Polychaete Bioturbation Alters the Taxonomic Structure, Co-occurrence Network, and Functional Groups of Bacterial Communities in the Intertidal Flat.

Polychaetes are important benthic macrofauna that lives in sediments, usually in intertidal flats with high organic content and high sulfide. It has been suggested that polychaete bioturbation could perform environmental remediation. During the process, the microbial community plays important roles. Here, we used high-throughput sequencing technology to study the bioturbation effects on the bacterial community in the polychaete (Perinereis aibuhitensis) burrows at different tidal positions in intertidal flat. The results showed that the bacterial communities were dramatically influenced by the polychaete bioturbation. The ACE, Chao, and Shannon indices of the polychaete burrows increased in summer. Dominant phyla in the polychaete burrows were Proteobacteria, Campilobacterota, Desulfobacterota, Chloroflexi, and Bacteroidota, and the dominant bacterial families were Sulfurvaceae, Flavobacteriaceae, Rhodobacteraceae, Woeseiaceae, Desulfobulbaceae, and Sulfurimonadaceae. Results of linear discriminant analysis effect size (LEfSe) showed that groups that include organic matter degraders, such as Bacteroidota, Flavobacteriaceae, Rhodobacteraceae, Woeseiaceae, and groups that include sulfur oxidizers, such as Campilobacterota, Sulfurovaceae, Rhodobacteraceae, Desulfobulbaceae, and Sulfurimonadaceae, were significantly increased due to the polychaete bioturbation. The polychaete bioturbation reduced the complexity of the bacterial co-occurrence network while increased its modularity and homogeneity. The polychaete bioturbation also changed the functional groups, which significantly enhanced in functional groups of aerobic nitrite oxidation, nitration, dark thiosulfate oxidation, dark sulfur oxidation, and dark sulfite oxidation, while nitrogen respiration and nitrate respiration decreased. These results provide insight into the impact of bacterial communities under the intertidal polychaete bioturbation.

CO2-Induced Ocean Acidification Alters the Burrowing Behavior of Manila Clam Ruditapes philippinarum by Reversing GABAA Receptor Function.

Biological burrowing behavior is an important driver shaping ecosystems that is being threatened by CO2-induced ocean acidification; however, the effects of ocean acidification on burrowing behavior and its neurological mechanism remain unclear. This study showed that elevated pCO2 significantly affected the burrowing behaviors of the Manila clam Ruditapes philippinarum, such as increased foot contraction, burrowing time, and intrabottom movement and decreased burrowing depth. Delving deeper into the mechanism, exposure to elevated pCO2 significantly decreased extracellular pH and increased [HCO3-]. Moreover, an indicator GABAA receptor, a neuroinhibitor for movement, was found to be closely associated with behavioral changes. In situ hybridization confirmed that the GABAA receptor was widely distributed in ganglia and foot muscles, and elevated pCO2 significantly increased the mRNA level and GABA concentration. However, the increase in GABAA receptor and its ligand did not suppress the foot movement, but rather sent "excitatory" signals for foot contraction. The destabilization of acid-base homeostasis was demonstrated to induce an increase in the reversal potential for GABAA receptor and an alteration in GABAA receptor function under elevated pCO2. This study revealed that elevated pCO2 affects the burrowing behavior of Manila clams by altering GABAA receptor function from inhibitory to excitatory.

Application of ultrasound markers measured at different time points of COH cycle in the prediction of ovarian response for individualised ovulation induction.

The purpose of this study was to evaluate the predictive value of ultrasound markers measured at different time points of the controlled ovarian hyperstimulation (COH) cycle on ovarian response and outcome indicators in the IVF-ET cycle. According to the oestrogen level and the number of retrieved oocytes, patients who planned for COH treatment were separated into low-response group, normal and high-response group. The ovarian stromal artery flow parameters on the day of pituitary down-regulation, day 1, day 7, day 10, and the day of hCG injection were collected prospectively. We also have collected the data of cumulus oophorus count on the day of hCG injection by transvaginal sonography. Compared with the low-response group, on the first day of the COH cycle PI, RI, and S/D were lower in the high-response group than they were in the low-response group (p < .05). PSV and EDV were significantly higher in the high-response group than they were in the low-response group (p < .01), and the PSV on the first day of the COH cycle have statistical significance in predicting the number of high-quality embryos. The number of cumulus oophorus on the day of hCG injection has statistical significance in predicting the number of oocytes retrieved and fertilised oocytes. We conclude that the ovarian stromal artery flow parameters on the first day of the COH cycle and cumulus oophorus count on hCG injection day can serve as efficient indicators for an early assessment of ovarian response and individualised ovulation induction.IMPACT STATEMENTWhat is already known on this subject? AMH, AFC, and the age of the patient are well-known effective parameters for the evaluation of ovarian response, but these are insufficient and full of individual differences. Some researchers have investigated the value of colour Doppler ultrasound and cumulus oophorus in assessing ovarian response, but no definitive conclusion has been reached.What do the results of this study add? The hemodynamic parameters of ovarian stromal artery on the first day of the COH cycle and the number of cumulus oophorus on the day of hCG injection detected by Transvaginal Colour Doppler Sonography (TV-CDS) could be used to predict the ovarian response.What are the implications of these findings for clinical practice and/or further research? Ovarian stromal artery flow parameters and cumulus oophorus detected by TV-CDS can potentially be offered as a complementary parameter for ovarian reserve.

Integrated transcriptomics and metabolomics analyses reveal benzo[a]pyrene enhances the toxicity of mercury to the Manila clam, Ruditapes philippinarum.

Mercury (Hg2+) and benzo[a]pyrene (BaP) are ubiquitous and persistent pollutants with multiple toxicities in bivalve molluscs. Here, the toxicological responses in the gills of Manila clams, Ruditapes philippinarum, to Hg2+ (10 µg L-1), BaP (3 µg L-1), and their mixture were analysed using transcriptomics and metabolomics approaches. Comparisons of the transcriptomes and metabolomes of Hg2+-and/or BaP-treated clams with control animals revealed the involvement of the detoxification metabolism, immune defence, energy-related pathways, and osmotic regulation in the stress response of R. philippinarum. Exposure to Hg2+ alone primarily enhanced the detoxification and energy metabolic pathways by significantly increasing the expression of genes associated with heat-shock proteins and oxidative phosphorylation. However, co-exposure to Hg2+ and BaP caused greater immunotoxicity and disrupted detoxification metabolism, the TCA cycle, glycolysis, and ATP generation. The expression levels of cytochrome P450 1A1 (CYP1A1), multidrug resistance-associated protein 1 (MRP1), and myosin (MYO), and the activity of electron transport system (ETS) in gills were detected, supporting the underlying toxic mechanisms of Hg2+ and BaP. We suggest that the presence of BaP enhances the toxicity of Hg2+ by 1) hampering the detoxification of Hg2+, 2) increasing the immunotoxicity of Hg2+, and 3) constraining energy availability for clams.

The effects of teflubenzuron on mortality, physiology and accumulation in Capitella sp.

The chitin synthesis inhibitor teflubenzuron (TFB) is a feed antiparasitic agents used to impede molting of the salmon lice, an ecto-parasite that severely affects the salmon industry. Low absorption of oral administered TFB may cause elevated concentrations in the feces discharged from the salmon into the benthic environment. The polychaete Capitella sp. are often dominant in such habitats and consume organic waste deposited on the sediment. In the present study, Capitella sp. were exposed to doses of TFB in salmon feed of 1, 2 and 4 g TFB kg-1 (0 g TFB kg-1 in control group) over an experimental period of 32 days. Cumulative mortality was 12%-15% in both treatment groups with 1 and 2 g TFB kg-1 and reached 27% in the group with 4 g TFB kg-1. Only the highest dose (4 g TFB kg-1) negatively affected feed intake, growth and respiration of the polychaetes while food conversion efficiency was not affected. At the end of the experiment, the concentrations of TFB in the Capitella sp. were high, in the range of 9.24-10.32 µg g-1 for the three treatment groups. It was suggested that a maximum level of absorption rate was reached, also for the lowest dose. High concentrations of TFB in the Capitella sp. might pose a risk to crustaceans that forage for polychaetes in the vicinity of fish farms. We conclude that the effects of TFB on Capitella sp. may therefore primarily be to the predators rather than the Capitella sp.

Transcriptome analysis of the Yesso scallop, Patinopecten yessoensis gills in response to water temperature fluctuations.

Water temperature fluctuations are considered to be a major factor affecting the immune functions and metabolic processes of scallops. To better understand the immune defense mechanisms of Yesso scallop, Patinopecten yessoensis following exposure to water temperature fluctuations, transcriptomic profiles in the gills from high-frequency fluctuations (HF_G), low-frequency fluctuations (LF_G), and no fluctuations (NF_G) groups were obtained using HiSeq™ 2500 (Illumina). For HF_G, scallops were transferred directly between 18 and 8 °C every 4 h and for 10 fluctuations, while scallops in LF_G were transferred between 18 and 13 °C every 12 h, for a total of 4 fluctuations. A total of 442,922,590 clean reads were generated in 9 libraries and then assembled into 210,780 unigenes with an average length of 705 bp and an N50 of 1253 bp. Based on sequence similarity, 54,529 unigenes (25.87%) were annotated in at least one database. Comparative analysis revealed that 696 unigenes differentially expressed in temperature stressed groups compared with the control, including 229 unigenes between HF_G and NF_G, and 548 unigenes between LF_G and NF_G, respectively. Additionally, among these differentially expressed genes (DEGs), there were 41 immune-related unigenes and 16 protein metabolism-related unigenes. These results provide fundamental information on the molecular defense mechanisms in the Yesso scallop gills after exposure to water temperature fluctuations.

A semi-supervised segmentation method for microscopic hyperspectral pathological images based on multi-consistency learning.

Pathological images are considered the gold standard for clinical diagnosis and cancer grading. Automatic segmentation of pathological images is a fundamental and crucial step in constructing powerful computer-aided diagnostic systems. Medical microscopic hyperspectral pathological images can provide additional spectral information, further distinguishing different chemical components of biological tissues, offering new insights for accurate segmentation of pathological images. However, hyperspectral pathological images have higher resolution and larger area, and their annotation requires more time and clinical experience. The lack of precise annotations limits the progress of research in pathological image segmentation. In this paper, we propose a novel semi-supervised segmentation method for microscopic hyperspectral pathological images based on multi-consistency learning (MCL-Net), which combines consistency regularization methods with pseudo-labeling techniques. The MCL-Net architecture employs a shared encoder and multiple independent decoders. We introduce a Soft-Hard pseudo-label generation strategy in MCL-Net to generate pseudo-labels that are closer to real labels for pathological images. Furthermore, we propose a multi-consistency learning strategy, treating pseudo-labels generated by the Soft-Hard process as real labels, by promoting consistency between predictions of different decoders, enabling the model to learn more sample features. Extensive experiments in this paper demonstrate the effectiveness of the proposed method, providing new insights for the segmentation of microscopic hyperspectral tissue pathology images.

Mapping Spatiotemporal Heterogeneity in Multifocal Breast Tumor Progression by Noninvasive Ultrasound Elastography-Guided Mass Spectrometry Imaging Strategy.

Spatiotemporal heterogeneity of tumors provides an escape mechanism for breast cancer cells, which can obstruct the investigation of tumor progression. While molecular profiling obtained from mass spectrometry imaging (MSI) is rich in biochemical information, it lacks the capacity for in vivo analysis. Ultrasound diagnosis has a high diagnostic accuracy but low chemical specificity. Here, we describe a noninvasive ultrasound elastography (UE)-guided MSI strategy (UEg-MSI) that integrates physical and biochemical characteristics of tumors acquired from both in vivo and in vitro imaging. Using UEg-MSI, both elasticity histopathology metabolism "fingerprints" and reciprocal crosstalk are revealed, indicating the intact, multifocal spatiotemporal heterogeneity of spontaneous tumorigenesis of the breast from early, middle, and late stages. Our results demonstrate a gradual increase in malignant degree of primary focus in cervical and thoracic mammary glands. This progression is characterized by increased stiffness according to elasticity scores, histopathological changes from hyperplasia to increased nests of neoplastic cells and necrotic areas, and regional metabolic heterogeneity and reprogramming at the spatiotemporal level. De novo fatty acid (FA) synthesis focused on independent (such as ω-9 FAs) and dependent (such as ω-6 FAs) dietary FA intake in the core cancerous nest areas in the middle and late stages of tumor or in the peripheral microareas in the early stage of the tumor. SM-Cer signaling pathway and GPs biosynthesis and degradation, as well as glycerophosphoinositol intensity, changed in multiple characteristic microareas. The UEg-MSI strategy holds the potential to expand MSI applications and enhance ultrasound-mediated cancer diagnosis. It offers new insight into early cancer discovery and the occurrence of metastasis.

Enhancing photothermal therapy of tumors with image-guided thermal control of gene-expressing bacteria.

Purpose: Bacteria-mediated tumor therapy has showed promising potential for cancer therapy. However, the efficacy of bacterial monotherapy treatment which can express and release therapeutic proteins in tumors has been found to be unsatisfactory. To date, synergistic therapy has emerged as a promising approach to achieve stronger therapeutic outcomes compared to bacterial monotherapy. It is a challenge to visualize these tumor-homing bacteria in vivo and guide them to express and release in situ therapeutic proteins. Procedure: We have developed a kind of engineered bacteria (named CGB@ICG) genetically incorporating acoustic reporter proteins and thermo-inducible ClyA expression gene circuit and chemically modified with indocyanine green on the bacterial surface. The presence of acoustic reporter proteins and indocyanine green facilitates the visualization of CGB@ICG via contrast-enhanced ultrasound imaging and optical imaging, making it possible to guide the sound wave or laser to irradiate precisely these bacteria for inducing the expression of ClyA protein via acoustic- or photothermal effects. The expression and secretion of ClyA protein in the tumor, combined with photothermal therapy, greatly enhanced the anti-tumor efficacy of the engineered bacteria and improved their biosafety. Results: We successfully performed multimodal imaging of CGB@ICG in vivo resulting in remoting control the expression of ClyA protein in tumor. In vivo experiments showed that bacteria-mediated therapy combined photothermal therapy exhibited a rapid decrease in tumor volume compared to other groups, while the tumor volume of the combination therapy group continued to decrease and even achieved complete healing. Thus, combination therapy not only reduced the rate of tumor growth but also prevented the proliferation of tumor cells for an extended period. Conclusion: Our study demonstrated that CGB@ICG serves as an efficacious imaging agent and delivery vector to combine engineered bacteria with photothermal therapy, holding great promise for tumor treatment.

Identification of a LPS-induced TNF-α factor (LITAF) from mollusk Solen grandis and its expression pattern towards PAMPs stimulation.

Lipopolysaccharide-induced TNF-α factor (LITAF) is one of the most important transcription factors mediating TNF-α transcription. In the present study, a LITAF gene (designated as SgLITAF) was identified from razor clams Solen grandis. The full-length cDNA of SgLITAF was of 1476 bp, encoding a polypeptide of 130 amino acids showed high similarity to other known LITAFs. SgLITAF encoded a LITAF domain and the Zn(2+)-binding motifs in the domain were well conserved. The mRNA transcripts of SgLITAF were detected in all tested tissues of healthy razor clams, including mantle, gill, gonad, hemocytes, muscle and hepatopancreas, and with the highest expression level in hepatopancreas. The expression level of SgLITAF in hemocytes was significantly up-regulated (P < 0.01) after razor clams were stimulated by LPS or ß-1, 3-glucan, but no obvious fluctuation of SgLITAF mRNA expression was observed after PGN stimulation. All the results indicated that there might be a LITAF-regulated TNF-α signaling pathway existing in S. grandis, which involved in the immune response not only against gram-negative bacteria but also towards fungi.

A food-web model as a tool for the ecosystem-level management of bivalves in an Atlantic coastal lagoon.

The Ria de Aveiro is an important coastal lagoon for wildlife in Portugal, where the production of bivalves reaches approximately 2700 tons annually. However, the illegal overfishing of bivalves is frequent in this lagoon, which causes critical changes in the ecosystem. In this study, using a developed food-web model (Ecopath model), the ecological carrying capacity (ECC) and maximum sustained yield (MSY) of the Manila clam, Ruditapes philippinarum were estimated, and the effects of further increases in clam biomass on other species were investigated. The results showed that 1) the current biomass and legal catch of R. philippinarum do not yet exceed the ECC (172.40 tons km-2) or the MSY (86.20 tons km-2 year-1) in Ria de Aveiro; 2) the harvested Manila clams of the MSY represent removing from the ecosystem âˆ¼ 581 tons carbon (C) and ∼83 tons nitrogen (N) annually, with substantial ecological and economic implications; and 3) a further increase in the biomass levels of this species may cause the ecotrophic efficiency of other groups to become unrealistic, potentially leading to decreases in ecosystem transfer efficiency, biodiversity and health. The results here are expected to guide the sustainable development and management of bivalve aquaculture in Ria de Aveiro and the protection of the local environment.

Imaging biomarker for quantitative analysis of endometrial injury based on optical coherence tomography/ultrasound integrated imaging mode.

Precise evaluation of endometrial injury is significant to clinical decision-making in gynecological disease and assisted reproductive technology. However, there is a lack of assessment methods for endometrium in vivo. In this research, we intend to develop quantitative imaging markers with optical coherence tomography (OCT)/ultrasound (US) integrated imaging system through intrauterine endoscopic imaging. OCT/US integrated imaging system was established as our previous research reported. The endometrial injury model was established and after treatment, OCT/US integrated imaging and uterus biopsy was performed to evaluate the endometrial thickness, number of superficial fold, and intrauterine area. According to the results, three quantitative indexes acquired from OCT/US image and HE staining have the same trend and have a strong relationship with the severity of the endometrial injury. Accordingly, we developed three imaging markers for quantitative analysis of endometrial injury in vivo, which provided a precise mode for endometrium evaluation in clinical practice.

A Nomogram Based on Conventional and Contrast-Enhanced Ultrasound for Pre-operative Prediction of Nipple-Areola Complex Involvement in Breast Cancer: A Prospective Study.

OBJECTIVE: Accurately predicting nipple-areola complex (NAC) involvement in breast cancer is essential for identifying eligible patients for a nipple-sparing mastectomy. This study was aimed at developing a pre-operative nomogram for NAC involvement in breast cancer using conventional ultrasound (US) and contrast-enhanced ultrasound (CEUS). METHODS: All patients with primary breast cancer confirmed by pre-operative biopsy underwent US and CEUS examinations. Post-operative pathology was used as the gold standard in assessing NAC involvement. Lasso regression was used to select the predictors most associated with NAC involvement. A nomogram was constructed to calculate the diagnostic efficacy. The data were internally verified with 500 bootstrapped replications, and a calibration curve was generated to validate the predictive capability. RESULTS: Seventy-six patients with primary breast cancer were included in this study, which included 16 patients (21.1%) with NAC involvement and 60 patients (78.9%) without NAC involvement. Among the 23 features of US and CEUS, Lasso regression selected one US feature and two CEUS features, namely, ductal echo extending from the lesion, ductal enhancement extending to the nipple and focal nipple enhancement. A nomogram was constructed, and the results revealed that the area under the curve, sensitivity, specificity and accuracy were 0.891, 81.3%, 86.7% and 85.5%, respectively. The calibration curve exhibited good consistency between the predicted probability and the actual probability. CONCLUSION: The nomogram developed based on US and CEUS had good performance in predicting NAC involvement in breast cancer before surgery, which may facilitate the selection of suitable patients for NAC preservation with greater oncological safety.

Cloning and transcriptional analysis of two sialic acid-binding lectins (SABLs) from razor clam Solen grandis.

Sialic acid-binding lectin (SABL) plays crucial role in both innate and adaptive immune responses benefiting from its predominant affinity toward glycan. In the present study, two SABLs from razor clam Solen grandis (designated as SgSABL-1 and SgSABL-2) were identified, and their expression patterns, both in tissues and towards microorganism glycan stimulation, were then characterized. The cDNA of SgSABL-1 and SgSABL-2 was 988 and 1281 bp, containing an open reading frame (ORF) of 744 and 570 bp, respectively, and deduced amino acid sequences showed high similarity to other invertebrates SABLs. Both SgSABL-1 and SgSABL-2 encoded a C1q domain. SgSABL-1 and SgSABL-2 were found to be constitutively expressed in a wide range of tissues with different levels, including mantle, gill, gonad, hemocyte, muscle, and hepatopancreas, and both of them were highly expressed in hepatopancreas. SgSABL-1 and SgSABL-2 could be significantly induced after razor clams were stimulated by acetylated subunits-containing glycan LPS and PGN, suggesting the two SgSABLs might perform potential function of glycan recognition. In addition, SgSABL-2 could also be induced by ß-1,3-glucan. All these results indicated that SgSABL-1 and SgSABL-2 might be involved in the immune response against microbe infection and contributed to the pathogens recognition.

Identification and transcriptional analysis of two types of lectins (SgCTL-1 and SgGal-1) from mollusk Solen grandis.

C-type lectin and galectin are two types of animal carbohydrate-binding proteins which serve as pathogen recognition molecules and play crucial roles in the innate immunity of invertebrates. In the present study, a C-type lectin (designated as SgCTL-1) and galectin (designated as SgGal-1) were identified from mollusk Solen grandis, and their expression patterns, both in tissues and toward three pathogen-associated molecular patterns (PAMPs) stimulation were characterized. The full-length cDNA of SgCTL-1 and SgGal-1 was 1280 and 1466 bp, containing an open reading frame (ORF) of 519 and 1218 bp, respectively. Their deduced amino acid sequences showed high similarity to other members of C-type lectin and galectin superfamily, respectively. SgCTL-1 encoded a single carbohydrate-recognition domain (CRD), and the motif of Ca(2+)-binding site 2 was EPN (Glu(135)-Pro(136)-Asn(137)). While SgGal-1 encoded two CRDs, and the amino acid residues constituted the carbohydrate-binding motifs were well conserved in CRD1 but partially conserved in CRD2. Although SgCTL-1 and SgGal-1 exhibited different tissue expression pattern, they were both constitutively expressed in all tested tissues, including hemocytes, gonad, mantle, muscle, gill and hepatopancreas, and they were both highly expressed in hepatopancreas and gill. Furthermore, the mRNA expression of two lectins in hemocytes was significantly (P < 0.01) up-regulated with different levels after S. grandis were stimulated by lipopolysaccharide (LPS), peptidoglycan (PGN) or ß-1,3-glucan. Our results suggested that SgCTL-1 and SgGal-1 from razor clam were two novel members of animal lectins, and they might function as pattern recognition receptors (PRRs) taking part in the process of pathogen recognition.

Two C-type lectins from shrimp Litopenaeus vannamei that might be involved in immune response against bacteria and virus.

C-type lectins play crucial roles in innate immunity to recognize and eliminate pathogens efficiently. In the present study, two C-type lectins from shrimp Litopenaeus vannamei (designated as LvLectin-1 and LvLectin-2) were identified, and their expression patterns, both in tissues and toward pathogen stimulation, were then characterized. The full-length cDNA of LvLectin-1 and LvLectin-2 was 567 and 625 bp, containing an open reading frame (ORF) of 471 and 489 bp, respectively, and deduced amino acid sequences showed high similarity to other members of C-type lectin superfamily. Both two C-type lectins encoded a single carbohydrate-recognition domain (CRD). The motif of Ca(2+) binding site 2 in CRD, which determined carbohydrate-binding specificity, was QPN (Gln(122)-Pro(123)-Asn(124)) in LvLectin-1, but QPD (Gln(128)-Pro(129)-Asp(130)) in LvLectin-2. Two C-type lectins exhibited similar tissue expression pattern, for their mRNA were both constitutively expressed in all tested tissues, including hepatopancreas, muscle, gill, hemocytes, gonad and heart, furthermore they were both mostly expressed in hepatopancreas, though the expression level of LvLectin-2 was much higher than LvLectin-1. The expression level of two C-type lectins mRNA in hemocytes varied greatly after the challenge of Listonella anguillarum or WSSV. After L. anguillarum challenge, the expression of both C-type lectins were significantly (P<0.01) up-regulated compared with blank group, and LvLectin-1 exhibited higher level than LvLectin-2; while after the stimulation of WSSV, the expression of LvLectin-2 was significantly up-regulated at 6 h (P<0.01) and 12 h (P<0.05), but the expression level of LvLectin-1 down-regulated significantly (P<0.01) to 0.4-fold at 6 and 12 h post-stimulation. The results indicated that the two C-type lectins might be involved in immune response toward pathogen infection, and they might perform different recognition specificity toward bacteria or virus.

Molecular cloning and mRNA expression of two peptidoglycan recognition protein (PGRP) genes from mollusk Solen grandis.

Peptidoglycan recognition proteins (PGRPs) play crucial role in innate immunity for both invertebrates and vertebrates, owing to their prominent ability in detecting and eliminating invading bacteria. In the present study, two short PGRPs from mollusk Solen grandis (designated as SgPGRP-S1 and SgPGRP-S2) were identified, and their expression patterns, both in tissues and toward three PAMPs stimulation, were then characterized. The full-length cDNA of SgPGRP-S1 and SgPGRP-S2 was 1672 and 1285 bp, containing an open reading frame (ORF) of 813 and 426 bp, respectively, and deduced amino acid sequences showed high similarity to other members of PGRP superfamily. Both SgPGRP-S1 and SgPGRP-S2 encoded a PGRP domain. The motif of Zn(2+) binding sites and amidase catalytic sites were well conserved in SgPGRP-S1, but partially conserved in SgPGRP-S2. The two PGRPs exhibited different tissue expression pattern. SgPGRP-S1 was highly expressed in muscle and hepatopancreas, while SgPGRP-S2 was highly in gill and mantle. The mRNA expression of SgPGRP-S1 could be induced acutely by stimulation of PGN, and also moderately by ß-1,3-glucan, but not by LPS, while expression of SgPGRP-S2 was significantly up-regulated (P < 0.01) when S. grandis was stimulated by all the three PAMPs, though the expression levels were relatively lower than SgPGRP-S1. Our results suggested SgPGRP-S1 and SgPGRP-S2 could serve as pattern recognition receptors (PRRs) involved in the immune recognition of S. grandis, and they might perform different functions in the immune defense against invaders.

A sigma-class glutathione S-transferase from Solen grandis that responded to microorganism glycan and organic contaminants.

Glutathione S-transferases (GSTs) are a superfamily of antioxidant enzymes, which play crucial roles in detoxification and protection of tissues from oxidative damage caused by reactive oxygen species (ROS). In this study, a sigma-class GST was identified from razor clam Solen grandis (designated as SgGST-S1), and its expression patterns, both in tissues and toward microorganism glycan as well as organic contaminants stimulation, were then characterized. The full-length cDNA of SgGST-S1 was of 1291 bp, containing a 5' untranslated region (UTR) of 27 bp, and a 3' UTR of 619 bp with a poly (A) tail. The open reading frame (ORF) was of 645 bp, encoding a polypeptide of 214 amino acids with the predicted molecular weight of 24.8 kDa, which shared 47% identity with GST from Ruditapes philippinarum. The analysis of conserved domain and phylogenetic relationship strongly suggested that SgGST-S1 was a member of sigma-class GST. The mRNA of SgGST-S1 was constitutively expressed in all tested tissues of healthy razor clam, including mantle, gill, gonad, hemocytes, muscle, and hepatopancreas, and it was highly expressed in hepatopancreas. The mRNA expression of SgGST-S1 in hemocytes was significantly up-regulated (P < 0.01) after razor clam was stimulated by peptidoglycan (PGN) or ß-1, 3-glucan, but not LPS. In addition, the SgGST-S1 transcript level was also significantly (P < 0.01) induced by exposure of benzo[a]pyrene (B[a]P) or Polybrominated Diphenyl Ethers (PBDE). All the results indicated that SgGST-S1 might serve as an antioxidant enzyme involving in the detoxification cause by both microorganism glycan and organic contaminants.

In vivo evaluation of endometrium through dual-modality intrauterine endoscopy.

Female infertilities are highly associated with poor endometrial receptivity. A receptive endometrium is generally characterized by the normal uterine cavity, intact endometrial surface, appropriate endometrial thickness, and echo pattern. Acquiring comprehensive structural information is the prerequisite of endometrium assessment, which is beyond the ability of any single-modality imaging method. In this paper, we introduce a custom-made intrauterine dual-modality (OCT/ultrasound) endoscopic imaging system and achieve in vivo imaging of rabbit uteri, for the first time to our knowledge. The endometrial features of the injured uteri in both ultrasonic and OCT images are consistent with their corresponding pathology. The quantified parameters, including uterine thickness and endometrial surface roughness, show the correlation with the endometrial injury degree but with poor performance for injury classification. The combination of these parameters was proved to assess the degrees of endometrial injury more accurately. Our work shows the potential of the dual-modality system to be translated into a clinical tool, providing multiple quantitative imaging information and helping evaluate the endometrial receptivity and diagnose infertility.

Microplastics influence physiological processes, growth and reproduction in the Manila clam, Ruditapes philippinarum.

Microplastics (<5 mm) are widely distributed in marine environments and pose a serious threat to bivalves. Here, the ingestion and accumulation of polystyrene microplastics (PS microplastics, diameters 5 and 10 µm) by the Manila clam, Ruditapes philippinarum, and their impacts on physiological processes, growth and reproduction were studied. The results showed that both PS microplastics were ingested by the Manila clam and accumulated in their gills, hepatopancreases and intestines. Furthermore, the accumulation of 5 and 10 µm PS microplastics significantly increased the rates of respiration and excretion while significantly decreasing feeding and absorption efficiency (AE), leading to a dramatically reduced amount of energy available for growth (SfG) and ultimately led to slower growth. The dynamic energy budget (DEB) model predicts that PS microplastic exposure for 200 days would cause lower shell/flesh growth rates and reproductive potentiality. Transcriptomic profiles support these results, as carbon and protein metabolism and oxytocin and insulin-related signaling pathways were significantly altered in clams in response to PS microplastics. This study provides evidence that microplastics strongly affect the physiological activities, energy allocation, growth and reproduction of filter-feeding bivalves.