Accelerated germination of aged recalcitrant seeds by K+-rich bulk oxygen nanobubbles.

Bulk nanobubbles, measuring less than 200 nm in water, have shown their salient properties in promoting growth in various species of plants and orthodox seeds, and as potential drug-delivery carriers in medicine. Studies of recalcitrant seeds have reported markedly increased germination rates with gibberellin treatment; however, neither the mechanism promoting germination nor the implication for food safety is well elucidated. In our study, recalcitrant wasabi (Eutrema japonicum) seeds treated with bulk oxygen nanobubbles (BONB) containing K+, Na+, and Cl- (BONB-KNaCl) showed significantly accelerated germination. As germination progressed, 99% of K+ ions in the BONB-KNaCl medium were absorbed by the seeds, whereas Ca2+ ions were released. These results suggest that the germination mechanism involves the action of K+ channels for migration of K+ ions down their concentration gradient and Ca2+ pumps for the movement of Ca2+ ions, the first potential discovery in germination promotion in recalcitrant seeds using nutrient solutions with BONB-KNaCl.

[Bleeding due to acquired factor V inhibitor successfully treated with platelet transfusion during treatment for recurrent prostate squamous cell carcinoma].

A 78-year-old man with prostate squamous cell carcinoma recurrence in his pelvis was admitted to our hospital. Rectal obstruction led to creation of an artificial anus on the transverse colon. Then, docetaxel and radiation therapies were started. A week later, severe hematuria and melena occurred. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were extremely prolonged. Cross-mixing test for APTT and PT revealed an inhibitor pattern, which was diagnosed as acquired factor V inhibitor. Fresh frozen plasma and vitamin K infusions were ineffective, but platelet transfusion successfully stopped the bleeding. Platelet factor V derived from megakaryocytes may affect local hemostasis. The patient received prednisolone (PSL), and the inhibitor disappeared on day 70 and was in remission. PSL could be stopped on day 100. Later, we demonstrated APTT and PT shortening of factor V deficient plasma by the supernatant of activated platelets with collagen.

Treating octasilanol [Si8O12][OH]8 with tetramethoxysilane and trimethoxyvinylsilane: a halogen-free synthetic route to alkoxysilyl-substituted double-four-ring siloxanes.

The synthesis of methoxysilyl-substituted siloxanes with a double-four-ring (D4R) framework [Si8O12][OSiR(OMe)2]8 (R = OMe or vinyl) via the dealcoholative coupling between silanol [Si8O12][OH]8 and alkoxysilane was accomplished and achieved a halogen-free synthetic process. The solid-state structure of [Si8O12][OSi(OMe)3]8 was determined via a single-crystal X-ray diffraction analysis.

Non-COVID-19 Patients with Life-threatening Diseases Who Visited a Fever Clinic: A Single-center, Observational Study in Tokyo, Japan.

Objective In fever clinics screening coronavirus disease (COVID-19), there could be patients with life-threatening diseases that physicians should not overlook. We exploratorily investigated the final diagnosis among non-COVID-19 hospitalized patients who visited the fever clinic. Methods This was a retrospective, observational, and single-centered study conducted in the National Center for Global Health and Medicine (NCGM), Tokyo, Japan. We conducted a retrospective chart review of patients who visited the fever clinic in the NCGM from 11 March 2020 to 24 April 2020. Patients Patients who met the following clinical criteria visited the fever clinic in the NCGM: (1) body temperature >37.5°C, (2) any symptoms consistent with COVID-19 or (3) referral from local healthcare facilities. In the fever clinic, all patients who met the above criteria had severe acute respiratory syndrome coronavirus 2 polymerase chain reaction test with nasopharyngeal swab specimens. Patients with severe symptoms or an unstable condition were sent to an outpatient clinic for infectious diseases for further evaluation and treatment. Results Among 1,470 patients who visited the fever clinic, 84 patients were hospitalized, and 45 of them were diagnosed as having COVID-19. Among the remaining 39 non-COVID-19 patients, there were nine patients with life-threatening diseases. The life-threatening diseases included acute heart failure, septic shock, pneumocystis pneumonia, peritonsillar abscess, and necrotizing fasciitis. Conclusion Physicians should evaluate each patient carefully while considering other life-threatening conditions even in such a COVID-19 pandemic era.

Prognostic value of volumetric FDG PET/CT parameters in patients with oral tongue squamous cell carcinoma who were treated by superselective intra-arterial chemoradiotherapy.

PURPOSE: This study aimed to evaluate the predictive and prognostic value of FDG PET/CT-based volumetric parameters in patients with oral tongue squamous cell carcinoma (OTSCC) treated by superselective intra-arterial chemoradiotherapy (IA-CRT). METHODS: We conducted a retrospective study including 33 patients with biopsy-proven OTSCC between May 2007 and February 2016. All of the patients were treated by IA-CRT. Pretreatment SUVmax and metabolic tumor volume (MTV) of the primary tumor were measured. The SUV thresholds of 2.5 and 5.0 were used. Progression-free survival (PFS) and overall survival (OS) were chosen as endpoints to evaluate prognosis. Univariate and multivariate analyses were performed to assess the potential independent effect of FDG PET/CT parameters. RESULTS: The median follow-up for surviving patients was 40.7 months (range 6.0-107.5 months). In univariate and multivariate analyses, SUVmax and MTV (5.0) were independent prognostic factors for PFS. In univariate analysis, SUVmax failed to predict OS. MTV (5.0) was a significant prognostic factor for OS, but multivariate analysis failed to show statistical independence because it could not exclude the possibility of an artifact due to N stage. CONCLUSIONS: FDG PET/CT-based volumetric parameters may be significant prognostic markers for survival of patients with OTSCC who are treated by IA-CRT.

Abnormal conformation and impaired degradation of propylthiouracil-induced neutrophil extracellular traps: implications of disordered neutrophil extracellular traps in a rat model of myeloperoxidase antineutrophil cytoplasmic antibody-associated vasculitis.

OBJECTIVE: Neutrophil extracellular traps (NETs) are composed of DNA and antimicrobial proteins, including myeloperoxidase (MPO). Recent studies have demonstrated that impaired regulation of NETs could trigger an autoimmune response. Propylthiouracil (PTU), an antithyroid drug, is associated with a risk of MPO antineutrophil cytoplasmic antibody (ANCA) production and MPO ANCA-associated vasculitis (MPO AAV). This study was undertaken to clarify the mechanism of MPO ANCA production, using the PTU-induced model of MPO AAV. METHODS: NETs were induced by treating human neutrophils with phorbol myristate acetate (PMA) in vitro. We examined whether the addition of PTU influenced the NET formation induced by PMA and the degradation of NETs by DNase I, which is regarded as a regulator of NETs. Furthermore, we examined whether NETs generated by the combination of PMA and PTU induced MPO ANCA and MPO AAV in vivo in rats. RESULTS: When NETs were induced by PMA with PTU using human neutrophils in vitro, abnormal conformation of NETs was observed. Interestingly, the abnormal NETs were hardly digested by DNase I. Moreover, rats immunized with the abnormal NETs, which had been induced by PMA with PTU using rat neutrophils, produced MPO ANCA and developed pulmonary capillaritis. When rats were given oral PTU with intraperitoneal injection of PMA, pauci-immune glomerulonephritis and pulmonary capillaritis occurred with MPO ANCA production in the serum. CONCLUSION: Our findings indicate that abnormal conformation and impaired degradation of NETs induced by PTU are involved in the pathogenesis of PTU-induced MPO ANCA production and MPO AAV. These findings suggest that disordered NETs can be critically implicated in the pathogenesis of MPO AAV.

Nucleocytoplasmic translocation of HDAC9 regulates gene expression and dendritic growth in developing cortical neurons.

Transcriptional regulation of gene expression is thought to play a pivotal role in activity-dependent neuronal differentiation and circuit formation. Here, we investigated the role of histone deacetylase 9 (HDAC9), which regulates transcription by histone modification, in the development of neocortical neurons. The translocation of HDAC9 from nucleus to cytoplasm was induced by an increase of spontaneous firing activity in cultured mouse cortical neurons. This nucleocytoplasmic translocation was also observed in postnatal development in vivo. The translocation-induced gene expression and cellular morphology was further examined by introducing an HDAC9 mutant that disrupts the nucleocytoplasmic translocation. Expression of c-fos, an immediately-early gene, was suppressed in the mutant-transfected cells regardless of neural activity. Moreover, the introduction of the mutant decreased the total length of dendritic branches, whereas knockdown of HDAC9 promoted dendritic growth. These findings indicate that chromatin remodeling with nucleocytoplasmic translocation of HDAC9 regulates activity-dependent gene expression and dendritic growth in developing cortical neurons.

A segmentectomy of the right upper lobe has an advantage over a right upper lobectomy regarding the preservation of the functional volume of the right middle lobe: analysis by perfusion single-photon emission computed tomography/computed tomography.

PURPOSE: To evaluate the advantages of a segmentectomy of the right upper lobe (RUL) over a right upper (RU) lobectomy regarding the preservation of the functional volume of the right middle lobe (RML), the postoperative forced expiratory volume in one second (FEV(1)) of the RML was compared between an RU lobectomy and an RUL segmentectomy, by using a coregistered perfusion single-photon emission computed tomography and computed tomography (SPECT/CT). METHODS: The pulmonary function tests and perfusion SPECT/CT were conducted before and after surgery (RU lobectomy: 7; RUL segmentectomy: 13). The FEV(1) of the RML before and after surgery was calculated from the data of the pulmonary function test and SPECT/CT. RESULTS: In the RU lobectomy group, the percentage change of FEV(1) was 71% +/- 12%, which was significantly lower in comparison to 92% +/- 9% in the RUL segmentectomy group (P = 0.001). In the lobectomy group, the preoperative FEV(1) of the RML was 0.17 +/- 0.10 l, which decreased significantly to 0.06 +/- 0.06l after surgery (P = 0.009). In the segmentectomy group, FEV(1) of the RML before and after the surgery were 0.23 +/- 0.10 l and 0.20 +/- 0.111, of which the difference was not significant. CONCLUSION: An RUL segmentectomy has an advantage over an RU lobectomy regarding the preservation of pulmonary function, due to a greater preservation of not only the lung tissue, but also the FEV(1) of the RML.

Three-dimensional cell culture of glioma and morphological comparison of four different human cell lines.

BACKGROUND: To explore the intracranial behaviors of glioma, a three-dimensional culture was devised and the morphology of four cell lines was examined. MATERIALS AND METHODS: Bioabsorbable and degradable gelatin was used as the scaffold and T98G, A172, KNS42, and U118MG representative standard malignant glioma cell lines were cultured three-dimensionally. RESULTS: When grown, the cells demonstrated characteristic conformations. The U118MG cells dispersed with numerous fiber formations. In contrast, the KNS42 and A172 cells aggregated, adhering to each other, resulting in the formation of balloon-like structures. The T98G cells demonstrated an intermediate character. CONCLUSION: The cell lines showed distinct characteristics in three-dimensional culture. This culture method may have a role in elucidating the fundamental character of cells in the human body.

Application of therapeutic insonation to malignant glioma cells and facilitation by echo-contrast microbubbles of levovist.

BACKGROUND: Malignancies affecting the central nervous system are intractable to conventional therapies thereby requiring an alternative strategy, such as ultrasound irradiation. MATERIALS AND METHODS: We originally designed a transducer for intracranial insonation and investigated the effect of 210.4 kHz ultrasound on malignant glioma cells. RESULTS: The insonation of 2.61 W/cm2 effectively disrupted the malignant cells. This effect was reinforced by the echo-contrast agent, Levovist. The condition was applied to tumor-bearing animals and external insonation inhibited subcutaneous tumor growth. It also repressed the growth of intracranially implanted tumors and prolonged survival of the animals. When Levovist was stereotactically injected into the tumors, the effect of insonation was significantly enhanced. CONCLUSION: A neuronavigation system or stereotactic device has been used commonly for patients with brain tumor. Administration of combination therapy consisting of insonation and a local echo-contrast agent will have a role in improving the treatment for malignant gliomas.

Quantification of the impact of segmentectomy on pulmonary function by perfusion single-photon-emission computed tomography and multidetector computed tomography.

OBJECTIVE: The impact of segmentectomy for preservation of pulmonary function was quantified by using a co-registered perfusion single-photon-emission computed tomography and multidetector computed tomography (SPECT/CT). METHODS: Pulmonary function tests and perfusion SPECT/CT were conducted before and after segmentectomy in 56 patients. Actual values of forced expiratory volume in 1 second (FEV(1)) after segmentectomy were compared with the FEV(1) after virtual lobectomy, which was calculated by SPECT/CT. The preoperative and postoperative FEV(1) of each lobe that had undergone segmentectomy was measured by SPECT/CT. RESULTS: The mean percent of FEV(1) preserved after segmentectomy was significantly higher than the value after virtual lobectomy (88% +/- 9% vs 77% +/- 7%; P < .001). Whereas the mean value of the preoperative FEV(1) of each lobe that was undergoing segmentectomy was 0.51+/-0.21 L, segmentectomy could preserve 41% +/- 24% of it. The FEV(1) of each lobe after the resection of more than three segments (n = 4) was preserved in 17% +/- 12% of the preoperative values, which was significantly less than 49% +/- 23% and 35% +/- 22% after the resection of one (n = 29) and two (n = 23) segments (P = .02 and .08, respectively). The FEV(1) of the left upper lobe after the upper division segmentectomy (n = 8) was preserved in 21% +/- 11% of the preoperative values, which was significantly less than 35% +/- 12% after the lingular segmentectomy (n = 7) (P = .03). CONCLUSION: Segmentectomy can preserve the pulmonary function more significantly than lobectomy, except for the resection of more than three segments or the left upper division segmentectomy.

Difference of sentinel lymph node identification between tin colloid and phytate in patients with non-small cell lung cancer.

BACKGROUND: The advantages and disadvantages of technetium Tc 99m tin colloid and technetium Tc 99m phytate as tracers for sentinel node (SN) identification in patients with clinical stage I non-small cell lung cancer were examined retrospectively. METHODS: Sentinel node identification was conducted using tin colloid and phytate, respectively, in 73 and 74 patients with clinical stage I non-small cell lung cancer. We compared these two tracers in terms of identification rates, numbers of SNs, characteristics of patients whose SNs could not be identified, and the pathologic results of SNs. RESULTS: The tin colloid tracer identified SNs in 54 of the 73 patients (74%), which was significantly lower than the 89% (66 of 74 patients) in the phytate group (p = 0.02). The number of SNs per patient was 1.7 +/- 0.8 in the tin colloid group, which was significantly less than the 2.4 +/- 1.5 in the phytate group (p = 0.002). Although patients in the tin colloid group whose SNs could not be identified had a significantly lower forced expiratory volume in 1 second to forced vital capacity ratio than those whose SNs could be identified (p = 0.04), the phytate group did not show such a difference. Eleven of 120 patients whose SNs could be identified had pathologic N1 or N2 disease, but neither group showed any false-negative results for SN identification. CONCLUSIONS: Both tin colloid and phytate are reliable tracers for identifying SNs in non-small cell lung cancer. The advantage of phytate is that SNs can be detected more frequently than with tin colloid, even in patients with a low forced expiratory volume in 1 second to forced vital capacity ratio. However, tin colloid requires fewer nodes than phytate to identify SNs.

Prediction of pulmonary function after lung lobectomy by subsegments counting, computed tomography, single photon emission computed tomography and computed tomography: a comparative study.

OBJECTIVE: The aim of the present study was to determine the optimal method of predicting postoperative pulmonary function (PPF) after lung lobectomy. METHODS: The forced expiratory volume in 1s (FEV(1)) was measured in 37 patients before and after lobectomy, and the following three methods of predicting the PPF were evaluated: (1) the number of functioning subsegments to be resected were counted (subsegments counting [SC]); (2) the volume of the functioning lung was calculated using CT images (quantitative CT); and (3) perfusion scintigraphy was performed using co-registered single photon emission computed tomography and CT imaging (SPECT/CT). The FEV(1) values predicted using these three methods were then compared with the measured postoperative FEV(1), and the correlations and differences were analyzed. RESULTS: While a paired t-test showed the SPECT/CT method to have the smallest difference between the measured and the predicted FEV(1) values (0.05 l, p=0.33), followed by the quantitative CT method (0.07 l, p=0.07), and finally the SC method (0.15 l, p<0.001), the difference between the two values was not significantly different between the quantitative CT and SPECT/CT method (p=0.22). CONCLUSIONS: While the SC method is inferior to both the quantitative CT and the SPECT/CT methods for predicting the PPF after lobectomy, the latter two methods are almost equally accurate.

Prospective clinical trial of magnetic-anchor-guided endoscopic submucosal dissection for large early gastric cancer (with videos).

BACKGROUND: The treatment of early gastric cancer (EGC) by endoscopic submucosal dissection (ESD) has been rapidly gaining popularity in Japan. However, the procedure needs a high quality of skill. To facilitate complicated ESD by using a single working-channel gastroscope ("one-hand surgery method"), the magnetic-anchor-guided ESD (MAG-ESD) controlled by an extracorporeal electromagnet was reported to be successful in a porcine model. OBJECTIVES: The purpose of this prospective clinical trial was to evaluate the feasibility of MAG-ESD for large EGC located on the gastric body in human beings. DESIGN: Prospective clinical trial at a single center. SETTING: National Cancer Center Hospital, Tokyo, Japan. SUBJECTS: From January 2005 to May 2006, 25 patients with EGC >20 mm in diameter, located in the gastric body, and intestinal-type histology were enrolled. Patients with a cardiac pacemaker, advanced malignancy in other organs, severe cardiac and/or pulmonary diseases, and uncontrolled hypertension and/or diabetes mellitus were excluded from this study. INTERVENTIONS: Similar to a standard ESD, the MAG-ESD procedure was performed with the patient under conscious sedation by intravenous injection of midazolam (3-5 mg) and pentazocine (15 mg). MAIN OUTCOME MEASUREMENTS: Unfavorable events and other intraoperative complications caused by the magnetic anchor or the magnetic force were recorded and evaluated. Two GI endoscopists (T.G., I.O.) assessed whether the magnetic anchor facilitated gastric ESD according to 2 criteria: "supportive" and "not supportive." The en bloc resection rate, complications, total operation time, bleeding, perforation, and recurrence rate were also evaluated. The total operation time was measured from insertion to withdrawal of the endoscope, including the retrieving of the magnetic anchor or anchors. RESULTS: All tumors were resected en bloc, without any perforations or severe uncontrollable bleeding. All magnetic anchors were safely retrieved. Two endoscopists assessed that the MAG system was supportive in 23 patients. None of the patients experienced physiologic and mental abnormalities as a result of long-term magnetic-field exposure. During a median follow-up of 20 months (15-32 months), neither delayed adverse effects nor allergies caused by the stainless steel of the magnetic anchor were observed. CONCLUSIONS: MAG-ESD is a feasible and safe method that allowed an excellent visualization by suitable tissue tension and facilitated gastric ESD in patients with EGC. The system should be miniaturized to make it applicable in daily clinical practice.

Epidermal growth factor receptor mutations in multicentric lung adenocarcinomas and atypical adenomatous hyperplasias.

BACKGROUND: The mechanisms of generation and progression of multicentric lung adenocarcinoma (AD), bronchioloalveolar carcinoma (BAC), and atypical adenomatous hyperplasia (AAH) in the peripheral lung is not well known. In this study, we analyzed epidermal growth factor receptor (EGFR) mutations in the cases of multicentric AD, BAC, and AAH to reveal the role of EGFR mutation in their generations and progressions. METHOD: Ninety-seven AAH, BAC, or AD lesions less than 3 cm in size in 26 patients were surgically resected. Of these, EGFR mutations of the nodules with the highest and the second highest grade of histologic malignancy were examined in each patient by using the peptide nucleic acid-locked nucleic acid polymerase chain reaction (PNA-LNA PCR) clamp method. RESULTS: EGFR mutations could be examined in 48 nodules in the 26 patients. The EGFR mutations were found more frequently in lesions with higher histologic malignancy, ie, 9 of 10 ADs (90%), 16 of 28 BACs (57%), and one of 10 AAHs (10%). In 22 patients who could be examined of EGFR mutations for the two lesions in each patient, only two patients (9%) had the same mutation patterns between the two lesions, whereas 15 patients (68%) had the different statuses and the remaining five (23%) had no mutations. CONCLUSION: Our data demonstrated that EGFR mutations seem to contribute to the acquisition of malignant potential in the AAH-AD sequence and occur independently in each lesion and in the cases of multicentric AD, BAC, and AAH.

Novel germline mutation: EGFR V843I in patient with multiple lung adenocarcinomas and family members with lung cancer.

A novel germline transmission of the epidermal growth factor receptor (EGFR) mutation V843I in a family with multiple members with lung cancer is reported. The proband was a 70-year-old woman with multiple adenocarcinomas who exhibited secondary EGFR mutations, either L858R or L861Q, in the specimens of resected tumors, in addition to a germline EGFR V843I mutation. These observations suggest that the germline EGFR V843I mutation might have altered EGFR signaling in the multicentric development of adenocarcinoma, bronchoalveolar carcinoma, and atypical adenomatous hyperplasia and also might have had a role in the development of lung cancer in multiple members of her family.

Sentinel node identification in clinical stage Ia non-small cell lung cancer by a combined single photon emission computed tomography/computed tomography system.

OBJECTIVE: A gamma probe can identify sentinel nodes before nodal dissection in the mediastinum but not in the hilum, owing to high radioactivity from primary tumors. We evaluated the utility of fused single photon emission computed tomography/computed tomography (SPECT/CT) images for the identification of sentinel nodes in the hilum for patients with clinical stage Ia non-small cell lung cancer. METHODS: Technetium-99m tin colloid was injected into the peritumoral region approximately 18 hours before surgery in 63 patients with clinical stage Ia non-small cell lung cancer. On the morning of the operation, approximately 16 hours after administration of tin colloid, sentinel nodes were identified by fused SPECT/CT; this was followed by intraoperative sentinel node identification in the dissected lymph nodes by gamma probe. Because the gamma probe is a standard method for sentinel node identification, the sensitivity of fused SPECT/CT images was examined on the basis of the data of the gamma probe. RESULTS: Fused SPECT/CT images could identify sentinel nodes at segmental and lobar lymph nodes with a sensitivity of 0.87 and 0.74, both of which were significantly higher than 0.40 in the mediastinum (P < .001 and P = .012, respectively). In 5 patients with pathologic N1 or N2 disease, both SPECT/CT and the gamma probe could identify sentinel nodes with metastases. CONCLUSIONS: SPECT/CT could identify sentinel nodes of the hilum especially in segmental and lobar lymph nodes but not in the mediastinum. Because the gamma probe can identify sentinel nodes before nodal dissection in the mediastinum but not in the hilum, a combination of SPECT/CT and the gamma probe can be used to identify sentinel nodes before nodal dissection in both the hilum and the mediastinum, which will enable sentinel node navigation surgery in non-small cell lung cancer.