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Accurate diagnosis of the periprosthetic joint infection (PJI) remains a challenge for surgeons. The purpose of this study was to assess the value of albumin to globulin ratio (AGR) and globulin (GLB) for diagnosing PJI. A total of 182 patients undergoing revision after arthroplasty were included and divided into 2 groups, 61 in knee group (PJI: 38; non-PJI: 23) and 121 in hip group (PJI: 26; non-PJI: 95). We used receiver operating characteristic curves to determine the diagnostic value of AGR, GLB, inflammatory markers (erythrocyte sedimentation rate [ESR] and C-reactive protein [CRP]). The receiver operating characteristic curves showed the areas under the curve of AGR, GLB, ESR, and CRP in the knee group were 0.940, 0.928, 0.867, and 0.848, respectively, and they were 0.855, 0.831, 0.886, and 0.912 in the hip group. The optimal predictive cut-off values for AGR in knee and hip groups were 1.375 and 1.295, respectively. The sensitivity and specificity of AGR, respectively, were 94.7% and 87.0% (knee group) and 84.6% and 75.8% (hip group) for diagnosing PJI. The sensitivity of "AGR or ESR" and specificity of "AGR and GLB" in the knee group were 99.6% and 98.9%, respectively. For knee or hip groups, the AGR exhibits good value for the diagnosis of PJI comparable with ESR and CRP. The AGR and GLB, together with CRP and ESR, should be used as the preferred indicators for diagnosing PJI. The "AGR or ESR" and "AGR and GLB" in the knee group have an excellent diagnostic value in sensitivity and specificity, respectively. [ABSTRACT FROM AUTHOR]

Lysozymes play an important role in the innate immune responses with which mollusks respond to bacterial invasion through its lytic activity. In the present study, two c-type lysozymes (designed as VpCLYZ-1 and VpCLYZ-2, respectively) were identified and characterized from the manila clam Venerupis philippinarum . The full-length cDNA of VpCLYZ-1 and VpCLYZ-2 was of 629 and 736 bp, encoding a polypeptide of 156 and153 amino acid residues, respectively. The deduced amino acid sequences of VpCLYZs showed high similarity to other known invertebrate c-type lysozymes. Multiple alignments and phylogenetic relationship strongly suggested that VpCLYZ-1 and VpCLYZ-2 belonged to the c-type lysozyme family. Both VpCLYZ-1 and VpCLYZ-2 transcripts were constitutively expressed in a wide range of tissues with different levels. The VpCLYZ-1 transcript was dominantly expressed in hepatopancreas and hemocytes, while VpCLYZ-2 transcript was mainly expressed in the tissues of hepatopancreas and gills. Both the mRNA expression of VpCLYZ-1 and VpCLYZ-2 was significantly up-regulated at 12 h post Vibrio anguillarum challenge. The recombinant VpCLYZ-1 and VpCLYZ-2 (designed as rVpCLYZ-1 and rVpCLYZ-2) exhibited lytic activity against all tested bacteria, and rVpCLYZ-1 showed higher activities than rVpCLYZ-2 in killing Micrococcus luteus and V. anguillarum . Overall, our results suggested that VpCLYZ-1 and VpCLYZ-2 belonged to the c-type lysozyme family, and played important roles in the immune responses of manila clam, especially in the elimination of pathogens. [ABSTRACT FROM AUTHOR]

An integrative biomarker approach was employed to evaluate the environmental quality of the north coast of Shandong Peninsula along the southern Bohai Sea of China, where pollution is an imminent threat due to rapid urbanization and industrialization. A battery of biomarkers and the metal bioaccumulation in tissues of native oyster Crassostrea gigas were measured under field conditions. Integrative biomarker index (IBR) and metal body burden were calculated to differentiate the pollution status of seven sampling sites. According to our results, Xinzhuang (XZ) site was the most severely contaminated, with the highest IBR value of 3.58, while the lowest IBR value (0.04) was obtained at Penglai (PL). Such an integrated biomarker approach was proved as a useful method for environmental quality assessment in the study area. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) are important mediators of the primary host defense system against microbial invasion. In the present study, we cloned and characterized a member of the invertebrate defensin from the clam Venerupis philippinarum , designated VpDef . Amino acid sequence analysis showed that VpDef was similar to defensins from marine mollusks and ticks. In non-stimulated clams, RT-PCR and immunohistochemical analysis revealed that both VpDef mRNA and the encoding peptide were constitutively expressed in hemocytes and mantles, as well as in other major tissues. VpDef transcripts were significantly induced in hemocytes at different time intervals post Vibrio anguillarum infection. The recombinant VpDef (rVpDef) showed the highest activity against Gram-positive bacteria Micrococcus luteus and less effective to Gram-negative bacteria. In addition, incubation of rVpDef with M. luteus at 1 × and 3 × MIC could induce an obvious decrease of the membrane potential and notable changes of membrane permeability in a dose-dependent manner. Membrane integrity and bacterial viability analysis also revealed that rVpDef increased the membrane permeability of M. luteus and then resulted in cell death at 2 × and 10 × MIC. Overall, these results suggest that VpDef has an important function in host defense against invasive pathogens, probably killing microbes by inducing membrane lesions. [ABSTRACT FROM AUTHOR]

The 14-3-3 proteins play important roles in various cellular processes by binding to different ligands, but little is known about these proteins in mollusks. In this study, two 14-3-3 cDNAs were identified from the Pacific abalone Haliotis discus hannai (designated 14-3-3ζ and 14-3-3ε), possessing 59.40% identity with each other. Both genes were predominantly expressed in the gills of unchallenged abalones, and their mRNA signals could also be detected in several other tissues, including the mantle, hepatopancreas and ovary. However, after Vibrio harveyi challenge, hemocytes were induced significantly (p < 0.01). Meanwhile, phagocytosis was inhibited, but apoptosis, reactive oxygen species formation, and caspase 3 expression were significantly induced (p < 0.01), and they were all suppressed with 14-3-3ζ knockdown (p < 0.01). The differences were that silencing 14-3-3ε reverted the decline in the phagocytic rate derived from bacterial infection, while ROS formation was not influenced significantly. In addition, the expression levels of several antimicrobial peptide and proinflammatory cytokine genes were also decreased with the silencing of 14-3-3 genes. However, with the knockdown of 14-3-3ζ , the expression of 14-3-3ε was further significantly increased (p < 0.01), and vice versa. Overall, our results suggested that 14-3-3ζ and 14-3-3ε should play important roles in innate immunity against V. harveyi infection. • Two abalone 14-3-3 cDNAs were identified. • They regulated cellular immunity. • They might exist a compensatory relationship. [ABSTRACT FROM AUTHOR]

Ocean acidification (OA) and pathogenic diseases pose a considerable threat to key species of marine ecosystem. However, few studies have investigated the combined impact of reduced seawater pH and pathogen challenge on the immune responses of marine invertebrates. In this study, Pacific oysters, Crassostrea gigas , were exposed to OA (~2000 ppm) for 28 days and then challenged with Vibrio splendidus for another 72 h. Hemocyte parameters showed that V. splendidus infection exacerbated the impaired oyster immune responses under OA exposure. An iTRAQ-based quantitative proteomic analysis revealed that C. gigas responded differently to OA stress and V. splendidus challenge, alone or in combination. Generally, OA appears to act via a generalized stress response by causing oxidative stress, which could lead to cellular injury and cause disruption to the cytoskeleton, protein turnover, immune responses and energy metabolism. V. splendidus challenge in oysters could suppress the immune system directly and lead to a disturbed cytoskeleton structure, increased protein turnover and energy metabolism suppression, without causing oxidative stress. The combined OA- and V. splendidus -treated oysters ultimately presented a similar, but stronger proteomic response pattern compared with OA treatment alone. Overall, the impaired oyster immune functions caused by OA exposure may have increased the risk of V. splendidus infection. These results have important implications for the impact of OA on disease outbreaks in marine invertebrates, which would have significant economic and ecological repercussions. [ABSTRACT FROM AUTHOR]

The pervasive presence of nanoplastics (NPs) in marine environments poses a threat to marine organisms. Gills, as the organ in direct contact with the environment in marine invertebrates, maybe the first to accumulate NPs. To date, the toxic effects of NPs on the gills of marine invertebrates are still largely unknown. In this study, the response of multiple biomarkers (i.e., total antioxidant capacity, the activity of acetylcholine, ion content and transport enzyme, metabolic enzymes, and lipids content) in mussels Mytilus galloprovincialis exposed to polystyrene nanoplastics (PS-NPs) for 7 days were evaluated. Significant inductions of total antioxidant capacity (T-AOC) and inhibition of acetylcholine (AChE) activity were detected after 7 days of PS-NPs exposure. PS-NPs also triggered significant alteration in ion content (Na+ and K+) and suppressed the activities of the ion transport enzyme (Na+/K+-ATPase). Moreover, we found the activity of metabolic enzymes (succinate dehydrogenase and pyruvate kinase) and lipids content (triacylglycerol and cholesterol) were significantly altered, suggesting the interference of PS-NPs on energy metabolism and lipid metabolism. This investigation provides substantial information to understand the physical responses of invertebrate gills to PS-NPs exposure. Given the crucial ecological roles of invertebrates, the presence of PS-NPs in the marine environment may have far-reaching impacts on population abundance, biodiversity, and stability of the marine ecosystem. • Exposure to PS-NPs induces redox unbalance in mussels. • PS-NPs induces decreased acetylcholinesterase activity. • PS-NPs induces ion permeation changes in mussels. • PS-NPs induces energy and lipid metabolism imbalance in mussels. [ABSTRACT FROM AUTHOR]

Highlights • Cu concentration was elevated in Cu-exposed oysters under OA. • Seawater acidification could exacerbate the toxicity caused by Cu in oysters. • Disturbed physiological functions were observed in oysters under Cu and/or OA. • IBR results suggested that co-exposure was the most stressful condition. Abstract Ocean acidification (OA) has been found to increase the release of free Cu2+ in seawater. However, only a handful of studies have investigated the influence of OA on Cu accumulation and cellular toxicity in bivalve species. In this study, Pacific oysters, Crassostrea gigas, were exposed to 25 μg/L Cu2+ at three pH levels (8.1, 7.8 and 7.6) for 14 and 28 days. Physiological and histopathological parameters [(clearance rate (CR), respiration rate (RR), histopathological damage and condition index (CI)), oxidative stress and neurotoxicity biomarkers [superoxide dismutase (SOD) and glutathione transferase (GST) activities, lipid peroxidation (LPO) and acetylcholinesterase (AChE) activity], combined with glycolytic enzyme activities [pyruvate kinase (PK) and hexokinase (HK)] were investigated in C. gigas. The bioconcentration of Cu was increased in soft tissues of Cu-exposed oysters under OA. Our results suggest that both OA and Cu could lead to physiological disturbance, oxidative stress, cellular damage, disturbance in energy metabolism and neurotoxicity in oysters. The inhibited CR, increased glycolytic enzymes activities and decreased CI suggested that the energy metabolism strategy adopted by oysters was not sustainable in the long term. Furthermore, integrated biomarker response (IBR) results found that OA and Cu exposure lead to severe stress to oysters, and co-exposure was the most stressful condition. Results from this study highlight the need to include OA in future environmental assessments of pollutants and hazardous materials to better elucidate the risks of those environmental perturbations. [ABSTRACT FROM AUTHOR]

Mounting evidence has demonstrated the combined effects of ocean acidification (OA) and other environmental stressors on marine organisms. Although metal pollution is widely distributed in coasts and estuaries, the combined effects of OA and metal pollution have received little attention until recent years. In this study, the accumulation and subcellular distribution of cadmium (Cd) and the physiological responses of the oyster Crassostrea gigas were investigated after 31 days of exposure to OA and Cd, either alone or in combination. Increased Cd accumulation was found both in gills (about 57% increase at pH 7.8, 22% increase at pH 7.6) and digestive glands (about 38% increase at pH 7.8, 22% increase at pH 7.6) of C . gigas under elevated p CO 2 exposure. Although a similar total Cd accumulation pattern was seen in oyster gills and digestive glands, a higher partition of Cd in the BIM (biologically inactive metal) fractions of gills (about 60%) was found in Cd-exposed treatments compared to the digestive glands (about 45%), which might correspond to the generally lower toxicity in gills. Moreover, synergetic effects of Cd and OA on the oxidative stresses, histopathological damage, and apoptosis of exposed oysters were observed in this study, which might be explained by significant interactions of these two factors on increased generation of ROS. These findings demonstrated that OA could aggravate the toxicity of metals in marine organisms, with significant implications for coastal benthic ecosystems regarding the widespread metal contamination and the concurrent increase of acidified seawater. [ABSTRACT FROM AUTHOR]

Highlights • Inorganic arsenic was apt to accumulate in digestive glands of R. philippinarum. • AsB and DMA accounted for the majority of total arsenic in R. philippinarum. • MTLPs and MRG fractions were the main subcellular distribution site of arsenic. • GSTΩ genes could enhance the arsenite tolerance of E. coli AW3110 (DE3) (Δ arsRBC). • GSTΩs may play an important role in the process of arsenic detoxification. Abstract The major hazard of arsenic in living organisms is increasingly being recognized. Marine mollusks are apt to accumulate high levels of arsenic, but knowledge related to arsenic detoxification in marine mollusks is still less than sufficient. In this study, arsenic bioaccumulation as well as the role of glutathione S-transferase omega (GSTΩ) in the process of detoxification were investigated in the Ruditapes philippinarum clam after waterborne exposure to As(III) or As(V) for 30 days. The results showed that the gills accumulated significantly higher arsenic levels than the digestive glands. Arsenobetaine (AsB) and dimethylarsenate (DMA) accounted for most of the arsenic found, and monomethylarsonate (MMA) can be quickly metabolized. A subcellular distribution analysis showed that most arsenic was in biologically detoxified metal fractions (including metal-rich granules and metallothionein-like proteins), indicating their important roles in protecting cells from arsenic toxicity. The relative mRNA expressions of two genes encoding GSTΩ were up-regulated after arsenic exposure, and the transcriptional responses were more sensitive to As(III) than As(V). The recombinant GSTΩs exhibited high activities at optimal conditions, especially at 37 °C and pH 4–5, with an As(V) concentration of 60 mM. Furthermore, the genes encoding GSTΩ significantly enhance the arsenite tolerance but not the arsenate tolerance of E. coli AW3110 (DE3) (Δ arsRBC). It can be deduced from these results that GSTΩs play an important role in arsenic detoxification in R. philippinarum. [ABSTRACT FROM AUTHOR]

Ocean acidification (OA) and antibiotic pollution pose severe threats to the fitness of keystone species in marine ecosystems. However, the combined effects of OA and antibiotic pollution on the intestinal microbiota of marine organisms are still not well known. In this study, we exposed the herbivorous gastropod Trochus niloticus , a keystone species to maintains the stability of coral reef ecosystems, to acidic seawater (pH 7.6) and/or sulfamethoxazole (SMX, 100 ng/L, 1000 ng/L) for 28 days and determined their impacts on (1) the accumulation of SMX in the intestine of T. niloticus ; (2) the characteristics of the intestinal microbiota in T. niloticus ; (3) the relative abundances of sulfonamide resistance genes (i.e., sul1 and sul2) and intI1 in the intestinal microbiota of T. niloticus. Our results show that OA exposure leads to dramatic microbiota dysbiosis in the intestine of T. niloticus , including changes in bacterial community diversity and structure, decreased abundances of dominant species, existences of characteristic taxa, and altered functional predictions. In addition, SMX exposure at environmentally relevant concentrations had little effect on the intestinal microbiota of T. niloticus , whether in isolation or in combination with OA. However, after exposure to the higher SMX concentration (1000 ng/L), the accumulation of SMX in the intestine of T. niloticus could induce an increase in the copies of sul2 in the intestinal microbiota. These results suggest that the intestinal health of T. niloticus might be affected by OA and SMX, which might lead to fitness loss of the keystone species in coral reef ecosystems. [Display omitted] • Exposure to OA leads to the microbiota dysbiosis in the intestine of T. niloticus. • Exposure to SMX barely affected the intestinal microbiota of T. niloticus. • Exposure to SMX accelerated spread of sulfonamide ARGs. [ABSTRACT FROM AUTHOR]

The formation of extracellular traps (ETs) is an important innate immune mechanism that serves to combat different invading pathogens. In this study, zymosan significantly induced the formation of ETs in the hemocytes of Ruditapes philippinarum , and this effect was accompanied by translocation of the mitochondria to the cell surface. Zymosan stimulation clearly induced an increase in intracellular ROS and MPO production and an overexpression of ROS-related genes (PI3K , AKT and HIF). In response to the ROS burst, the mitochondrial membrane potential decreased, and the mitochondrial permeability transition pore opened. Conversely, mitochondrial superoxide inhibitor (Mito-TEMPO) significantly inhibited the formation of ETs, suggesting that mitochondrial ROS were necessary for the formation of ETs. In addition, we found that zymosan-induced ETs showed antibacterial activities against gram-negative and gram-positive bacteria, such as Vibrio anguillarum , Vibrio harveyi , Escherichia coli and Micrococcus luteus. Taken together, these findings elucidated a new antibacterial approach for R. philippinarum and highlighted the role of mitochondria in the formation of zymosan-induced ETs. • Zymosan induced extracellular trap (ET) formation in hemocytes of Ruditapes philippinarum. • Zymosan increased the production of ROS and MPO in ETosis. • Mitochondrial ROS are essential in the formation of zymosan-triggered ETs. • Zymosan-induced ETs exhibited antibacterial activities against both gram-negative and gram-positive bacteria. [ABSTRACT FROM AUTHOR]

Extracellular traps (ETs) have been found to be an important strategy of mammals to immobilize and kill invading microorganisms. In the present study, we observed the formation of ETs in the hemocytes of marine mollusks Ruditapes philippinarum in response to challenge from bacteria Vibrio anguillarum , and examined the potential factors and signaling pathways underling this process. We detected an increase of reactive oxygen species (ROS) and myeloperoxidase (MPO) production during ETosis, accompanied by significantly up-regulated expression of ROS-related and MPO genes. The suppression of ETs structures by nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor (diphenyleneiodonium chloride, DPI) and MPO inhibitor (aminobenzoic acid hydrazide, ABAH) further confirmed the essential roles ROS and MPO played in ETosis. Furthermore, ET production could be inhibited by phosphotidylinsitol-3-kinase (PI3K) inhibitor (LY294002) and extracellular regulated protein kinase (ERK) inhibitor (U0126), suggesting the idea that both the PI3K and ERK pathways were suggested to function during ETosis. In addition, the ETosis process was accompanied by enhancement of glycolysis-related enzymatic activities, e.g., pyruvate kinase (PK) and hexokinase (HK), and over-expression of the glycolysis-related genes, e.g., PK, HK and glucose transport protein (GLUT), indicating high involvement of glycolysis in the ETosis process. Furthermore, our scanning electron microscopy (SEM) observation and antibacterial activities test successfully showed the patterns how clam ETs entrapped and killed the invading V. anguillarum. Taken together, our results revealed that ETosis with bactericidal effect increased ROS, MPO and glycolysis level and carried out in a ROS-, MPO-, PI3K-ERK-dependent manner. • Vibrio anguillarum induced extracellular trap (ET) formation in clam hemocytes. • The production of reactive oxygen species (ROS) and myeloperoxidase (MPO) was significantly increased in clam ETosis. • Glycolysis was involved during the production of V. anguillarum -induced ETs. • ETosis was carried out in a ROS-, MPO-, PI3K-ERK-dependent manner. [ABSTRACT FROM AUTHOR]