Identification, characterization, and structural analyses of a fungal endo-ß-1,2-glucanase reveal a new glycoside hydrolase family.

endo-ß-1,2-Glucanase (SGL) is an enzyme that hydrolyzes ß-1,2-glucans, which play important physiological roles in some bacteria as a cyclic form. To date, no eukaryotic SGL has been identified. We purified an SGL from Talaromyces funiculosus (TfSGL), a soil fungus, to homogeneity and then cloned the complementary DNA encoding the enzyme. TfSGL shows no significant sequence similarity to any known glycoside hydrolase (GH) families, but shows significant similarity to certain eukaryotic proteins with unknown functions. The recombinant TfSGL (TfSGLr) specifically hydrolyzed linear and cyclic ß-1,2-glucans to sophorose (Glc-ß-1,2-Glc) as a main product. TfSGLr hydrolyzed reducing-end-modified ß-1,2-gluco-oligosaccharides to release a sophoroside with the modified moiety. These results indicate that TfSGL is an endo-type enzyme that preferably releases sophorose from the reducing end of substrates. Stereochemical analysis demonstrated that TfSGL is an inverting enzyme. The overall structure of TfSGLr includes an (α/α)6 toroid fold. The substrate-binding mode was revealed by the structure of a Michaelis complex of an inactive TfSGLr mutant with a ß-1,2-glucoheptasaccharide. Mutational analysis and action pattern analysis of ß-1,2-gluco-oligosaccharide derivatives revealed an unprecedented catalytic mechanism for substrate hydrolysis. Glu-262 (general acid) indirectly protonates the anomeric oxygen at subsite -1 via the 3-hydroxy group of the Glc moiety at subsite +2, and Asp-446 (general base) activates the nucleophilic water via another water. TfSGLr is apparently different from a GH144 SGL in the reaction and substrate recognition mechanism based on structural comparison. Overall, we propose that TfSGL and closely-related enzymes can be classified into a new family, GH162.

[Extreme Obese Patients who Underwent Cardiac Surgery after Preoperative Weight Reduction;Report of Two Cases].

Obesity is a risk factor of postoperative complications. We experienced 2 extremely obese patients:a 32-year-old male with coronary artery disease and a 75-year-old female with aortic valve stenosis. Their initial body weights were 133 kg and 88.5 kg, respectively, and their initial body mass indexes (BMIs) were both 41. Their BMIs were reduced to 35.5 and 35, respectively, after preoperative weight reduction. Off-pump coronary artery bypass grafting and aortic valve replacement were performed, respectively. After surgery, the non-invasive positive pressure ventilation( NPPV) support was effective, and their postoperative courses were uneventful. Preoperative weight reduction and NPPV are useful for extremely obese patients who undergo cardiac surgery.

Biochemical and structural analyses of a bacterial endo-ß-1,2-glucanase reveal a new glycoside hydrolase family.

ß-1,2-Glucan is an extracellular cyclic or linear polysaccharide from Gram-negative bacteria, with important roles in infection and symbiosis. Despite ß-1,2-glucan's importance in bacterial persistence and pathogenesis, only a few reports exist on enzymes acting on both cyclic and linear ß-1,2-glucan. To this end, we purified an endo-ß-1,2-glucanase to homogeneity from cell extracts of the environmental species Chitinophaga arvensicola, and an endo-ß-1,2-glucanase candidate gene (Cpin_6279) was cloned from the related species Chitinophaga pinensis The Cpin_6279 protein specifically hydrolyzed linear ß-1,2-glucan with polymerization degrees of ≥5 and a cyclic counterpart, indicating that Cpin_6279 is an endo-ß-1,2-glucananase. Stereochemical analysis demonstrated that the Cpin_6279-catalyzed reaction proceeds via an inverting mechanism. Cpin_6279 exhibited no significant sequence similarity with known glycoside hydrolases (GHs), and thus the enzyme defines a novel GH family, GH144. The crystal structures of the ligand-free and complex forms of Cpin_6279 with glucose (Glc) and sophorotriose (Glc-ß-1,2-Glc-ß-1,2-Glc) determined up to 1.7 Å revealed that it has a large cavity appropriate for polysaccharide degradation and adopts an (α/α)6-fold slightly similar to that of GH family 15 and 8 enzymes. Mutational analysis indicated that some of the highly conserved acidic residues in the active site are important for catalysis, and the Cpin_6279 active-site architecture provided insights into the substrate recognition by the enzyme. The biochemical characterization and crystal structure of this novel GH may enable discovery of other ß-1,2-glucanases and represent a critical advance toward elucidating structure-function relationships of GH enzymes.

Investigations on the charge transfer mechanism at donor/acceptor interfaces in the quest for descriptors of organic solar cell performance.

Herein, we theoretically and experimentally investigated the mechanisms of charge separation processes of organic thin-film solar cells. PTB7, PTB1, and PTBF2 have been chosen as donors and PC71BM has been chosen as an acceptor considering that effective charge generation depends on the difference between the material combinations. Experimental results of transient absorption spectroscopy show that the hot process is a key step for determining external quantum efficiency (EQE) in these systems. From the quantum chemistry calculations, it has been found that EQE tends to increase as the transferred charge, charge transfer distance, and variation of dipole moments between the ground and excited states of the donor/acceptor complexes increase; this indicates that these physical quantities are a good descriptor to assess the donor-acceptor charge transfer quality contributing to the solar cell performance. We propose that designing donor/acceptor interfaces with large values of charge transfer distance and variation of dipole moments of the donor/acceptor complexes is a prerequisite for developing high-efficiency polymer/PCBM solar cells.

[A Case of Inflammatory Breast Cancer with Very Severe Obesity].

A 39-year-old woman with very sever obesity was admitted to our hospital for a right breast redness and hardness. Her height, weight and BMI were 166 cm, 145 kg and 52.6 kg/m2. Her breast had peau d'orange. CT scan showed swelling of whole right breast and Level I , II lymph node. We performed core needle biopsy and diagnosed as the inflammatory breast cancer with ER and HER2 positive. We introduced chemotherapy(pertuzumab, trastuzumab and paclitaxel)and nutrition counseling in order to reduce her body weight. After 4 courses of chemotherapy, the clinical complete response was obtained and her body weight decreased to 125 kg. We performed mastectomy and axillary node resection and confirmed pathological complete response. Adjuvant chemotherapy(5-FU, epirubicin and cyclophosphamide), adjuvant trastuzumab therapy, postmastectomy radiation therapy and adjuvant hormonal therapy were administered. There have been no signs of recurrence as of 2 years after the operation.

A Novel Tricyclic Polyketide, Vanitaracin A, Specifically Inhibits the Entry of Hepatitis B and D Viruses by Targeting Sodium Taurocholate Cotransporting Polypeptide.

UNLABELLED: Anti-hepatitis B virus (HBV) drugs are currently limited to nucleos(t)ide analogs (NAs) and interferons. A challenge of drug development is the identification of small molecules that suppress HBV infection from new chemical sources. Here, from a fungus-derived secondary metabolite library, we identify a structurally novel tricyclic polyketide, named vanitaracin A, which specifically inhibits HBV infection. Vanitaracin A inhibited the viral entry process with a submicromolar 50% inhibitory concentration (IC50) (IC50 = 0.61 ± 0.23 µM), without evident cytotoxicity (50% cytotoxic concentration of >256 µM; selectivity index value of >419) in primary human hepatocytes. Vanitaracin A did not affect the HBV replication process. This compound was found to directly interact with the HBV entry receptor sodium taurocholate cotransporting polypeptide (NTCP) and impaired its bile acid transport activity. Consistent with this NTCP targeting, antiviral activity of vanitaracin A was observed with hepatitis D virus (HDV) but not hepatitis C virus. Importantly, vanitaracin A inhibited infection by all HBV genotypes tested (genotypes A to D) and clinically relevant NA-resistant HBV isolate. Thus, we identified a fungal metabolite, vanitaracin A, which was a potent, well-tolerated, and broadly active inhibitor of HBV and HDV entry. This compound, or its related analogs, could be part of an antiviral strategy for preventing reinfection with HBV, including clinically relevant nucleos(t)ide analog-resistant virus. IMPORTANCE: For achieving better treatment and prevention of hepatitis B virus (HBV) infection, anti-HBV agents targeting a new molecule are in great demand. Although sodium taurocholate cotransporting polypeptide (NTCP) has recently been reported to be an essential host factor for HBV entry, there is a limited number of reports that identify new compounds targeting NTCP and inhibiting HBV entry. Here, from an uncharacterized chemical library, we isolated a structurally new compound, named vanitaracin A, which inhibited the process of entry of HBV and hepatitis D virus (HDV). This compound was suggested to directly interact with NTCP and inhibit its transporter activity. Importantly, vanitaracin A inhibited the entry of all HBV genotypes examined and of a clinically relevant nucleos(t)ide analog-resistant HBV isolate.

Isolation and structure of vanitaracin A, a novel anti-hepatitis B virus compound from Talaromyces sp.

Two new tricyclic polyketides, vanitaracin A (1) and B (2), together with three novel compounds 3, 4 and 5, were isolated from a culture broth of a fungus, Talaromyces sp. The chemical structures of these compounds were determined from spectroscopic data (1D/2D NMR, MS and IR). The five isolated compounds were then tested for anti-hepatitis B virus (HBV) activity and vanitaracin A was found to exhibit an IC50 value of 10.5 µM using a HBV-susceptible cell line. By contrast, the derivative 2 displayed weak anti-HBV action, which suggested that the substituents at C-9 in 1 are likely to be important for its antiviral activity. We believe the two vanitaracin derivatives constitute a new class of anti-HBV agents.

Acute necrotizing colitis due to sigmoid colon cancer.

When obstructive colitis becomes fulminant, it is known as 'acute necrotizing colitis'. We report a rare case of acute necrotizing colitis due to sigmoid colon cancer, in which shock status occurred within ten hours of onset. A 79-year-old female with acute abdominal pain was transported to our hospital with acute shock. Abdominal enhanced computed tomography revealed thickening of the wall of the sigmoid colon and marked dilation of the proximal colon. Emergency surgery was performed with the intraoperative findings of severe sigmoid colon stenosis and proximal dilation of the colon without perforation, and a large volume of putrid ascitic fluid. The intestine was proximally dilated and black in color, from the sigmoid colon to the ileum 60 cm proximal to the terminal ileum, suggesting acute necrosis. Total colectomy with 80 cm resection of terminal ileum and ileostomy was performed. Whereas acute necrotizing colitis is a rare condition and its etiology remains obscure, the chronic ischemic state must play some role. Our patient was of advanced age and had diabetes mellitus and hypertension. These factors might lead to a chronic ischemic state of the bowel due to arteriosclerosis. In addition to the underlying condition, massive bacterial reflux into the ileum from the colon might cause the capillary vasoconstriction of the bowel that led to her critical state.

Advancing availability of papillary muscle sandwich plasty from functional to organic mitral valve regurgitation.

An original papillary muscle heads approximation procedure, commonly known as a sandwich plasty, has been successfully used for the treatment of functional mitral regurgitation (MR) associated with ischemic cardiomyopathy (ICM) and aortic valve disease (AVD). In this study, we evaluated the availability of this method as the concomitant procedure for the plasty of organic MR in comparison with the operative results in patients with functional MR. Fifty-six patients who underwent sandwich plasty were reviewed, including 45 functional MR (29 ICM and 16 AVD) patients and 11 organic MR patients. The mean age of patients was 67 years. In the organic MR patients, predominant cardiac diseases were solitary MR in six patients and combined valve diseases including aortic valve stenosis in five. Mitral valve changes included prolapse in six patients and moderate cusp thickening with calcification in five. Two heads of the papillary muscle connecting to the choldae of both the anterior and posterior leaflets are fixed with two teflon-pledgeted 3-0 TiCron™ (Covidien, Dublin, Ireland) sutures in order to achieve coaptation of the two leaflets. Prominent MR was observed in a patient with functional MR after surgery, however residual MR was not detected in organic MR patients. The tenting height (coaptation distance) of mitral valve significantly decreased after surgery from 11±1 to 7±2mm in the organic MR patients, which was similar to the results in the functional MR patients (from 12±2 to 7±2 mm). The postoperative mean mitral orifice area in the organic MR patients was 4.3±0.1cm2 without stenosis. Sandwich plasty reduces the distance of choldae connecting to anterior and posterior leaflets and achieves the better coaptation of two leaflets. This procedure is effective in the treatment of both functional and organic MR.

[Locoregional recurrence of HER2-positive breast cancer - treatment after complete remission and long-term clinical benefits].

We report 7 cases of locoregional recurrence in human epidermal growth factor receptor 2 (HER2)-positive breast cancer that we treated. An early complete response (CR) and long-term response was achieved in 5 cases. There were 4 HER2- subtype and 3 Luminal HER2-type cases. Metastasis and recurrence were detected in the residual breast tissue and the supraclavicular, axillary, and parasternal lymph nodes. Chemotherapy consisting of trastuzumab was administered as first-line treatment. A CR was observed 3-4 months after the initiation of therapy in 4 cases, and the time to progression was 27.6- 65.8 months. After achieving a CR, 3 patients terminated treatment and 2 patients continued to take trastuzumab. However, due to adverse effects associated with the chemotherapy, 1 patient changed to endocrine therapy. A second, long-term, CR was achieved in 2 relapsed CR patients by re-challenging with the same chemotherapy regimen. Two patients did not achieve CR and died due to distant metastases. For a better quality of life, it is advisable to continue treatment after a clinical CR for solitary or more complex locoregional recurrences. Following the first-line therapy and a so-called chemoholiday, the patient's disease can be re-challenged using the previously sensitive regimen with careful observation.

[Two cases of radiotherapy-induced sarcoma after breast cancer surgery].

We report 2 cases of radiotherapy-induced sarcoma of the residual breast after breast cancer surgery. In 1 case, the patient was a 64-year-old woman. She underwent breast-conserving surgery and axillary lymph node dissection followed by irradiation to the residual breast in July 2001. A 1.1 × 1.0-cm tumor was noted in the residual breast 7 years 5 months after radiotherapy. An excisional biopsy was performed, and a histological diagnosis of angiosarcoma was made. She died of lung and peritoneal metastases 3 years 2 months after the diagnosis. In the other case, the patient was also 64 years old. She underwent breast-conserving surgery and sentinel lymph node biopsy followed by irradiation to the residual breast in October 2006. A 5.7 × 3.9-cm induration was noted in the residual breast 3 years 5 months after radiotherapy. A core needle biopsy was performed, and a histological diagnosis of sarcoma was made. Mastectomy was performed, and the histological diagnosis was malignant fibrous histiocytoma. She died of chest wall and intrapleural tumor recurrence 3 months after the mastectomy. Although radiotherapy-induced sarcoma is rare, early detection of the tumor in the irradiation area is important, as radiotherapy is often performed for breast cancer patients.

[The efficacy of early diagnosis of brain metastasis and systemic treatment after radiotherapy in patients with metastatic breast cancer].

We treated 9 patients diagnosed with brain metastasis from breast cancer. Although 1 patient was initially diagnosed as having Stage IV disease, 5 had Stage I/II early breast cancer. All patients had defined brain metastasis after chemotherapy. Brain metastasis was symptomatic in 7 patients, 4 of whom had brain edema, and asymptomatic in 2 patients. The median survival time from breast cancer metastasis was 23 days for patients who did not receive radiotherapy and 19.6 months for those who received radiotherapy. Among the patients treated with radiotherapy, the median survival time was 4.3 months for patients who did not receive further treatment and 19.7 months for those who received chemotherapy or chemotherapy with trastuzumab. One patient with a solitary brain metastasis underwent stereotactic radiosurgery, and treatment is being continued for 1 of the 2 patients who received systemic therapy after whole-brain radiotherapy and additional stereotactic radiosurgery at recurrence to control brain disease. Systemic treatment after radiotherapy is important for brain metastasis from breast cancer, and early diagnosis of brain metastasis facilitates the use of various available treatments.

[A case of coexisting borderline phyllodes tumor and non-invasive ductal carcinoma].

A 36-year-old woman with benign phyllodes tumor of the left breast had undergone lumpectomy 1 year ago and was admitted to our hospital because of a left breast mass on the operation scar. Ultrasonography showed a 35 mm low-echoic, elliptical mass with a high depth to width( D/W) ratio in the C area and a 10 mm low-echoic, polygonal mass with a high D/W ratio in the E area. Histological examination of an ultrasonography-guided vacuum-assisted biopsy specimen indicated recurrent phyllodes tumor. Since both tumors were assumed to be recurrent phyllodes tumors, quadrantectomy was performed. Finally, the mass in the C area was diagnosed as a recurrent phyllodes tumor and the mass in the E area was diagnosed as a fibroadenoma. A non-invasive ductal carcinoma was incidentally detected between the 2 tumors, and the surgical margin was negative. Radiotherapy was performed on the remnant breast tissue.

An automated screening model for aortic emergencies using convolutional neural networks and cropped computed tomography angiography images of the aorta.

PURPOSE: Patients with aortic emergencies, such as aortic dissection and rupture, are at risk of rapid deterioration, necessitating prompt diagnosis. This study introduces a novel automated screening model for computed tomography angiography (CTA) of patients with aortic emergencies, utilizing deep convolutional neural network (DCNN) algorithms. METHODS: Our model (Model A) initially predicted the positions of the aorta in the original axial CTA images and extracted the sections containing the aorta from these images. Subsequently, it predicted whether the cropped images showed aortic lesions. To compare the predictive performance of Model A in identifying aortic emergencies, we also developed Model B, which directly predicted the presence or absence of aortic lesions in the original images. Ultimately, these models categorized patients based on the presence or absence of aortic emergencies, as determined by the number of consecutive images expected to show the lesion. RESULTS: The models were trained with 216 CTA scans and tested with 220 CTA scans. Model A demonstrated a higher area under the curve (AUC) for patient-level classification of aortic emergencies than Model B (0.995; 95% confidence interval [CI], 0.990-1.000 vs. 0.972; 95% CI, 0.950-0.994, respectively; p = 0.013). Among patients with aortic emergencies, the AUC of Model A for patient-level classification of aortic emergencies involving the ascending aorta was 0.971 (95% CI, 0.931-1.000). CONCLUSION: The model utilizing DCNNs and cropped CTA images of the aorta effectively screened CTA scans of patients with aortic emergencies. This study would help develop a computer-aided triage system for CT scans, prioritizing the reading for patients requiring urgent care and ultimately promoting rapid responses to patients with aortic emergencies.

[A case of liposarcoma of the abdominal wall complicated by thrombocytpenia as a paraneoplastic syndrome].

A 59-year-old woman was admitted to our hospital because of right chest pain. CT scan showed a mass on the right abdominal wall and bilateral pleural effusion. The histological diagnosis following core needle biopsy was undifferentiated sarcoma. The right pleural effusion gradually increased despite negative cytology. Although we planned chemotherapy for the clinically diagnosed pleural invasion, thrombocytopenia as a paraneoplastic syndrome appeared. The minimum thrombocyte count was 4,000/mm3. While transfusion was not effective, per os dexamethasone at 2.0 mg/day kept the thrombocyte count at around 6×10 4/mm3. Anti-thrombocyte antibody was negative. Tumor resection surgery with partial diaphragm resection and 11th and 12th rib resection, and abdominal wall plasty with mesh was performed. The final histological diagnosis was dedifferentiated liposarcoma. The thrombocyte count returned to the normal range just after the operation. However, she died of pleural dissemination, peritoneal dissemination, and local recurrence 69 days after the operation.

A new screening model for quantitative risk assessment of blunt thoracic aortic injury.

PURPOSE: Early identification of blunt thoracic aortic injury is vital for preventing subsequent aortic rupture. However, risk factors for blunt thoracic aortic injury remain unclear, and a prediction rule remains to be established. We developed and internally validated a new nomogram-based screening model that allows clinicians to quantify blunt thoracic aortic injury risk. METHODS: Adult patients (age ≥ 18 years) with blunt injury were selected from a nationwide Japanese database (January 2004-May 2019). Patients were randomly divided into training and test cohorts. A new nomogram-based blunt thoracic aortic injury-screening model was constructed using multivariate logistic regression analysis to quantify the association of potential predictive factors with blunt thoracic aortic injury in the training cohort. RESULTS: Overall, 305,141 patients (training cohort, n = 152,570; test cohort, n = 152,571) were eligible for analysis. Median patient age was 65 years, and 60.9% were men. Multivariate analysis in the training cohort revealed that 13 factors (positive association: age ≥ 55 years, male sex, high-energy impact, hypotension on hospital arrival, Glasgow Coma Scale score < 9 on hospital arrival, diaphragmatic injuries, hepatic injuries, pulmonary injuries, cardiac injuries, renal injuries, sternum fractures, multiple rib fractures, and pelvic fractures) were significantly associated with blunt thoracic aortic injury and included in the screening model. In the test cohort, the new screening model had an area under the curve of 0.87. CONCLUSIONS: Our novel nomogram-based screening model aids in the quantitative assessment of blunt thoracic aortic injury risk. This model may improve tailored decision-making for each patient.

[Radio-frequency ablation therapy to the liver metastasis from colon cancer which neighbors a hepatic-cyst].

We report a case of liver metastasis from colon cancer successfully treated with radio-frequency ablation therapy. The patient was a 75-year-old man. We performed sigmoidectomy in February 2006, but a metastatic-focus in S5 segment of the liver with the size of 28 mm was discovered by November 2006. Chemotherapy was enforced to the patient but we could not control the metastasis. There were multiple hepatic-cysts in his liver and one with size of 60 mm surrounded the metastatic-focus. RFA therapy was performed after enforcing a percutaneous ethanol sclerosing therapy to the neighboring hepatic-cyst. After 4 months from RFA therapy, the CT showed a good cauterization effect on the metastasis. We thought that RFA therapy after enforcing a percutaneous ethanol sclerosing therapy to the neighboring hepatic-cyst was as good as hepatectomy with low invasiveness.

[A case of liver metastasis from gastric cancer successfully treated with radio-frequency ablation therapy].

We report a case of liver metastasis from gastric cancer successfully treated with radio-frequency ablation therapy. The patient was a 63-year-old man whom we performed distal gastrectomy with D2 lymphnode dissection for type II gastric cancer on April 2004. A post operative pathological finding was pT2N1M0, Stage II. Therefore, the operation was evaluated to be as curative A. No adjuvant therapy was performed. After 11 months computed tomography showed a space occupied lesion (SOL) in S7 segment of the liver, we thought it to be as liver metastasis. We performed radio-frequency ablation therapy, and complete response was obtained. He is alive without a sign of recurrence for 30 months. We thought that radio -frequency ablation therapy was one of the useful treatments for liver metastasis from gastric cancer.

[A case of gastrointestinal stromal tumor of the stomach with rapid growth in a short term].

A 75-year-old man was admitted to our hospital with massive hematemesis. Upper gastrointestinal endoscopy revealed a submucosal gastric tumor with ulcer lesion, about 30 mm in diameter, in the anterior wall of the cardia. Pathological findings of biopsy specimens showed a gastrointestinal stromal tumor (GIST) of the stomach. Abdominal computed tomography (CT) showed an extramural low density mass, measuring 27 mm, in the cardia. From these findings, we thought we should be performing a wedge resection of the stomach for this tumor. After one month from the first examination, preoperative upper gastrointestinal endoscopy revealed a growth of the tumor. Abdominal CT also showed an obvious growth of the tumor, measuring 50 mm. Thus, laparoscopy-assisted proximal gastrectomy was performed, because the tumor was close to the cardia. Postoperative diagnosis of the tumor was pathologically confirmed to be a GIST with a high malignant potential. The case report of GIST that shows rapid growth for a short term is rare. We herein describe a case of GIST with rapid growth during one month before the operation.

[Radiofrequency ablation of malignant lung tumors--preliminary report of 12 cases].

UNLABELLED: Percutaneous CT fluoroscopy-guided radiofrequency ablation (RFA) was performed in 12 patients with 4 primary and 8 metastatic lung cancers as a pilot study. The main complication was a pneumothorax, which occurred in six cases and 2 of them required a chest tube drainage. Four patients died of extrapulmonary tumor progression at 3-17 months after RFA. The remaining 8 patients were alive with the disease and their median follow-up period was 12 months (range, 6-39 months). An overall survival rate was 75% at 6 months, 63% at 1 year and 43% at 2 years. A local progression occurred in 4 cases at 4-17 months after RFA and the cause of relapse was considered as insufficient ablation due to tumor localization close to the trachea, SVC or intrapulmonary vessels and bronchi. A local control rate was 92% at 6 months, 64% at 1 year and 35% at 2 years. CONCLUSION: RFA of lung tumors is a safe and minimally invasive therapeutic intervention with a good local control for sufficiently ablated lesions. So it appears to be a favorable treatment option for high-risk patients or those with multiple lesions.