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Accurate species identification, based on DNA barcoding, can be achieved when sufficient sequence variations are present amongst species in the sampled marker. In general, the ability to discriminate species decreases with shorter sequences; however, shorter regions have a merit in amplification success by the polymerase chain reaction. In either case, it is important to investigate sequence variations amongst species before barcoding to understand its reliability and limitations. In this study, we investigate how accurately short, but hypervariable portion of the mitochondrial 12S ribosomal RNA (12S) gene (MiFish region with approximately 180 bp) is used to identify each species in diversified pomacentrid fishes compared with the longer region of the same gene (approximately 750 bp). We prepared three datasets with 301 sequences of the MiFish region for 150 species, the same 301 of sequences of the longer 12S region and 476 sequences of the MiFish region for 183 species. Neighbour-joining (NJ) analyses and genetic distance analyses revealed several indistinguishable pairs of species in these DNA regions. Although the number of such pairs was larger in the MiFish region, 83.6% (153 of 183) of species possessed respective unique sequences even in the MiFish region (versus 96.0% [144 of 150 species] in the longer 12S region). A part of indistinguishable pairs of species might have caused by mitochondrial DNA introgressions and taxonomically unresolved problems. Our analysis clarified the effectiveness and limitations of species identification using DNA barcoding for Pomacentridae and the sequences we provided here contribute to the expansion of references for pomacentrid mitochondrial 12S sequences.

Summary: Lenalidomide (LEN) is commonly used as an effective therapeutic agent for multiple myeloma (MM). However, in some patients, primary resistance to LEN is observed, the mechanisms of which remain poorly understood. In this study, we combined a LEN sensitivity assay with proteomics data from 15 MM cell lines to identify protein expression profiles associated with primary LEN resistance. Our findings revealed that CSN5 expression is lower in LEN-resistant cell lines than in LEN-sensitive lines. Moreover, we established that CSN5 is degraded via the cullin-RING ubiquitin ligase (CRL)-mediated ubiquitin-proteasome pathway through ubiquitination at lysine 194. Our data suggest that reduced CSN5 expression leads to abnormalities in the ubiquitination cycle of CRL4A, resulting in the inhibition of LEN-mediated degradation of IKZF1 and IKZF3. These findings delineate an additional mechanism of LEN resistance in MM cells and may contribute to the development of alternative therapeutic strategies to overcome LEN resistance.

Objectives: Adipose-derived stem cells (ADSCs) are widely used in wound care because they release a variety of cytokines. However, the molecular mechanism of paracrine action remains unclear. It has been reported that basic fibroblast growth factor (bFGF) enhances the therapeutic potential of ADSCs. In this study, we searched for cytokines whose release from ADSCs is enhanced by bFGF stimulation. Results: Quantitative RT-PCR and ELISA analyses revealed that bFGF upregulates CXCL-1 and IL-8 mRNA synthesis and secretion from ADSCs. Both cytokines showed the ability to promote important processes for wound healing, including tube formation of vascular and lymphatic endothelial cells and cell migration of fibroblasts in vitro. Conclusions: These results suggest that bFGF stimulation increases the secretion of CXCL-1 and IL-8 from ADSCs and that these cytokines may promote angiogenesis, lymphangiogenesis, and cell migration, leading to enhanced efficiency of wound healing.

Mixed-valence complexes contain two metals with different formal oxidation numbers and, therefore, show mixed properties that are influenced by the electronic coupling between the two metals, which is, in turn, regulated by a bridging ligand. This is an attractive point for many researchers. Oxalate is widely used as a bridging ligand for preparing polynuclear complexes. More than 1000 complexes have been reported until now. However, dithiooxamidate, which is an oxalate analog, is less popular as a bridging ligand. Here, a new dithiooxamidate-bridged Ni-diphosphine dinuclear complex with the formula [(μ2-toxa){Ni(dppe)}2](BF4)2 (toxa = dithiooxamidate; dppe = 1,2-bis(diphenylphosphino)ethane) was prepared and characterized via single-crystal X-ray diffraction. When using 1,3-bis(diphenylphosphino)propane (dppp) instead of dppe, dinuclear, trinuclear, and tetranuclear complexes were obtained, i.e., [(μ2-toxa){Ni(dppp)}2](BF4)2, [{μ2-Ni(toxa)2}{Ni(dppp)}2](BF4)2, and [{μ3-Ni(toxa)3}{Ni(dppp)}3](BF4)2, respectively. Bidentate toxa ligands in dinuclear complexes coordinate each Ni atom as κ(S,N). However, the trinuclear and the tetranuclear complexes have the toxa ligands with κ(N,N) and κ(S,S) coordination. The [(μ2-toxa){Ni(dppe)}2](BF4)2 complex undergoes four reversible redox processes, whose analysis via a controlled-potential absorption spectrum reveals the presence of a Ni(II)-Ni(I) mixed-valence state at ∆E1/2 = 0.22 V with a comproportionation constant of 6.1 × 103.

Nicotinamide adenine dinucleotide (NAD+) plays a pivotal role in various physiological processes within mammalian cells, including energy metabolism, redox homeostasis, and genetic regulation. In the majority of mammalian cellular contexts, NAD+ biosynthesis primarily relies on vitamin B3, including nicotinamide (NAM) and nicotinic acid (NA). The concept of NAD+ augmentation therapy has recently emerged as a promising strategy to mitigate aging-associated phenomena, termed rejuvenation. Despite the involvement of diverse enzymatic cascades in NAD+ biosynthesis, certain cellular environments exhibit deficiencies in specific enzymes, suggesting cell type-dependent variability in optimal NAD+ precursor selection. However, the optimization of NAD+ precursors for topical formulations has received scant attention thus far. In the present investigation, we sought to delineate the most efficacious precursor for augmenting NAD+ levels in human skin keratinocytes. Remarkably, NA supplementation led to a significant 1.3-fold elevation in intracellular NAD+ levels, even in the presence of nicotinamide phosphoribosyltransferase inhibition by FK866. Additionally, NA mononucleotide demonstrated a 1.5-fold increase (but not significant) in NAD+ levels following 100 μM application. Conversely, NAM and its derivatives failed to elicit a NAD+ response in keratinocytes. Notably, NA supplementation elicited up-regulation of mitochondrial superoxide dismutase (SOD2) and sirtuin 3 (SIRT3), indicative of its beneficial impact on mitochondrial function. Furthermore, NA mitigated rotenone-induced mitochondrial reactive oxygen species (ROS) accumulation. Collectively, these findings advocate for the potential utility of NA in topical applications aimed at skin rejuvenation.

This study established a rapid estimation method for the fatigue limit of laser welds of aluminum alloys in order to optimize welding conditions, an important factor in ensuring the safety and reliability of laser welding. This study focused on an estimation method of fatigue limit based on the dissipated energy. Dissipated energy is the temperature change caused by local plastic deformation. The stress amplitude at which the temperature change due to dissipated energy rapidly increased in the laser welds was defined as the estimated fatigue limit, and it was almost consistent with the fatigue limit of the joint.

Heated tobacco products are devices that deliver nicotine into the body via inhalation of the mainstream aerosols generated during direct and/or indirect heating of tobacco leaf material. Ammonia in aerosols potentially increases the alkalinity and, therefore, the proportion of free nicotine for easy absorption. Meanwhile, ammonia can be a cause of adverse health effects when involved in the aerosols. This study aimed to grasp the emission behaviour of ammonia in the mainstream aerosols generated from four kinds of devices that employ different heating temperatures from 40 to 350 °C. The aerosols were generated by a vaping machine following the CRM 81 puffing protocol. Ammonia in the forms of gas and particles was trapped in 5 mM oxalic acid and subsequently determined by ion chromatography. The results showed that the total emission amount of ammonia increased with an increase in the heating temperature regardless of the device used. The gas-particle distribution of ammonia also depended on the heating temperature; gaseous ammonia was only found in the device with 40 °C of the heating temperature. These results show that ammonia in the mainstream aerosols was emitted from a common thermal process, probably thermal extraction in water vapour from a tobacco leaf.

This study aimed to clarify the cause of rugby head and spinal cord injuries through a network centrality analysis of 14-year (2004-2018) longitudinal data in Japan. The study hypothesis is that understanding the causal relationship among the occurrence of serious injuries, the quality of player experience and play situation as a network structure could be possible to obtain practical knowledge on injury prevention. In this study, bipartite graphs are used to make it easier to understand the situation of players and injuries. This would also help to elucidate more characteristic subgroup. A network bipartite graph and subgroup (cluster) analyses were performed to clarify the injured players' experience and the cause of injury. We used the algorithm of R program, IGRAPH, clustering edge betweenness. For subgroup extraction, the modularity Q value was used to determine which step to cut. The Japanese rugby population was 93,873 (2014-2018 average), and 27% were high school students. The data showed that careful attention would be particularly needed for groups of inexperienced Japanese high school players. Our study suggests that we should consider introducing rules that prohibit "head-on collisions" in youth rugby.

Background:. Free flaps have evolved from musculocutaneous flaps to perforator-based cutaneous flaps. The subdermal plexus is now thought to play a significant role in skin paddle perfusion. We propose a new concept, the “intradermal plexus,” allowing survival of dermis flaps, according to our study. Methods:. A dermis flap was used in 6 cases to reconstruct small defects. The superficial branch of the superficial circumflex iliac artery was traced distally using an ultrasound device with a 70-MHz linear array transducer until the artery’s branch entered the dermis. The location of the dermis entry site was marked and the vessels running inside the dermis were observed and video-recorded. A flap was elevated above the superficial fascia, and the adipose tissue was removed using scissors after confirmation of the vessels’ dermis entry point. Results:. The use of 70-MHz ultrasonography permitted observation in all patients of small arteries entering the dermis layer. The artery was observed to give off branches after entering the dermis, in effect constituting an “intradermal plexus.” Small veins entering the dermis were similarly visualized using 70 MHz ultrasonography. All flaps survived completely. Conclusions:. Small arteries and veins entering and running inside the dermis were visualized for the first time with 70 MHz real-time ultrasonography. Knowledge of the existence of this “intradermal plexus” made it possible to discard nearly all subdermal adipose tissue quickly and safely, without resorting to the elaborate measures described in previous reports.

Summary:. Indocyanine green (ICG) lymphography is a useful imaging modality for evaluation of lymphedema and detection of lymphatic vessels. It also allows us to ensure patency of the anastomosed vessels intraoperatively. However, strong light from the operating microscope usually disturbs ICG fluorescence imaging. Only some built-in ICG camera systems with specific operating microscopes make real-time ICG lymphography possible in lymphaticovenular anastomosis (LVA). We applied a new high-resolution ICG videolymphography system, which is separated from the operating microscope. Because the system can divide near-infrared fluorescence light of ICG from visible light of the operating microscope, real-time ICG videolymphography-navigated LVA under operating microscope illumination is possible regardless types of operating microscopes. The study involved 10 patients with upper extremity lymphedema characterized by International Society of Lymphology stage 2 and treated by 3 lymphaticovenular anastomoses at the forearm (30 lymphaticovenular anastomoses incorporating 30 lymphatic vessels) under real-time ICG videolymphography. The rate of intraoperative detection of lymphatic vessels using real-time ICG videolymphography was 86.7% (0.25–0.85 mm in diameter), and that of lymph flow through the lymphaticovenular anastomoses was 76.7%. None of lymphatic vessels and no flow were detected under the microscope light by means of another non-built-in ICG lymphography camera. Real-time ICG videolymphography in LVA is beneficial, because the surgeon could find lymphatic vessels easily by checking dual images of original view and ICG fluorescent view and ensure accuracy of the LVA in a suture by a suture without any pauses of the surgical procedures.

Background:. In free-flap reconstruction of the midface, options for the recipient artery are quite limited; the superficial temporal artery and the facial artery are the most commonly used arteries. We report our approach for the use of the angular artery (the terminal branch of the facial artery) as the recipient artery in free-flap reconstruction of the midface. Methods:. Nine patients with midface defects underwent free-flap reconstructions using the angular artery as the recipient artery. Identification and marking of the facial artery were performed preoperatively using handheld Doppler ultrasound. The angular artery was located through an incision made on the side of the nose. When present, a vena comitans of the facial artery or any subcutaneous vein in the vicinity of the defect was used as the recipient vein. In other cases, the facial vein in the submandibular region was chosen as the recipient vein, using a vein graft. Results:. The average diameter of the angular artery was 0.9 mm (range, 0.7–1.0 mm). In all cases, arterial anastomosis was performed in an end-to-end fashion, and flaps survived completely. In 4 cases, a vein graft was used to bridge the pedicle vein and the facial vein. Conclusions:. Although supermicrosurgical skills may be required for its anastomosis, the angular artery is an anatomically consistent artery, which is suitable for use as the recipient artery in free-flap reconstruction of the midface. Use of the angular artery as the recipient artery allows shorter flap pedicles and decreases the number of vein grafts necessary.

BackgroundClearSight is a noninvasive arterial blood pressure monitor, but it remains unknown whether it is affected by the state of perfusion to the fingers. We investigated whether the lower perfusion index (PI) measured with a pulse oximeter, which reflects finger perfusion, would affect the agreement between arterial pressures measured with ClearSight versus those obtained with an arterial catheter.MethodsPaired arterial pressure data (ClearSight and arterial catheter-based pressures) and PI values were prospectively obtained from 30 patients undergoing major abdominal surgery. The primary outcome was standard deviation (SD) of the bias (precision) of blood pressure between ClearSight and arterial catheter. The ratio of the adjusted SD of the bias between PI≤1 and PI>1 was calculated using the linear mixed-effects model. The secondary outcomes were the bias and the limits of agreement (LOA) between the two devices (repeated measures Bland-Altman analysis).ResultsWe analyzed 6312 paired data points. The adjusted SD of bias in PI ≤1 compared with those in PI >1 was 1.4-fold (95% confidence interval: 1.3- to 1.4-fold) for systolic arterial pressure, 1.5-fold (95% confidence interval: 1.3- to 1.6-fold) for diastolic arterial pressure, and 1.3-fold (95% confidence interval: 1.2- to 1.5-fold) for mean arterial pressure. The bias (LOA) were as follows: systolic arterial pressure in the PI ≤1 and PI >1 groups, -3.5 (-35.4 to 28.4) mmHg and 2.2 (-19.9 to 24.3) mmHg, respectively; diastolic arterial pressure in the PI ≤1 and PI >1 groups, 13.1 (-5.1 to 31.3) mmHg and 9.0, (-2.6 to 20.6) mmHg, respectively; and mean arterial pressure in the PI ≤1 and PI >1 groups, 8.7 (-11.3 to 28.7) mmHg and 7.6 (-6.2 to 21.3) mmHg, respectively.ConclusionsPI ≤1 was associated with a large SD of the bias between the devices. The PI value could be a real-time indicator of ClearSight precision.

Background:. Identification and localization of functional lymphatic vessels are important for lymphaticovenular anastomosis. Conventional high-frequency ultrasound (CHFUS) has been reported to be useful for them, but it has some disadvantages. In this article, we present new capabilities of ultra high-frequency ultrasound (UHFUS) for imaging of the lymphatic vessels, which may overcome the weakness of CHFUS. Methods:. Thirty unaffected extremities in 30 unilateral secondary lymphedema patients (13 upper limbs and 17 lower limbs) were examined. Identification of the lymphatic vessels using UHFUS and CHFUS were performed at 3 sites in each unaffected extremity. Number and diameter of the detected lymphatic vessels were compared between UHFUS and CHFUS groups. At the same time, new characteristics of the lymphatic vessels seen with UHFUS were investigated. Results:. One hundred sixty-nine lymphatic vessels were detected with UHFUS, and 118 lymphatic vessels with CHFUS. The number of lymphatic vessels found in upper and lower extremities was significantly larger with UHFUS than with CHFUS. The diameter of lymphatic vessels found in upper and lower extremities was significantly smaller with UHFUS than with CHFUS. All lymphatic vessels that were detected in UFHUS were less likely to collapse when the transducer was against the skin of the examined sites. Conclusions:. Detection rate of the lymphatic vessels in nonlymphedematous extremities with UHFUS was higher than that with CHFUS. UHFUS provides images with extremely high resolution, demonstrating new characteristics of the lymphatic vessels.

Background:. Surgical treatment of lower extremity lymphedema (LEL) remains challenging. Application of the superior-edge-of-the-knee incision method for lymphaticovenular anastomosis (LVA) is reported to have a strong therapeutic effect in patients with LEL because lymph-to-venous flow at the anastomosis is enhanced by knee joint movement during normal walking. We investigated whether a single LVA created by this method is adequate for early LEL. Methods:. The study involved 10 patients with LEL characterized by stage 2 or 3 leg dermal backflow and treated by a single LVA at the thigh via the superior-edge-of-the-knee incision method. The lymphatic vessel and direction of flow were assessed intraoperatively, and reduction in lymphedema volume was assessed postoperatively. Results:. Use of our incision method yielded a single anastomosis in all patients with stage 2 leg dermal backflow and in all patients with stage 3 leg dermal backflow. The lymphatic vessel was 0.65 ± 0.08 mm in diameter (0.65 ± 0.09 and 0.65 ± 0.09 mm, respectively; P = 1.000). No venous reflux occurred in any patient. Mean follow-up was 7.70 ± 3.30 months (9.60 ± 3.29 and 5.80 ± 2.17 months, respectively; P = 0.068). Mean reduction in the LEL index was 20.160 ± 9.892 (22.651 ± 12.272 and 17.668 ± 7.353, respectively; P = 0.462). Conclusion:. A single LVA created by the superior-edge-of-the-knee incision method can be expected to have a strong therapeutic effect in patients with stage 2 or 3 leg dermal backflow.

Summary:. Bone perfusion evaluation methods in cadaver studies have yet to be established. The aim of this report was to introduce and validate the feasibility of indocyanine green (ICG) fluorescence angiography for evaluation of bone perfusion in the femoral medial condyle in cadavers. In 4 fresh nonembalmed cadavers (2 female), the descending genicular artery was dissected and carefully cannulated bilaterally. A 10 mL solution containing 5 mL ICG solution and 5 mL methylene blue solution was injected into the descending genicular artery. After the injection, the medial femoral condyle was cut with an oscillating saw. A photograph was taken of the cut ends of the bone. The cut ends of the bones were observed using a near-infrared camera. Images corresponding to the previously taken photographs of the cut ends were captured for comparative analysis. After injection of methylene blue and ICG, the blue dye could be seen in the periosteum in all specimens, but not inside the cortex or the cancellous region of the bone. When observed with ICG fluorescence angiography, however, the cancellous region was highlighted through small perforators penetrating the periosteum. Perfusion inside the medial femoral condyle in cadavers was confirmed using ICG fluorescence angiography. Our method can be especially beneficial in confirming the bone perfusion of a new bone flap based on a particular artery, both in cadavers as well as in patients, because ICG can be injected into specific arteries.

Abstract.: Recently, we showed that a methanol extract of Ganoderma lucidum inhibits scratching, an itch-related response, induced by intradermal injections of some pruritogens in mice. The present study investigated whether G. lucidum extract would inhibit allergic itch. In mice sensitized with an extract of salivary gland of mosquito (ESGM), an intradermal injection of ESGM elicited scratching, which was suppressed by oral administration of G. lucidum extract (100 and 300 mg/kg). The scratching was inhibited by the H1 histamine–receptor antagonist azelastine, but not by the peripherally acting H1-antagonist terfenadine, at the oral dose of 30 mg/kg. In sensitized mice, ESGM increased the activity of cutaneous nerve, which was suppressed by G. lucidum extract (300 mg/kg). Although terfenadine (30 mg/kg) inhibited plasma extravasation induced by ESGM in the sensitized mice, G. lucidum extract (300 mg/kg) was without effect. These results suggest that G. lucidum extract relieves allergic itch through a peripheral action. The results support the idea that mast cells and H1 histamine receptors are not the primary sites of the antipruritic action of G. lucidum extract. Keywords:: Ganoderma lucidum, itch and scratch, mosquito allergy, cutaneous nerve, plasma extravasation

ObjectiveThe number of bypasses is the most important factor in lymphaticovenular anastomosis (LVA) for lymphedema treatment. Side-to-end (S-E) LVA, which can bypass bidirectional lymph flows via one anastomosis, is considered to be the most efficient bypass, but creation of lateral window to a small lymphatic vessel is technically demanding. To overcome the difficulty, we introduced S-E anastomosis through temporary lymphatic expansion (SEATTLE) procedure in S-E LVA.MethodsThis was a retrospective observational study set in a teaching hospital. Forty eight lower extremity lymphedema (LEL) patients underwent LVA. S-E LVAs were performed with (SEATTLE group) or without (non-SEATTLE group) temporary lymphatic expansion. S-E LVAs were evaluated to compare anastomosis result in SEATTLE and non-SEATTLE groups.ResultsS-E LVAs resulted in 44 anastomoses in SEATTLE group (n = 25) and 37 anastomoses in non-SEATTLE group (n = 23). LEL index reduction in SEATTLE group was significantly greater than that in non-SEATTLE group (16.5±14.5 vs. 10.9±11.8, P = 0.041). Success rate of S-E LVA in SEATTLE group was significantly higher than that in non-SEATTLE group (95.5% vs 81.1%, P = 0.040). Thirty seven of 44 (84.1%) lymph vessels in SEATTLE group were successfully dilated by temporary lymphatic expansion maneuver. All of 9 failed S-E LVAs used a lymphatic vessel with diameter of 0.35 mm or smaller.ConclusionsThe SEATTLE procedure facilitates S-E LVA by a simple and easy maneuver. When the diameter of the lymphatic vessel is 0.35 mm or smaller even after the temporary lymphatic expansion maneuver, S-E LVA is not recommended due to relatively high failure rate.

BACKGROUND: Secondary lymphedema causes swelling in limbs due to lymph retention following lymph node dissection in cancer therapy. Initiation of treatment soon after appearance of edema is very important, but there is no method for early diagnosis of lymphedema. In this study, we compared the utility of four diagnostic imaging methods: magnetic resonance imaging (MRI), computed tomography (CT), lymphoscintigraphy, and Indocyanine Green (ICG) lymphography. PATIENTS AND METHODS: Between April 2010 and November 2011, we examined 21 female patients (42 arms) with unilateral mild upper limb lymphedema using the four methods. The mean age of the patients was 60.4 years old (35-81 years old). Biopsies of skin and collecting lymphatic vessels were performed in 7 patients who underwent lymphaticovenous anastomosis. RESULTS: The specificity was 1 for all four methods. The sensitivity was 1 in ICG lymphography and MRI, 0.62 in lymphoscintigraphy, and 0.33 in CT. These results show that MRI and ICG lymphography are superior to lymphoscintigraphy or CT for diagnosis of lymphedema. In some cases, biopsy findings suggested abnormalities in skin and lymphatic vessels for which lymphoscintigraphy showed no abnormal findings. ICG lymphography showed a dermal backflow pattern in these cases. CONCLUSIONS: Our findings suggest the importance of dual diagnosis by examination of the lymphatic system using ICG lymphography and evaluation of edema in subcutaneous fat tissue using MRI.

INTRODUCTION: To date, an electron microscopy study of the collecting lymphatic vessels has not been conducted to examine the early stages of lymphedema. However, such histological studies could be useful for elucidating the mechanism of lymphedema onset. The aim of this study was to clarify the changes occurring in collecting lymphatic vessels after lymphadenectomy. METHODS: The study was conducted on 114 specimens from 37 patients who developed lymphedema of the lower limbs after receiving surgical treatment for gynecologic cancers and who consulted the University of Tokyo Hospital and affiliated hospitals from April 2009 to March 2011. Lymphatic vessels that were not needed for lymphatico venous anastomosis surgery were trimmed and subsequently examined using electron microscopy and light microscopy. RESULTS: Based on macroscopic findings, the histochemical changes in the collecting lymphatic vessels were defined as follows: normal, ectasis, contraction, and sclerosis type (NECST). In the ectasis type, an increase in endolymphatic pressure was accompanied by a flattening of the lymphatic vessel endothelial cells. In the contraction type, smooth muscle cells were transformed into synthetic cells and promoted the growth of collagen fibers. In the sclerosis type, fibrous elements accounted for the majority of the components, the lymphatic vessels lost their transport and concentrating abilities, and the lumen was either narrowed or completely obstructed. CONCLUSIONS: The increase in pressure inside the collecting lymphatic vessels after lymphadenectomy was accompanied by histological changes that began before the onset of lymphedema.