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Enzymes have been known to impact the biofilm forming capacity. However, how the enzymes mediate the biofilm formation and macrofouling remains little known. Here, we investigated the effects of the three kinds of proteases, four kinds of glycosidases and one kind of lipase on the detachment of biofilms of Shewanella marisflavi ECSMB14101, identified biofilm total proteins response to enzyme treatments, and then tested the effects of biofilms treated with enzymes on the settlement of the mussel Mytilus coruscus plantigrades. The results showed that the cell density of bacteria in biofilms formed at different initial bacterial density were noticeably reduced after treating with all tested enzymes, and Neutrase and α-Amylase exhibited best removing efficiency of > 90%. Bacterial total proteins in S. marisflavi biofilm noticeably reduced or disappeared after treated by Alcalase. For the settlements of the mussel M. coruscus plantigrades, inducing capacities of S. marisflavi biofilm were noticeably suppressed and downregulation was > 75% at the initial density of 5 × 10 6 cells/cm 2 . Thus, the tested enzymes could effectively remove the adhered bacterial cell, inhibit the biofilm formation and finally suppress the mussel settlement. Our findings extend novel knowledge to developing eco-friendly approach to control micro- and macro-fouling.

PurposeThis study evaluated the results of provocative angiography performed to identify the site of hemorrhage in lower gastrointestinal bleeding to enable transcatheter arterial embolization.Materials and methodsThe subjects of this study were 11 patients with acute lower gastrointestinal bleeding who underwent provocative angiography, after contrast agent extravasation could not be identified during conventional angiography. One patient underwent provocative angiography twice, making 12 cases of provocative angiography that were analyzed in this study. Urokinase was used in all cases. Heparin was administered in 2 cases, nicardipine in 3 cases, and alprostadil and isosorbide in 1 case each.ResultsContrast agent extravasation as a result of provocative angiography was observed in 6/12 cases (50%). Selective transcatheter arterial embolization was performed in all 6 cases in which extravasation was apparent, and it was technically successful in all six. Clinical success was achieved in 5 (83.3%) of the 6 cases in which technical success was achieved after provocative angiography.ConclusionProvocative angiography enabled the site of lower gastrointestinal bleeding to be identified in 6 of 12 cases (50.0%) when it could not be identified by conventional angiography, and in all 6 cases hemostasis was effectively achieved by embolization.

Purpose Spinal bone metastases directly affect quality of life, and patients with lytic-dominant lesions are at high risk for neurological symptoms and fractures. To detect and classify lytic spinal bone metastasis using routine computed tomography (CT) scans, we developed a deep learning (DL)-based computer-aided detection (CAD) system. Methods We retrospectively analyzed 2125 diagnostic and radiotherapeutic CT images of 79 patients. Images annotated as tumor (positive) or not (negative) were randomized into training (1782 images) and test (343 images) datasets. YOLOv5m architecture was used to detect vertebra on whole CT scans. InceptionV3 architecture with the transfer-learning technique was used to classify the presence/absence of lytic lesions on CT images showing the presence of vertebra. The DL models were evaluated via fivefold cross-validation. For vertebra detection, bounding box accuracy was estimated using intersection over union (IoU). We evaluated the area under the curve (AUC) of a receiver operating characteristic curve to classify lesions. Moreover, we determined the accuracy, precision, recall, and F1 score. We used the gradient-weighted class activation mapping (Grad-CAM) technique for visual interpretation. Results The computation time was 0.44 s per image. The average IoU value of the predicted vertebra was 0.923 ± 0.052 (0.684–1.000) for test datasets. In the binary classification task, the accuracy, precision, recall, F1-score, and AUC value for test datasets were 0.872, 0.948, 0.741, 0.832, and 0.941, respectively. Heat maps constructed using the Grad-CAM technique were consistent with the location of lytic lesions. Conclusion Our artificial intelligence-aided CAD system using two DL models could rapidly identify vertebra bone from whole CT images and detect lytic spinal bone metastasis, although further evaluation of diagnostic accuracy is required with a larger sample size.

Biofilms are critical components of most marine systems and provide biochemical cues that can significantly impact overall community composition. Although progress has been made in the bacteria–animal interaction, the molecular basis of modulation of settlement and metamorphosis in most marine animals by bacteria is poorly understood. Here, Pseudoalteromonas marina showing inducing activity on mussel settlement and metamorphosis was chosen as a model to clarify the mechanism that regulates the bacteria–mussel interaction. We constructed a flagellin synthetic protein gene fliP deletion mutant of P. marina and checked whether deficiency of fliP gene will impact inducing activity, motility, and extracellular polymeric substances of biofilms. Furthermore, we examined the effect of flagellar proteins extracted from bacteria on larval settlement and metamorphosis. The deletion of the fliP gene caused the loss of the flagella structure and motility of the ΔfliP strain. Deficiency of the fliP gene promoted the biofilm formation and changed biofilm matrix by reducing β-polysaccharides and increasing extracellular proteins and finally reduced biofilm-inducing activities. Flagellar protein extract promoted mussel metamorphosis, and ΔfliP biofilms combined with additional flagellar proteins induced similar settlement and metamorphosis rate compared to that of the wild-type strain. These findings provide novel insight on the molecular interactions between bacteria and mussels.

Uncoordinate-45B (UNC-45B) is a myosin chaperone in striated muscles. Heat shock leads to myosin denaturation, and UNC-45B is involved in myosin refolding. Pacific bluefin tuna (PBT) was chosen for this study because of its high value and severe “burnt meat” problem in summer. cDNA cloning and expression analysis were carried out to evaluate the heat stress response of PBT UNC-45B. The cDNA of PBT unc-45b was cloned, and two isoforms were detected successfully. Isoform 1 consisted of 2994 bp encoding 931 amino acids, and isoform 2 consisted of 3071 bp encoding 930 amino acids. To clarify the effects of heat stress on UNC-45B, cultured PBT were sampled in summer (29.2 °C) and winter (20.9 °C), and analyzed by qPCR and immunoblotting. Compared with winter, both transcriptional and translational expression of UNC-45B in summer were dramatically higher. In addition, from the mRNA expression results, we found that PBT UNC-45B isoform 2 mainly responded to heat stress. Our findings indicate that an increase in UNC-45B expression levels can sensitively reflect the heat stress response in PBT. Therefore, UNC-45B may be considered as a candidate biomarker to evaluate heat stress tolerance in PBT. This will improve aquaculture development by enabling the selection of heat stress-tolerant juveniles.

The present work aims to clarify the mechanism of gel weakening (modori phenomenon) on surimi-based products and provide information on how to prevent it from happening in threadfin bream. We investigated the distribution of endogenous proteinases in the normal and belly muscles and their effects on the quality of the surimi gel. There was severe degradation of myofibrillar proteins in the belly muscle during heating at 50 °C, which was not found in the normal muscle. The surimi gel prepared from the belly muscle also showed a low gel strength as compared to that of the normal muscle. The myofibrillar proteins in the belly muscle were found to be degraded by a sarcoplasmic serine proteinase (SSP). The SSP activity was detected in the belly muscle and hepatopancreas but not in the normal muscle, while the SSP mRNA was detected only in the hepatopancreas. It is suggested that SSP in the hepatopancreas leaks to the belly muscle during post-harvest storage and is responsible for the modori phenomenon. These results imply that high-quality surimi gel of the threadfin bream can be produced by preventing protease leakage to the muscle during post-harvest storage.

This study aimed to examine the influence of dosimetric factors on gastrointestinal toxicity after radical re-irradiation for lymph node recurrence in the abdominopelvic region using a composite plan. Between January 2008 and March 2017, 33 patients underwent radical re-irradiation for lymph node recurrence in the abdominopelvic region with a complete overlap with previous radiation therapy (RT) with the median prescription dose of the second RT of 71.7 Gy . Re-irradiation planning protocol for target volume and organs at risk (OARs) (duodenum, small and large intestines) was decided as follows: more than equal to 97% of the prescription dose was administered to the D95 (percentage of the minimum dose that covered 95% of the target volume) of planning target volume (PTV); minimal dose to the maximally irradiated doses delivered to 1cc [D1 cc] and 5cc [D5 cc] of OARs was set below 70 Gy and 50 Gy , respectively; and D1 cc and D5 cc in the cumulative plans to OARs were 120 Gy and 100 Gy . Kaplan-Meier analyses were performed to evaluate overall survival (OS) and univariate log-rank and multivariate Cox proportional hazards model analyses were performed to explore predictive factors. Using dose summation of the first and re-irradiation plans, we conducted a dosimetric analysis for grade ≥ 3 toxicities of the duodenum and intestine. With a median follow-up of 18 months, the two-year OS rate was 45.5%. The number of RT fields (localized or multiple) was a significant predisposing factor for OS rate with a hazard ratio of 0.23 (95% confidence interval 0.07-0.73). The two-year OS of the patients with a localized RT field was 63.6% and 9.1% for multiple RT fields (p= 0.00007). Four patients experienced grade ≥3 gastrointestinal toxicity related to re-irradiation (4/33=12.1%). We could not find any predisposing dosimetric value in the comparisons with and without toxicity. The dose constraints presented in this study are relatively low rates of toxicity, which may be useful when planning re-irradiation. Especially, for the patients who could be treated with localized RT field, radical re-irradiation with a high curative dose is a good option. No dosimetric predisposing factor was found for radical re-irradiation of abdominopelvic lesions in the composite plan.

R-phycoerythrin prepared from red algae ( Bangia fusco - purpurea ) was hydrolyzed by pepsin followed by trypsin to produce angiotensin I-converting enzyme (ACE) inhibitory peptides. The IC 50 of the hydrolysate of R-phycoerythrin (HRPE) was 191.1 ± 4.1 μg/mL, and the molecular weight of most products (89.9 %) was below 2 kDa. After sequential gel permeation and reversed-phase chromatography steps to purify the hydrolysate, two peptides with the sequences of ALLAGDPSVLEDR and VVGGTGPVDEWGIAGAR were obtained and their IC 50 values were 57.2 ± 5.0 and 66.2 ± 4.2 μg/mL, respectively. The ALLAGDPSVLEDR and VVGGTGPVDEWGIAGAR peptides were derived from the β- and α-subunit of R-phycoerythrin from Polysiphonia urceolata , with a 92.3 % (12/13) and 94.1 % (16/17) match, respectively. Both peptides were resistant to digestion by proteinases common in the gastrointestinal tract. Therefore, the identified novel peptides derived from R-phycoerythrin may be used as potential nutraceuticals for development of functional foods.

The molecular mechanisms underlying the conspecific cue-mediated larval settlement in Crassostrea gigas is not yet fully understood. In this study, we described and compared the transcriptomes of competent pediveligers (Pedi) and conspecific cue-induced postlarvae (PL). A total of 2383 candidate transcripts were identified: 740 upregulated and 1643 downregulated transcripts, after settlement. Gene Ontology analysis revealed active chitin binding, calcium ion binding, and extracellular region processes in both stages. Results showed that the differential expression trend of six candidate transcripts were consistent between the quantitative real-time PCR and transcriptome data. The differential transcript expression related to shell formation showed closely linked dynamics with a gene regulatory network that may involve the interplay of various hormone receptors, neurotransmitters, and neuropeptide receptors working together in a concerted way in the Pedi and PL stages. Our results highlight the transcriptome dynamics underlying the settlement of oysters on conspecific adult shells and demonstrate the potential use of this cue as an attractant for wild and hatchery-grown oyster larval attachment on artificial substrates. It also suggests the possible involvement of an ecdysone signal pathway that may be linked to a neuroendocrine-biomineralization crosstalk in C. gigas settlement.

Modori-inducing proteinases (MIPs) in horse mackerel and the effect of sodium citrate on the modori phenomenon were investigated. Cysteine protease caused the modori phenomenon only at 60 ℃, while metalloproteinase caused it at both 60 ℃ and 70 ℃. Moreover, the autolysis of myofibrillar proteins was mainly inhibited by ethylene diamine tetra-acetic acid (EDTA), verifying the existence of insoluble metalloproteinase. From the results comparing thermal gel properties and protease activities, we confirmed that increased activity of insoluble metalloproteinase corresponded to the weakening of the thermal gel. Thus, insoluble metalloproteinase was the major MIP in horse mackerel. Moreover, sodium citrate could markedly inhibit insoluble metalloproteinase activity at 60 ℃. The addition of sodium citrate significantly improved the breaking force of the thermal gel at 60 ℃ and 70 ℃, suggesting that sodium citrate could suppress the modori phenomenon by inhibiting insoluble metalloproteinase. Therefore, it is desirable to use sodium citrate in horse mackerel surimi-based products to prevent the modori phenomenon.