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Although plasmonic palladium (Pd) nanospheres are thermodynamically stable and have high photothermal conversion due to the free and bound electron coupling associated with the intrinsic high interband transition, they have not attracted attention as a photothermal conversion material for next-generation photothermal cancer therapy. This is because the Pd nanospheres generate the localized surface plasmon resonance (LSPR) intrinsically in the ultraviolet region, which is far away from the biological transparent window (750-900 nm). In this study, we controlled the LSP wavelength of Pd nanospheres by coating with high refractive index TiO 2 shells taking advantage of the Pd LSPR which is highly sensitive to changes in the local refractive index around the nanospheres. Our calculations indicated that the absorption cross section at 808 nm (corresponding to the wavelength used for photothermal treatment) was increased by 4.5 times by redshifting the LSPR and increasing the extinction intensity associated with the coating with TiO 2 shell. Experiments confirmed the theoretical prediction in that the LSPR of the synthesized Pd nanospheres with a diameter of 81 nm was significantly redshifted by coating with amorphous TiO 2 shell, resulting in significant large extinction intensity at 808 nm. The photothermal conversion efficiency was estimated to be 50%. In vitro cell tests, HeLa cells incubated with 100-300 μg/mL TiO 2 -coated Pd nanospheres were efficiently killed by irradiating 808 nm laser (1.8 W) even though the nanospheres with the same concentrations showed little cytotoxicity. These results indicate that the Pd nanospheres coated with high refractive index shells can be promising as a photothermal therapy agent., (© 2022. The Author(s).)

Purpose: This study evaluated the reproductive potential of premature ovarian insufficiency (POI) patients with abnormal karyotypes undergoing infertility treatments., Methods: A retrospective analysis of infertility treatments in POI patients with an abnormal karyotype treatment. Clinical and laboratory data were analyzed., Results: The study group was forty-nine POI patients. Follicular growth was achieved in 29% (89/307) controlled ovarian stimulation (COS) cycles in 57% (28/49) of patients. Oocyte retrieval was attempted in 47% (23/49) of patients with a proportion of successful oocyte retrieval per oocyte pick-up (OPU) of 59.4% (41/69). The average number of retrieved oocytes was 2.4 ± 2.7 per patient and fertilization rate was 70.7% (29/41). Embryo transfer (ET) performed in eight patients with a total of nine ET attempts, resulting in 33.3% (3/9) of live birth rate per ET. Three patients delivered a healthy baby (6.1% (3/49) of live birth rate per patient). Mosaic Turner syndrome patients had a longer duration of amenorrhea and lower chances of successful follicular growth with OPU in 35.7% (5/14) of patients, whereas 47XXX had shorter duration of amenorrhea and COS with follicle growth with OPU in 83.3% (5/6)., Conclusion: COS might provide an opportunity for POI women with abnormal karyotypes to conceive a biological offspring., Competing Interests: All authors declare they did not receive funding for this study, and declare no conflicts of interest., (© 2022 The Authors. Reproductive Medicine and Biology published by John Wiley & Sons Australia, Ltd on behalf of Japan Society for Reproductive Medicine.)

Collecting duct carcinoma (CDC) is a rare subset of high-grade renal cell carcinoma (RCC). To diagnose CDC, it is necessary to rule out other renal tumors including renal medullary carcinoma and fumarate hydratase (FH)-deficient RCC. However, there is overlap in the morphology of these three tumors, which all have poor outcomes. There is also still a need to sufficiently examine the therapeutic strategies for each of these tumors. In this study, we retrospectively reclassified invasive/infiltrating high-grade RCC and investigated its pathological features. We reviewed 18 cases previously diagnosed as "CDC," "FH-deficient RCC," and "unclassified RCC," which were reclassified as SMARCB1/INI1-deficient RCC, FH-deficient RCC, and CDC by SMARCB1/INI1, FH, and 2SC immunohistochemistry (IHC) and FH gene mutational status. As the result, 18 cases were reclassified into 2 cases of SMARCB1/INI1-deficient RCC, 7 cases of FH-deficient RCC, and 9 cases of CDC. The morphological features of each group overlapped, and no specific immunohistochemical expression except for SMARCB1/INI1, FH, and 2SC was detected. These results suggest that invasive/infiltrating high-grade RCC should be diagnosed by the combination of immunohistochemistry and molecular biological technique., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Our understanding of how each hereditary kidney cancer adapts to its tissue microenvironment is incomplete. Here, we present single-cell transcriptomes of 108,342 cells from patient specimens including from six hereditary kidney cancers. The transcriptomes displayed distinct characteristics of the cell of origin and unique tissue microenvironment for each hereditary kidney cancer. Of note, hereditary leiomyomatosis and renal cell carcinoma (HLRCC)-associated kidney cancer retained some characteristics of proximal tubules, which were completely lost in lymph node metastases and present as an avascular tumor with suppressed T cells and TREM2-high macrophages, leading to immune tolerance. Birt-Hogg-Dubé (BHD)-associated kidney cancer exhibited transcriptomic intratumor heterogeneity (tITH) with increased characteristics of intercalated cells of the collecting duct and upregulation of FOXI1-driven genes, a critical transcription factor for collecting duct differentiation. These findings facilitate our understanding of how hereditary kidney cancers adapt to their tissue microenvironment., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)

A 10-month-old, intact male Toy Poodle was referred for a postural abnormality. Blood biochemical tests revealed a marked increase in plasma creatine phosphokinase (CPK) concentration. The isoenzyme test showed that 99% of serum CPK consisted of CPK-MM. Histopathological evaluation of muscle biopsy samples confirmed scattered degeneration and necrosis of myofibers. Immunohistochemistry for dystrophin showed an absence of staining in muscle cells. Based on these findings, the dog was diagnosed with dystrophin-deficient muscular dystrophy. Whole genome sequencing using genomic DNA extracted from blood revealed a single base pair insertion in exon 45 of the Duchenne muscular dystrophy (DMD) gene. This is the first report on muscular dystrophy in Toy Poodles and identified a novel mutation in the DMD gene.

Antibacterial materials are widely used to prevent hospital-acquired infections. In our previous report, metal (calcium, copper or zinc)-doped raw silk fabrics were shown to possess strong antibacterial activities against Escherichia coli . However, antibacterial materials may occasionally be harmful to the human body; thus, in this study, we investigated the cytotoxicities of extracts from metal-doped raw silk fabrics with respect to fibroblasts and osteoblasts indirectly. Calcium-doped raw silk fabric demonstrated cytocompatibility with fibroblasts. Contrarily, copper- and zinc-doped raw silk fabrics remarkably decreased the cell densities of fibroblasts, indicating their cytotoxic effects. This observation could be attributed to the high concentrations of the released copper or zinc ions. However, calcium-, copper- and zinc-doped raw silk fabrics did not demonstrate any cytotoxic effects on osteoblasts because a high concentration of the serum alleviated the effects of these metal ions released from the fabrics. Thus, calcium-doped raw silk fabric is a promising antibacterial material that does not induce strong cytotoxicity. This study will facilitate the design of materials that are both antibacterial and safe.

Bone metabolism is regulated by the cooperative activity between bone-forming osteoblasts and bone-resorbing osteoclasts. However, the mechanisms mediating the switch between the osteoblastic and osteoclastic phases have not been fully elucidated. Here, we identify a specific subset of mature osteoblast-derived extracellular vesicles that inhibit bone formation and enhance osteoclastogenesis. Intravital imaging reveals that mature osteoblasts secrete and capture extracellular vesicles, referred to as small osteoblast vesicles (SOVs). Co-culture experiments demonstrate that SOVs suppress osteoblast differentiation and enhance the expression of receptor activator of NF-κB ligand, thereby inducing osteoclast differentiation. We also elucidate that the SOV-enriched microRNA miR-143 inhibits Runt-related transcription factor 2, a master regulator of osteoblastogenesis, by targeting the mRNA expression of its dimerization partner, core-binding factor β. In summary, we identify SOVs as a mode of cell-to-cell communication, controlling the dynamic transition from bone-forming to bone-resorbing phases in vivo., (© 2022. The Author(s).)

L-type amino acid transporter 1 (LAT1), also referred to as SLC7A5, is believed to regulate tumor metabolism and be associated with tumor proliferation. In invasive breast cancer, we clinicopathologically investigated the utility of LAT1 expression. LAT1 expression was evaluated via immunohistochemistry analyses in 250 breast cancer patients undergoing long-term follow-up. We assessed the relationships between LAT1 expression and patient outcomes and clinicopathological factors. Breast cancer-specific survival stratified by LAT1 expression was assessed. Human epidermal growth factor receptor 2 (HER2)-positive patients with metastasis received trastuzumab therapy. The density of tumor-infiltrating lymphocytes (TILs) was evaluated according to the International Working Group guidelines. In the current study, high LAT1 expression was significantly correlated with estrogen receptor (ER) negativity, progesterone receptor negativity, high histological grade, increased TILs, and programmed death ligand 1 positivity. Among the ER-positive and HER2-negative patients, high LAT1 was an independent indicator of poor outcomes (hazard ratio (HR) = 2.97; 95% confidence interval (CI), 1.16-7.62; p = 0.023). Moreover, high LAT1 expression was an independent poor prognostic factor in luminal B-like breast cancer with aggressive features (HR = 3.39; 95% CI 1.35-8.52; p = 0.0094). In conclusion, high LAT1 expression could be used to identify a subgroup of invasive breast cancer characterized by aggressive behavior and high tumor immunoreaction. Our findings suggest that LAT1 might be a candidate therapeutic target for breast cancer patients, particularly those with luminal B-like type breast cancer., (© 2022. The Author(s).)

Type B insulin resistance syndrome (TBIR) is a rare autoimmune disease characterised by autoantibodies targeting insulin receptors. TBIR is often complicated by systemic lupus erythematosus (SLE). We describe the case of a 59-year-old Japanese man with TBIR complicated with lupus nephritis (LN), who presented with nephrotic syndrome and severe hypoglycaemia. Treatment with prednisolone (PSL), mycophenolate mofetil (MMF), and tacrolimus (TAC) resulted in improved SLE activity and glucose intolerance with the reduction of anti-insulin receptor autoantibodies. To the best of our knowledge, this is the first reported case of TBIR complicated with LN that was successfully treated using multitarget therapy with PSL, MMF, and TAC., (© Japan College of Rheumatology 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We developed a method for passively controlling microdroplet rotation, including interior rotation, using a parallel flow comprising silicone and sesame oils. This device has a simple 2D structure with a straight channel and T-junctions fabricated from polydimethylsiloxane. A microdroplet that forms upstream moves into the sesame oil. Then, the largest flow velocity at the interface of the two oil layers applies a rotational force to the microdroplet. A microdroplet in the lower oil rotates clockwise while that in the upper oil rotates anti-clockwise. The rotational direction was controlled by a simple combination of sesame and silicone oils. Droplet interior flow was visualized by tracking microbeads inside the microdroplets. This study will contribute to the efficient creation of chiral molecules for pharmaceutical and materials development by controlling rotational direction and speed.

Background: The most common presentation of symptomatic Meckel's diverticulum (MD) are intestinal obstruction, gastrointestinal hemorrhage, and inflammation of the MD with or without perforation. Intraperitoneal hemorrhage because of MD is extremely rare. We report a case of MD with intraperitoneal hemorrhage in a child detected with screening laparoscopy., Case Presentation: An 11-year-old girl presented to another hospital with lower abdominal pain and vomiting that lasted for 2 days. Acute appendicitis was suspected, and she was referred to our department. Abdominal enhanced computed tomography showed an abscess in the lower abdomen with ascites in the pelvis. She was diagnosed with a localized intra-abdominal abscess and the decision was made to treat with antibiotics. However, her abdominal pain worsened, with abdominal distension, tenderness and guarding. She was diagnosed with panperitonitis and the decision was made for surgery 5 h after admission. During surgery, laparoscopic observation from the umbilical region revealed 200 ml of fresh blood throughout the peritoneal cavity, originating from the mesentery of the ileum. MD was observed with bleeding from the surrounding mesentery. Small bowel resection was performed, and the patient was discharged on the 5th postoperative day. Pathological findings revealed an MD containing ectopic gastric mucosa and small intestinal ulcer perforation at the base of the MD., Conclusions: We report an extremely rare case of an MD with intraperitoneal hemorrhage in a child. In pediatric cases, it is possible that perforation with ectopic gastric mucosa may cause massive bleeding because of rupture of the surrounding mesenteric blood vessels., (© 2021. The Author(s).)

We analyzed data from 466 patients with premature ovarian insufficiency (POI) who wished to have a biological child and were followed up while undergoing hormone replacement (HR) therapy with or without ovarian stimulation (OS) between April 2014 and December 2020. OS was conducted in 6891 cycles in 429 patients (Group OS), whereas only HR (Group HR) was conducted in 1117 cycles in 37 patients. The follicle growth rate was 48.3% (207/429) per patient in Group OS and 5.4% (2/37) in Group HR (p<0.01). There were 51 live births (LBs) in 50 patients during follow-up. In Group OS, the LB rate was 5.8% (47/807) in cycles where in vitro fertilization (IVF) and embryo transfer were attempted (Group IVF), and 1.3% (3/236) in cycles where intrauterine insemination/timed intercourse was attempted (p<0.01). No pregnancies occurred in Group HR. Among the patients in Group IVF, the LB rate was significantly higher in patients aged <35 years at the initiation of follow-up than in patients who started at later ages (p<0.01). Among the cases who achieved an LB, 39 were patients with idiopathic POI (Group IVF-1, n=297) and seven were patients who had undergone surgical treatment for benign ovarian tumors (Group IVF-2, n=50); however, no LBs occurred in patients who had undergone treatment for malignancy (n=17), and only one in patients with chromosomal abnormalities (n=22). The LB rate per case in the patients in Group IVF-1 and those aged <35 years at the start of follow-up (Group IVF-1-a) was 24.1% (26/108), which was higher than those of the other age groups. The LB rate per case in the patients in Group IVF-1-a with <4 years of amenorrhea was 37.3% (19/51), and that in the patients in Group IVF-2 with <4 years of amenorrhea was 21.2% (7/33). These results suggest that infertility treatment is possible in some patients with POI, especially those that can be classified in Group IVF-1-a and Group IVF-2 with <4 years of amenorrhea. Therefore, OS combined with HR therapy should be considered for such patients before attempts at oocyte donation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ishizuka, Furuya, Kimura, Kamioka and Kawamura.)

A green method of the oxidation of benzylamines to imines was developed using a novel binary system of Au/C-CuO. This system was evaluated under atmospheric oxygen, and the corresponding imines were obtained in up to 100% yields by loading 0.006 mol % of Au/C and 1.25 mol % of CuO under mild conditions. This system was also successfully applied to the syntheses of N -containing functional molecules, as well as that of imines on the scale of several grams. Furthermore, the turnover number of the system (more than 8000 times on a gold basis) as well as its ability to be reused more than 10 times for benzylamine oxidation demonstrates the excellent durability and recyclability of the developed system., Competing Interests: The authors declare no competing financial interest., (© 2021 The Authors. Published by American Chemical Society.)

Most of the diseases for which apheresis therapy is indicated are intractable and rare, and each patient has a different background and treatment course prior to apheresis therapy initiation. Therefore, it is difficult to conduct large-scale randomized controlled trials to secure high-quality evidence. Under such circumstances, the American Society for Apheresis (ASFA) issued its guidelines in 2007, which were repeatedly revised until the latest edition in 2019. The ASFA guidelines are comprehensive. However, in the United States, a centrifugal separation method is mainly used for apheresis, whereas the mainstream procedure in Japan is the membrane separation method. The target diseases and their backgrounds are different from those in Japan. Due to these differences, the direct adoption of the ASFA guidelines in Japanese practice creates various problems. One of the features of apheresis in Japan is the development of treatment methods using hollow-fiber devices such as double filtration plasmapheresis (DFPP) and selective plasma exchange and adsorption-type devices such as polymyxin B-immobilized endotoxin adsorption columns. Specialists in emergency medicine, hematology, collagen diseases/rheumatology, respiratory medicine, cardiovascular medicine, gastroenterology, neurology, nephrology, and dermatology who are familiar with apheresis therapy gathered for this guideline, which covers 86 diseases. In addition, since apheresis therapy involves not only physicians but also clinical engineers, nurses, dieticians, and many other medical professionals, this guideline was prepared in the form of a worksheet so that it can be easily understood at the bedside. Moreover, to the clinical purposes, this guideline is designed to summarize apheresis therapy in Japan and to disseminate and further develop Japanese apheresis technology to the world. As diagnostic and therapeutic techniques are constantly advancing, the guidelines need to be revised every few years. In order to ensure the high quality of apheresis therapy in Japan, both the Japanese Society for Apheresis Registry and the guidelines will be inseparable., (© 2021 International Society for Apheresis, Japanese Society for Apheresis, and Japanese Society for Dialysis Therapy.)

The purpose of this study was to determine the optimal imaging protocol for contrast-enhanced computed tomography (CECT) using micro-CT (μ-CT) for the posterior cardinal vein (PCV), dorsal aorta (DA), hepatic portal vein (HPV), kidney, liver, cephalic arteries (CAs), and gills of Cloudy Catsharks Scyliorhinus torazame. Additionally, we examined the availability of CECT screening for the coelomic organs. Different doses of iopamidol (100, 300, 500, and 700 mg iodine [mgI]/kg) were administered intravenously for 20 s in six sharks. The CT scans from the pectoral girdle to the pelvic girdle were performed at 0-600 s after administration. Contrast-enhanced CT imaging of the CAs, gills, and coelomic organs was examined. Assessment of the signal enhancement value revealed that the PCV was easily visualized with all contrast doses at 25 s. The CAs, gills, and DA were visible at a slightly higher dose (CAs and gills: 200 mgI/kg at 40 s; DA: 300 mgI/kg at 50 s). The HPV was obvious at a dose of at least 500 mgI/kg after a 150-s delay. The parenchyma of the kidney had a contrast effect at 300 mgI/kg, 150 s after the contrast effect of the renal portal system disappeared. The liver, which stores a lot of lipids, had poor overall contrast enhancement that was optimized at the highest dose of 700 mgI/kg. Contrast-enhanced CT screening at 700 mgI/kg and 150 s is likely to obtain the optimal imaging of the reproductive organs, such as the ovary, oviducal gland, uterus, and testis. The present findings can be applied not only to clinical practice but also to academic research and education on elasmobranchs in aquariums., (© 2021 American Fisheries Society.)

We performed proteomic analysis of rat serum proteins adsorbed on hydroxyapatite (HAp) and α-alumina (α-Al 2 O 3 ) in order to identify proteins that specifically adsorb onto HAp and control cellular responses. Proteins with either or both molecular weight of 22-32 kDa and computed isoelectric point of 5.0-5.5 were preferentially adsorbed on HAp. In total, 182 proteins were adsorbed on both HAp and α-Al 2 O 3 , of which 14 were highly enriched on HAp, whereas 68 were adsorbed only on HAp. Therefore, 82 (14+68) proteins were further evaluated by bioinformatics and literature-based analyses. We predicted that hepatocyte growth factor and angiopoietin-like protein 3 (ANGPTL3) are candidate proteins responsible for the osteoconductivity of HAp. Although ANGPTL3 promoted the attachment and spreading of MC3T3-E1 cells, it did not promote their proliferation and differentiation. Our results suggest that specific adsorption of ANGPTL3 on HAp induced osteoconductivity by enhancing the attachment and spreading of osteoblasts.

Magnetic nanoparticles are widely studied for their use in various therapeutic and diagnostic purposes. As biomaterials, their biocompatibility is as important as their magnetic properties. Iron nitride (Fe x N y ) has excellent magnetic properties, and thus Fe x N y nanoparticles could be useful as potential biomaterials. However, the biocompatibility of Fe x N y nanoparticles is yet to be investigated. In this study, we assessed the biocompatibility of Fe x N y nanoparticles by evaluating their direct-contact cytotoxicity compared with that of magnetite nanoparticles (MNPs). Rat fibroblasts were incubated with the nanoparticle samples dispersed in culture medium at concentrations of 10, 50, and 100 μg/ml. The DNA concentration measurement, MTT assay, and trypan blue exclusion test were conducted after days 1 and 3 of incubation. After day 1, the cell viability decreased, and cell death increased with increasing sample concentration when compared with the control. However, after day 3, there were no significant differences when compared with the control, irrespective of the sample concentrations. Further, there were no significant differences between the Fe x N y nanoparticles and MNPs at the same concentrations in all the cytotoxicity evaluation tests. Therefore, it is suggested that Fe x N y nanoparticles might be as cytocompatible as the conventional MNPs., (© 2021 Wiley Periodicals LLC.)

Objective: Only a few studies have addressed clinical outcomes of revision surgery for adjacent segment disease. The purpose of this study was to elucidate clinical outcomes of second (repeat) posterior lumbar interbody fusion (PLIF) by focusing on the relationship between clinical outcomes and spinopelvic parameters and predisposing factors requiring subsequent corrective long fusion after repeat PLIF., Methods: We analyzed the data of 47 patients ≥40 years old who underwent repeat PLIF after single-segment PLIF owing to adjacent segment disease. The correlation between clinical outcomes and radiographic parameters was investigated. Patient demographics and radiographic parameters were compared between patients with and without subsequent corrective long fusion., Results: Japanese Orthopaedic Association score at final follow-up was 13.4, and the recovery rate was 37.2%. All sagittal parameters except pelvic tilt and C7-central sacral vertical line at final follow-up showed weak to moderate (|r| = 0.30-0.56) correlation with clinical scores. Finally, 11% of patients required subsequent long corrective fusion. Pelvic incidence minus lumbar lordosis (PI-LL) mismatch (cutoff value of 27.5°) and thoracic kyphosis (cutoff value of 12.5°) before repeat PLIF were identified as predisposing factors for subsequent long corrective fusion., Conclusions: The clinical outcomes of repeat PLIF were inferior to outcomes of primary PLIF. Once PI-LL mismatch occurs after initial PLIF, it will be difficult to resolve the PI-LL mismatch during the second PLIF. To stop the chain of reoperations in patients whose preoperative PI-LL exceeds 27.5° before repeat PLIF, corrective long fusion may be a surgical option to consider., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Introduction: In quantitative assays for hepatitis B virus (HBV) DNA, although the amplification reaction signal is detected for low-positive cases, quantification remains challenging. HBV reactivation has been reported in many studies, but only a few have focused on HBV low-positive cases. This study aimed to determine the reactivation rate and risk factors for HBV reactivation in low-positive cases., Methods: In this retrospective cohort study, we analyzed 7498 patients who had their HBV DNA measured at Sapporo Medical University Hospital between April 2008 and November 2020. Patient selection criteria were defined as follows: hepatitis B surface antigen was negative; HBV DNA was detectable but not quantifiable at least once. HBV DNA was monitored according to the guidelines for HBV reactivation., Results: In total, 49,086 HBV DNA quantitative tests were performed. HBV DNA levels of 2578 tests were detectable but not quantifiable. Eighty patients met the criteria in this study. The median observation period was 497 days, and the 2-year reactivation rate was 15%. Ten patients had low HBV DNA positivity at baseline. Malignant lymphoma was observed in 15 patients; chemotherapy was used to treat other solid tumors in 35 patients, and immunosuppressive therapy was used in 30 patients. Multivariate analysis revealed that HBV DNA detected below the quantification level at baseline was an independent risk factor for HBV reactivation (adjusted hazard ratio 5.82; P = 0.010)., Conclusions: Patients with low HBV DNA positivity, especially at baseline, are at high risk for HBV reactivation and therefore require closer monitoring., (Copyright © 2021 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Background: Autologous repair using costal cartilage grafts remains the most widely accepted method of microtia reconstruction. A major complication of current techniques is loss of ear shape caused by scarring, contracture and cartilage absorption. We present a new surgical technique utilizing the posterior auricular artery free flap in microsurgical reconstruction of the retroauricular sulcus in microtia., Method: Reconstruction is performed in two stages. In the first stage, a fabricated costal cartilage framework is inserted into a skin pocket as described by Nagata. In the second stage, the ear framework is elevated from the scalp and held by an additional cartilage wedge. Following indocyanine green angiography perforator mapping, a posterior auricular artery perforator flap is harvested from the contralateral (normal) ear and used to reconstruct the posterior auricular sulcus covering the cartilage framework and elevating wedge., Results: The technique was applied to three patients aged 11-15 years with a follow-up time of 8 months to 3 years. The average flap artery diameter was 0.73 mm and the vein was 0.7 mm. Venous congestion occurred in one case and was resolved with a vein graft leading to complete flap recovery. Good ear shape, elevation, projection, skin color and texture were achieved in all the cases., Conclusion: Posterior auricular artery flap reconstruction of the retroauricular sulcus in microtia repair is a useful alternative to the current skin graft and tissue expander-based techniques. It provides the ideal skin color and texture match and may improve the overall results of microtia reconstruction by enhancing vascularity., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interest., (Copyright © 2021 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.)

Objective: Although several reports have described adjacent-segment disease (ASD) after posterior lumbar interbody fusion (PLIF), there have been only a few reports focusing on early-onset ASD occurring within 3 years after primary PLIF. The purpose of this study was to investigate the prevalence and postoperative pathologies of early-onset ASD and its relation with radiological parameters such as segmental lordosis (SL)., Methods: The authors reviewed a total of 256 patients who underwent single-segment PLIF at L4-5 for degenerative lumbar spondylolisthesis (DLS) and were followed up for at least 5 years. The definition of ASD was a symptomatic condition requiring an additional operation at the adjacent fusion segment in patients who had undergone PLIF. ASD occurring within 3 years after primary PLIF was categorized as early-onset ASD. As a control group, 54 age- and sex-matched patients who had not suffered from ASD for more than 10 years were selected from this series., Results: There were 42 patients with ASD at the final follow-up. ASD prevalence rates at 3, 5, and 10 years postoperatively and at the final follow-up were 5.0%, 8.2%, 14.1%, and 16.4%, respectively. With respect to ASD pathologies, lumbar disc herniation (LDH) was significantly more common in early-onset ASD, while lumbar spinal stenosis and DLS occurred more frequently in late-onset ASD. Significant differences were detected in the overall postoperative range of motion (ROM) and in the changes in ROM (ΔROM) at L3-4 (the cranial adjacent fusion segment) and changes in SL (ΔSL) at L4-5 (the fused segment), while there were no significant differences in other pre- and postoperative parameters. In stepwise logistic regression analysis, ΔSL was identified as an independent variable (p = 0.008) that demonstrated significant differences, especially in early-onset ASD (control 1.1° vs overall ASD -2.4°, p = 0.002; control 1.1° vs early-onset ASD -6.6°, p = 0.00004)., Conclusions: The study results indicated that LDH was significantly more common as a pathology in early-onset ASD and that ΔSL was a major risk factor for ASD, especially early-onset ASD.

Surgical treatment has improved the prognosis of canine idiopathic chylothorax, although a recurrence of the disease occurs occasionally after the procedure. An improved understanding of possible causes for this recurrence would be helpful for prognosis and treatment planning in affected patients. In this retrospective case series study, we described the detailed pre- and postoperative computed tomographic lymphography (CTLG) imaging characteristics for a group of dogs with surgically confirmed idiopathic chylothorax. Preoperative CTLG was performed in 12 of 14 dogs diagnosed with idiopathic chylothorax. Thoracic ducts were present on the right side in 10 dogs, left side in one dog, and bilaterally in one dog. All the 14 dogs received a combination therapy of pericardiectomy and thoracic duct ligation (TDL) by video-assisted thoracoscopic surgery. One week after surgery, a postoperative CTLG was performed, and the thoracic ducts were apparent in seven of 14 dogs. Three dogs had an unchanged course of the thoracic duct, which could have resulted from a missed duct. Four dogs were identified as having a bypass formation: the oblique duct originated at the ligation site and connected to the duct on the other side. Our findings indicated that one of the possible causes for postoperative recurrence of chylothorax in dogs could be "invisible or sleeping" fine ducts that are collapsed and not visible in preoperative CTLG scans. After TDL causes a change in the pressure of lymphatic flow, these fine thoracic ducts may become apparent using postoperative CTLG., (© 2021 American College of Veterinary Radiology.)

In recent years, research on the application of microdroplets in the fields of biotechnology and chemistry has made remarkable progress, but the technology for the stable generation of single-micrometer-scale microdroplets has not yet been established. In this paper, we developed an efficient and stable single-micrometer-scale droplet generation device based on the fragmentation of droplet tails, called "tail thread mode", that appears under moderate flow conditions. This method can efficiently encapsulate microbeads that mimic cells and chemical products in passively generated single-micrometer-scale microdroplets. The device has a simple 2D structure; a T-junction is used for droplet generation; and in the downstream, multi-branch channels are designed for droplet deformation into the tail. Several 1-2 µm droplets were successfully produced by the tail's fragmentation; this continuous splitting was induced by the branch channels. We examined a wide range of experimental conditions and found the optimal flow rate condition can be reduced to one-tenth compared to the conventional tip-streaming method. A mold was fabricated by simple soft lithography, and a polydimethylsiloxane (PDMS) device was fabricated using the mold. Based on the 15 patterns of experimental conditions and the results, the key factors for the generation of microdroplets in this device were examined. In the most efficient condition, 61.1% of the total droplets generated were smaller than 2 μm.

Osteoclastic bone resorption and osteoblastic bone formation/replenishment are closely coupled in bone metabolism. Anabolic parathyroid hormone (PTH), which is commonly used for treating osteoporosis, shifts the balance from osteoclastic to osteoblastic, although it is unclear how these cells are coordinately regulated by PTH. Here, we identify a serine protease inhibitor, secretory leukocyte protease inhibitor (SLPI), as a critical mediator that is involved in the PTH-mediated shift to the osteoblastic phase. Slpi is highly upregulated in osteoblasts by PTH, while genetic ablation of Slpi severely impairs PTH-induced bone formation. Slpi induction in osteoblasts enhances its differentiation, and increases osteoblast-osteoclast contact, thereby suppressing osteoclastic function. Intravital bone imaging reveals that the PTH-mediated association between osteoblasts and osteoclasts is disrupted in the absence of SLPI. Collectively, these results demonstrate that SLPI regulates the communication between osteoblasts and osteoclasts to promote PTH-induced bone anabolism.

Methotrexate (MTX) is an anticancer and anti-rheumatoid arthritis drug that is considered to block nucleotide synthesis and the cell cycle mainly by inhibiting the activity of dihydrofolate reductase (DHFR). Using affinity-matrix technology and X-ray analysis, the present study shows that MTX also interacts with macrophage migration inhibitory factor (MIF). Fragment molecular-orbital calculations quantified the interaction between MTX and MIF based on the structure of the complex and revealed the amino acids that are effective in the interaction of MTX and MIF. It should be possible to design new small-molecule compounds that have strong inhibitory activity towards both MIF and DHFR by structure-based drug discovery.

The emergence and global spread of extended-spectrum or AmpC β-lactamase (ESBL/AmpC)-producing Enterobacteriaceae in companion animals have led to the hypothesis that companion animals might be reservoirs for cross-species transmission because of their close contact with humans. However, current knowledge in this field is limited; therefore, the role of companion animals in cross-species transmission remains to be elucidated. Herein, we studied ESBL/AmpC-producing Enterobacteriaceae, Escherichia coli in particular, isolated from extraintestinal sites and feces of companion dogs. Whole-genome sequencing analysis revealed that (i) extraintestinal E. coli isolates were most closely related to those isolated from feces from the same dog, (ii) chromosomal sequences in the ST131/C1-M27 clade isolated from companion dogs were highly similar to those in the ST131/C1-M27 clade of human origin, (iii) certain plasmids, such as IncFII/pMLST F1:A2:B20/blaCTX-M-27, IncI1/pMLST16/blaCTX-M-15, or IncI1/blaCMY-2 from dog-derived E. coli isolates, shared high homology with those from several human-derived Enterobacteriaceae, (iv) chromosomal blaCTX-M-14 was identified in the ST38 isolate from a companion dog, and (v) eight out of 14 tested ESBL/AmpC-producing E. coli isolates (i.e., ST131, ST68, ST405, and ST998) belonged to the human extraintestinal pathogenic E. coli (ExPEC) group. All of the bla-coding plasmids that were sequenced genome-wide were capable of horizontal transfer. These results suggest that companion dogs can spread ESBL/AmpC-producing ExPEC via their feces. Furthermore, at least some ESBL/AmpC-producing ExPECs and bla-coding plasmids can be transmitted between humans and companion dogs. Thus, companion dogs can act as an important reservoir for ESBL/AmpC-producing E. coli in the community., Competing Interests: The authors have declared that no competing interests exist.

Objective: Although concomitant foraminotomy has been reported to increase the risk of postoperative upper limb palsy (ULP) in cervical laminoplasty, the specific effects of concomitant foraminotomy on ULP remain uncertain. This study aimed to clarify the effect of concomitant foraminotomy on ULP in cervical laminoplasty., Methods: We identified 19 patients who developed ULP after laminoplasty with concomitant foraminotomy for radiculomyelopathy with nerve root impingement (laminoplasty with concomitant foraminotomy group [F-group]) from 4080 patients who underwent primary cervical laminoplasty at 27 affiliated institutions between 2012 and 2018. An age- and sex-matched control group comprised patients who developed ULP after laminoplasty without concomitant foraminotomy (n = 76, 4:1 ratio with F-group). Collected data included the time of onset and distribution of ULP (side and level). The site of foraminotomy was recorded in the F-group., Results: The F-group showed a significantly higher incidence of ULP than the candidates for the control group (15.1% vs. 3.1%, P < 0.001). The site of foraminotomy was consistent with the distribution of ULP in 79% (15 of 19 patients) of the F-group. The F-group showed a significantly higher proportion of preoperative upper-limb muscle weakness (74% vs. 37%, P = 0.005) and early-onset ULP occurring by postoperative day 1 (63% vs. 33%, P = 0.02) compared with the control group., Conclusions: Our results indicate that the foraminotomy procedure in the stenotic foramen is directly involved in ULP. Combined with a previous report suggesting that early-onset ULP is associated with thermal nerve damage, our results indicate that thermal nerve damage partly explains the increased incidence of ULP in the F-group., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Heat shock proteins (HSPs) play critical roles as molecular chaperones, thereby promoting cellular homeostasis. HSPs are overexpressed in many types of human tumors and their serum concentration is elevated in cancer patients. Recent studies have suggested that HSPs may promote tumorigenesis via interactions with tumor-related proteins. There are only a few studies that address the expression of HSPs in canine tumors. In our previous study, we identified elevated levels of HSP110 expression in canine mammary gland tumors (cMGTs). In this study, we examined both serum concentrations and tissue expression of HSP110 in dogs with cMGT. We found that serum HSP110 concentrations were not significantly different in a comparison between dogs with cMGT (3.44 ± 1.27 μg/mL) and healthy controls (3.23 ± 1.18 μg/mL). By contrast, significant differences in levels of HSP110 expression were identified in comparisons between simple carcinoma and benign mixed tumor (p = 0.001), simple carcinoma and non-neoplastic lesions (p < 0.001), complex carcinoma and benign mixed tumor (p = 0.015), complex carcinoma and non-neoplastic lesions (p < 0.001), simple adenoma and benign mixed tumor (p = 0.041), and simple adenoma and non-neoplastic lesions (p = 0.007). Similarly, significantly different levels of HSP110 expression were identified when comparing grade Ⅲ with non-neoplastic lesion (p = 0.026), grade Ⅱ with benign tumor (p = 0.015), grade Ⅱ with non-neoplastic lesion (p < 0.001), and grade Ⅰ with non-neoplastic lesion (p < 0.001). Taken together, our results indicate that expression of HSP110 correlates with the malignancy in this cohort of dogs diagnosed with cMGT. These findings also suggest that HSP110 is associated with tumorigenesis and the relative malignancy of cMGT., (Copyright © 2020 Elsevier B.V. All rights reserved.)

We experienced a case of fumarate hydratase (FH) -deficient renal cell carcinoma (RCC) suspected of hereditary leiomyomatosis renal cell carcinoma (HLRCC) and herein report our findings. A 42-year-old man with an unremarkable medical history was referred to our hospital with an initial impression of renal cancer, cT3aN2M0. He underwent a right radical nephrectomy with lymph node dissection and showed a pathological diagnosis of FH-deficient RCC, pT3aN2. Clinicopathologic features indicated the possibility of HLRCC; however,-associated RCC. genetic testing showed negative for pathogenic FH mutation.HLRCC is an autosomal dominant condition caused by an FH gene mutation on chromosome 1q43. It is also a syndrome that develops in the smooth muscles of the skin and uterus, and has a renal cancer risk of 10-16%. HLRCC-associated RCC tends to metastasize early and shows poor prognosis. In FH-deficient RCC, the possibility of HLRCC-related RCC should be considered; thus, if patients fulfill the clinical diagnostic criteria, genetic counseling and screening of HLRCC are needed. Even if genetic testing does not confirm HLRCC, FH-deficient RCC still has a poor prognosis and careful follow-up is required.

Hereditary leiomyomatosis and renal cell cancer (HLRCC) is a rare autosomal dominant disorder that results from a germline mutation in the fumarate hydratase gene (FH). Individuals with FH mutations are at risk of developing renal cell carcinoma (RCC). Patients with HLRCC-associated RCC (HLRCC-RCC) have aggressive clinical courses, but there is as yet no standardized therapy for advanced HLRCC-RCC. We report an aggressive RCC case in a 49-year-old man. Nine weeks after undergoing a total nephroureterectomy of the right kidney, he had a metastasectomy at port site. Within 14 weeks of the initial surgery, multiple recurrent tumors developed in the right retroperitoneal space. The pathological diagnosis was FH-deficient RCC. Genetic testing identified a heterozygous germline mutation of FH (c.641&#95;642delTA), which confirmed the diagnosis of HLRCC-RCC. He received combination therapy with the immune checkpoint inhibitors (ICIs) nivolumab and ipilimumab as the first-line therapy. After 31 weeks of ICI treatment, a complete response was achieved. The disease-free condition has been prolonged for 24 months since the initial surgical treatment. This is the first case report of successful treatment of HLRCC-RCC with nivolumab plus ipilimumab. This combination immunotherapy is expected to be an effective approach to treat patients with advanced-stage HLRCC-RCC.

Background: Autoimmune hepatitis (AIH) is a disorder of unknown etiology in which immune-mediated liver injury progress to cirrhosis or hepatocellular carcinoma (HCC). The aim of the present study was to determine whether circulating soluble TIM3 (sTIM3) is elevated in patients with AIH patients and whether sTIM-3 levels are associated with clinical parameters of AIH., Methods: We enrolled 123 Japanese patients with AIH who were identified from the National Hospital Organization-AIH-liver-network database, as well as 32 patients with chronic hepatitis C (CHC), 30 patients with primary biliary cholangitis (PBC) and healthy control subjects. Serum sTIM-3 concentrations were quantified by ELISA., Results: Serum levels of sTIM-3 were significantly higher in AIH patients (median 4865 pg/ml; [interquartile range (IQR); 3122-7471]) compared to those in CHC (1026 pg/ml [IQR: 806-1283] p<0.001), PBC (2395 pg/ml [IQR: 2012-3422] p<0.001) or healthy controls (1285 pg/ml [IQR: 1098-1812] p<0.001). In AIH group, serum sTIM-3 were correlated with alanine aminotransferase (ALT), or total bilirubin (TB) and negatively correlated with serum levels of albumin (Alb). Serum levels of sTIM-3 were also strongly correlated with Mac-2 binding protein glycosylation isomer (M2BPGi) levels, but did not correlate with the histological grade of liver fibrosis. Steroid treatment of AIH patients significantly reduced serum sTIM-3 levels (2147±623pg/ml versus 1321±378pg/ml, p<0.001)., Conclusions: Circulating sTIM-3 levels were elevated in AIH patients and are associated with AIH disease activity and AIH-related liver damage. These findings indicate that serum sTIM-3 correlated with disease status of AIH and could be useful biomarkers to detect autoimmune-mediated liver injury. Our data suggest a possible link between the TIM-3/GAL-9 pathway and AIH severity or phenotype, and further investigations of the TIM-3 pathway and AIH pathophysiology is warranted., Competing Interests: KM has received grants from Chugai, Pfizer and AbbVie. Funds from these grants were not used for this study. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Blood and lymphatic vessels structurally bear a strong resemblance but never share a lumen, thus maintaining their distinct functions. Although lymphatic vessels initially arise from embryonic veins, the molecular mechanism that maintains separation of these two systems has not been elucidated. Here, we show that genetic deficiency of Folliculin, a tumor suppressor, leads to misconnection of blood and lymphatic vessels in mice and humans. Absence of Folliculin results in the appearance of lymphatic-biased venous endothelial cells caused by ectopic expression of Prox1, a master transcription factor for lymphatic specification. Mechanistically, this phenotype is ascribed to nuclear translocation of the basic helix-loop-helix transcription factor Transcription Factor E3 (TFE3), binding to a regulatory element of Prox1, thereby enhancing its venous expression. Overall, these data demonstrate that Folliculin acts as a gatekeeper that maintains separation of blood and lymphatic vessels by limiting the plasticity of committed endothelial cells.

A green method for the oxidation of alcohols to carboxylic acids was developed using a novel co-catalytic system based on gold, silver, and copper catalysts. This reaction system was conducted under atmospheric oxygen in water and mild conditions to selectively oxidize 5-hydroxymethylfurfural to 2,5-furandicarboxylic acid, as a building block for polyethylene furanoate, which is a 100% bio-based, future alternative to the petroleum-based polyethylene terephthalate. Furthermore, various primary alcohols were conveniently oxidized to their corresponding carboxylic acids in up to quantitative yields., Competing Interests: The authors declare no competing financial interest., (© 2020 American Chemical Society.)

Background: Delirium is associated with high mortality after cardiac surgery. However, evidence on the epidemiology of delirium in patients with acute decompensated heart failure (ADHF) is limited. This study aimed to assess the incidence and prognostic impact of delirium in patients with ADHF., Methods: This single-center prospective observational study enrolled 132 consecutive patients with ADHF. We utilized the Diagnostic and Statistical Manual of Mental Disorders, fifth edition and classified the patients into two groups according to the presence or absence of delirium. The primary endpoint was 90-day all-cause mortality. The prognostic impact and risk factors of delirium were evaluated using multivariable Cox and logistic regression analyses, respectively., Results: The median patient age was 83 (interquartile range, 75-87) years. Approximately 51.5% were men. Delirium occurred in 36 (27.3%) patients, and hyperactive delirium was the most frequent type (86.1%). The 90-day all-cause mortality was higher in the patients with delirium than in those without (21.6% versus 3.9%, log-rank p = 0.002). Delirium was associated with higher mortality with an adjusted hazard ratio of 6.8 (95% confidence interval, 1.1-42.6, p = 0.042). The risk factors associated with delirium included advanced age, male sex, higher clinical frailty scale score, and dementia., Conclusions: Delirium was associated with a higher 90-day all-cause mortality in the older adult patients with ADHF. Hyperactive delirium was the most common subtype.

Aims: Hereditary leiomyomatosis and renal cell cancer (HLRCC) is an autosomal dominant disorder caused by germline mutations in fumarate hydratase (FH). Affected families have an increased risk of renal cell carcinoma (RCC). HLRCC-associated RCC (HLRCC-RCC) is highly aggressive. Clinicopathological information of genetically diagnosed patients with HLRCC-RCC contributes to the establishment of effective therapies., Methods: Ten Japanese patients with HLRCC-RCC were enrolled in the study. Genetic testing for FH was carried out. Somatic mutations in FH and immunohistochemical analyses of FH and B7 family ligands (PD-L1 and B7-H3) were investigated in 13 tumours. Copy number variations were evaluated in two tumours., Results: All patients had FH germline mutations. Regarding histology, most tumours had type 2 papillary architecture or tubulocystic pattern or both. All tumours were FH deficient by immunohistochemistry. Ten tumours were positive for PD-L1, and 12 tumours were positive for B7-H3. Somatic mutation analysis demonstrated loss of heterozygosity of FH in 10 tumours. Copy number variation analysis revealed uniparental disomy between 1q24.2 and 1q44 encompassing FH ; gain of chromosome 2 p was also common. All patients had either metastases or residual tumours. Three patients died of HLRCC-RCC and one of colon cancer, whereas the other six are currently alive, including two without recurrence., Conclusions: HLRCC-RCCs appear to have unique molecular profiles, including PD-L1 expression. One patient had complete response to immunotherapy, which may be an option for HLRCC-RCC., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Objective: To elucidate tumor mutation profiles associated with outcomes of uterine cervical cancer (UCC) patients treated with definitive radiotherapy., Methods: Ninety-eight patients with newly diagnosed and pathologically confirmed UCC (82 squamous cell carcinomas, 12 adenocarcinomas, and four adenosquamous carcinomas) who were treated with definitive radiotherapy were analyzed. DNA was extracted from pre-treatment tumor biopsy specimens. The exons of 409 cancer-related genes were sequenced using a next-generation sequencer. Genetic mutations were identified and analyzed for correlations with clinical outcome., Results: Recurrent mutations were observed in PIK3CA (35.7%), ARID1A (25.5%), NOTCH1 (19.4%), FGFR3 (16.3%), FBXW7 (19.4%), TP53 (13.3%), EP300 (12.2%), and FGFR4 (10.2%). The prevalence of mutations in FGFR family genes (i.e., FGFR1-4) was almost as high (24.5%) as that in PIK3CA and ARID1A, both of which are well-studied drivers of UCC. Fifty-five percent (21 of 38) of the identified FGFR mutations were located in the FGFR protein tyrosine kinase domain. Five-year progression-free survival (PFS) rates for FGFR mutation-positive patients (n = 24) were significantly worse than those for FGFR mutation-negative patients (n = 74) (43.9% vs. 68.5%, respectively; P = 0.010). Multivariate analysis identified FGFR mutations as significant predictors of worse 5 year PFS (P = 0.005), independent of clinicopathological variables., Conclusions: FGFR mutations are associated with worse PFS in UCC patients treated with definitive radiotherapy. These results warrant further validation in prospective studies., Competing Interests: Declaration of Competing Interest Yoshiyuki Suzuki holds a research grant from Chugai Pharmaceutical Co., Ltd. for unrelated work, as well as receiving consulting fees from Kyowa Kirin Co., Ltd. Yuya Yoshimoto has received lecture fees from AstraZeneca K.K. Yasushi Sasaki has received research grants from Ono Pharmaceutical Co., Ltd., Daiichi Sankyo Company Limited, Merck Sharp & Dohme, Yuasa Kinenkai, and the Ono Cancer Research Fund for unrelated research, as well as lecture fees from Chugai Pharmaceutical Co., Ltd., (Copyright © 2020 The Author(s). Published by Elsevier Inc. All rights reserved.)

The aryl hydrocarbon receptor (AhR) is a very unstable protein. AhR binds to the molecular chaperone complex (HSP90-p23-XAP2) to maintain a stable structure in the cytoplasm. After binding to ligands, such as dioxin, AhR translocates from the cytoplasm to the nucleus with a molecular chaperone complex. The protein forms a heterodimer with Arnt after nuclear transfer, functions as a transcription factor by binding to a xenobiotic responsive element (XRE), and induces the cytochrome P450 1A1 (CYP1A1). Because of the unstable protein, expression of the full-length AhR in the E. coli expression system is very difficult. Many studies investigated AhR using AhR domains in vitro. We expressed and purified the human full-length AhR in E. coli expression system. Furthermore, specific antibodies were prepared. Purified full-length AhR could bind to ligand. In the presence of ligand, α-helix and random coil of AhR increased and β-sheet decreased on CD spectrum. Full-length AhR could bind to HSP90, XAP2 and p23 in the presence or absence of ligand. We now show the biochemical properties of full-length AhR., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.)

Human patients with inflammatory bowel disease may have poor prognosis with hypozincemia. However, there are limited data on zinc concentrations in the blood of dogs with lymphocytic-plasmacytic enteritis (LPE). The purpose of this study was to investigate the serum zinc concentration in dogs with LPE and its influence on disease severity and prognosis. Thirty-five dogs with LPE were recruited. Serum zinc concentration was measured using atomic absorption spectrometry. Hypozincemia was observed in 18/35 (51%) dogs with LPE. Serum zinc concentration was inversely correlated with histological and clinical severities. Overall survivals were significantly shorter in dogs with hypozincemia than in those without it. These findings suggest that serum zinc concentration is a useful biomarker for LPE severity and prognosis in dogs.

Background: Innate immune cells play a crucial role in the pathophysiology of rheumatoid arthritis (RA) via release of cytokines. Small-molecule inhibitors of Janus kinases (JAKi) are clinically efficacious in patients with RA. However, the isoform-specific action of each JAKi is difficult to assess, since JAKs form heterodimeric complexes with cytokine receptors. We assessed the effects of several JAKi on GM-CSF-primed human innate immune cells., Results: Treatment with JAKi (tofacitinib, baricitinib, upadacitinib) prevented GM-CSF-induced JAK2/STAT5 phosphorylation at higher concentrations (400 nM) in THP-1 cells. Whereas compared with baricitinib or upadacitinib, the inhibitory effects of tofacitinib on the GM-CSF-induced JAK2/STAT5 phosphorylation were weak at lower concentrations (≤ 100 nM). All JAKi inhibited GM-CSF-induced IL-1β production by human neutrophils. However, the inhibitory effects of baricitinib on IL-1β production were larger compared to those of tofacitinib or upadacitinib at lower concentrations (≤ 100 nM). Similarly, all JAKi inhibited GM-CSF-induced caspase-1(p20) production by human neutrophils., Conclusion: We conclude that incubation with JAKi prevents GM-CSF-mediated JAK2/STAT5 activation in human innate immune cells. Although baricitinib and upadacitinib almost completely blocked GM-CSF-mediated JAK2/STAT5 signaling, the inhibitory effects of tofacitinib were weaker at lower concentrations suggesting that variation exists among these JAKi in the inhibition of JAK2 signaling pathways.

Objective: To compare the outcomes of pericardiectomy performed with conventional clipping thoracic duct ligation (C-TDL) to those with en bloc thoracic duct ligation (EB-TDL) using video-assisted thoracoscopic surgery (VATS) for canine idiopathic chylothorax., Study Design: Retrospective consecutive case series., Animals: Thirteen client-owned dogs with idiopathic chylothorax., Methods: Medical records of dogs treated with pericardiectomy in combination with TDL by VATS without intraoperative contrast were reviewed. Five and seven dogs underwent C-TDL and EB-TDL, respectively, and 11 dogs were evaluated by preoperative and 7- to 10-days-postoperative computed tomography-lymphography (CTLG). No clinical symptoms with absent or minimal pleural effusion was defined as clinical improvement. Long-term remission (LTR) was defined as rapid resolution of pleural effusion and no recurrence for more than 1 year. Anesthesia time, operation time, the duration of hospitalization, and time until pleural effusion resolution were compared., Results: Clinical improvement was achieved in 91.7% of the cases (C-TDL, 4/5; EB-TDL, 7/7), excluding one case of intraoperative death. The LTR rate was significantly higher with EB-TDL (6/7 [85.7%]) than with C-TDL (1/5 [20%]). Anesthesia time, operation time, and time until pleural effusion resolution were significantly better with EB-TDL than with C-TDL. The rates of thoracic ducts visualization by postoperative CTLG were 100% (5/5) with C-TDL and 42.9% (3/7) with EB-TDL., Conclusion: En bloc TDL was an effective treatment for canine idiopathic chylothorax in this patient population. It compared favorably to C-TDL, although missed branches at the time of surgery may explain the difference between C-TDL and EB-TDL in this small population of cases., Clinical Significance: En bloc TDL by VATS was an effective minimally invasive treatment for canine idiopathic chylothorax. Computed tomography-lymphography can be used for surgical planning and postoperative evaluation., (© 2019 The American College of Veterinary Surgeons.)

Objective: Posterior lumbar interbody fusion (PLIF) is a widely accepted procedure for degenerative lumbar diseases, and there have been many reports concerning adjacent-segment disease (ASD) after PLIF. In the reports of ASD in which the fusion level was limited to 1 segment, all reports describe ASD of the L3-4 segment after L4-5 PLIF. On the basis of these reports, it is thought that ASD mainly occurs at the cranial segment. However, no report has covered ASD after L3-4 PLIF. Therefore, the authors investigated ASD after L3-4 PLIF., Methods: In conducting a retrospective case series analysis, the authors reviewed a surgical database providing details of all spine operations performed between 2006 and 2017 at a single institution. During that period, PLIF was performed to treat 632 consecutive patients with degenerative lumbar diseases. Of these patients, 71 were treated with L3-4 PLIF alone, and 67 who were monitored for at least 2 years (mean 5.8 years; follow-up rate 94%) after surgery were enrolled in this study. Radiological ASD (R-ASD), symptomatic ASD (S-ASD), and operative ASD (O-ASD) were evaluated. These types of ASD were defined as follows: R-ASD refers to radiological degeneration adjacent to the fusion segment as shown on plain radiographs; S-ASD is a symptomatic condition due to neurological deterioration at the adjacent-segment degeneration; and O-ASD refers to S-ASD requiring revision surgery., Results: All patients had initial improvement of neurological symptoms after primary PLIF. R-ASD was observed in 32 (48%) of 67 patients. It occurred at the cranial segment in 12 patients and at the caudal segment in 24; R-ASD at both adjacent segments was observed in 4 patients. Thus, the occurrence of R-ASD was more significant in the caudal segment than in the cranial segment. S-ASD was observed in 10 patients (15%), occurring at the cranial segment in 3 patients and at the caudal segment in 7. O-ASD was observed in 6 patients (9%): at the cranial segment in 1 patient and at the caudal segment in 5. Thus, the rate of involvement of the caudal segment was 67% in R-ASD, 70% in S-ASD, and 83% in O-ASD., Conclusions: The incidences of R-ASD, S-ASD, and O-ASD were 48%, 15%, and 9%, respectively, after L3-4 PLIF for degenerative lumbar diseases. In contrast to ASD after L4-5 PLIF, ASD after L3-4 PLIF was more frequently observed at the caudal segment than at the cranial segment. In follow-up for patients with L3-4 PLIF, surgeons should pay attention to ASD in the caudal segment.

Raw silk has the potential to be a flexible, osteoconductive material because it forms bone-like apatite on its surface in acellular simulated body fluid with ion concentrations nearly 1.5 times greater than that of human plasma (1.5SBF). It has been reported that silk-which has many similarities to raw silk-develops antibacterial properties when heated in inert gas, which may be advantageous in preventing bacterial infection. Hence, raw silk heated in inert gas may be a flexible, osteoconductive material with antibacterial activity. Thus, we examined the effect of the heat treatment of raw silk fabric on its apatite-forming ability in 1.5SBF and on the growth of Escherichia coli. Raw silk fabric was heated in argon gas at several temperatures, to a maximum of 500 °C. The results of soaking tests in 1.5SBF indicate that the apatite-forming ability of raw silk decreases with increasing temperature. This may be because favourable structures for apatite formation, such as carboxyl groups, are thermally decomposed. The results of bacterial tests indicate that raw silk fabrics heated to 300 °C or 500 °C exhibit reduced bacterial growth compared to those that were not heated or were heated only to 100 °C. This might be because hydrophobic surfaces inhibit bacterial adhesion, or because the thermal decomposition of sericin-a component of raw silk-leads to a lack of available nutrients for the bacteria. Although this study did not demonstrate the expected material properties needed for clinical applications, this research contributes to a better understanding of silk biomaterials.

Bone morphogenetic protein (BMP)-2 plays a central role in bone-tissue engineering because of its potent bone-induction ability. However, the process of BMP-induced bone formation in vivo remains poorly elucidated. Here, we aimed to establish a method for intravital imaging of the entire process of BMP-2-induced ectopic bone formation. Using multicolor intravital imaging in transgenic mice, we visualized the spatiotemporal process of bone induction, including appearance and motility of osteoblasts and osteoclasts, angiogenesis, collagen-fiber formation, and bone-mineral deposition. Furthermore, we investigated how PTH1-34 affects BMP-2-induced bone formation, which revealed that PTH1-34 administration accelerated differentiation and increased the motility of osteoblasts, whereas it decreased morphological changes in osteoclasts. This is the first report on visualization of the entire process of BMP-2-induced bone formation using intravital imaging techniques, which, we believe, will contribute to our understanding of ectopic bone formation and provide new parameters for evaluating bone-forming activity.

Objective: Surgical management of massive ossification of the posterior longitudinal ligament (OPLL) is challenging. To reduce surgical complications, the authors have performed anterior selective stabilization combined with laminoplasty (antSS+LP) for massive OPLL since 2012. This study aimed to elucidate the short-term outcome of the antSS+LP procedure., Methods: The authors' analysis was based on data from 14 patients who underwent antSS+LP for cervical myelopathy caused by massive OPLL and were followed up for at least 2 years after surgery (mean follow-up duration 3.3 years). Clinical outcome was evaluated preoperatively, at 6 months and 1 year postoperatively, and at the final follow-up using the Japanese Orthopaedic Association (JOA) scoring system for cervical myelopathy and the recovery rate of the JOA score. The following radiographic parameters were measured preoperatively, immediately after surgery, at 1 year after surgery, and at the final follow-up: the C2-7 angle, measured on lateral plain radiographs, and the segmental lordosis angle (SLA), measured on sagittal CT scans. The correlation between radiographic parameters and clinical outcomes was evaluated., Results: The mean JOA score increased from 10.4 before surgery to 13.6 and 13.8 at 6 months and 1 year after surgery, respectively; at the final follow-up the mean score was 13.4. This postoperative recovery was significant (p = 0.004) and was maintained until the final follow-up. No patient required revision surgery due to postoperative neurological deterioration. However, the C2-7 angle gradually deteriorated postoperatively. Similarly, the SLA was significantly increased immediately after surgery, but the improvement was not maintained. The recovery rate at the final follow-up correlated positively with the change in C2-7 angle (r = 0.60, p = 0.03) and the change in SLA (r = 0.72, p < 0.01)., Conclusions: AntSS+LP is safe and effective and may be an alternative to anterior decompression and fusion for the treatment of patients with massive OPLL. No postoperative neurological complications or significant postoperative exacerbation of neck pain were observed in our case series. Not only reducing intervertebral motion and decompressing the canal at the maximal compression level but also acquiring segmental lordosis at the maximal compression level are crucial factors for achieving successful outcomes of antSS+LP.