Leaf Gas Film 1 promotes glycerol ester accumulation and formation of a tight root barrier to radial O2 loss in rice.

Rice (Oryza sativa L.) and many other wetland plants form an apoplastic barrier in the outer parts of the roots to restrict radial O2 loss to the rhizosphere during soil flooding. This barrier facilitates longitudinal internal O2 diffusion via gas-filled tissues from shoot to root apices, enabling root growth in anoxic soils. We tested the hypothesis that Leaf Gas Film 1 (LGF1), which influences leaf hydrophobicity in rice, plays a crucial role in tight outer apoplastic barriers formation in rice roots. We examined the roots of a rice mutant (dripping wet leaf 7, drp7) lacking functional LGF1, its wild type, and an LGF1 overexpression line for their capacity to develop outer apoplastic barriers that restrict radial O2 loss. We quantified the chemical composition of the outer part of the root and measured radial O2 diffusion from intact roots. The drp7 mutant exhibited a weak barrier to radial O2 loss compared to the wild type. However, introducing functional LGF1 into the mutant fully restored tight barrier function. The formation of a tight barrier to radial O2 loss was associated with increased glycerol ester levels in exodermal cells, rather than differences in total root suberization or lignification. These results demonstrate that, in addition to its role in leaf hydrophobicity regulation, LGF1 plays an important role in controlling the function of the outer apoplastic barriers in roots. Our study suggests that increased deposition of glycerol esters in the suberized root exodermis establishes a tight barrier to radial O2 loss in rice roots.

Percutaneous sclerotherapy with OK-432 for lymphocele after pelvic or para-aortic lymphadenectomy: preliminary results.

BACKGROUND: Lymphoceles can result from disruption of lymphatic vessels after surgical procedures in areas with extensive lymphatic networks. Percutaneous catheter drainage with sclerotherapy can be performed for the treatment of lymphoceles. OK-432 has been used to treat benign cysts, such as lymphangioma and ranula. Therefore, we aimed to report the efficacy and safety of sclerotherapy using OK-432 for postoperative lymphoceles. This study retrospectively analyzed 16 patients who underwent sclerotherapy using OK-432 for postoperative pelvic and para-aortic lymphoceles between April 1, 2012, and March 31, 2020. All the patients underwent percutaneous drainage before sclerotherapy. The indications for sclerotherapy were persistent drainage tube output of greater than 50 mL per day and recurrent lymphoceles after percutaneous drainage. If less than 20 mL per day was drained after sclerotherapy, the tube was removed. When the drainage tube output did not decrease to less than 20 mL per day after the first sclerotherapy, the second sclerotherapy was performed 1 week later. Technical success was defined as the completion of drainage and sclerotherapy procedures. Clinical success was defined as the resolution of the patient's symptoms resulting from lymphoceles without surgical intervention. This study also evaluated the complications of sclerotherapy and their progress after sclerotherapy. RESULTS: The mean initial lymphocele size and drainage duration after sclerotherapy were 616 mL and 7.1 days, respectively. The technical success rate and clinical success rate were 100% and 93%, respectively. Thirteen patients were treated by one-session sclerotherapy and three patients were treated by two-session sclerotherapy. Minor complications (fever) were observed in eight patients (50%). A major complication (small bowel fistula) was observed in one patient (7%). No recurrence of lymphoceles was observed during the mean follow-up period of 17 months. CONCLUSION: Sclerotherapy with OK-432 is an effective therapeutic method for postoperative lymphoceles. Although most complications are minor, a small bowel fistula was observed in one patient.

Noninvasive imaging of hollow structures and gas movement revealed the gas partial-pressure-gradient-driven long-distance gas movement in the aerenchyma along the leaf blade to submerged organs in rice.

Rice (Oryza sativa) plants have porous or hollow organs consisting of aerenchyma, which is presumed to function as a low-resistance diffusion pathway for air to travel from the foliage above the water to submerged organs. However, gas movement in rice plants has yet to be visualized in real time. In this study involving partially submerged rice plants, the leaves emerging from the water were fed nitrogen-13-labeled nitrogen ([13 N]N2 ) tracer gas, and the gas movement downward along the leaf blade, leaf sheath, and internode over time was monitored. The [13 N]N2 gas arrived at the bottom of the plant within 10 min, which was 20 min earlier than carbon-11 photoassimilates. The [13 N]N2 gas movement was presumably mediated by diffusion along the aerenchyma network from the leaf blade to the root via nodes functioning as junctions, which were detected by X-ray computed tomography. These findings imply the diffusion of gas along the aerenchyma, which does not consume energy, has enabled plants to adapt to aquatic environments. Additionally, there were no major differences in [13 N]N2 gas movement between paddy rice and deepwater rice plants, indicative of a common aeration mechanism in the two varieties, despite the difference in their response to flooding.

Local Efficacy and Safety of Transarterial Chemoembolization for Hepatocellular Carcinoma: Epirubicin-Loaded DC Beads vs. Epirubicin-Lipiodol Emulsion with Gelatin Sponge (cTACE).

Purpose: To compare the efficacy and safety between epirubicin-loaded DC Beads (DCB-TACE) and conventional TACE (cTACE) used in transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC). Materials and Methods: This retrospective study enrolled 64 patients (mean age, 73.3 years; 44 men, 20 women) who underwent initial DCB-TACE between 2014 and 2015, and 66 patients (mean age, 71.3 years; 38 men, 28 women) who underwent initial cTACE between 2011 and 2013 as historical controls. Treatment effects on the target lesions at 3 months after TACE, the period until re-treatment of the target lesion, and adverse events after TACE were compared between the groups. Univariate and multivariate analyses were also performed to estimate the factors influencing the treatment effects. Results: Based on the Response Evaluation Criteria in Cancer of the Liver version 2015, treatment response of the target lesions equivalent to a complete response and termed as TE4, was 51.0% (53/104) in the DCB-TACE group and 74.4% (64/86) in the cTACE group (p<0.001). Multivariate analysis revealed that the TACE procedure, Child-Pugh score, serum aspartate aminotransferase (AST) level, alpha fetoprotein level, and tumor size were independent significant predictors of TE4. The frequencies of elevated serum AST and alanine transaminase levels after TACE were significantly lower in patients in the DCB-TACE group (p<0.001 each). No significant difference in biliary/liver damage was evident between the groups. Conclusion: The local efficacy of cTACE was higher than that of DCB-TACE. Adverse events were milder after DCB-TACE than after cTACE.

Functional Domain Order of an Anti-EGFR × Anti-CD16 Bispecific Diabody Involving NK Cell Activation.

Bispecific antibodies (bsAbs) have emerged as promising therapeutics. A bispecific diabody (bsDb) is a small bsAb consisting of two distinct chimeric single-chain components, with two possible arrangements of the domains. We previously reported the effect of domain order on the function of a humanized bsDb targeting the epidermal growth factor receptor (EGFR) on cancer cells, and CD3 on T cells. Notably, the co-localization of a T-cell receptor (TCR) with CD3 is bulky, potentially affecting the cross-linking ability of bsDbs, due to steric hindrance. Here, we constructed and evaluated humanized bsDbs, with different domain orders, targeting EGFR and CD16 on natural killer (NK) cells (hEx16-Dbs). We predicted minimal effects due to steric hindrance, as CD16 lacks accessory molecules. Interestingly, one domain arrangement displayed superior cytotoxicity in growth inhibition assays, despite similar cross-linking abilities for both domain orders tested. In hEx16-Dbs specifically, domain order might affect the agonistic activity of the anti-CD16 portion, which was supported by a cytokine production test, and likely contributed to the superiority of one of the hEx16-Dbs. Our results indicate that both the target antigen and mode of action of an antibody must be considered in the construction of highly functional bsAbs.

Efficacy of prophylactic selective arterial embolization for renal angiomyolipomas: identifying predictors of 50% volume reduction.

BACKGROUND: Transcatheter arterial embolization (TAE) has been widely performed for renal angiomyolipomas (AMLs) as prophylaxis or emergency treatment. On the other hand, mammalian target of rapamycin (mTOR) inhibitors have recently been used for tuberous sclerosis (TSC)-related AMLs, and no comparison between the effectiveness of mTOR inhibitors versus prophylactic selective TAE has yet been performed. Therefore, the purpose of this study was to evaluate the efficacy of TAE for AML tumor volume reduction and predictors of tumor volume decrease over 50%, with reference to the EXIST-2 trial. METHODS: A total of 44 patients who underwent 48 prophylactic embolization procedures for 50 AMLs in a single institution between 2004 and 2018 were included. Indications for TAE of AMLs were tumor size ≥4 cm or aneurysm ≥5 mm in diameter on contrast-enhanced computed tomography (CECT). Microspheres, ethanol, and micro-coils were used as embolic agents. The percentage volume reduction from before TAE to the minimum volume during follow-up after TAE was calculated, and predictors for 50% volume reduction were identified by univariate and multivariate binary logistic regression analyses. RESULTS: The technical success rate was 100% (50 of 50). No severe acute complications related to the procedure were encountered. Tumor volume reduction of ≥50% was observed in 35/50 AMLs. There was a significant difference in the rate of tumor volume reduction of 50% between the presence and absence of an aneurysm ≥5 mm and between tumor diameter ≥ 70 mm and < 70 mm on univariate analysis. On multivariate analysis, tumor diameter < 70 mm was the only independent predictor of significant tumor volume reduction after TAE. CONCLUSION: Prophylactic selective TAE for AMLs has good tumor-reduction effects, especially for AMLs with tumor diameter < 70 mm.

Diel O2 Dynamics in Partially and Completely Submerged Deepwater Rice: Leaf Gas Films Enhance Internodal O2 Status, Influence Gene Expression and Accelerate Stem Elongation for 'Snorkelling' during Submergence.

Deepwater rice has a remarkable shoot elongation response to partial submergence. Shoot elongation to maintain air-contact enables 'snorkelling' of O2 to submerged organs. Previous research has focused on partial submergence of deepwater rice. We tested the hypothesis that leaf gas films enhance internode O2 status and stem elongation of deepwater rice when completely submerged. Diel patterns of O2 partial pressure (pO2) were measured in internodes of deepwater rice when partially or completely submerged, and with or without gas films on leaves, for the completely submerged plants. We also took measurements for paddy rice. Deepwater rice elongated during complete submergence and the shoot tops emerged. Leaf gas films improved O2 entry during the night, preventing anoxia in stems, which is of importance for elongation of the submerged shoots. Expressions of O2 deprivation inducible genes were upregulated in completely submerged plants during the night, and more so when gas films were removed from the leaves. Diel O2 dynamics showed similar patterns in paddy and deepwater rice. We demonstrated that shoot tops in air enabled 'snorkelling' and increased O2 in internodes of both rice ecotypes; however, 'snorkelling' was achieved only by rapid shoot elongation by deepwater rice, but not by paddy rice.

Ethylene-gibberellin signaling underlies adaptation of rice to periodic flooding.

Most plants do poorly when flooded. Certain rice varieties, known as deepwater rice, survive periodic flooding and consequent oxygen deficiency by activating internode growth of stems to keep above the water. Here, we identify the gibberellin biosynthesis gene, SD1 (SEMIDWARF1), whose loss-of-function allele catapulted the rice Green Revolution, as being responsible for submergence-induced internode elongation. When submerged, plants carrying the deepwater rice-specific SD1 haplotype amplify a signaling relay in which the SD1 gene is transcriptionally activated by an ethylene-responsive transcription factor, OsEIL1a. The SD1 protein directs increased synthesis of gibberellins, largely GA4, which promote internode elongation. Evolutionary analysis shows that the deepwater rice-specific haplotype was derived from standing variation in wild rice and selected for deepwater rice cultivation in Bangladesh.

Structural basis of the correct subunit assembly, aggregation, and intracellular degradation of nylon hydrolase.

Nylon hydrolase (NylC) is initially expressed as an inactive precursor (36 kDa). The precursor is cleaved autocatalytically at Asn266/Thr267 to generate an active enzyme composed of an α subunit (27 kDa) and a ß subunit (9 kDa). Four αß heterodimers (molecules A-D) form a doughnut-shaped quaternary structure. In this study, the thermostability of the parental NylC was altered by amino acid substitutions located at the A/D interface (D122G/H130Y/D36A/L137A) or the A/B interface (E263Q) and spanned a range of 47 °C. Considering structural, biophysical, and biochemical analyses, we discuss the structural basis of the stability of nylon hydrolase. From the analytical centrifugation data obtained regarding the various mutant enzymes, we conclude that the assembly of the monomeric units is dynamically altered by the mutations. Finally, we propose a model that can predict whether the fate of the nascent polypeptide will be correct subunit assembly, inappropriate protein-protein interactions causing aggregation, or intracellular degradation of the polypeptide.

Rice leaf hydrophobicity and gas films are conferred by a wax synthesis gene (LGF1) and contribute to flood tolerance.

Floods impede gas (O2 and CO2 ) exchange between plants and the environment. A mechanism to enhance plant gas exchange under water comprises gas films on hydrophobic leaves, but the genetic regulation of this mechanism is unknown. We used a rice mutant (dripping wet leaf 7, drp7) which does not retain gas films on leaves, and its wild-type (Kinmaze), in gene discovery for this trait. Gene complementation was tested in transgenic lines. Functional properties of leaves as related to gas film retention and underwater photosynthesis were evaluated. Leaf Gas Film 1 (LGF1) was identified as the gene determining leaf gas films. LGF1 regulates C30 primary alcohol synthesis, which is necessary for abundant epicuticular wax platelets, leaf hydrophobicity and gas films on submerged leaves. This trait enhanced underwater photosynthesis 8.2-fold and contributes to submergence tolerance. Gene function was verified by a complementation test of LGF1 expressed in the drp7 mutant background, which restored C30 primary alcohol synthesis, wax platelet abundance, leaf hydrophobicity, gas film retention, and underwater photosynthesis. The discovery of LGF1 provides an opportunity to better understand variation amongst rice genotypes for gas film retention ability and to target various alleles in breeding for improved submergence tolerance for yield stability in flood-prone areas.

Structural considerations for functional anti-EGFR × anti-CD3 bispecific diabodies in light of domain order and binding affinity.

We previously reported a functional humanized bispecific diabody (bsDb) that targeted EGFR and CD3 (hEx3-Db) and enhancement of its cytotoxicity by rearranging the domain order in the V domain. Here, we further dissected the effect of domain order in bsDbs on their cross-linking ability and binding kinetics to elucidate general rules regarding the design of functional bsDbs. Using Ex3-Db as a model system, we first classified the four possible domain orders as anti-parallel (where both chimeric single-chain components are variable heavy domain (VH)-variable light domain (VL) or VL-VH order) and parallel types (both chimeric single-chain components are mixed with VH-VL and VL-VH order). Although anti-parallel Ex3-Dbs could cross-link the soluble target antigens, their cross-linking ability between soluble targets had no correlation with their growth inhibitory effects. In contrast, the binding affinity of one of the two constructs with a parallel-arrangement V domain was particularly low, and structural modeling supported this phenomenon. Similar results were observed with E2x3-Dbs, in which the V region of the anti-EGFR antibody clone in hEx3 was replaced with that of another anti-EGFR clone. Only anti-parallel types showed affinity-dependent cancer inhibitory effects in each molecule, and E2x3-LH (both components in VL-VH order) showed the most intense anti-tumor activity in vitro and in vivo. Our results showed that, in addition to rearranging the domain order of bsDbs, increasing their binding affinity may be an ideal strategy for enhancing the cytotoxicity of anti-parallel constructs and that E2x3-LH is particularly attractive as a candidate next-generation anti-cancer drug.

Computed tomography-guided preoperative localization of small lung nodules with indocyanine green.

Background Small, deep-seated lung nodules and sub-solid nodules are often difficult to locate without marking. Purpose To evaluate the success and complication rates associated with the use of indocyanine green (ICG) to localize pulmonary nodules before resection. Material and Methods This retrospective study was approved by our institutional review board. Informed consent for performing preoperative localization using ICG marking was obtained from all patients. Thirty-seven patients (14 men, 23 women; mean age = 63.1 years; age range = 10-82 years) with small peripheral pulmonary nodules underwent computed tomography (CT)-guided ICG marking immediately before surgery between March 2007 and June 2016. The procedural details and complication rates associated with ICG marking are described. Results The average nodule size and depth were 9.1 mm (range = 2-22 mm) and 9.9 mm (range = 0-33 mm), respectively. Marking was detected at the pleural surface in 35 patients (95%). Three cases of mild pneumothorax (8%), five cases of cough (14%), and one case of mild bloody sputum (3%) with no clinical significance were noted. There were no severe complications. The average duration required to perform the marking was 19.4 min (range = 12-41 min). Conclusion Our results indicate that CT-guided ICG marking is safe and useful for detecting the location of small pulmonary nodules preoperatively.

Pulmonary tumor thrombotic microangiopathy caused by prostate carcinoma.

Pulmonary tumor thrombotic microangiopathy (PTTM) is a fatal malignancy-related condition that involves rapidly progressing hypoxia and pulmonary hypertension. We report a case of PTTM caused by prostate carcinoma, which was diagnosed before autopsy in an 81-year-old man. Computed tomography showed diffuse ground-glass opacities, consolidation, and small nodules in the peripheral regions of the lung. Autopsy showed adenocarcinoma cells embolizing small pulmonary arteries with fibrocellular intimal proliferation, which was consistent with PTTM caused by prostate carcinoma.

Domain order of a bispecific diabody dramatically enhances its antitumor activity beyond structural format conversion: the case of the hEx3 diabody.

The domains of bispecific diabodies (BsDbs) can be ordered in four different ways; however, the influence of domain order on the cytotoxicity of BsDbs that retarget immune cells against tumor cells had not been addressed. We previously reported the marked antitumor effects of a humanized BsDb that targets epidermal growth factor receptor and CD3 (hEx3-Db). Here, we rearranged the domains of hEx3-Db to examine the influence of domain order on the function of BsDbs. We successfully prepared homogenous dimers of hEx3-Db in all four domain configurations. Interestingly, all three rearranged hEx3s inhibited cancer growth more effectively than did the original hEx3-Db, in which both components were in variable heavy domain (VH)-variable light domain (VL) order (redesignated as hEx3-HL), and the highest effects were observed with hEx3-LH (hEx3-Db with both components in VL-VH order). In addition, hEx3-LH had comparable in vitro growth inhibitory effects to those of the tandem single-chain variable fragment (scFv) format of hEx3-Db (hEx3-tandem scFv (taFv)), which we previously showed to have greater cytotoxicity than does hEx3-HL. Flow cytometry suggested that the enhanced cytotoxicity of hEx3-LH is attributable to structural superiority for cross-linking, similar to that of hEx3-taFv. Furthermore, hEx3-LH inhibited cancer growth in mice more effectively than did hEx3-taFv; this difference may be due to differences in antibody stability. Our results show that merely rearranging the domain order of BsDbs can enhance their effects beyond those with structural format conversion.

The ethylene response factors SNORKEL1 and SNORKEL2 allow rice to adapt to deep water.

Living organisms must acquire new biological functions to adapt to changing and hostile environments. Deepwater rice has evolved and adapted to flooding by acquiring the ability to significantly elongate its internodes, which have hollow structures and function as snorkels to allow gas exchange with the atmosphere, and thus prevent drowning. Many physiological studies have shown that the phytohormones ethylene, gibberellin and abscisic acid are involved in this response, but the gene(s) responsible for this trait has not been identified. Here we show the molecular mechanism of deepwater response through the identification of the genes SNORKEL1 and SNORKEL2, which trigger deepwater response by encoding ethylene response factors involved in ethylene signalling. Under deepwater conditions, ethylene accumulates in the plant and induces expression of these two genes. The products of SNORKEL1 and SNORKEL2 then trigger remarkable internode elongation via gibberellin. We also demonstrate that the introduction of three quantitative trait loci from deepwater rice into non-deepwater rice enabled the latter to become deepwater rice. This discovery will contribute to rice breeding in lowland areas that are frequently flooded during the rainy season.