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The present report describes the first known, case of a pseudoaneurysm of the perforating branch of the deep femoral artery following anterior cruciate ligament (ACL) reconstruction. A 19-year-old man underwent ACL reconstruction using the outside-in femoral tunnel-creation method. Seven days after the surgery, he complained of abnormal thigh pain and had swelling with local heat on the distal lateral thigh. Magnetic resonance imaging, computed tomography, and color Doppler ultrasonography showed the pseudoaneurysm in the thigh. Resection surgery was successfully performed by a vascular surgeon 12 days after ACL reconstruction. Careful examination and awareness of postoperative symptoms such as thigh pain and swelling after ACL reconstruction were critical for the early diagnosis of pseudoaneurysm.

Objectives: Because therapeutic options for severe diabetes are currently limited, there is a continuing need for new therapeutic strategies, especially in the field of regenerative medicine. Collaborative efforts across the fields of tissue engineering technology and islet biology may be able to create functionally engineered islets capable of restoring endocrine function in patients with insulin-dependent diabetes., Methods: This engineered scaffold was seeded with isolated primary porcine islets via the pancreatic duct using a multi-step infusion technique. Endocrine function of perfusion-cultured islets in the native scaffold was analyzed by immunohistochemical staining of insulin and glucagon as well as by the insulin stimulation test., Results: The pancreas in this large animal could be uniformly decellularized by perfusion with trypsin and TritonX-100 via the pancreatic duct, as shown by positive staining of extracellular matrix (ECM) components. These scaffolds derived from porcine pancreas were able to maintain the cellular integrity of islets that had repopulated the parenchymal space, which is fundamental for the restoration of endocrine function. Insulin release up to four days after islet infusion was maintained., Conclusions: This scaffold from a large animal maintained islet survival and function in the short-term, retaining the cells as a solid organ in the parenchymal space after infusion through the pancreatic duct. These results suggest that this scaffold is suitable for further fabrication of fully functional bioengineered endocrine pancreases when implanted in vivo. Therefore, it may represent a key improvement in the field of beta-cell replacement therapy. Nonetheless, the facilitation of longer-term islet survival and studies of implantation in vivo is required for successful clinical translation., (Copyright © 2016 IAP and EPC. Published by Elsevier B.V. All rights reserved.)

We report a patient with vascular-type Ehlers-Danlos syndrome (vEDS) who developed pneumothorax and was treated with a total pleural covering technique (TPC). A 24-year-old man developed repeat pneumothorax with intermittent hemo-sputum. Based on unusual radiological manifestations of lung lesions and physical findings, EDS was suspected as an underlying cause of the pneumothorax. Surgical treatment was performed using a mediastinal fat pad and TPC, and no relapse was seen up to 2 years after surgery. TPC is a less invasive surgical approach for selected patients with vEDS. Accurate underlying diagnosis of vEDS and systemic evaluation of vascular complications are necessary before planning surgery.

Plasma membrane-localized pattern recognition receptors (PRRs) such as FLAGELLIN SENSING2 (FLS2), EF-TU RECEPTOR (EFR), and CHITIN ELICITOR RECEPTOR KINASE1 (CERK1) recognize microbe-associated molecular patterns (MAMPs) to activate pattern-triggered immunity (PTI). A reverse genetics approach on genes responsive to the priming agent β-aminobutyric acid (BABA) revealed IMPAIRED OOMYCETE SUSCEPTIBILITY1 (IOS1) as a critical PTI player. Arabidopsis thaliana ios1 mutants were hypersusceptible to Pseudomonas syringae bacteria. Accordingly, ios1 mutants showed defective PTI responses, notably delayed upregulation of the PTI marker gene FLG22-INDUCED RECEPTOR-LIKE KINASE1, reduced callose deposition, and mitogen-activated protein kinase activation upon MAMP treatment. Moreover, Arabidopsis lines overexpressing IOS1 were more resistant to bacteria and showed a primed PTI response. In vitro pull-down, bimolecular fluorescence complementation, coimmunoprecipitation, and mass spectrometry analyses supported the existence of complexes between the membrane-localized IOS1 and BRASSINOSTEROID INSENSITIVE1-ASSOCIATED KINASE1 (BAK1)-dependent PRRs FLS2 and EFR, as well as with the BAK1-independent PRR CERK1. IOS1 also associated with BAK1 in a ligand-independent manner and positively regulated FLS2-BAK1 complex formation upon MAMP treatment. In addition, IOS1 was critical for chitin-mediated PTI. Finally, ios1 mutants were defective in BABA-induced resistance and priming. This work reveals IOS1 as a novel regulatory protein of FLS2-, EFR-, and CERK1-mediated signaling pathways that primes PTI activation., (© 2016 American Society of Plant Biologists. All rights reserved.)

Aims/introduction: Resistin, secreted from adipocytes, causes insulin resistance in mice. In humans, the resistin gene is mainly expressed in monocytes and macrophages. Tunicamycin is known to induce endoplasmic reticulum (ER) stress, and reduce resistin gene expression in 3T3-L1 mouse adipocytes. The aim of the present study was to examine whether ER stress affects resistin gene expression in human monocytes., Materials and Methods: The relationship between resistin messenger ribonucleic acid (mRNA) and ER stress markers mRNA was analyzed by reverse transcription polymerase chain reaction in isolated monocytes of 30 healthy volunteers. The effect of endotoxin/lipopolysaccharides or tunicamycin on resistin gene expression was analyzed in THP-1 human monocytes. Signaling pathways leading to resistin mRNA were assessed by the knockdown using small interfering RNA or overexpression of key molecules involved in unfolded protein response., Results: Resistin mRNA was positively associated with immunoglobulin heavy chain-binding protein (BiP) or CAAT/enhancer binding protein-α homologous protein (CHOP) mRNA in human isolated monocytes. In THP-1 cells, lipopolysaccharides increased mRNA of BiP, pancreatic endoplasmic reticulum eukaryotic initiation factor 2α kinase (PERK) and CHOP, as well as resistin. Tunicamycin also increased resistin mRNA. This induction appeared to be dose- and time-dependent. Tunicamycin-induced resistin mRNA was inhibited by chemical chaperone, 4-phenylbutyric acid. The knockdown of either PERK, activating transcription factor 4 (ATF4) or CHOP reduced tunicamycin-induced resistin mRNA. Conversely, overexpression of ATF4 or CHOP increased resistin mRNA., Conclusions: Endoplasmic reticulum stress induced by tunicamycin increased resistin mRNA through the PERK-ATF4-CHOP pathway in THP-1 human monocytes. ER stress could lead to insulin resistance through enhanced resistin gene expression in human monocytes.

Plants use receptor kinases (RKs) and receptor-like proteins (RLPs) as pattern recognition receptors (PRRs) to sense pathogen-associated molecular patterns (PAMPs) that are typical of whole classes of microbes. After ligand perception, many leucine-rich repeat (LRR)-containing PRRs interact with the LRR-RK BRI1-ASSOCIATED KINASE 1 (BAK1). BAK1 is thus expected to interact with unknown PRRs. Here, we used BAK1 as molecular bait to identify a previously unknown LRR-RLP required for the recognition of the csp22 peptide derived from bacterial cold shock protein. We established a method to identify proteins that interact with BAK1 only after csp22 treatment. BAK1 was expressed transiently in Nicotiana benthamiana and immunopurified after treatment with csp22. BAK1-associated proteins were identified by mass spectrometry. We identified several proteins including known BAK1 interactors and a previously uncharacterized LRR-RLP that we termed RECEPTOR-LIKE PROTEIN REQUIRED FOR CSP22 RESPONSIVENESS (NbCSPR). This RLP associates with BAK1 upon csp22 treatment, and NbCSPR-silenced plants are impaired in csp22-induced defense responses. NbCSPR confers resistance to bacteria in an age-dependent and flagellin-induced manner. As such, it limits bacterial growth and Agrobacterium-mediated transformation of flowering N. benthamiana plants. Transgenic expression of NbCSPR into Arabidopsis thaliana conferred responsiveness to csp22 and antibacterial resistance. Our method may be used to identify LRR-type RKs and RLPs required for PAMP perception/responsiveness, even when the active purified PAMP has not been defined.

Both the α1- and the α2-isomers of mono-ruthenium (Ru)-substituted Dawson-type phosphotungstates with terminal aqua ligands, [α1-P2W17O61Ru(III)(H2O)](7-) (α1-RuH2O) and [α2-P2W17O61Ru(III)(H2O)](7-) (α2-RuH2O), were prepared in pure form by cleavage of the Ru-S bond of the corresponding DMSO derivatives, [α1-P2W17O61Ru(DMSO)](8-) (α1-RuDMSO) and [α2-P2W17O61Ru(DMSO)](8-) (α2-RuDMSO), respectively. Redox studies indicated that α1-RuH2O and α2-RuH2O show proton-coupled electron transfer (PCET), and the Ru(III)(H2O) species was reversibly reduced to Ru(II)(H2O) species and oxidized to Ru(IV)([double bond, length as m-dash]O) species and further to Ru(V)([double bond, length as m-dash]O) species in aqueous solution depending on the pH. Their redox potentials and thermal stabilities were compared with those of the corresponding α-Keggin-type derivatives ([α-XW11O39Ru(H2O)](n-); X = Si(4+) (n = 5), Ge(4+) (n = 5), or P(5+) (n = 4)). The basic electronic and redox features of Ru(L)-substituted Keggin- and Dawson-type heteropolytungstates (with L = H2O or O(2-)) were analyzed by means of density functional calculations. Similar to the corresponding α-Keggin-type derivatives, both α1-RuH2O and α2-RuH2O show catalytic activity for water oxidation.

Helicobacter pylori divides in the human stomach resulting in persistent infections and causing various disorders. Bacterial cell division is precisely coordinated by many molecules, including FtsZ and Min proteins. However, the role of Min proteins in H. pylori division is poorly understood. We investigated the functional characteristics of Min proteins in wild-type HPK5 and five HPK5-derivative mutants using morphological and genetic approaches. All mutants showed a filamentous shape. However, the bacterial cell growth and viability of three single-gene mutants (minC, minD, minE) were similar to that of the wild-type. The coccoid form number was lowest in the minE-disruptant, indicating that MinE contributes to the coccoid form conversion during the stationary phase. Immunofluorescence microscopic observations showed that FtsZ was dispersedly distributed throughout the bacterial cell irrespective of nucleoid position in only minD-disruptants, indicating that MinD is involved in the nucleoid occlusion system. A chase assay demonstrated that MinC loss suppressed FtsZ-degradation, indicating that FtsZ degrades in a MinC-dependent manner. Molecular interactions between FtsZ and Min proteins were confirmed by immunoprecipitation (IP)-western blotting (WB), suggesting the functional cooperation of these molecules during bacterial cell division. This study describes the intrinsic characteristics of Min proteins and provides new insights into H. pylori cell division., (© FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Plant NADPH oxidases, also known as respiratory burst oxidase homologues (RBOHs), produce reactive oxygen species (ROS) that perform a wide range of functions. RbohD and RbohF, two of the 10 Rboh genes present in Arabidopsis, are pleiotropic and mediate diverse physiological processes including the response to pathogens. We hypothesized that the spatio-temporal control of RbohD and RbohF gene expression might be critical in determining their multiplicity of functions. Transgenic Arabidopsis plants with RbohD and RbohF promoter fusions to β-glucuronidase and Luciferase reporter genes were generated. Analysis of these plants revealed a differential expression pattern for RbohD and RbohF throughout plant development and during immune responses. RbohD and RbohF gene expression was differentially modulated by pathogen-associated molecular patterns. Histochemical stains and in vivo expression analysis showed a correlation between the level of RbohD and RbohF promoter activity, H2O2 accumulation and the amount of cell death in response to the pathogenic bacterium Pseudomonas syringae pv. tomato DC3000 and the necrotrophic fungus Plectosphaerella cucumerina. A promoter-swap strategy revealed that the promoter region of RbohD was required to drive production of ROS by this gene in response to pathogens. Moreover, RbohD promoter was activated during Arabidopsis interaction with a non-virulent P. cucumerina isolate, and susceptibility tests with the double mutant rbohD rbohF uncovered a new function for these oxidases in basal resistance. Altogether, our results suggest that differential spatio-temporal expression of the Rboh genes contributes to fine-tune RBOH/NADPH oxidase-dependent ROS production and signaling in Arabidopsis immunity., (© The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Heat shock protein 20 (Hsp20) is a small molecule heat shock protein that plays an important role in plant growth, development, and stress resistance. Little is known about the function of Hsp20 family genes in apple (Malus domestica). Here, we performed a genome-wide analysis of the apple Hsp20 gene family, and a total of 49 Hsp20s genes were identified from the apple genome. Phylogenetic analysis revealed that the 49 genes were divided into 11 subfamilies, and MdHsp18.2b, a member located in the CI branch, was selected as a representative member for functional characterization. Treatment with NaCl and Botryosphaeria dothidea (B. dothidea), the causal agent of apple ring rot disease, significantly induced MdHsp18.2b transcription level. Further analysis revealed that overexpressing MdHsp18.2b reduced the resistance to salt stress but enhanced the resistance to B. dothidea infection in apple calli. Moreover, MdHsp18.2b positively regulated anthocyanin accumulation in apple calli. Physiology assays revealed that MdHsp18.2b promoted H 2 O 2 production, even in the absence of stress factors, which might contribute to its functions in response to NaCl and B. dothidea infection. Hsps usually function as homo- or heterooligomers, and we found that MdHsp18.2b could form a heterodimer with MdHsp17.9a and MdHsp17.5, two members from the same branch with MdHsp18.2b in the phylogenetic tree. Therefore, we identified 49 Hsp20s genes from the apple genome and found that MdHsp18.2b was involved in regulating plant resistance to salt stress and B. dothidea infection, as well as in regulating anthocyanin accumulation in apple calli., (© 2024 Scandinavian Plant Physiology Society.)

Attempts to determine whether class II transactivator (CIITA) is responsible for the impaired up-regulation of major histocompatibility complex II molecules. Evaluation of the surface marker expression on thymic epithelial cells; Relationship between CIITA and the phenotypic characteristics of lymphocytes; Characteristics of thymoma.

The branched-chain α-ketoacid dehydrogenase (BCKDH) complex regulates branched-chain amino acid (BCAA) catabolism by controlling the second step of this catabolic pathway. In the present study, we examined the in vivo effects of treatment with an mTORC1 inhibitor, rapamycin, on cardiac BCKDH complex activity in mice. Oral administration of leucine in control mice significantly activated the cardiac BCKDH complex with an increase in cardiac concentrations of leucine and α-ketoisocaproate. However, rapamycin treatment significantly suppressed the leucine-induced activation of the complex despite similar increases in cardiac leucine and α-ketoisocaproate levels. Rapamycin treatment fully inhibited mTORC1 activity, measured by the phosphorylation state of ribosomal protein S6 kinase 1. These results suggest that mTORC1 is involved in the regulation of cardiac BCAA catabolism.

Problem: A number of endogenous and external factors influence the development of obesity. However, the factors responsible for these differences in obesity pathogenesis between males and females are largely unknown., Methods: We investigated the expression of 35 genes related to lipid and glucose metabolism and to receptors for insulin signaling in white adipose tissue (WAT) of 8-week-old 129/Sv mice and mice fed standard diet (STD) or high fat diet (HFD) for 35 weeks in males and females., Results: At 8 weeks, the expression levels of two genes for fatty acid synthesis, Acaca and Fasn, were higher in females than in males. Female mice fed a STD for 35 weeks also had higher expression levels of an additional four genes related to glucose transporters (Slc2a1 and Slc2a4) and adipokines (Adipoq and Nampt). The expression levels of these six genes were also higher in females than in males fed a HFD for 35 weeks. At 43 weeks old, the female-to-male expression ratio of these six genes was similar for the STD and HFD groups. Furthermore, glucose tolerance testing showed that the half-life for the elimination of elevated blood glucose was shorter in females than males, although blood glucose parameters were generally similar between females and males., Conclusions: These findings suggest that sex and aging may cause diet-independent differences in gene expression levels in female and male mice, and that higher expression of these genes in females could contribute to higher metabolic activity and resistance to obesity compared with males., (Copyright © 2015 Asia Oceania Association for the Study of Obesity. Published by Elsevier Ltd. All rights reserved.)

Objective: To investigate the prevalence and the risk factors of surgical-site infection (SSI) and delayed wound healing (DWH) in patients with rheumatoid arthritis (RA) underwent orthopedic surgery., Methods: We reviewed the records of 1036 elective orthopedic procedures undertaken in RA patients. Risk factors for SSI and DWH were assessed by logistic regression analysis using age, body mass index, disease duration, pre-operative laboratory data, surgical procedure, corticosteroid use, co-morbidity, and use of conventional synthetic disease-modifying anti-rheumatic drugs (csDMARDs) and biological DMARDs (bDMARDs) as variables., Results: SSI and DWH were identified in 19 cases and 15 cases, respectively. One case of SSI and three cases of DWH were recorded among 196 procedures in patients using bDMARDs. Foot and ankle surgery was associated with an increased risk of SSI (odds ratio (OR), 3.167; 95% confidence interval (CI), 1.256-7.986; p = 0.015). Total knee arthroplasty (TKA; OR, 4.044; 95% CI, 1.436-11.389; p = 0.008) and disease duration (OR, 1.004; 95% CI, 1.000-1.007; p = 0.029) were associated with an increased risk of DWH., Conclusions: Our results indicated foot and ankle surgery, and TKA and disease duration as risk factors for SSI and DWH, respectively. bDMARDs was not associated with an increased risk of SSI and DWH.

Membrane proteins are difficult to study for numerous reasons. The surface of membrane proteins is relatively hydrophobic and sometimes very unstable, additionally requiring detergents for their extraction from the membrane. This leads to challenges at all levels, including expression, solubilization, purification, identification of associated proteins, and the identification of post-translational modifications. However, recent advances in immunoprecipitation technology allow to isolate membrane proteins efficiently, facilitating the study of protein-protein interactions, the identification of novel associated proteins, and to identify post-translational modifications, such as phosphorylation. Here, we describe an optimized immunoprecipitation protocol for plant plasma membrane receptor-like kinases.

Introduction: We previously introduced video-assisted thoracoscopic ET (VATS-ET) as a therapeutic option for MG with acceptable results. We have conducted further investigations to improve the procedure without deterioration of operative results, including myasthenia gravis (MG) remission rate and palliation rate. Here, we report the details of our current procedure, as well as surgical results and patient outcomes as compared with the original VATS-ET procedure., Material and Methods: From January 2002 to September 2013, we performed a VATS-ET procedure with an anterior chest wall lifting method for 77 patients who had MG with or without a thymoma. During that period, we investigated the appropriate indications and improved the procedure., Results: Our current indication for this procedure is MG with the anti-acetylcholine receptor antibody or sero-negative type, or MG with a thymoma <5 cm in diameter without invasion to adjacent organs. With our procedure, the thymus and surrounding tissue are sufficiently resected using a bilateral thoracoscopic surgical method without neck incision. Remission and palliation rates were found to be equivalent to those obtained with the original VATS-ET procedure., Conclusion: VATS-ET is suitable for select patients with MG with or without a thymoma. In addition, our current method has shown to be effective while also offering cosmetic advantages as compared with the original, neck incision needed, VATS-ET method.

Objective: It has been reported that administration of octanoic acid, one of medium-chain fatty acids (MCFAs), promoted leucine oxidation in vitro and in vivo, but it remained unclear how octanoic acid stimulated leucine oxidation. Therefore, the aim of this study was to elucidate the mechanism that octanoic acid facilitates branched-chain amino acid (BCAA) catabolism., Materials/methods: In in vivo experiments, male rats were orally administered MCFAs as free fatty acids or triacylglycerol (trioctanoin), and then activities of hepatic branched-chain α-ketoacid dehydrogenase (BCKDH) complex (BCKDC) and BCKDH kinase (BDK) and alterations in the concentration of blood components were analyzed. In in vitro experiments, purified BCKDC associated with BDK (BCKDH-BDK complex) was reacted with various concentrations of hexanoic, octanoic, and decanoic acids., Results: Oral administration of trioctanoin in rats activated hepatic BCKDC via down-regulation of BDK activity in association with a decrease in plasma BCAA concentration and an increase in serum ketone body concentration. In vitro experiments using purified BCKDH-BDK complex showed that MCFAs (hexanoic, octanoic, and decanoic acids) inhibited BDK activity and that this inhibition was higher in hexanoic and octanoic acids than in decanoic acid. Oral administration of octanoic acid, but not decanoic acid, in rats activated hepatic BCKDC via down-regulation of BDK activity by decreasing the amount of BDK bound to the complex. The serum ketone body level was elevated by both administration of octanoic acid and decanoic acid., Conclusion: These results suggest that octanoic acid promotes BCAA catabolism in vivo by activation of BCKDC via decreasing the bound form of BDK., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Pathogen recognition induces the production of reactive oxygen species (ROS) by NADPH oxidases in both plants and animals. ROS have direct antimicrobial properties, but also serve as signaling molecules to activate further immune outputs. However, ROS production has to be tightly controlled to avoid detrimental effects on host cells, but yet must be produced in the right amount, at the right place and at the right time upon pathogen perception. Plant NADPH oxidases belong to the respiratory burst oxidase homolog (RBOH) family, which contains 10 members in the model plant Arabidopsis thaliana. The perception of pathogen-associated molecular patterns (PAMPs) by pattern recognition receptors (PRRs) leads to a rapid, specific and strong production of ROS, which is dependent on RBOHD. RBOHD is mainly controlled by Ca(2+) via direct binding to EF-hand motifs and phosphorylation by Ca(2+)-dependent protein kinases. Recent studies have, however, revealed a critical role for a Ca(2+)-independent regulation of RBOHD. The plasma membrane-associated cytoplasmic kinase BIK1 (BOTRYTIS-INDUCED KINASE1), which is a direct substrate of the PRR complex, directly interacts with and phosphorylates RBOHD upon PAMP perception. Impairment of these phosphorylation events completely abolishes the function of RBOHD in immunity. These results suggest that RBOHD activity is tightly controlled by multilayered regulations. In this review, we summarize recent advances in our understanding of the regulatory mechanisms controlling RBOHD activation., (© The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Interactions between stromal and epithelial cells play important roles in the development, homeostasis, and pathological conditions of the cornea. Soluble cytokines are critical factors in stromal-epithelial interactions, and growth factors secreted from corneal stromal cells contribute to the regulation of proliferation and differentiation of corneal epithelial cells (CECs). However, the manner in which the expression of growth factors is regulated in stromal cells has not been completely determined. To study stromal-epithelial cell interactions, we used an organotypic culture model. Human or rabbit CECs (HCECs or RCECs) were cultured on amniotic membranes placed on human corneal fibroblasts (HCFs) embedded in a collagen gel. The properties of the organotypic culture were examined by hematoxylin-eosin staining and immunofluorescence. In the organotypic culture, HCECs or RCECs were stratified into two-three layers after five days and five-seven layers after nine days. However, stratification was not observed when the HCECs were seeded on a collagen gel without fibroblasts. K3/K12 were expressed on day 9. The HCF-embedded collagen gels were collected on days 3, 5, or 9 after seeding the RCECs, and mRNA expression of growth factors FGF7, HGF, NGF, EGF, TGF-α, SCF, TGF-β1, TGF-β2, and TGF-β3 were quantified by real-time PCR. mRNA expression of the growth factors in HCFs cultured with RCECs were compared with those cultured without RCECs, as well as in monolayer cultures. mRNA expression of TGF-α was markedly increased in HCFs cultured with RCECs. However, mRNA expression of the TGF-β family was suppressed in HCFs cultured with RCECs. Principal component analysis revealed that mRNA expression of the growth factors in HCFs were generally similar when they were cultured with RCECs. In organotypic cultures, the morphological changes in the CECs and the expression patterns of the growth factors in the stromal cells clearly demonstrated stromal-epithelial cell interactions, and the results suggest that stromal cells and epithelial cells may act in concert in the cornea., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Objectives: The purpose of this study is to assess the clinicopathological characteristics of non-small-cell lung cancer (NSCLC) occurring in the left lingular division (LLD) in association with a proposal of the LLD-specific regional lymph node stations., Methods: Medical records of patients, who underwent complete tumour resection with mediastinal lymph node dissection (MLND) for LLD-NSCLC from 2000 to 2009 in multiple institutions, were retrospectively examined. We analysed patient clinicopathological characteristics and obtained the LLD-specific regional lymph node stations, and then the validity of intraoperative navigation in lymphadenectomy for LLD-NSCLC was investigated., Results: One hundred and eighty-four LLD-NSCLC patients (97 males and 87 females, and 128 adenocarcinomas and 56 non-adenocarcinomas) were studied. The 5-year overall survival (OS) and disease-free survival (DFS) rates for all LLD-NSCLC patients were 72.9 and 58.3%, respectively. We examined the lymph node metastasis patterns in 42 node-positive tumours. The frequent metastatic lymph node stations were #12u lobar node (n = 22), #5 subaortic node (n = 15) and #11 interlobar node (n = 13) in order. These three node stations were also single metastatic sites in some patients. Metastases to sub-carinal (#7) or inferior mediastinal nodes (#8) were rare. Thus, we assigned the three stations (#5, #11, #12u) as the regional lymph node stations for LLD-NSCLC. If these regional lymph node stations had been examined pathologically during surgery for a total of 160 LLD-NSCLC patients with c-T2N1M0 or lower stage disease, 125 p-N0 and 5 p-N1 patients diagnosed with no metastasis would have been subjected to selective MLND, while 14 p-N1 and all 16 p-N2 patients diagnosed with metastasis would have had complete MLND carried out. As a result, these regional lymph node stations could accurately predict the existence of p-N2 metastasis, and appropriately lead to a selective or complete MLND., Conclusions: An intraoperative pathological examination using our proposed LLD-specific regional lymph node stations may accurately diagnose the status of node metastasis, and appropriately lead to selective or complete MLND in LLD-NSCLC patients with c-T2N1M0 or lower stage disease., (© The Author 2015. Published by Oxford University Press on behalf of the European Association for Cardio-Thoracic Surgery. All rights reserved.)

Metallothionein (MT) family proteins are small molecular weight and cysteine-rich proteins that regulate zinc homeostasis and have potential protective effects against oxidative stress and toxic metals. To investigate whether MTs play a role in longevity determination in mammals, we measured the lifespans of wild-type (WT) and MT-1 and -2 gene knockout (MTKO) mice in a 129/Sv genetic background. MTKO mice of both sexes had shorter lifespans than WT mice. In particular, male MTKO mice living beyond the mean lifespan exhibited signs of weight loss, hunchbacked spines, lackluster fur and an absence of vigor. These results suggest that lifespan is shortened due to accelerated senescence in the absence of MT genes., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Thymectomy is regarded as a useful therapeutic option for myasthenia gravis (MG), though perioperative management in MG patients is largely empirical. While evidence-based medicine is limited in the perioperative management of MG patients, treatment guidelines are required as a benchmark. We selected issues faced by physicians in clinical practice in the perioperative management of extended thymectomy for MG, and examined them with a review of the literature. The present guidelines have reached the stage of consensus within the Japanese Association for Chest Surgery.

Introduction: Non-exposed endoscopic wall-inversion surgery (NEWS) is a novel technique of endoscopic full-thickness resection without transluminal access mainly designed to treat gastric cancer. Here, we report a successful case of NEWS with sentinel node basin dissection (SNBD) for early gastric cancer (EGC) with the risk of lymph node metastasis., Patient and Methods: A 55-year-old female patient with a 2-cm, diffuse-type intramucosal EGC with ulceration was referred to our hospital for a less invasive gastrectomy based on sentinel node navigation surgery. After obtaining informed consent, NEWS with SNBD was applied. After placing mucosal markings, indocyanine green solution was injected endoscopically into the submucosa around the lesion to examine sentinel nodes (SNs). The SN basin (the area of the left gastric artery), including three stained SNs(#3), was dissected, and an intraoperative pathological diagnosis confirmed that no metastasis had occurred. Subsequently, NEWS was performed for the primary lesion. Serosal markings were placed laparoscopically, submucosal injection was added endoscopically, and circumferential sero-muscular incision and suturing were performed laparoscopically, with the lesion inverted toward the inside of the stomach. Finally, the circumferential mucosal incision was performed, and the lesion was retrieved perorally., Results: The operation was finished in 270 min without complications. The patient was uneventfully discharged 10 days after the procedure. The final pathological diagnosis was coincident with the pre- and intraoperative assessment., Conclusions: We demonstrated the feasibility and safety of NEWS with SNBD with a favorable result. This surgical concept is expected to become a promising, minimally invasive, function-preserving surgery to cure cases of EGC that are possibly node-positive.

Introduction: Plants detect the invasion of microbial pathogens through pathogen-associated molecular patterns (PAMPs). Cold shock proteins (CSPs) are a class of PAMPs specifically recognized by Solanales plants. While peptide inoculation studies have revealed the effects of CSPs, their in vivo roles remain poorly understood. Methods: A model system involving the interactions between Pseudomonas fluorescens and P. syringae pv. tomato DC3000 with Nicotiana benthamiana has been widely used to investigate the molecular mechanism of plant-microbe interactions. Here, we employed this model system to explore the in vivo roles of CSPs in modulating plant immunity by multiple genetic approaches. Results: Our findings revealed that three highly-conserved CSPs were identified in Pseudomonas strains. Transient expression of these CSPs neither induced reactive oxygen species (ROS) production nor suppressed the hypersensitive response (HR) in N. benthamiana , however, it restricted bacterial effector translocation. Genetic analysis revealed that these CSPs did not contribute to the ROS burst or HR inhibition in vivo but were functionally redundant in suppressing effector translocation in a flagellin (FliC)-independent manner. Furthermore, we demonstrated that the suppression of effector translocation mediated by CSPs was less pronounced compared to that triggered by FliC. Additionally, inoculation with csp15 and csp22 epitopes triggered the pattern-triggered immunity-associated suppression of effector translocations. Discussion: This study revealed the redundant roles of CSPs in suppressing bacterial effector translocation in vivo , providing deep insights into the PTI elicited by cytoplasmic bacterial proteins. [ABSTRACT FROM AUTHOR]

Osteoarthritis is a degenerative disease of cartilage, and exosome derived from mesenchymal stem cells (MSCs) are considered promising for treating inflammatory musculoskeletal disorders, although their mechanisms are not fully understood. This study aimed to investigate the effects of exosomes derived from canine bone marrow mesenchymal stem cells (cBMSCs-Exos) on the expression of inflammatory factors and genes related cartilage matrix metabolism in IL-1β-induced canine chondrocytes. Canine BMSCs were isolated and characterized for surface markers and trilineage differentiation. Exosomes were then extracted and performed surface labeling detection. Canine chondrocytes were exposed to IL-1β to mimic osteoarthritis in vitro. Subsequently, the chondrocytes were treated with exosomes from BMSCs, and the expression levels of related genes and IL-6 protein were assessed. The mesenchymal stem cells isolated from bone marrow and cultured exhibited positive CD44 and CD90, negative expression of CD45 and HLA, and demonstrated potential to differentiate into adipocytes, osteoblasts and chondrocytes. Exosomes from BMSCs exhibited positivity expression of CD9, CD63 and CD81. Treatment with exosomes significantly reduced IL-6 and TNF-α mRNA levels induced by IL-1β, as well as IL-6 protein expression. Additionally, a significant decrease was observed in the mRNA levels catabolic marker genes MMP-13, ADAMTS-5, and COX2. Conversely, there was a significant increase in the mRNA levels of anti-inflammatory cytokines IL-4, IL-10, and anabolic marker genes, such as COL2A1, ACAN, and SOX9. cBMSCs-Exos play a vital role in cartilage protection by suppressing the expression of pro-inflammatory and anabolic genes while simultaneously enhancing the expression of genes involved in synthesis metabolism. [ABSTRACT FROM AUTHOR]

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Indole-3-acetic acid (IAA) is the most common form of the phytohormone auxin found in plants. IAA has been previously reported from mammals, and has recently been observed to be widespread in arthropods, particularly insects. Several pathways for the biosynthesis of IAA from tryptophan (Trp) have been mapped and documented in plants, bacteria, and fungi, and a new pathway has been proposed from an insect. The widespread distribution of IAA in insects also argues that synthesis rather than consumption and sequestration accounts for its presence. We used exemplar enzymes from all known plant and bacterial Trp-based IAA biosynthesis pathways to search the 1KITE database of 670 hexapod and insect transcriptomes for evidence of a complete biosynthetic pathway for IAA. We first aligned exemplar enzymes to transcriptomes and then aligned transcripts with alignments against a database of well-annotated insect and hexapod genomes to ensure that we were not identifying paralogs. We found that none of the currently recognized IAA biosynthesis pathways were widespread in Hexapoda and Insecta (occur in > 80% species). However, transcripts encoding proteins homologous to enzymes in the pathway that converts Trp → TAM → IAAld → IAA via tryptamine (TAM) and indole–3-acetaldehyde (IAALD) were detected in the transcriptomes of most species within the Hexapoda, Palaeoptera, and Polyneoptera, but were also detected in genomes largely from the Holometabola. Transcripts encoding proteins homologous to two enzymes in the TAM pathway, aromatic-L-amino-acid/L-tryptophan decarboxylase (EC: 4.1.1.28 and EC: 4.1.1.105) and aldehyde dehydrogenase/indole-3-acetaldehyde oxidase (EC: 1.2.1.3, EC: 1.2.3.7) were found to be widespread in Hexapoda and Insecta. A newly proposed pathway for the biosynthesis of IAA in insects based on an examination of Euura sp. ‘Pontania’ and Bombyx mori, suggests that an aromatic aldehyde synthase (PonAAS2) is responsible for the conversion of Trp → IAAld and an aldehyde oxidase (BmIAO1) is responsible for the conversion of IAAld → IAA in Bombyx mori. We detected aromatic aldehyde synthases (AAS) and aldehyde oxidases (AO) in about 33% of our transcriptomes with RNA extracted largely from adults, which would imply that these enzymes are not widespread in Insects and Hexapoda. However, when we examined a sample of 167 insect genomes, we detected at least one aromatic aldehyde synthase and at least one aldehyde oxidase in 80.8 and 94% of the genomes, respectively, which suggest that these enzymes are more widespread in insect genomes even if undetected in transcriptomes of adult insects and hexapods. However, it is likely that some of these putative aromatic aldehyde synthase proteins are homologs of 3, 4-dihydroxyphenylacetaldehyde synthase, which is involved in cuticular hardening, rather than being homologs of the sole aromatic aldehyde synthase as yet identified to convert Trp to IAAld. So, both the TAM pathway and the newly proposed insect pathway based on Euura sp. ‘Pontania’ could account in part for the biosynthesis of IAA in insects and hexapods. Our results also suggest that an aldehyde dehydrogenase could contribute to the biosynthesis of IAA by the enzymatic conversion of IAAld to IAA. Further careful biochemical experiments and efforts to characterize additional AAS enzymes capable of converting Trp to IAAld might guide a refined assessment of the variety of enzymes involved and the breadth and distribution of IAA biosynthesis pathway in Hexapods and Insecta. [ABSTRACT FROM AUTHOR]

Objectives: Complete removal of the tumor and surrounding tissue is the most important prognostic factor such as survival after surgery. When the tumor invades the phrenic nerve, the impact of intraoperative phrenic nerve sacrifice on the short- and long-term prognosis of patients is not clear. This study aims to explore the differences in prognosis between patients with malignant thymoma with and without phrenic nerve sacrifice during surgery, as well as analyze related factors. Methods: A total of 209 patients who underwent thymoma resection in the Department of Thoracic Surgery at our hospital from February 2006 to November 2022 were collected for retrospective analysis. The groups were divided into two according to the presence or absence of intraoperative phrenic nerve sacrifice. A comparative analysis was conducted on postoperative complications, long-term survival recurrence between the two groups. Cox regression was used to analyze the factors related to the differences in short- and long-term prognosis between two groups. Results: 29.6% of patients developed phrenic nerve sacrifice during thymoma surgery. Compared to patients without phrenic nerve sacrifice, the long-term survival rate was lower (P = 0.031). The independent risk factors for reduced long-term survival were intraoperative phrenic nerve sacrifice, secondary postoperative complications, and modified Masaoka staging III/IV. Conclusion: Our data show that nearly one-third of patients develop phrenic nerve sacrifice during complete resection of thymoma. Phrenic nerve sacrifice has significant impact on short-term complications and long-term survival. Secondary postoperative complications and modified Masaoka staging III/IV are also risk factors for reduced long-term survival. [ABSTRACT FROM AUTHOR]