A Regulatory Circuit Controlling the Dynamics of NFκB cRel Transitions B Cells from Proliferation to Plasma Cell Differentiation.

Humoral immunity depends on efficient activation of B cells and their subsequent differentiation into antibody-secreting cells (ASCs). The transcription factor NFκB cRel is critical for B cell proliferation, but incorporating its known regulatory interactions into a mathematical model of the ASC differentiation circuit prevented ASC generation in simulations. Indeed, experimental ectopic cRel expression blocked ASC differentiation by inhibiting the transcription factor Blimp1, and in wild-type (WT) cells cRel was dynamically repressed during ASC differentiation by Blimp1 binding the Rel locus. Including this bi-stable circuit of mutual cRel-Blimp1 antagonism into a multi-scale model revealed that dynamic repression of cRel controls the switch from B cell proliferation to ASC generation phases and hence the respective cell population dynamics. Our studies provide a mechanistic explanation of how dysregulation of this bi-stable circuit might result in pathologic B cell population phenotypes and thus offer new avenues for diagnostic stratification and treatment.

Direct observation correlates NFκB cRel in B cells with activating and terminating their proliferative program.

Antibody responses require the proliferative expansion of B cells controlled by affinity-dependent signals. Yet, proliferative bursts are heterogeneous, varying between 0 and 8 divisions in response to the same stimulus. NFκB cRel is activated in response to immune stimulation in B cells and is genetically required for proliferation. Here, we asked whether proliferative heterogeneity is controlled by natural variations in cRel abundance. We developed a fluorescent reporter mTFP1-cRel for the direct observation of cRel in live proliferating B cells. We found that cRel is heterogeneously distributed among naïve B cells, which are enriched for high expressors in a heavy-tailed distribution. We found that high cRel expressors show faster activation of the proliferative program, but do not sustain it well, with population expansion decaying earlier. With a mathematical model of the molecular network, we showed that cRel heterogeneity arises from balancing positive feedback by autoregulation and negative feedback by its inhibitor IκBε, confirmed by mouse knockouts. Using live-cell fluorescence microscopy, we showed that increased cRel primes B cells for early proliferation via higher basal expression of the cell cycle driver cMyc. However, peak cMyc induction amplitude is constrained by incoherent feedforward regulation, decoding the fold change of cRel activity to terminate the proliferative burst. This results in a complex nonlinear, nonmonotonic relationship between cRel expression and the extent of proliferation. These findings emphasize the importance of direct observational studies to complement gene knockout results and to learn about quantitative relationships between biological processes and their key regulators in the context of natural variations.

Starter feed for carnivorous species as a practical replacement of bloodworms for a vertebrate model organism in ageing, the turquoise killifish Nothobranchius furzeri.

The absence of a controlled diet is unfortunate in a promising model organism for ageing, the turquoise killifish (Nothobranchius furzeri Jubb, 1971). Currently captive N. furzeri are fed bloodworms but it is not known whether this is an optimal diet. Replacing bloodworms with a practical dry feed would reduce diet variability. In the present study, we estimated the nutritional value of the diet ingested by wild fish and determined the fish-body amino acid profile as a proxy for their nutritional requirements. We compared the performance of fish fed four commercial feeds containing 46%-64% protein to that achieved with bloodworms and that of wild fish. Wild fish target a high-protein (60%) diet and this is supported by their superior performance on high-protein diets in captivity. In contrast, feeds for omnivores led to slower growth, lower fecundity and unnatural liver size. In comparison to wild fish, a bloodworm diet led to lower body condition, overfeeding and male liver enlargement. Out of the four dry feeds tested, the fish fed Aller matched wild fish in body condition and liver size, and was comparable to bloodworms in terms of growth and fecundity. A starter feed for carnivorous species appears to be a practical replacement for bloodworms for N. furzeri. The use of dry feeds improved performance in comparison to bloodworms and thus may contribute to reducing response variability and improving research reproducibility in N. furzeri research.

Icy affairs: Understanding recent advancements in the freezing and frozen storage of fish.

Freezing methods have evolved over the last 30 years. This review states the effect of various freezing methods on the quality of fish and seafood. Freezing temperatures, freezing, and frozen storage temperatures were also analyzed and reviewed. The changes in the ice crystal, protein, and lipid affect the fish quality and nutritional value during freezing and frozen storage. Freezing methods when combined with various additives or preprocessing approaches help improve the efficacy of freezing and frozen storage. Several experimental or emerging methods also have positive effects on the products' quality. According to the metadata reanalysis of quality markers, freshly frozen fish using different freezing methods may vary much in terms of ice diameter, but not others. High pressure freezing or immersion freezing-derived fish retains the best quality through frozen storage. More data are required on freezing methods (electrical-assisted freezing, microwave-assisted freezing, magnetic-assisted freezing, radiofrequency-assisted freezing, and the commercial's application and investment should be considered in the future. This review sheds light on finding a balanced initial shear force during freezing and the use of certain additives to control freezing-related damages. Focusing on ice diameter alone may be futile (e.g., liquid N2 freezing). Future optimization of technologies should be in a way that several processes along the farm to fork such as freezing, frozen storage, thawing, thermal processing of fish, and even refabrication of food should mutually complement each other's needs to deliver safe and high-quality fish to the consumer's plate, even after a prolonged shelf-life.

Nongenetic origins of cell-to-cell variability in B lymphocyte proliferation.

Rapid antibody production in response to invading pathogens requires the dramatic expansion of pathogen-derived antigen-specific B lymphocyte populations. Whether B cell population dynamics are based on stochastic competition between competing cell fates, as in the development of competence by the bacterium Bacillus subtilis, or on deterministic cell fate decisions that execute a predictable program, as during the development of the worm Caenorhabditis elegans, remains unclear. Here, we developed long-term live-cell microscopy of B cell population expansion and multiscale mechanistic computational modeling to characterize the role of molecular noise in determining phenotype heterogeneity. We show that the cell lineage trees underlying B cell population dynamics are mediated by a largely predictable decision-making process where the heterogeneity of cell proliferation and death decisions at any given timepoint largely derives from nongenetic heterogeneity in the founder cells. This means that contrary to previous models, only a minority of genetically identical founder cells contribute the majority to the population response. We computationally predict and experimentally confirm nongenetic molecular determinants that are predictive of founder cells' proliferative capacity. While founder cell heterogeneity may arise from different exposure histories, we show that it may also be due to the gradual accumulation of small amounts of intrinsic noise during the lineage differentiation process of hematopoietic stem cells to mature B cells. Our finding of the largely deterministic nature of B lymphocyte responses may provide opportunities for diagnostic and therapeutic development.

Bi-FPNFAS: Bi-Directional Feature Pyramid Network for Pixel-Wise Face Anti-Spoofing by Leveraging Fourier Spectra.

The emergence of biometric-based authentication using modern sensors on electronic devices has led to an escalated use of face recognition technologies. While these technologies may seem intriguing, they are accompanied by numerous implicit drawbacks. In this paper, we look into the problem of face anti-spoofing (FAS) on a frame level in an attempt to ameliorate the risks of face-spoofed attacks in biometric authentication processes. We employed a bi-directional feature pyramid network (BiFPN) that is used for convolutional multi-scaled feature extraction on the EfficientDet detection architecture, which is novel to the task of FAS. We further use these convolutional multi-scaled features in order to perform deep pixel-wise supervision. For all of our experiments, we performed evaluations across all major datasets and attained competitive results for the majority of the cases. Additionally, we showed that introducing an auxiliary self-supervision branch tasked with reconstructing the inputs in the frequency domain demonstrates an average classification error rate (ACER) of 2.92% on Protocol IV of the OULU-NPU dataset, which is significantly better than the currently available published works on pixel-wise face anti-spoofing. Moreover, following the procedures of prior works, we performed inter-dataset testing, which further consolidated the generalizability of the proposed models, as they showed optimum results across various sensors without any fine-tuning procedures.

Modeling pre-spawning fitness and optimal climate of spotted snakehead Channa punctata (Bloch, 1793) from a Gangetic floodplain wetland of West Bengal, India.

The spawning and well-being of fish in an ecosystem are closely linked to climatic cues, viz., temperature and rainfall. Reduced fitness can affect the reproductive performance and lead to skipped spawning. Benchmarking the threshold fitness required for a fish population to achieve readiness for spawning, and understanding how climatic parameters influence the fitness will aid in predicting the fate of its reproductive success in future climatic conditions. This study determined the threshold condition factor pre-spawning fitness (Kspawn50) at which 50% of the female Channa punctata population can be deemed fit for spawning. The optimal climate within which pre-spawning fitness is attained by this species under Indian climatic conditions was also identified. The study was conducted from June 2015 to September 2016, covering two spawning seasons (June-August) in a Gangetic floodplain wetland of West Bengal, India. The non-parametric Kaplan-Meier method (survival fit) was used for estimation of pre-spawning fitness. "Ready to spawn" females were classified based on binary coding of the gonadal maturity stages. The thermal and precipitation range within which spawning fitness is achieved was identified by using the locally weighted smoothing technique. Female C. punctata pre-spawning fitness (Kspawn50) ranged from 1.26 to 1.39 with an estimated median of 1.29 units. Temperatures between 29 and 32 °C and rainfall above 100 mm were conducive to attaining the requisite pre-spawning fitness in C. punctata. This is the first study benchmarking the pre-spawning fitness and optimal climate for C. punctata. Understanding spawning requirements can inform the climate change-induced impacts on reproductive plasticity and evolutionary adaptations of snakeheads in the Ganga river basin.

Pattern of the state of eutrophication in the floodplain wetlands of eastern India in context of climate change: a comparative evaluation of 27 wetlands.

The floodplain wetlands in different regional settings vary with time and space in terms of function and geomorphological diversity. In recent decades, these eco-sensitive waterbodies have been exposed to a wide range of anthropogenic threats and climatic changes. Therefore, assessment of these ecological and environmental threats is prerequisite to understand the state of ecosystem and to develop a sustainable management strategy for conservation of wetland biodiversity and fisheries enhancement. This paper discusses the region-specific pattern of trophic state index (TSI) of the 27 floodplain wetlands in West Bengal, India. Carlson TSI and Lamparelli TSI methods were used to determine a better approach based on historical and continuous dataset and to delineate the interrelationship among historical climatic variability for sustainable management of the resources. The study revealed that agro-climatic divisions do not unveil any significant impact on the TSI calculated using Carlson TSI as well as Lamparelli TSI method. The TSI scores for the two methods were significantly different (p < 0.01) for different zones based on wetland habitat types. The TSI scores revealed most of the wetlands to be in mesotrophic state. Principal component analysis (PCA) indicated that TSI scores were having similar pattern of variation with rainfall and water temperature. The present study also conveys fundamental information on ecological status based on the trophic state, which will aid to develop region-specific strategies for sustainable fisheries enhancement.

Climato-environmental influence on breeding phenology of native catfishes in River Ganga and modeling species response to climatic variability for their conservation.

The main objectives of the present study were to quantify the environmental, especially temperature and rainfall, effects on breeding phenology of selected catfish species and to predict changes in breeding phenology of the selected species in relation to climatic variability for the Ganga River Basin. The study showed that changes in rainfall pattern may have the most profound effect on gonad maturation and breeding of Mystus tengara and Mystus cavasius followed by the effect of increased water temperature due to rising air temperature. Indication of region-specific adaptation was noticed in reproductive phenology of Eutropiichthys vacha based on local trends of warming climate. The other habitat parameters, such as dissolved oxygen, alkalinity, nitrate, and phosphate, were correlated with gonad maturity and spawning. Climatic variability may bring region-specific changes in breeding phenology of fish species in the Ganga River. Under a warming climate, changes in precipitation pattern manifested into riverine flow pulse may be the key driver in dictating breeding phenology. Our study indicates E. vacha as a climate sensitive species that may be selected as a target species for climate change impact studies.

Understanding nutrient throughput of operational RAS farm effluents to support semi-commercial aquaponics: Easy upgrade possible beyond controversies.

The present research attempted to address a key industry-level question amidst Recirculating Aquaculture System (RAS) waste throughput and aquaponics limitations controversies. Nutrient throughput of three operational RAS farms with progressive size proportions (16, 130, 1400 m3), aquaculture intensity (24, 62, 86 kg stock m-3) were studied. Results suggest - daily total efflux and potency of nutrients in effluents should not be generalized, extreme variability exists. Consistencies of nutrients in wastewater (except N, Ca and Na) are higher than in sludge. Asynchrony between patterns of nutrient loading and effluent nutrient concentrations exist for secondary macronutrients and micronutrients (S, Mg, Fe, Cu, Zn, B, Mo). Macronutrient output generally increases with increasing farm size and culture intensity but same cannot be said for micronutrients. Deficiency in wastewater can be completely masked using raw or mineralized sludge, usually containing 3-17 times higher nutrient concentrations. RAS effluents (wastewater and sludge combined) contain adequate N, P, Mg, Ca, S, Fe, Zn, Cu, Ni to meet most aquaponic crop needs. K is generally deficient requiring a full-fledged fertilization. Micronutrients B, Mo are partly sufficient and can be easily ameliorated by increasing sludge release. The presumption surrounding 'definite' phyto-toxic Na levels in RAS effluents should be reconsidered - practical solutions available too. No threat of heavy metal accumulation or discharge was observed. Most of the 'well-known' operational influences failed to show any significant predictable power in deciding nutrient throughput from RAS systems. Calibration of nutrient output from operational RAS farms may be primarily focused around six predictors we identified. Despite inherent complexity of effluents, the conversion of RAS farms to semi-commercial aquaponics should not be deterred by nutrient insufficiency or nutrient safety arguments. Incentivizing RAS farm wastes through semi-commercial aquaponics should be encouraged - sufficient and safe nutrients are available.

Benchmarking pre-spawning fitness, climate preferendum of some catfishes from river Ganga and its proposed utility in climate research.

The concept of threshold condition factor (Fulton), beyond which more than 50% of the female fish population may attain readiness for spawning coined as pre-spawning fitness (K spawn50), has been proposed in the present article and has been estimated by applying the non-parametric Kaplan-Meier method for fitting survival function. A binary coding strategy of gonadal maturity stages was used to classify whether a female fish is "ready to spawn" or not. The proposed K spawn50 has been generated for female Mystus tengara (1.13-1.21 units), M. cavasius (0.846-0.945 units), and Eutropiichthys vacha (0.716-0.799 units). Information on the range of egg parameters (fecundity, egg weight, egg diameter) expected at the pre-spawning stage was also generated. Additional information on species-specific thermal and precipitation window (climate preferendum) within which K spawn50 is attained was also generated through the LOESS smoothing technique. Water temperatures between 31 and 36 °C (M. tengara), 30 and 32 °C (M. cavasius), and 29.5 and 31 °C (E. vacha) and monthly rainfall between 200 and 325 mm (M. tengara), > 250 mm (M. cavasius), and around 50 mm and between 350 and 850 mm (E. vacha) were found to be optimum for attainment of K spawn50. The importance of parameterization and benchmarking of K spawn50 in addition to other conventional reproductive biology parameters has been discussed in the present article. The purposes of the present study were fulfilled by generating baseline information and similar information may be generated for other species replicating the innovative methodology used in this study.

Class II MHC/peptide interaction in Leishmania donovani infection: implications in vaccine design.

We show that Leishmania donovani-infected macrophages (MΦs) are capable of stimulating MHC class II (MHC-II)-restricted T cells at 6 h of infection. At 48 h, infected MΦs (I-MΦs) failed to stimulate MHC-II-restricted T cells but not MHC class I-restricted ones, in contrast to normal MΦs. Such I-MΦs could stimulate T cells at a higher Ag concentration, indicating that general Ag processing and trafficking of peptide-MHC-II complexes are not defective. Analysis of the kinetic parameters, like "kon" and "koff," showed that peptide-MHC-II complex formation is compromised in I-MΦs compared with normal MΦs. This indicates interference in loading of the cognate peptide to MHC-II, which may be due to the presence of a noncognate molecule. This notion received support from the finding that exposure of I-MΦs to low pH or treatment with 2-(1-adamantyl)-ethanol, a molecule that favors peptide exchange, led to T cell activation. When treated with 2-(1-adamantyl)-ethanol, splenocytes from 8 wk-infected BALB/c mice showed significantly higher antileishmanial T cell expansion in vitro compared with untreated controls. Hence, it is tempting to speculate that high, but not low, concentrations of cognate peptide may favor peptide exchange in I-MΦs, leading to expansion of the antileishmanial T cell repertoire. The results suggest that a high Ag dose may overcome compromised T cell responses in visceral leishmaniasis, and this has an important implication in therapeutic vaccine design.

Enterotoxigenic Escherichia coli EtpA mediates adhesion between flagella and host cells.

Adhesion to epithelial cells and flagella-mediated motility are critical virulence traits for many Gram-negative pathogens, including enterotoxigenic Escherichia coli (ETEC), a major cause of diarrhoea in travellers and children in developing countries. Many flagellated pathogens export putative adhesins belonging to the two-partner secretion (TPS) family. However, the actual function of these adhesins remains largely undefined. Here we demonstrate that EtpA, a TPS exoprotein adhesin of enterotoxigenic E. coli, mimics and interacts with highly conserved regions of flagellin, the major subunit of flagella, and that these interactions are critical for adherence and intestinal colonization. Although conserved regions of flagellin are mostly buried in the flagellar shaft, our results suggest that they are at least transiently exposed at the tips of flagella where they capture EtpA adhesin molecules for presentation to eukaryotic receptors. Similarity of EtpA to molecules encoded by other motile pathogens suggests a potential common pattern for bacterial adhesion, whereas participation of conserved regions of flagellin in adherence has implications for development of vaccines for Gram-negative pathogens.

Contribution of the highly conserved EaeH surface protein to enterotoxigenic Escherichia coli pathogenesis.

Enterotoxigenic Escherichia coli (ETEC) strains are among the most common causes of diarrheal illness worldwide. These pathogens disproportionately afflict children in developing countries, where they cause substantial morbidity and are responsible for hundreds of thousands of deaths each year. Although these organisms are important targets for enteric vaccines, most development efforts to date have centered on a subset of plasmid-encoded fimbrial adhesins known as colonization factors and heat-labile toxin (LT). Emerging data suggest that ETEC undergoes considerable changes in its surface architecture, sequentially deploying a number of putative adhesins during its interactions with the host. We demonstrate here that one putative highly conserved, chromosomally encoded adhesin, EaeH, engages the surfaces of intestinal epithelial cells and contributes to bacterial adhesion, LT delivery, and colonization of the small intestine.

Simultaneous inhibition of key growth pathways in melanoma cells and tumor regression by a designed bidentate constrained helical peptide.

Protein-protein interactions are part of a large number of signaling networks and potential targets for drug development. However, discovering molecules that can specifically inhibit such interactions is a major challenge. S100B, a calcium-regulated protein, plays a crucial role in the proliferation of melanoma cells through protein-protein interactions. In this article, we report the design and development of a bidentate conformationally constrained peptide against dimeric S100B based on a natural tight-binding peptide, TRTK-12. The helical conformation of the peptide was constrained by the substitution of α-amino isobutyric acid--an amino acid having high helical propensity--in positions which do not interact with S100B. A branched bidentate version of the peptide was bound to S100B tightly with a dissociation constant of 8 nM. When conjugated to a cell-penetrating peptide, it caused growth inhibition and rapid apoptosis in melanoma cells. The molecule exerts antiproliferative action through simultaneous inhibition of key growth pathways, including reactivation of wild-type p53 and inhibition of Akt and STAT3 phosphorylation. The apoptosis induced by the bidentate constrained helix is caused by direct migration of p53 to mitochondria. At moderate intravenous dose, the peptide completely inhibits melanoma growth in a mouse model without any significant observable toxicity. The specificity was shown by lack of ability of a double mutant peptide to cause tumor regression at the same dose level. The methodology described here for direct protein-protein interaction inhibition may be effective for rapid development of inhibitors against relatively weak protein-protein interactions for de novo drug development.

Considerations for protein and amino acids in standardized reference diet for parthenogenetic marbled crayfish Procambarus virginalis model organism.

The concept of a standardized reference diet (SRD) is used in laboratory model organisms to ensure nutritional control between studies and laboratories. Although models using the genetically identical, all female parthenogenetic marbled crayfish (Procambarus virginalis) are growing in popularity, research into nutrition in this species still has many knowledge gaps. To fast track the development of a SRD in terms of protein and amino acids (SRDprotein) for this species, we first analyzed the composition of its body amino acids to determine the ideal protein concept (IPC) of indispensable amino acids in wild-caught P. virginalis (which had an unusually high preponderance of leucine and arginine). Then, we strategically evaluated three common clusters of types of fish feed: (1) ornamental fish feed (SER) fortified with a naturally occurring alga (Spirulina). This type of feed was protein-high in arginine and leucine (SER + SPI) that fulfils the species' IPC for iso-protein (~ 40%), iso-phosphorus (~ 0.8%) and near iso-energetic (~ 475 kcal 100 g-1); (2) freeze-dried live feed consisting of chironomid larvae (CHI) fortified with Spirulina (CHI + SPI) that fulfils the IPC for iso-protein (~ 46%), iso-phosphorus (~ 0.7%) and near iso-energetic (~ 405 kcal 100 g-1); and (3) a commercially standardized 'starter diet' for carnivorous fish larvae (FISH) and post-larval shrimps (SHRIMP) with iso-protein (~ 56%) and iso-phosphorus (~ 1.6%). A total of six diets, embracing a diverse range of proteinaceous feeds, were used in a 100-day ad libitum feeding and growth trial. The FISH group outperformed all the other groups (p < 0.05) and our exploratory multivariate analysis revealed an ideal demand of > 44% protein (tailored to deliver high arginine 3% and leucine 4%, followed by the usual lysine > 3.5% and methionine 1.2%) but also the lowest carbohydrate level (21%). For SRDprotein, our findings show that the FISH diet is ideal and suggest the possibilities of using a CHI + SPI diet for further optimization (more economic use of protein and phosphorus).

Considerations for fatty acids in standardized reference diet for parthenogenetic marbled crayfish Procambarus virginalis model organism.

Fatty acid accumulation was studied in the parthenogenetic all-female marbled crayfish Procambarus virginalis using six arbitrarily designed experimental feeds and related to individuals with glair glands (sexual maturity) after 100 days of ad libitum feeding at 21 °C, including gravid females from the wild as a reference. Fatty acids 16:0 and 18:1n-9 comprised 40% of the total amount of fatty acids and tended to up-concentrate in bodies. Shorter chain 14:0 depleted from feed to body. Across diets, there was a concomitant decrease in precursor fatty acid and increase in product fatty acid, such as reinforcements in monounsaturated fatty acid (18:1n-9), eicosanoid precursors 20:4n-6 (arachidonic acid, ARA) and 20:5n-3 (eicosapentaenoic acid, EPA) in-vivo, but not 22:6n-3 (docosahexaenoic acid, DHA) except when deficient in CHI or CHI + SPI diets. Saturation kinetics modeling (R2 0.7-0.9, p < 0.05) showed that when the ARA share is ~ 1%, the EPA share is ~ 8%, and the DHA share is ~ 2% in the food lipids, the accumulation of fatty acids in body lipids levels off. The lowest DHA in the CHI (0% glair glands) or CHI + SPI (0-3.9% glair glands) diets, and the lowest ARA in SER (0% glair glands) or SER + SPI (0-3% glair glands) diets, were synchronous with negligible sexual maturity despite a wide range of observed specific growth rates (2.77-3.60% per day), body size (0.44-0.84 g), ≤ 5% crude lipid and 40-46% crude protein feed. The FISH and SHRIMP diets (56% protein, 11-14% lipid) with the highest ARA, EPA, and DHA together seem to be the most conducive diets for sexual maturity (up to 20% of individuals with glair glands). We propose a fatty acid profile mimicking the FISH or SHRIMP diets as a starting point for designing the lipid content required in the marbled crayfish standardized reference diet.

Fish stocks as phosphorus sources or sinks: Influenced by nutritional and metabolic variations, not solely by dietary content and stoichiometry.

The study provides a descriptive understanding of when fish (Cyprinus carpio model) are the source or sink of phosphorus. Dissolved reactive phosphorus (DRP; PO4-P) losses (51.1 ± 5.9 % of intake-P) increase at excess of bioavailable P (>0.83 g 100 g-1 dry matter, DM fed) or when food (digestible) N:P mass ratio (≤4.4:1) approaches organismal storage threshold (~4:1). This is known, however, even at a sub-threshold food P content (0.57 g 100 g-1 DM) and food N:P mass ratio (7.3:1), DRP losses (57.8 ± 4.5 % of intake-P) may be extraordinary if two indispensable amino acids are biologically insufficient (lysine ≤1.43 g, methionine ≤0.39 g 100 g-1 DM fed). Given that methionine and lysine are sufficient, DRP losses cease (≈0 %) and even some P from water is absorbed, given there is support from non-protein energy (NPE). Insufficient NPE (<180 kcal 100 g-1 DM fed) may drive DRP losses (81.6 ± 4.3 % of intake-P) beyond expected levels (46-59 % of intake-P) at a given food P content (0.91 g 100 g-1 DM). Natural food seldom fulfills low P, high lysine + methionine, and high NPE contents simultaneously, thus keeping fish in a perpetual P recycling for algae (scaleless carp > scaly carp). Such P recycling ceases only during basal metabolism. During feeding state, the richness of lysine + methionine bound N and lipid + carbohydrate bound C in the food base may enhance the fishes' threshold of P storage. P storage can be diminished when they are insufficient. We show that for fish, the decision of P recycling or not recycling (for algae) may change based on the supply of specific fractions of N or C from the food web or metabolic variations (basal metabolism, presence of scales). NOVELTY STATEMENT: The ecological stoichiometry theory is better connected to fish nutritional bioenergetics for better understanding and biomanipulation of eutrophication processes.

Synergistic digestibility effect by planktonic natural food and habitat renders high digestion efficiency in agastric aquatic consumers.

A digestibility enhancing effect of natural food on stomachless fish model (Cyprinus carpio) was verified by fluorogenic substrate assays of enzymatic activities in experimental pond carp gut flush and planktonic food over a full vegetative season. Then compared with size-matched conspecific grown artificially (tank carp) and an advanced omnivore species possessing true stomach (tilapia, Oreochromis niloticus). Results suggested activities of digestive enzymes (except amylolytic) were significantly higher in pond carp (p ≤ 0.05) than in the size-matched tank carp. Even compared to tilapia, pond carp appeared superior (p < 0.05; proteolytic or chitinolytic activities) or comparable (p > 0.05; phosphatase or cellulolytic activities). Amylolytic, chitinolytic, and phosphatases activities in pond carp gut significantly increased (p ≤ 0.01) over season. Several orders-of-magnitude higher enzymatic activities were detected in planktonic natural food than expressed in carp gut. Amino acid markers in planktonic food revealed a higher share of zooplankton (microcrustaceans), but not phytoplankton, synchronized with higher activities of complex polysaccharide-splitting enzymes (cellulolytic and chitinolytic) in fish gut. Periods of clear water phase low in chlorophyll-a and nutrients, but high in certain zooplankton (preferably cladocerans), may create a synergistic digestibility effect in pond carp. We conclude aquatic ecosystem components (natural food, water, microbiota) enhance fishes' hydrolyzing capabilities of C/N/P macromolecules and even their complex polymers such as cellulose, chitin, and maybe phytate (to be validated), to the extent that being stomachless is not an issue. Aquatic nutritional ecologists may consider that laboratory-based understandings of digestibility may underestimate digestion efficiency of free-ranging fish in ponds or lakes.

Cholesterol lowering drug may influence cellular immune response by altering MHC II function.

Major histocompatibility complex class II (MHC II) expressed on the surface of antigen-presenting cells (APCs) displays peptides to CD4⁺ T cells. Depletion of membrane cholesterol from APCs by methyl ß-cyclodextrin treatment compromises peptide-MHC II complex formation coupled with impaired binding of conformational antibody, which binds close to the peptide binding groove of MHC II. Interestingly, the total cell surface of MHC II remains unaltered. These defects can be corrected by restoring membrane cholesterol. In silico docking studies with a three-dimensional model showed the presence of a cholesterol binding site in the transmembrane domain of MHC II (TM-MHC-II). From the binding studies it was clear that cholesterol, indeed, interacts with the TM-MHC-II and alters its conformation. Mutation of cholesterol binding residues (F240, L243, and F246) in the TM-MHC-II decreased the affinity for cholesterol. Furthermore, transfection of CHO cells with full-length mutant MHC II, but not wild-type MHC II, failed to activate antigen-specific T cells coupled with decreased binding of conformation-specific antibodies. Thus, cholesterol-induced conformational change of TM-MHC-II may allosterically modulate the peptide binding groove of MHC II leading to T cell activation.