Sustained activation of NF-κB through constitutively active IKKß leads to senescence bypass in murine dermal fibroblasts.

Although the transcription factor nuclear factor κB (NF-κB) plays a central role in the regulation of senescence-associated secretory phenotype (SASP) acquisition, our understanding of the involvement of NF-κB in the induction of cellular senescence is limited. Here, we show that activation of the canonical NF-κB pathway suppresses senescence in murine dermal fibroblasts. IκB kinase ß (IKKß)-depleted dermal fibroblasts showed ineffective NF-κB activation and underwent senescence more rapidly than control cells when cultured under 20% oxygen conditions, as indicated by senescence-associated ß-galactosidase (SA-ß-gal) staining and p16INK4a mRNA levels. Conversely, the expression of constitutively active IKKß (IKKß-CA) was sufficient to drive senescence bypass. Notably, the expression of a degradation-resistant form of inhibitor of κB (IκB), which inhibits NF-κB nuclear translocation, abolished senescence bypass, suggesting that the inhibitory effect of IKKß-CA on senescence is largely mediated by NF-κB. We also found that IKKß-CA expression suppressed the derepression of INK4/Arf genes and counteracted the senescence-associated loss of Ezh2, a catalytic subunit of the Polycomb repressive complex 2 (PRC2). Moreover, pharmacological inhibition of Ezh2 abolished IKKß-CA-induced senescence bypass. We propose that NF-κB plays a suppressive role in the induction of stress-induced senescence through sustaining Ezh2 expression.

A Case of a Chronic Expanding Hematoma Caused by an Epidermal Cyst.

Chronic expanding hematoma (CEH) is a rare type of hematoma that expands slowly and continuously without compromising coagulation. Its etiology is often unknown. However, we experienced a rare case of CEH, in which an epidermal cyst was thought to be the cause. A 57-year-old man had developed a painless soft-tissue tumor in his left buttock 45 years earlier, which slowly grew to 11 cm in diameter. Preoperative magnetic resonance imaging suggested a large cyst containing several masses. Surgery revealed a hematoma and keratin debris surrounded by a white fibrous cyst wall and a fibrous nodule measuring 4 cm in diameter. On histopathological examination of the white fibrous wall, an epidermal component was observed contralateral to the superficial punctum, but the epidermal component was absent from most of the wall, including the fibrous nodule. Based on a pathological examination, the CEH was suggested to have been caused by partial rupturing and inflammation of an epidermal cyst. To the best of our knowledge, there are no reports of epidermal cysts causing CEH. In addition, the large fibrous nodule protruding from the CEH cyst wall was considered to be rare. This was considered to be a rare CEH that may have originated from an epidermal cyst.

A Sublatissimus Dorsi Abscess Due to Pasteurella multocida Caused by a Cat Scratch.

We encountered a 51-year-old male patient who was not immunocompromised. Thirteen days before his admission, his right forearm was scratched by his pet cat. Swelling, redness, and purulent discharge appeared at the site, but he did not seek medical attention. He developed a high fever and was hospitalized with a diagnosis of septic shock, respiratory failure, and cellulitis on plain computed tomography. After admission, the swelling on his forearm was relieved with empirical antibiotics, but the symptoms spread from his right axilla to his waist. We suspected necrotizing soft tissue infection and made a trial incision in the lateral chest up to the latissimus dorsi, but were unable to prove it. However, an abscess was later found under the muscle layer. Second incisions were made to allow the abscess to drain. The abscess was relatively serous, and no tissue necrosis was observed. The patient's symptoms improved rapidly. In retrospect, the patient probably already had the axillary abscess on admission. It may have been detected at this point if contrast-enhanced computed tomography had been performed, and early axillary drainage may have accelerated the patient's recovery, which could also have prevented the formation of the latissimus dorsi muscle abscess. In conclusion, the Pasteurella multocida infection on the patient's forearm induced a very unusual presentation and caused an abscess to form under the muscle, unlike necrotizing soft tissue infections. Early contrast-enhanced computed tomography may aid earlier and more appropriate diagnosis and treatment in such cases.

Deletion of IKKß in activated fibroblasts promotes tumor progression in melanoma.

Cancer-associated fibroblasts (CAFs) are a major component of the tumor microenvironment and play critical roles in tumorigenesis. CAFs consists of multiple subpopulations, which have diverse functions. The detailed mechanism, including the role of NF-κB, a critical transcription factor for inflammation and cell survival, in CAFs has not been adequately explored. In this study, we examined the roles of IKKß, a key kinase for NF-κB activation, in activated CAFs by using mice (KO mice) with deletion of IKKß in activated fibroblasts (aFbs). We found that melanoma cells implanted in KO mice showed significantly more growth than those implanted in control mice. To exclude the effects of deletion of IKKß in cells other than aFbs, we implanted a mixture of melanoma cells and IKKß-deleted aFbs in wild-type mice and observed that the mixture showed greater growth than a mixture of melanoma cells and normal aFbs. In exploring the mechanisms, we found that conditioned medium from IKKß-deleted aFbs promotes the proliferation of melanoma cells, and the expression of growth arrest-specific 6 (GAS6) and hepatocyte growth factor (HGF), which are major tumor-promoting factors, was upregulated in IKKß-deleted aFbs. These results indicated the tumor-suppressing function of IKKß in activated CAFs.

Effects of constitutively active IKKß on cardiac development.

NF-κB is a major transcription factor regulating cell survival, organ development and inflammation, but its role in cardiac development has been inadequately explored. To examine this function, we generated mice in which IKKß, an essential kinase for NF-κB activation, was constitutively activated in embryonic cardiomyocytes. For this purpose, we used smooth muscle-22α (SM22α)-Cre mice, which are frequently used for gene recombination in embryonic cardiomyocytes. Embryonic hearts of SM22αCre-CA (constitutively active) IKKßflox/flox mice revealed remarkably thin, spongy and hypoplastic myocardium. In exploring the mechanism, we found that the expression of bone morphogenetic protein 10 (BMP10) and T-box transcription factor 20 (Tbx20), major regulators of cardiac development, was significantly downregulated and upregulated, respectively, in the SM22αCre-CAIKKßflox/flox mice. We also generated NK2 homeobox 5 (Nkx2.5) Cre-CAIKKßflox/wt mice since Nkx2.5 is also expressed in embryonic cardiomyocytes and confirmed that the changes in these genes were also observed. These results implicated that the activation of NF-κB affects cardiac development.

Costal Cartilage Graft to Prevent Drooping after Free Flap Reconstruction of the Lower Lip.

For large lower lip defects, a thin flap combined with a tendon is the standard reconstructive option. However, this method can result in flap ptosis, which occurred in two of our patients. To correct the ptosis, we transplanted costal cartilage into the reconstructed lower lips, which produced good or moderate results. We report our experience based on long-term follow-up. In case 1, reconstruction was performed with a latissimus dorsi myocutaneous flap. Within 10 years of the first cartilage transplant, two additional surgeries were required due to cartilage/screw breakage. These problems may have been triggered by the bulkiness of the flap and/or the angle at which the cartilage was anchored in place. There have not been any further problems for 3 years. In case 2, reconstruction was performed with a free anterolateral thigh flap. The skin around the flap had poor extensibility, and the patient had marked Class II occlusion. We grafted cartilage without fixing it to the mandible. However, temporary interference with the maxillary dentition was observed. In conclusion, costal cartilage grafts are effective against flap ptosis after free flap reconstruction of the lower lip in patients without Class II occlusion. To achieve long-term stability, the optimal angle and positioning of the cartilage and the extensibility of the skin must be thoroughly investigated before surgery, and a thick piece of cartilage must be firmly fixed in place.

A Ventral Hernia-repair-related Mycobacterium mageritense Mesh Infection Treated with NPWT without Mesh Removal.

Abdominal hernias are often repaired using prosthetic mesh, which is susceptible to infections. Normally, it is necessary to remove the mesh. However, successful mesh salvation with negative-pressure wound therapy (NPWT) has recently been reported. We encountered Mycobacterium(M) mageritense infection after hernia repair using the mesh. M. mageritense is classified as a fast-growing nontuberculous mycobacterium, but few cases have been reported. Nontuberculous mycobacterium can cause rare chronic infections. Skin and soft-tissue infections by nontuberculous mycobacterium involving localized abscess formation and chronic abscesses under various situations have been reported. We report an 85-year-old woman in whom a ventral hernia repair-related M. mageritense mesh infection was treated with NPWT without mesh removal. The hernia was repaired using Bard Ventralex mesh. Pus discharge was seen on the seventh postoperative day, and there was a small area of necrosis under the mesh. From the 13th postoperative day, NPWT was performed for 4 weeks. On the 29th postoperative day, a M. mageritense infection was diagnosed, which was resistant to multiple drugs. After the NPWT, most of the wound showed good granulation tissue formation. In conclusion, the mesh used to repair a hernia became infected with M. mageritense, but NPWT was able to salvage it. In cases of mesh infection involving small necrotic areas, performing NPWT under the guidance of an infectious disease expert may make it possible to preserve the mesh.

A Case of Dissecting Cellulitis which Was Initially Suspected to Be a Trichilemmal Cyst.

We treat infected cysts on a daily basis, but it is difficult to diagnose similar lesions produced by inflammatory conditions that are not primarily caused by bacteria. Dissecting cellulitis of the scalp (DCS) is a chronic inflammatory disease that results in disfiguring, painful, and purulent lesions. It often takes a long time to diagnose. The pathophysiology of DCS remains unclear. Various treatments for DCS have been proposed, depending on the severity of the disease. However, none of these treatments are clearly superior to the others. If DCS spreads to the entire occipital region, aggressive surgical treatment may be beneficial in terms of the patient's quality of life. However, surgical interventions, such as drainage, are not effective at preventing the progression of the disease. Herein, we report the case of a young female patient who developed a cyst in the occipital region. We initially suspected that the lesion was a normal infected trichilemmal cyst. However, DCS was subsequently suspected because the lesion exhibited an unusual course after drainage and debridement. We consider that we made a diagnosis relatively early, but if we had sufficient knowledge about DCS we could have made a diagnosis even earlier by performing debridement sooner. Minocycline was administered for 5 months, which caused the lesion to disappear. After 2 years, no recurrence had been observed.

A case of cryptococcal necrotizing fasciitis and immune reconstitution inflammatory syndrome in a renal transplantation recipient.

INTRODUCTION AND IMPORTANCE: Immunocompromised patients are at high risk of unexpectedly serious infections caused by uncommon bacteria or fungi. We experienced a case of Cryptococcus neoformans-induced necrotizing fasciitis (NF) of the lower extremities. The progress so far has been reported by the urology department [1]. Moreover, after the NF had been treated, the patient developed immune reconstitution inflammatory syndrome (IRIS). We report from surgeon's view point. CASE PRESENTATION: A 51-year-old male renal transplant patient complained of pain in both lower extremities (LE). After the initial debridement, periodic acid-Schiff after diastase digestion (D-PAS) staining confirmed the diagnosis. No symptoms were seen in the lungs or cerebrospinal system. The patient was reluctant to undergo surgical treatment but several debridement improved patient's condition. After the LE wound healed, prednisolone was discontinued, then painful nodules appeared on both LE. Based on the negative culture results and the fact that the patient had been treated with flucytosine and fluconazole, we suspected that the nodules had been caused by IRIS. CLINICAL DISCUSSION: It was difficult to diagnose Cryptococcus-induced NF and paradoxical IRIS. Cooperation from other specialists was essential. CONCLUSION: We think this patient needed earlier and more definitive debridement. Fortunately, we were able to save the patient's life and maintain his LE function. In immunocompromised patients, cryptococcus can be a pathogen. In addition, IRIS can occur during treatment. Management of IRIS is the capital point of sepsis management, careful anti-inflammatory drug control by specialists is required.

A radiation ulcer that required partial lung resection and recurred in a small residual area of ectopic calcification.

INTRODUCTION AND IMPORTANCE: Surgery for chest radiation ulcers must involve appropriate wide margins, but it is not usually possible to remove all radiation-damaged tissue. Therefore, it is difficult to determine how extensive such surgery should be. There have not been any reports about the recurrence of such ulcers years after surgery. In addition, how ectopic calcification should be treated and the need for partial lung resection in such cases have not been fully elucidated. We report the case of a patient who had a large severe radiation ulcer. CASE PRESENTATION: A 46-year-old patient underwent cancer resection and received postoperative radiotherapy. Seventeen years later, a chest ulcer developed. Computed tomography showed a depression of the lung parenchyma, which exhibited old radiation pneumonitis, and pathological fractures of the ribs around the ulcer. We excised a region of skin that exhibited a clear change in color together with an additional 1 cm around this area including 4 ribs and grossly calcified area. The lung was partially resected because of strong adhesion, and the chest wall was reconstructed. Two small calcifications remained and which required additional surgery several years later. CLINICAL DISCUSSION: Since multiple surgeries were required, we consider that more generous resection margins were necessary from the beginning. CONCLUSION: In such cases, it might be necessary to perform more extensive surgery that includes asymptomatic calcified areas.

Successful salvage of an infected breast prothesis by changing from continuous to intermittent suction under continuous irrigation.

INTRODUCTION: Implant-based breast reconstruction is a widely performed procedure. However, prostheses are susceptible to infection and there are currently no established guidelines on treatment. In the present case, a prosthesis was salvaged by changing from continuous irrigation and suction to continuous irrigation and intermittent suction. This case report has been reported in line with the SCARE criteria [1]. PRESENTATION OF CASE: A 50-year-old female patient underwent implant-based breast reconstruction following surgery for breast cancer. One month later, the left breast prosthesis was infected with abscesses. Surgical treatment and continuous irrigation were performed as postoperative therapy. However, recurrent infection was detected a few days after surgery. Continuous irrigation was changed to continuous irrigation with intermittent aspiration, which successfully controlled the infection. DISCUSSION: Factors that limit the effectiveness of continuous irrigation and aspiration have not yet been identified. Inflow/discharge shunt routes may be established in continuous aspiration, and, thus, sufficient cleaning may not be possible. On the other hand, the storage of water throughout the wound in intermittent aspiration may facilitate cleaning. CONCLUSION: Intermittent suction worked well in this patient and, thus, warrants further study.

Squamous cell carcinoma developed in a chronic radiation-induced chest wall ulcer that is difficult to undergo thorough preoperative histological examination.

INTRODUCTION: We report a case in which squamous cell carcinoma (SCC) developed in a large chronic radiation-induced thoracic ulcer after flap surgery in areas where preoperative histological examinations are difficult. PRESENTATION OF CASE: The patient was a 75-year-old female. She had undergone resection and radiotherapy for left breast cancer 15 years earlier. Six years ago, the ulcer expanded from the subclavian to xiphoid levels, exposing the lung and pericardium. A histopathological examination, which avoided the lung and pericardium, was performed. Inflammation was diagnosed. We reconstructed the chest wall with a pedicled rectus abdominis flap. Eighteen months later, three verrucous tissue-lined fistulas formed. A histological examination revealed well-differentiated SCC. Six months later, the patient died of massive bleeding from a fistula. DISCUSSION: It is unclear exactly when the SCC occurred. As three fistulas formed at the margins of the flap around the pericardium, we suspect that the cancer developed within or near the pericardial region. We need to reflect on the lack of a thorough biopsy. As no pericardial biopsy was performed, we should have asked a thoracic/cardiac surgeon to conduct a biopsy during the debridement operation. If the tumor had been localized to the pericardium, it could have been removed. CONCLUSION: It is necessary to consider the best method for performing the most thorough histological examination possible, even in areas where histological examinations are difficult, as all ulcers can contain tumors.

Improvement of insulin signalling rescues inflammatory cardiac dysfunction.

Inflammation resulting from virus infection is the cause of myocarditis; however, the precise mechanism by which inflammation induces cardiac dysfunction is still unclear. In this study, we investigated the contribution of insulin signalling to inflammatory cardiac dysfunction induced by the activation of signalling by NF-κB, a major transcriptional factor regulating inflammation. We generated mice constitutively overexpressing kinase-active IKK-ß, an essential kinase for NF-κB activation, in cardiomyocytes (KA mice). KA mice demonstrated poor survival and significant cardiac dysfunction with remarkable dilation. Histologically, KA hearts revealed increased cardiac apoptosis and fibrosis and the enhanced recruitment of immune cells. By molecular analysis, we observed the increased phosphorylation of IRS-1, indicating the suppression of insulin signalling in KA hearts. To evaluate the contribution of insulin signalling to cardiac dysfunction in KA hearts, we generated mice with cardiac-specific suppression of phosphatase and tensin homologue 10 (PTEN), a negative regulator of insulin signalling, in the KA mouse background (KA-PTEN). The suppression of PTEN successfully improved insulin signalling in KA-PTEN hearts, and interestingly, KA-PTEN mice showed significantly improved cardiac function and survival. These results indicated that impaired insulin signalling underlies the mechanism involved in inflammation-induced cardiac dysfunction, which suggests that it may be a target for the treatment of myocarditis.

Function Preservation of the Upper Lip after Tumor Resection Using Residual Orbicularis Oris Muscle and Attached Levator Labii Superioris Alaeque Nasi.

We report a case, function preservation of the upper lip after tumor resection was possible using residual orbicularis oris muscle and attached levator labii superioris alaeque nasi. Patient was 67-year-old male with squamous cell carcinoma at the vermilion border. The tumor was resected with an 8-mm margin, leaving the oral mucosa as intact as possible. To reconstruct the red lip, we used the oral mucosa as a rotational transposition flap. The white lip was reconstructed with a cheek rotation flap. A levator labii superioris alaque nasi muscle flap, which was attached to the remaining orbicularis oris muscle, was used to increase marginal lip volume. The movement of the reconstructed lip was good. At 9 postoperative months, induration of the red lip was palpable, and we suspected that the blood supply to the levator labii superioris alaque nasi was borderline insufficient. Slight drooping of the reconstructed lip occurred. We dissected this was caused by dissection of mid facial muscles from orbicularis oris muscle to ease downward rotation of the cheek flap and obscure the original nasolabial fold. Although some drooping and induration of the lip occurred, the white and red lip were reconstructed in a single-stage procedure, which resulted in good movement and preserved the function of the orbicularis oris muscle.

Efficient de novo assembly of highly heterozygous genomes from whole-genome shotgun short reads.

Although many de novo genome assembly projects have recently been conducted using high-throughput sequencers, assembling highly heterozygous diploid genomes is a substantial challenge due to the increased complexity of the de Bruijn graph structure predominantly used. To address the increasing demand for sequencing of nonmodel and/or wild-type samples, in most cases inbred lines or fosmid-based hierarchical sequencing methods are used to overcome such problems. However, these methods are costly and time consuming, forfeiting the advantages of massive parallel sequencing. Here, we describe a novel de novo assembler, Platanus, that can effectively manage high-throughput data from heterozygous samples. Platanus assembles DNA fragments (reads) into contigs by constructing de Bruijn graphs with automatically optimized k-mer sizes followed by the scaffolding of contigs based on paired-end information. The complicated graph structures that result from the heterozygosity are simplified during not only the contig assembly step but also the scaffolding step. We evaluated the assembly results on eukaryotic samples with various levels of heterozygosity. Compared with other assemblers, Platanus yields assembly results that have a larger scaffold NG50 length without any accompanying loss of accuracy in both simulated and real data. In addition, Platanus recorded the largest scaffold NG50 values for two of the three low-heterozygosity species used in the de novo assembly contest, Assemblathon 2. Platanus therefore provides a novel and efficient approach for the assembly of gigabase-sized highly heterozygous genomes and is an attractive alternative to the existing assemblers designed for genomes of lower heterozygosity.

NMR studies on effects of temperature, pressure, and fluorination on structures and dynamics of alcohols in liquid and supercritical states.

We measured 1H NMR chemical shifts (delta H) and 1H and 2H NMR spin-lattice relaxation times (1H- and 2H-T1) of methanol, ethanol, 2-propanol, 2,2,2-trifluoroethanol, and 1,1,1,3,3,3-hexafluoro-2-propanol in the temperature range from 298 to 673 K at reduced pressures ( Pr = P/ Pc) of 1.22 and 3.14. The delta H values showed that the degree (X HB) of hydrogen bonding decreased in the order of methanol > ethanol >2-propanol > H2O, and that the hydrogen bonding was much affected by fluorination, because of the intramolecular H-F interactions in supercritical (sc) states. Moreover, 1H- T 1 measurements revealed that the relaxation processes of OH groups in nonfluoroalcohols are controlled by dipole-dipole (DD) and spin-rotation (SR) mechanisms below and above the critical temperature (Tc), while the cross-correlation effects connected with intramolecular DD interactions between a carbon atom and an adjacent proton played an important role for hydrocarbon groups (CHn, n = 1-3) under sc conditions. This interpretation was also supported by two other results. The first is that the intramolecular H-F interactions strongly inhibit the internal rotation of CH and CH2 groups of sc fluoroalcohols, and the second is that the molecular reorientational correlation times (tauc(D)) obtained from 2H- T 1 values of deuterated hydrocarbon groups (CDn ) at temperatures above T c have significantly less temperature dependence than those of OD groups. Actually, the apparent activation energy (DeltaEa) for molecular reorientational motions in sc alcohols was smaller compared with liquid alcohols, being about 1 order of magnitude.