), etc. MCU, which is the pore-forming subunit, has 2 highly conserved coiled-coil domains (CC1 and CC2); however, their functional roles are unknown. The yeast expression system of mammalian MCU and EMRE enables precise reconstitution of the properties of the mammalian MCU complex in yeast mitochondria. Using the yeast expression system, we here showed that, when MCU mutant lacking CC1 or CC2 was expressed together with EMRE in yeast, their mitochondrial Ca2+-uptake function was lost. Additionally, point mutations in CC1 or CC2, which were expected to prevent the formation of the coiled coil, also disrupted the Ca2+-uptake function. Thus, it is essential for the Ca2+ uptake function of MCU that the coiled-coil structure be formed in CC1 and CC2. The loss of function of those mutated MCUs was also observed in the mitochondria of a yeast strain lacking the yeast MCUR1 homolog. Also, in the D. discoideum MCU, which has EMRE-independent Ca2+-uptake function, the deletion of either CC1 or CC2 caused the loss of function. These results indicated that the critical functions of CC1 and CC2 were independent of other regulatory subunits such as MCUR1 and EMRE, suggesting that CC1 and CC2 might be essential for pore formation by MCUs themselves. Based on the tetrameric structure of MCU, we discussed the functional roles of the coiled-coil domains of MCU.

Two cases of numb chin syndrome diagnosed as malignant disease.

Numb chin syndrome (NCS) is defined as reduced or absent sensation in an area of the chin and lower lip within the distribution of the mental or inferior alveolar nerves. The causes of NCS may be neoplastic, traumatic, dental, toxic, drug-induced, inflammatory, autoimmune or infectious. NCS may be the preliminary symptom of malignancy or recurrence/metastasis in patients with cancer. Therefore, the occurrence of NCS warrants careful examination and monitoring of such patients. This article presents two cases of NCS reported in a patient with prostate cancer and in a patient with Burkitt lymphoma/leukaemia.

Dynamic measurement of surface strain distribution on the foot during walking.

To clarify the mechanism underlying the development of foot disorders such as diabetic ulcers and deformities, it is important to understand how the foot surface elongates and contracts during gait. Such information is also helpful for improving the prevention and treatment of foot disorders. We therefore measured temporal changes in the strain distribution on the foot surface during human walking. Five adult male participants walked across a glass platform placed over an angled mirror set in a wooden walkway at a self-selected speed and the dorsolateral and plantar surfaces of the foot were filmed using two pairs of synchronized high-speed cameras. Three-dimensional (3D) digital image correlation was used to quantify the spatial strain distribution on the foot surface with respect to that during quiet standing. Using the proposed method, we observed the 3D patterns of foot surface strain distribution during walking. Large strain was generated around the ball on the plantar surface of the foot throughout the entire stance phase, due to the windlass mechanism. The dorsal surface around the cuboid was stretched in the late stance phase, possibly due to lateral protruding movement of the cuboid. It may be possible to use this technique to non-invasively estimate movements of the foot bones under the skin using the surface strain distribution. The proposed technique may be an effective tool with which to analyze foot deformation in the fields of diabetology, clinical orthopedics, and ergonomics.

Birth of Cloned Microminipigs Derived from Somatic Cell Nuclear Transfer Embryos That Have Been Transiently Treated with Valproic Acid.

In our previous study, we found that treatment of miniature pig somatic cell nuclear transfer (SCNT) embryos with 4 mM valproic acid (VPA), a histone deacetylase inhibitor, for 48 hours after activation enhanced blastocyst formation rate and octamer-binding transcription factor-3/4 (Oct-3/4) gene expression at the late blastocyst stage; however, the production of viable cloned pups failed, when those VPA-treated SCNT embryos were transferred to recipients. This failure suggests that the present VPA treatment is suboptimal. In the present study, we explored the optimal conditions for VPA to have beneficial effects on the development of SCNT embryos. When miniature pig SCNT embryos were treated with 8 mM VPA for 24 hours after activation, both the rates of blastocyst formation and blastocysts expressing the Oct-3/4 gene were significantly (p < 0.05) improved. A similar increase in blastocyst formation was also observed when microminipig-derived cells were used as SCNT donors. Five cloned piglets were obtained after the transfer of 152 microminipig SCNT embryos that had been treated with 8 mM VPA for 24 hours. The results indicated that a short duration of treatment with VPA improves the development of both miniature pig and microminipig SCNT embryos, possibly via an enhanced reprogramming mechanism.

Logic operations of chemically assembled single-electron transistor.

Double-gate single-electron transistors (SETs) were fabricated by chemical assembling using electroless gold-plated nanogap electrodes and chemisorbed chemically synthesized gold nanoparticles. The fabricated SET showed periodic and stable Coulomb oscillations under application of voltages of both gates. The sole SET also exhibited all two-input logic operations-XOR, XNOR, NAND, OR, NOR, and AND-with an on/off ratio of 10(2). This demonstrates the potential of chemical assembling to give highly stable SETs exhibiting all logic operations.

Ideal discrete energy levels in synthesized Au nanoparticles for chemically assembled single-electron transistors.

Ideal discrete energy levels in synthesized Au nanoparticles (6.2 ± 0.8 nm) for a chemically assembled single-electron transistor (SET) are demonstrated at 300 mK. The spatial structure of the double-gate SET is determined by two gate and drain voltages dependence of the stability diagram, and electron transport to the Coulomb box of a single, nearby Coulomb island of Au nanoparticles is detected by the SET. The SET exhibits discrete energy levels, and the excited energy level spacing of the Coulomb island is evaluated as 0.73 meV, which well corresponds to the expected theoretical value. The discrete energy levels show magnetic field evolution with the Zeeman effect and dependence on the odd-even electron number of a single Au nanoparticle.