In vivo CD8+ T cell CRISPR screening reveals control by Fli1 in infection and cancer.

Improving effector activity of antigen-specific T cells is a major goal in cancer immunotherapy. Despite the identification of several effector T cell (TEFF)-driving transcription factors (TFs), the transcriptional coordination of TEFF biology remains poorly understood. We developed an in vivo T cell CRISPR screening platform and identified a key mechanism restraining TEFF biology through the ETS family TF, Fli1. Genetic deletion of Fli1 enhanced TEFF responses without compromising memory or exhaustion precursors. Fli1 restrained TEFF lineage differentiation by binding to cis-regulatory elements of effector-associated genes. Loss of Fli1 increased chromatin accessibility at ETS:RUNX motifs, allowing more efficient Runx3-driven TEFF biology. CD8+ T cells lacking Fli1 provided substantially better protection against multiple infections and tumors. These data indicate that Fli1 safeguards the developing CD8+ T cell transcriptional landscape from excessive ETS:RUNX-driven TEFF cell differentiation. Moreover, genetic deletion of Fli1 improves TEFF differentiation and protective immunity in infections and cancer.

ZMYND8-regulated IRF8 transcription axis is an acute myeloid leukemia dependency.

The transformed state in acute leukemia requires gene regulatory programs involving transcription factors and chromatin modulators. Here, we uncover an IRF8-MEF2D transcriptional circuit as an acute myeloid leukemia (AML)-biased dependency. We discover and characterize the mechanism by which the chromatin "reader" ZMYND8 directly activates IRF8 in parallel with the MYC proto-oncogene through their lineage-specific enhancers. ZMYND8 is essential for AML proliferation in vitro and in vivo and associates with MYC and IRF8 enhancer elements that we define in cell lines and in patient samples. ZMYND8 occupancy at IRF8 and MYC enhancers requires BRD4, a transcription coactivator also necessary for AML proliferation. We show that ZMYND8 binds to the ET domain of BRD4 via its chromatin reader cassette, which in turn is required for proper chromatin occupancy and maintenance of leukemic growth in vivo. Our results rationalize ZMYND8 as a potential therapeutic target for modulating essential transcriptional programs in AML.

DLBCL-associated NOTCH2 mutations escape ubiquitin-dependent degradation and promote chemoresistance.

Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma. Up to 40% of patients with DLBCL display refractory disease or relapse after standard chemotherapy treatment (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone [R-CHOP]), leading to significant morbidity and mortality. The molecular mechanisms of chemoresistance in DLBCL remain incompletely understood. Using a cullin-really interesting new gene (RING) ligase-based CRISPR-Cas9 library, we identify that inactivation of the E3 ubiquitin ligase KLHL6 promotes DLBCL chemoresistance. Furthermore, proteomic approaches helped identify KLHL6 as a novel master regulator of plasma membrane-associated NOTCH2 via proteasome-dependent degradation. In CHOP-resistant DLBCL tumors, mutations of NOTCH2 result in a protein that escapes the mechanism of ubiquitin-dependent proteolysis, leading to protein stabilization and activation of the oncogenic RAS signaling pathway. Targeting CHOP-resistant DLBCL tumors with the phase 3 clinical trial molecules nirogacestat, a selective γ-secretase inhibitor, and ipatasertib, a pan-AKT inhibitor, synergistically promotes DLBCL destruction. These findings establish the rationale for therapeutic strategies aimed at targeting the oncogenic pathway activated in KLHL6- or NOTCH2-mutated DLBCL.

Prognostic Value of a Combined Nomogram Model Integrating 3-Dimensional Deep Learning and Radiomics for Head and Neck Cancer.

OBJECTIVE: The preoperative prediction of the overall survival (OS) status of patients with head and neck cancer (HNC) is significant value for their individualized treatment and prognosis. This study aims to evaluate the impact of adding 3D deep learning features to radiomics models for predicting 5-year OS status. METHODS: Two hundred twenty cases from The Cancer Imaging Archive public dataset were included in this study; 2212 radiomics features and 304 deep features were extracted from each case. The features were selected by univariate analysis and the least absolute shrinkage and selection operator, and then grouped into a radiomics model containing Positron Emission Tomography /Computed Tomography (PET/CT) radiomics features score, a deep model containing deep features score, and a combined model containing PET/CT radiomics features score +3D deep features score. TumorStage model was also constructed using initial patient tumor node metastasis stage to compare the performance of the combined model. A nomogram was constructed to analyze the influence of deep features on the performance of the model. The 10-fold cross-validation of the average area under the receiver operating characteristic curve and calibration curve were used to evaluate performance, and Shapley Additive exPlanations (SHAP) was developed for interpretation. RESULTS: The TumorStage model, radiomics model, deep model, and the combined model achieved areas under the receiver operating characteristic curve of 0.604, 0.851, 0.840, and 0.895 on the train set and 0.571, 0.849, 0.832, and 0.900 on the test set. The combined model showed better performance of predicting the 5-year OS status of HNC patients than the radiomics model and deep model. The combined model was shown to provide a favorable fit in calibration curves and be clinically useful in decision curve analysis. SHAP summary plot and SHAP The SHAP summary plot and SHAP force plot visually interpreted the influence of deep features and radiomics features on the model results. CONCLUSIONS: In predicting 5-year OS status in patients with HNC, 3D deep features could provide richer features for combined model, which showed outperformance compared with the radiomics model and deep model.

The impact of the combat method on radiomics feature compensation and analysis of scanners from different manufacturers.

BACKGROUND: This study investigated whether the Combat compensation method can remove the variability of radiomic features extracted from different scanners, while also examining its impact on the subsequent predictive performance of machine learning models. MATERIALS AND METHODS: 135 CT images of Credence Cartridge Radiomic phantoms were collected and screened from three scanners manufactured by Siemens, Philips, and GE. 100 radiomic features were extracted and 20 radiomic features were screened according to the Lasso regression method. The radiomic features extracted from the rubber and resin-filled regions in the cartridges were labeled into different categories for evaluating the performance of the machine learning model. Radiomics features were divided into three groups based on the different scanner manufacturers. The radiomic features were randomly divided into training and test sets with a ratio of 8:2. Five machine learning models (lasso, logistic regression, random forest, support vector machine, neural network) were employed to evaluate the impact of Combat on radiomic features. The variability among radiomic features were assessed using analysis of variance (ANOVA) and principal component analysis (PCA). Accuracy, precision, recall, and area under the receiver curve (AUC) were used as evaluation metrics for model classification. RESULTS: The principal component and ANOVA analysis results show that the variability of different scanner manufacturers in radiomic features was removed (P˃0.05). After harmonization with the Combat algorithm, the distributions of radiomic features were aligned in terms of location and scale. The performance of machine learning models for classification improved, with the Random Forest model showing the most significant enhancement. The AUC value increased from 0.88 to 0.92. CONCLUSIONS: The Combat algorithm has reduced variability in radiomic features from different scanners. In the phantom CT dataset, it appears that the machine learning model's classification performance may have improved after Combat harmonization. However, further investigation and validation are required to fully comprehend Combat's impact on radiomic features in medical imaging.

YY1 and CTCF orchestrate a 3D chromatin looping switch during early neural lineage commitment.

CTCF is an architectural protein with a critical role in connecting higher-order chromatin folding in pluripotent stem cells. Recent reports have suggested that CTCF binding is more dynamic during development than previously appreciated. Here, we set out to understand the extent to which shifts in genome-wide CTCF occupancy contribute to the 3D reconfiguration of fine-scale chromatin folding during early neural lineage commitment. Unexpectedly, we observe a sharp decrease in CTCF occupancy during the transition from naïve/primed pluripotency to multipotent primary neural progenitor cells (NPCs). Many pluripotency gene-enhancer interactions are anchored by CTCF, and its occupancy is lost in parallel with loop decommissioning during differentiation. Conversely, CTCF binding sites in NPCs are largely preexisting in pluripotent stem cells. Only a small number of CTCF sites arise de novo in NPCs. We identify another zinc finger protein, Yin Yang 1 (YY1), at the base of looping interactions between NPC-specific genes and enhancers. Putative NPC-specific enhancers exhibit strong YY1 signal when engaged in 3D contacts and negligible YY1 signal when not in loops. Moreover, siRNA knockdown of Yy1 specifically disrupts interactions between key NPC enhancers and their target genes. YY1-mediated interactions between NPC regulatory elements are often nested within constitutive loops anchored by CTCF. Together, our results support a model in which YY1 acts as an architectural protein to connect developmentally regulated looping interactions; the location of YY1-mediated interactions may be demarcated in development by a preexisting topological framework created by constitutive CTCF-mediated interactions.

5C-ID: Increased resolution Chromosome-Conformation-Capture-Carbon-Copy with in situ 3C and double alternating primer design.

Mammalian genomes are folded in a hierarchy of compartments, topologically associating domains (TADs), subTADs, and looping interactions. Currently, there is a great need to evaluate the link between chromatin topology and genome function across many biological conditions and genetic perturbations. Hi-C can generate genome-wide maps of looping interactions but is intractable for high-throughput comparison of loops across multiple conditions due to the enormous number of reads (>6 Billion) required per library. Here, we describe 5C-ID, a new version of Chromosome-Conformation-Capture-Carbon-Copy (5C) with restriction digest and ligation performed in the nucleus (in situ Chromosome-Conformation-Capture (3C)) and ligation-mediated amplification performed with a double alternating primer design. We demonstrate that 5C-ID produces higher-resolution 3D genome folding maps with reduced spatial noise using markedly lower cell numbers than canonical 5C. 5C-ID enables the creation of high-resolution, high-coverage maps of chromatin loops in up to a 30 Megabase subset of the genome at a fraction of the cost of Hi-C.

Predation experience underlies the relationship between locomotion capability and survival.

The positive relationship between locomotion performance and survival under predation has long been suggested yet seldom demonstrated with direct evidence. We investigate the effects of predator exposure on locomotion capacity (both fast-start escape and critical swimming performance), survival under predation and the relationships between these factors in juvenile Chinese bream (Parabramis pekinensis). This study aims to test whether there is a positive relationship between the above factors and whether such relationships are context dependent (i.e., with or without 20 d of predator exposure). We found that predator-exposed Chinese bream showed higher rates of survival under predation and improved fast-start swimming performance compared with individuals not exposed to predation. At individual level, no relationship was found between survival and any locomotion performance component in the no-predator group, but mean fast-start swimming speed, maneuverability and responsiveness were all positively related to survival in the predator group after 20 d of exposure. This finding indicates that the recognition of and vigilance for predators achieved through predation experience can be crucial preconditions for prey to employ the fast-start escape response, especially to escape ambush predators. Furthermore, a tradeoff was observed between the critical and fast-start swimming performances in the predator group, but not in the no-predator group, which may have been due to the intensified competition throughout the entire locomotion-support system (e.g., energy, proportions of slow- and fast-twitch muscle fibers) between critical and fast-start swimming because the increased demand for fast-start escape capacity constrains (or compromises) critical swimming performance under the threat of predation.

Deciphering the biodegradation of petroleum hydrocarbons using FTIR spectroscopy: application to a contaminated site.

The chemical composition of groundwater in a petroleum-contaminated site is determined by the present functional groups and these play a vital role in a feasibility remediation technique. Based on the in situ investigation of a contaminated shallow groundwater in an oilfield, Fourier transform infrared (FTIR) spectroscopy associated with chemometric treatments, principal component analysis (PCA), and simple-to-use interactive self-modeling mixture analysis (SIMPLISMA), were used to decipher the biodegradation process by analyzing the conversion of functional groups. Environmental factors that can influence microbial metabolism were also evaluated for a comprehensive explanation. FTIR spectroscopy and PCA results showed that the contamination in the study area can be divided into three parts based on FTIR spectra: (1) regular contamination plume distribution and biodegradation level to fresh oil, (2) moderate biodegradation area, and (3) intensive biodegradation area. FTIR spectra further revealed the present functional groups as aliphatic, aromatic, and polar family compounds. SIMPLISMA was used to discuss the degree of biodegradation along the flow path quantitatively and qualitatively and elucidated that the aliphatic and aromatic compounds were mainly metabolized into polar compounds with nitrogen, sulfur, and oxygen via microbes. During metabolism, microbial indices, such as the Shannon-Weaver, Simpson, and Pielou indices, indicated that microbial diversity did not greatly change; hence, hydrocarbons were constantly consumed to feed dominant microbes. Dissolved oxygen concentrations decreased from 4.58 ± 0.31 mg/L (in monitoring well Z1) to 3.21 ± 0.26 mg/L (in monitoring well Z16) and then became constant in the down-gradient area, demonstrating that aerobic biodegradation was the dominant process at the up-gradient plume. Results were in accordance with the oxidation index, which continuously increased from 0.028 ± 0.013 (in monitoring well Z1) to 0.669 ± 0.047 (in monitoring well Z10), showing that oxygen was consumed along the flow path. Similarly, concentration changes in Fe2+, Mn2+, and SO4 2- proved that the down-gradient area was in reduction condition.

Regulation of Active DNA Demethylation by a Methyl-CpG-Binding Domain Protein in Arabidopsis thaliana.

Active DNA demethylation plays crucial roles in the regulation of gene expression in both plants and animals. In Arabidopsis thaliana, active DNA demethylation is initiated by the ROS1 subfamily of 5-methylcytosine-specific DNA glycosylases via a base excision repair mechanism. Recently, IDM1 and IDM2 were shown to be required for the recruitment of ROS1 to some of its target loci. However, the mechanism(s) by which IDM1 is targeted to specific genomic loci remains to be determined. Affinity purification of IDM1- and IDM2- associating proteins demonstrated that IDM1 and IDM2 copurify together with two novel components, methyl-CpG-binding domain protein 7 (MBD7) and IDM2-like protein 1 (IDL1). IDL1 encodes an α-crystallin domain protein that shows high sequence similarity with IDM2. MBD7 interacts with IDM2 and IDL1 in vitro and in vivo and they form a protein complex associating with IDM1 in vivo. MBD7 directly binds to the target loci and is required for the H3K18 and H3K23 acetylation in planta. MBD7 dysfunction causes DNA hypermethylation and silencing of reporter genes and a subset of endogenous genes. Our results suggest that a histone acetyltransferase complex functions in active DNA demethylation and in suppression of gene silencing at some loci in Arabidopsis.

Digesting or swimming? Integration of the postprandial metabolism, behavior and locomotion in a frequently foraging fish.

Fish that are active foragers usually perform routine activities while digesting their food; thus, their postprandial swimming capacity and related behavior adjustments might be ecologically important. To test whether digestion affect swimming performance and the relationships of digestion with metabolism and behavior in an active forager, we investigated the postprandial metabolic response, spontaneous swimming activities, critical swimming speed (Ucrit), and fast-start escape performance of both fasted and digesting (3h after feeding to satiation) juvenile rose bitterling (Rhodeus ocellatus). Feeding to satiation elicited a 50% increase in the oxygen consumption rate, which peaked at 3h after feeding and returned to the prefeeding state after another 3h. However, approximately 50% and 90% of individuals resumed feeding behavior at 2 and 3h postfeeding, respectively, although the meal size varied substantially. Digestion showed no effect on either steady swimming performance as suggested by the Ucrit or unsteady swimming performance indicated by the maximum linear velocity in fast-start escape movement. However, digesting fish showed more spontaneous activity as indicated by the longer total distance traveled, mainly through an increased percentage of time spent moving (PTM). A further analysis found that fasting individuals with high swimming speed were more inclined to increase their PTM during digestive processes. The present study suggests that as an active forager With a small meal size and hence limited postprandial physiological and morphological changes, the swimming performance of rose bitterling is maintained during digestion, which might be crucial for its active foraging mode and anti-predation strategy.

A comparison of constant acceleration swimming speeds when acceleration rates are different with critical swimming speeds in Chinese bream under two oxygen tensions.

To investigate the effect of acceleration rates on the constant acceleration test speed (U cat) and to compare U cat with the critical swimming speed (U crit) in Chinese bream (Parabramis pekinensis), the U cat test at acceleration rates of 0.05, 0.1, 0.2, 0.4 and 0.8 cm s(-2) and the U crit test in juvenile fish at 20 °C in either normoxia (>90 % saturation oxygen tension) or hypoxia (30 % saturation) were compared. The lactate concentration ([lactate]) of white muscle, liver and plasma and the glycogen concentration ([glycogen]) of white muscle and liver were also measured to identify whether tissue substrate depletion or tissue lactate accumulation correlated with exhaustion. The U cat decreased with the acceleration rate, and there was no significant difference between U crit and U cat at lower acceleration rates. Hypoxia resulted in lower U cat and U crit, and the difference increased with decreased acceleration rates of the U cat test, possibly due to the increased contribution of aerobic components in U crit or U cat at low acceleration rates. Hypoxia elicited a significant decrease in muscle [glycogen] and an increase in muscle and liver [lactate] in resting fish. All post-exercise fish had similar muscle [lactate], suggesting that tissue lactate accumulation may correlate with exercise exhaustion. Unlike hypoxia, exercise induced an increase in muscle [lactate] and a significant increase in plasma [lactate], which were worthy of further investigation. The similar swimming speed and biochemical indicators after exercise in the U crit and U cat groups at low acceleration rates suggested that U cat can be an alternative for the more frequently adopted protocols in U crit in Chinese bream and possibly in other cyprinid fish species.

Predator-driven intra-species variation in locomotion, metabolism and water velocity preference in pale chub (Zacco platypus) along a river.

Fish inhabit environments that vary greatly in terms of predation intensity, and these predation regimes are generally expected to be a major driver of divergent natural selection. To test whether there is predator-driven intra-species variation in the locomotion, metabolism and water velocity preference of pale chub (Zacco platypus) along a river, we measured unsteady and steady swimming and water velocity preference among fish collected from both high- and low-predation habitats in the Wujiang River. We also measured the routine metabolic rate (RMR), maximum metabolic rate (MMR) and cost of transport (COT) and calculated the optimal swimming speed (Uopt). The fish from the high-predation populations showed a shorter response latency, elevated routine metabolism, lower swimming efficiency at low swimming speed and lower water velocity preference compared with those from the low-predation populations. Neither of the kinematic parameters fast-start and critical swimming speed (Ucrit) showed a significant difference between the high- and low-predation populations. The fish from the high-predation populations may improve their predator avoidance capacity primarily through an elevated routine metabolism and shorter response latency to achieve advanced warning and escape, rather than an improved fast-start swimming speed or acceleration. Thus, the cost of this strategy is an elevated RMR, and no trade-off between unsteady and steady swimming performance was observed in the pale chub population under various predation stresses. It was interesting to find that the high-predation fish showed an unexpected lower velocity preference, which might represent a compromise between predation avoidance, foraging and energy saving.

Variations in temperature acclimation effects on glycogen storage, hypoxia tolerance and swimming performance with seasonal acclimatization in juvenile Chinese crucian carp.

The aim of this study was to test whether temperature acclimation (10 vs 20 °C) effects on tissue glycogen content, hypoxia tolerance, and swimming performance of Chinese crucian carp (Carassius auratus) varied with seasonal acclimatization (winter vs spring) and potential combined interactions. Both the routine metabolic rate (MO(2rout)) and critical oxygen tension (P(crit)) of the MO(2rout) increased significantly with temperature, whereas the seasonal acclimatization showed no significant effect. Only the high temperature group that acclimatized in spring showed a significantly higher aquatic surface respiration (ASR(crit)) value compared with the other three groups. Fish in spring tended to show ASR behavior at higher oxygen tension compared with those in winter, which might have been caused by a more active lifestyle. Time to show LOE prolonged by 25-34% under low temperature. Spring fish showed 20% shorter LOE duration at 10 °C, whereas the difference tended to vanish at 20 °C. Glycogen contents in both liver and muscle were higher in winter than spring. The liver and muscle glycogen content decreased by 5-42% after exposure to anoxic conditions, whereas the magnitude was much smaller in spring. When fish swam in normoxic conditions, fish in higher temperatures showed higher critical swimming speed (Ucrit) than low temperature (5.49 vs 3.74 BL s(-1) in winter and 4.27 vs 3.21 BL s(-1) in spring), whereas fish in winter also showed higher U(crit) than fish in spring for each temperature. However, when fish swam in hypoxic waters, fish in higher temperatures showed a more profound decrease (52-61%) in U(crit) compared to those in lower temperature (25-27%). Fish in lower temperatures that had acclimatized in winter showed the highest U(crit), which might have been caused by higher glycogen storage. The present study suggested that both glycogen storage and alterations in lifestyle had profound effects on hypoxia tolerance and swimming performance, which resulted in a profound difference between seasons and acclimation temperatures.

Effect of temperature on hypoxia tolerance and its underlying biochemical mechanism in two juvenile cyprinids exhibiting distinct hypoxia sensitivities.

It is increasingly important to investigate the effect of temperature on hypoxia tolerance in fish species, as worldwide hypoxia worsens with increases in global warming. We selected the hypoxia-tolerant crucian carp (Carassius carassius) and the hypoxia-sensitive Chinese bream (Parabramis pekinensis) as model fish and investigated their hypoxia tolerance based on the critical oxygen tension of the routine metabolic rate (M˙O2rout) (Pcrit), aquatic surface respiration (ASRcrit) and loss of equilibrium (LOEcrit) after two weeks of acclimation at either 10, 20 or 30 °C. We also measured the tissue substrate (glycogen and glucose of muscle and liver) and lactate levels of both normoxia- and hypoxia-treated fish (post-LOE). Crucian carp exhibited significantly lower Pcrit and LOEcrit but not ASRcrit. Crucian carp possessed higher hypoxia tolerance, partially due to a higher tissue glycogen reserve, which provides cellular fuel under severe hypoxia, as well as higher lactate tolerance and clearance ability than Chinese bream. The hypoxia tolerance was maintained in crucian carp but was decreased in Chinese bream as the temperature increased. The difference between the two species is based on the greater recruitment of tissue glycogen, resulting in an increased level of cellular fuel during hypoxia in crucian carp than in Chinese bream. In addition, crucian carp possessed the greater liver lactate clearance capacity, and the smaller increase in the M˙O2rout at higher temperatures compared to Chinese bream. Furthermore, substrate shortage and decreased lactate tolerance at high temperatures in Chinese bream might also contribute to the difference in hypoxia tolerance between the two species.

The effect of temperature on repeat swimming performance in juvenile qingbo (Spinibarbus sinensis).

To investigate the effect of temperature on the repeat constant acceleration swimming performance and on the metabolic recovery capacity in juvenile qingbo (Spinibarbus sinensis), their constant acceleration test speed (U(CAT)) and excess post-exercise oxygen consumption (EPOC) recovery process were measured twice with 1-h intervals at different acclimation temperatures (10, 15, 20, 25 and 30 °C). Temperature significantly affected U(CAT), the pre-exercise metabolic rate (MO(2)), metabolic peak values (MO(2peak)), the metabolic scope (MS, MO(2peak)--pre-exercise MO(2)) and the magnitude of the EPOC (P < 0.05). These parameters significantly increased as the temperature increased from 15 to 25 °C and significantly decreased (U(CAT) and EPOC magnitude) or did not change (MO(2peak) and MS) when the temperature increased from 25 to 30 °C in the first test (P < 0.05). The relationships between temperature (T) and these parameters (U(CAT), MO(2peak), MS and EPOC magnitude) in the first test were as follows: U(CAT) = 62.14/{1 + [(T - 25.1)/21.1](2)} (r = 0.847, P < 0.001, n = 40); MO(2peak) = 1,052.11/{1 + [(T - 29.2)/18.9](2)} (r = 0.901, P < 0.001, n = 39); MS = 753.74/{1 + [(T - 27.1)/18.6](2)} (r = 0.768, P < 0.001, n = 39); and EPOC = 195.42/{1 + [(T - 25.6)/8.7](2)} (r = 0.752, P < 0.001, n = 39). The optimal temperatures for U(CAT), MO(2peak), MS and EPOC magnitude in juvenile qingbo were 25.1, 29.2, 27.1 and 28.6 °C, respectively. Repeat exercise had different effect on U(CAT) and EPOC magnitude at different temperature (interaction effect, P < 0.05). There was no difference in U(CAT) and in EPOC magnitude between the first and second tests at low temperatures (10-20 °C). However, both U(CAT) and EPOC magnitude decreased significantly during the second test compared with the first test at high temperatures (25 and 30 °C) (P < 0.05). The present study showed that the recovery of the constant acceleration swimming performance was poorer at higher temperatures than at low temperatures in juvenile qingbo. These differences may be related to larger anaerobic metabolism, a lower pH value in the blood, larger ionic fluids and/or higher levels of hormones present at high temperatures.

Behavior, metabolism and swimming physiology in juvenile Spinibarbus sinensis exposed to PFOS under different temperatures.

The harmful effects of perfluorooctane sulfonate (PFOS) are of growing international concern. This paper aimed to gain an integrated understanding of fitness-related ecological end points, such as behavior, metabolism and swimming physiology, in juvenile Spinibarbus sinensis in response to PFOS toxicity at different temperatures. The fish were exposed to a range of PFOS concentrations (0, 0.32, 0.8, 2 and 5 mg/L) at different temperatures (18 and 28 °C) for 30 days. The effects on fish behavior, metabolic characteristics and aerobic swimming performance caused by PFOS at different temperatures were investigated. Our results showed that both PFOS and temperature had important influences on spontaneous swimming behavior, social interactions, routine metabolic rate (RMR), net energetic cost of transport (COTnet) and critical swimming speed (U crit) in fish. The lowest observed effect concentration for both U crit and RMR was 5 and 0.8 mg/L at 18 and 28 °C, respectively. We found that PFOS affected various behavioral and social end points and also appeared to affect metabolic rates and reduced U crit, likely as a result of increased COTnet, and that many of these effects also changed with respect to temperature. Our results further the understanding of the metabolic and behavioral toxicity of PFOS to aquatic organisms.

Interspecific variation in hypoxia tolerance, swimming performance and plasticity in cyprinids that prefer different habitats.

This study quantified and compared hypoxia tolerance and swim performance among cyprinid fish species from rapid-, slow- and intermediate-flow habitats (four species per habitat) in China. In addition, we explored the effects of short-term acclimation on swim performance, maximum metabolic rate (M(O2,max)) and gill remodelling to detect habitat-associated patterns of plastic response to hypoxia. Indices of hypoxia tolerance included oxygen threshold for loss of equilibrium (LOE50) and aquatic surface respiration (ASR50), and critical oxygen tension for routine metabolic rate (Pcrit). Critical swimming speed (Ucrit) and M(O2,max) were measured under normoxic and hypoxic conditions after 48 h acclimation to normoxia and hypoxia, and gill remodelling was estimated after 48 h of hypoxia exposure. Both traditional ANCOVA and phylogenetically independent contrast (PDANOVA) analyses showed that fish species from rapid-flow habitats exhibited lower LOE50 compared with fish from intermediate- and slow-flow habitats. Habitat-specific differences in Pcrit and Ucrit were detected using PDANOVA but not traditional ANCOVA analyses, with fish species from rapid-flow habitats exhibiting lower Pcrit but higher Ucrit values compared with fish from intermediate- and slow-flow habitats. Fish species from rapid-flow habitats were also characterized by less plasticity in swim performance and gill morphology in response to hypoxia acclimation compared with species from slow-flow habitats, but a greater drop in swim performance in response to acute hypoxia exposure. The study detected a habitat-specific difference in hypoxia tolerance, swimming performance and its plasticity among fish from habitats with different flow conditions, possibly because of the long-term adaptation to the habitat caused by selection stress. The PDANOVA analyses were more powerful than traditional statistical analyses according to the habitat effects in both hypoxia tolerance and swimming performance in this study.

The effects of constant and diel-fluctuating temperature acclimation on the thermal tolerance, swimming capacity, specific dynamic action and growth performance of juvenile Chinese bream.

We investigated the effects of constant and diel-fluctuating temperature acclimation on the thermal tolerance, swimming capacity, specific dynamic action (SDA) and growth performance of juvenile Chinese bream (Parabramis pekinensis). The critical thermal maxima (CTmax), critical thermal minima (CTmin), lethal thermal maxima (LTmax), lethal thermal minima (LTmin), critical swimming speed (Ucrit) and fast-start escape response after 30 d acclimation to three constant temperatures (15, 20 and 25 °C) and one diel-fluctuating temperature (20±5 °C) were measured. In addition, feeding rate (FR), feeding efficiency (FE) and specific growth rate (SGR) were measured. The diel-fluctuating temperature group showed lower CTmin than the 20 °C group but a similar CTmax, indicating a wider thermal scope. SDA linearly increased with the temperature. Temperature variation between 20 and 25 °C had little effect on either swimming or growth performance. However, fish in the 15 °C group exhibited much poorer swimming and growth performance than those in the 20 °C group. Ucrit decreased slightly under low acclimation temperature due to the pronounced improvement in swimming efficiency under cold temperature. Fish in the diel-fluctuating temperature group fed more but exhibited similar SGR compared to 20 °C group, possibly due in part to an increase in energy expenditure to cope with the temperature fluctuation. The narrower thermal scope and lower CTmax of Chinese bream together with the conservation of CTmax with temperature acclimation, suggests that local water temperature elevations may have more profound effects on Chinese bream than on other fish species in the Three Gorges Reservoir.

The effects of fasting on swimming performance in juvenile qingbo (Spinibarbus sinensis) at two temperatures.

We measured the following variables to investigate the effects of fasting and temperature on swimming performance in juvenile qingbo (Spinibarbus sinensis): the critical swimming speed (Ucrit), resting metabolic rate (MO2rest) and active metabolic rate (MO2active) of fish fasting for 0 (control), 1, 2 and 4 weeks at low and high acclimation temperatures (15 and 25°C). Both fasting treatment and temperature acclimation had significant effects on all parameters measured (P<0.05). Fasting at the higher temperature had a negative effect on all measured parameters after 1 week (P<0.05). However, when acclimated to the lower temperature, fasting had a negative effect on Ucrit until week 2 and on (MO2rest), (MO2active) and metabolic scope (MS, (MO2active)-(MO2rest)) until week 4 (P<0.05). The values of all parameters at the lower temperature were significantly lower than those at the higher temperature in the identical fasting period groups except for (MO2rest) of the fish that fasted for 2 weeks. The relationship between fasting time (T) and Ucrit was described as Ucrit(15)=-0.302T(2)-0.800T+35.877 (r=0.781, n=32, P<0.001) and Ucrit(25)=0.471T(2)-3.781T+50.097 (r=0.766, n=32, P<0.001) at 15 and 25°C, respectively. The swimming performance showed less decrease in the early stage of fasting but more decrease in the later stage at the low temperature compared to the high temperature, which might be related to thermal acclimation time, resting metabolism, respiratory capacity, energy stores, enzyme activity in muscle tissue and energy substrate utilization changes with fasting between low and high temperatures. The divergent response of the swimming performance to fasting in qingbo at different temperatures might be an adaptive strategy to seasonal temperature and food resource variation in their habitat.