They display a characteristic ruffled border where proteases and

They display a characteristic ruffled border where proteases and acid are secreted, allowing for bone resorption and formation of ‘resorption pits’ in the bone surface [25]. Osteoclast morphology varies between mammals and teleosts (bony fishes), and also between different groups of teleosts [20]. In the skeleton of young zebrafish for example, osteoclast activity is carried out by both mononucleated and multinucleated cells [26]. In fact, there is an ontogenetic progression from mono- towards multinucleated osteoclasts. In juvenile zebrafish, bone resorbing cells in the developing lower jaw are

at first mononucleated. In thin skeletal tissues such as the neural arch, mononucleated cells are even predominant in adults [26]. In rainbow trout, scale resorption find more Androgen Receptor Antagonist is predominantly carried out by mononucleated osteoclasts [27]. Although in mammals these mononucleated cells are often just regarded as osteoclast precursors, in fish mononucleated osteoclasts are active bone resorbing cells [28] and [29]. One family of osteoclast proteases

is the matrix metalloproteinases (MMPs). They are involved in the breakdown of extracellular matrix by osteoclasts, but also by other cell types like fibroblasts [30]. MMPs are multi-domain enzymes that require zinc as cofactor for proteolytic activity. Extracellular matrix turnover occurs in a wide range of physiological processes, including embryonic development and morphogenesis, bone resorption and tissue regeneration. Moreover, MMP-mediated breakdown of the extracellular matrix has been implicated in disease processes including cartilage destruction in osteoarthritis [31]. The importance of MMPs in bone development is underlined by studies on mmp2 and mmp9 null mice, which suffer from bone abnormalities, osteoporosis and osteopetrosis respectively [32]. In view of their role in physiological and pathological 3-mercaptopyruvate sulfurtransferase processes, MMPs are important targets in pharmaceutical research and drug development. In bone turnover, secreted MMPs participate in the breakdown of collagen, which in turn allows osteoclast attachment [33]. Furthermore, MMP-9 is associated with osteoclast migration through the collagen

matrix [34]. Matrix metalloproteinases may also break down residual collagen left by cathepsin K after the pH rises in the resorption pit [35]. MMP-2 and MMP-9 (gelatinases A and B, respectively) are particularly active against gelatins (denatured collagens) and intact collagen types I and IV. In bone of dermal origin, matrix degradation is thought to rely more on MMPs and less on cathepsin K [36]. MMP-2 has also been described to play a crucial role in formation and maintenance of the osteocytic canalicular network, whereas MMP-9 is active in early calvarian bone development and in orthodontic tooth movement [37], [38] and [39]. Regenerating fins of adult zebrafish expresses mmp-2 and regeneration can be inhibited by the MMP inhibitor GM6001 [40].

The atmospheric model COSMO-CLM is a non-hydrostatic regional cli

The atmospheric model COSMO-CLM is a non-hydrostatic regional climate model. The model setup complies with CORDEX-EU in the CORDEX framework (Coordinated Regional climate Downscaling Experiment) (Giorgi et al. 2006). The domain covers the whole of Europe, North

Africa, the Atlantic Ocean and the Mediterranean Sea (Figure 1a). The horizontal resolution is 0.44° (approximately 50 km) and the time step is 240 seconds; it has 40 vertical levels. COSMO-CLM PLX4032 solubility dmso applies a ‘mixed’ advection scheme, in which a positive-definite advection scheme is used to approximate the horizontal advection while vertical advection and diffusion are calculated with a partially implicit Crank-Nicholson scheme. In COSMO-CLM, several turbulence schemes are available; in our experiments, we used the so-called 1-D TKE-based diagnostic closure, which is a prognostic

turbulent kinetic energy (TKE) scheme. It includes the interaction of air with solid objects at the surface (roughness elements). We modified the model code to adapt it to the coupled mode. Originally, COSMO-CLM did not have sub-grid scale ice; a grid over the ocean is either fully covered with ice or fully open-water. Thus, a grid size of 50 × 50 km2 implies a rather coarse approximation of real ocean conditions. In addition, COSMO-CLM does not have an ice mask over the ocean; an ocean grid is handled as sea ice or open water depending on the SST. If the temperature is below the freezing point of water, which is −1.7 °C GKT137831 order in COSMO-CLM, the surface is considered to be sea ice. When the temperature is equal to or higher than the freezing point, COSMO-CLM Fenbendazole handles the surface as open water. However, a freezing point of water of −1.7 °C is applicable to sea water with a salinity of approximately 35 PSU

(Practical Salinity Units). In contrast, brackish sea water like the Baltic Sea has a much lower salinity than the average salinity of the World Ocean. At the centre of the Baltic Sea, the Baltic Proper, the salinity is only 7–8 PSU, and this decreases even further northwards to the Bothnian Sea, Bothnian Bay and Gulf of Riga (Gustafsson 1997). The freezing point of this brackish water should therefore be higher than −1.7 °C. When the freezing point is so low, the sea ice cover in the Baltic Sea in COSMO-CLM will be substantially underestimated. Therefore, when coupling COSMO-CLM with the ocean model NEMO, the sea ice treatment is modified in the surface roughness and surface albedo schemes. In the current albedo calculation scheme, COSMO-CLM attributes fixed albedo values to the water surface (0.07) and the sea ice surface (0.7) for the whole grid cell. In the coupled mode, as COSMO-CLM receives the ice mask from NEMO, it can now calculate the weighted average of the albedo based on the fraction of ice and open water in a grid cell. The surface roughness length of the sea ice and open-water grid is calculated in the turbulence scheme of COSMO-CLM.

Note that the transformed model rainfall values preserve the obse

Note that the transformed model rainfall values preserve the observed mean rainfall Enzalutamide order over the 20th century while the simulated inflows preserve the observed mean inflow. Fig. 8 summarizes the results for projected inflows. It compares the observed 31-year average inflows with the full ensemble results based on the rainfall simulations from all the models. It can be seen that ensemble maximum values match the observations for the early part of the 20th century but

it is not possible to match the relatively low observed values over the latter part. This is not caused solely because of differences in rainfall, since these are reasonably well estimated during the first part and are only moderately overestimated during the second part. This is further demonstrated by comparing the results from the seven selected models whose rainfall time series partly

match the observed time series. None of the simulated inflows from these models matches the relatively selleck chemical extreme decline in observed inflows after 1960. The most likely explanation is that the rainfall inflow relationship used does not adequately represent the real relationship that appears to apply over recent decades, i.e. there appears to be another (effectively unknown) factor involved. As a consequence, it is likely that any long-term inflow projections will tend to be overestimates. Using the median values as a rough guide, Fig. 7 and Fig. 8 indicate that an approximate 25% reduction in rainfall between 1916 and 2085 translates into an approximate 72% reduction in inflows. The ratio (2.9) or “elasticity” factor is consistent with estimates based on analyses of earlier model projections heptaminol and detailed hydrologic modeling. For example, Islam et al. (2013) estimated a reduction (for later this century) of 74%

in runoff associated with a decrease in rainfall of 24% for single catchment within SWWA – a ratio of 3.0. Silberstein et al. (2012) investigated the effect of projected rainfall changes on 13 basins within SWWA – a key feature being that the percentage change in runoff can be up to a factor of three times the percentage change in annual rainfall. However, if the relationship between rainfall and inflows has recently changed, it is quite feasible that, assuming the rainfall projections are realistic, the actual declines could be greater than those simulated here. It is apparent that the protracted dry episode experienced by SWWA since the 1970s has continued up to the present (2013). Secondly, it is also apparent that it is possible to use large-scale average (i.e. SWWA) rainfall to estimate total inflows to Perth dams. This is particularly useful since it implies that climate model results, which are typically only meaningful at these scales, can be directly used to estimate the impacts of projected rainfall changes on inflows, i.e.

Mice were sacrificed by cervical dislocation, and the liver, kidn

Mice were sacrificed by cervical dislocation, and the liver, kidneys, and brain were quickly removed, placed on ice, and homogenized in 10 volumes of cold, Tris buffer (10 mM, pH 7.4). The homogenates were centrifuged at 4000×g at 4 °C for 10 min to yield a low-speed supernatant fraction (S1) for each tissue (liver, kidney and brain) that was used for SNP-induced lipid peroxidation

and H2DCF-DA assays. The antioxidant effect of the PCs was evaluated against production of SNP (5 μM)-induced thiobarbituric acid reactive substances (TBARS), using vehicle, dimethyl sulfoxide (DMSO), or PCs (1–100 μM). The S1 was pre-incubated for 1 h at 37 °C in a buffered medium with the PCs in the presence or absence of SNP. TBARS formation was determined spectrophotometrically

selleckchem at 532 nm, using malondialdehyde (MDA) as a standard, according to Ohkawa et al. (1979). In this work we used the SNP as a mechanism of toxicity, in a concentration of 5 μM according to previously described (Puntel et al., 2009). In fact, sodium nitroprusside (SNP) is a good chemical inducer of lipid peroxidation in mice tissues (Rauhala et al., 1998), since it release in a short-lasting time NO in tissue preparations. The antioxidant effect of the PCs was evaluated against basal production of thiobarbituric acid reactive substances (TBARS), using vehicle, dimethyl sulfoxide (DMSO), selleck compound library or PCs (1–100 μM). The S1 was pre-incubated for 1 h at 37 °C in a buffered medium with the PCs. TBARS formation was determined spectrophotometrically at 532 nm, using malondialdehyde (MDA) as a standard, according to Ohkawa et al. (1979). S1 was used for the 2,7-dichlorodihydrofluorescein diacetate (H2DCF-DA)

oxidation assay to evaluate levels of RS (reactive species). S1, in Tris buffer (10 mM, pH 7.4) was incubated with different PCs at concentrations of 1, 5, 10, 50, and 100 μM at 37 °C. After 1 h, aliquots were removed, H2DCF-DA (7 μM) was added to the medium, and incubation was continued for 1 h in the dark. Fluorescence was determined using 488 nm for excitation and 520 nm for emission. A standard curve was created using increasing concentrations of 2,6-dicloroindophenol sodium salt hydrate (DCF) incubated in parallel (Pérez-Severiano GNAT2 et al., 2004). The results were analyzed as a percentage value in relation to the control group. The protein content was determined according to Lowry et al. (1951), using bovine serum albumin (BSA) as a standard. The scavenging of NO was assessed by incubating SNP (5 mM, in potassium buffer) with different PC concentrations at 25 °C. After 120 min, 0.5 mL of incubation solution was sampled and mixed with 0.5 mL of Griess reagent (Green et al., 1981), and the absorbance was measured at 550 nm. The amount of nitrite was calculated using different concentrations of sodium nitrite.

g Koop et al (1990) and Mort et al (2010) The simulation of a

g. Koop et al. (1990) and Mort et al. (2010). The simulation of ammonium generated from organic matter is split into pathways of nitrification of ammonium, which intensifies with increasing oxygen concentration, and the release of ammonium to the water column. The check details deep parts of the Baltic Sea, such as the Gotland Deep and the Gdańsk Deep, are on occasion characterised by anoxic sediments. Under such conditions nitrification is highly dependent on the dynamics of the redoxcline, which determines the mixing of ammonium-rich waters with oxygenated ones (Hietanen et al. 2012). In the Gulf of Riga, long-term average and minimal oxygen

concentrations rarely reach hypoxic levels and never anoxic levels (Müller-Karulis & Aigars 2011). Furthermore, organic nitrogen mineralisation in the Gulf of Riga delivers large amounts of ammonium (Henriksen & Kemp 1988, Tuominen et al. 1998, Savchuk 2002). Therefore, both ammonium and oxygen supplies should be appropriate for continuous nitrification. However, despite the suitable conditions for nitrification and the reasonable correlation between the simulated and observed ammonium fluxes (Table 1), the dynamics of observed 3Methyladenine ammonium and thus its modelling approach contains some issues that need clarification, for example, the high observed experimental values of NH4+ flux at an O2 concentration

of 2 mg l−1 (Figure 4). This oxygen concentration marks the borderline between hypoxic and oxygenated conditions, as well as the oxygen level needed to sustain most animal life (Hansson et al. 2011). According to Henriksen & Kemp (1988), the higher observed ammonium flux at oxygen concentrations of 2 mg l−1 may be related to the less efficient activity of nitrifying bacteria, which 5-FU in vivo are outcompeted by

heterotrophic bacteria at low oxygen concentrations. Moreover, McCarthy et al. (2008) indicate that the hypoxia threshold provides good conditions for dissimilatory nitrate reduction to ammonium (DNRA). The findings of these authors, as well as the 108% higher NH4+ flux at oxygen concentrations of 2 mg l−1 as compared to ammonium fluxes at lower and higher concentrations (Figure 4), lead us to the conclusion that studies of DNRA and the processes driving it in the Gulf of Riga should be undertaken and that the biogeochemical model should be expanded to include DNRA. Compared to the previously reported results of the biogeochemical model of the Gulf of Riga (Müller-Karulis & Aigars 2011), the simulation of the nitrate flux has been improved in the current study. Here, the simulated nitrate flux increases with oxygen concentration. It is formed as the sum of nitrate diffusion, which marks the nitrate inflow in sediments from the overlying water and thus Dw, and the portion of nitrified nitrate that escapes denitrification, which represents the outward flux from sediments.

, 2003 and Schnoor et al , 2009) Recently, others have demonstra

, 2003 and Schnoor et al., 2009). Recently, others have demonstrated the successful transfection of mRNA into primary murine and human monocytes using mouse macrophage Nucleofector and Human Monocyte Nucleofector kits, INK128 respectively (Zimmermann et al., 2012). Although our methods (specifically

the kit used) are similar to the published study, our unsuccessful attempts could illustrate the need for a cell-type specific Nucleofector kit, optimized for primary rat monocytes, in order to achieve effective transfection. It could also be possible that mRNA transfection is more potent for primary monocyte nucleofections. Despite exhaustive optimization attempts, classical transfection methods (i.e. lipid-based reagents, electroporation,

and nucleofection) were unable to generate stable NGF expression in primary rat monocytes. Although nucleofection generated some NGF expression in monocytes (0.8 ± 0.2 ng/ml NGF per 24 h per 1 million cells), reproducibility was highly variable (21% successful). Even after exhaustive attempts at optimizing the transfection conditions (i.e. plasmid purity, cell purity, various incubation and culture conditions, etc.), we were unable to achieve Selleck LDK378 better reproducible results. Since our interest is to later administer NGF-secreting monocytes in vivo, we concluded that this method would not serve as an attractive method for future experiments. In this study, we demonstrated that lentiviral vectors and Bioporter were the most efficient methods for generating NGF-secreting monocytes. Others have also reported success using viral transduction methods in these cells. Herold et al. (2006) demonstrated that adenoviral infection transduced approximately 95%

of primary monocytes and Mordelet et al. (2002) demonstrated the success and efficiency of lentiviral transduction mafosfamide for monocyte/macrophage gene delivery in rats. In this study, monocytes transduced with lentiviral vector pHR-ba-NGF or pHR-SFFV-NGF produced 15.6 ± 2.5 or 9.1 ± 2.6 ng/ml per 1 million cells, respectively. Both exhibited high reproducibility at 100% and 86%, respectively. Thus, our data is in line with others supporting the use of viral transduction for successful DNA delivery to primary monocytes. Since the use of lentiviral vectors still remains controversial due their immunogenicity properties and we ultimately plan on using these cells for in vivo studies, we also investigated Bioporter as a nonviral approach for generating NGF-secreting monocytes. Bioporter is a protein delivery system that relies on lipid complexes to translocate proteins into target cells. Previous investigations have established that this system can effectively deliver functional recombinant proteins to a wide variety of cell types (Böttger et al., 2010). In the present study, we demonstrate that Bioporter is an efficient nonviral method to deliver NGF to primary rat monocytes, in which monocytes secrete 0.6 ± 0.

e to very short echo times 2τ) The PROJECT (Periodic Refocusing

e. to very short echo times 2τ). The PROJECT (Periodic Refocusing Of J Evolution by Coherence Transfer) approach uses a CPMG sequence with quadrature 90° pulses inserted in the middle of each double spin echo, and is based on the so-called

perfect echo [32] and [33]. The extra 90° pulses refocus J-evolution, for arbitrary τ in AX spin systems and for all spin systems if τJ ≪ 1. If diffusion weighting is added, for example by including field gradient pulses in each echo as in the PROJECTED (PROJECT Extended to DOSY) sequences of Fig. 2, then spin echo DOSY spectra may be obtained free of both exchange effects and J modulation if τk, τJ ≪ 1, where k is the exchange rate constant. The DOSY spectrum of Fig. 1a was acquired for click here a mixture of flavone and catechin. At first sight there appear to be two impurities present. In fact these signals Akt inhibitor are simply the flavone hydroxyl resonances, but their diffusion coefficients are increased by exchange with the small amount of (protio-) water present in the sample. As noted above, such signals are typically better dispersed than backbone proton signals, but serve only to confuse in the spectrum of Fig. 1a. If exchange effects are suppressed using the PROJECTED sequence (Fig.

2, first and last gradient pulses omitted, no 45° pulse), however, the assignment of the signals becomes obvious: hydroxyl and backbone signals alike align correctly in the diffusion domain, as shown in Fig. 1b. Of course the effect is not

limited to exchanging OH signals (which can, if appropriate, be suppressed by addition of D2O), but is general (and extends to magnetization exchange through the Overhauser effect). The use of PROJECT-based DOSY experiments is not limited to cases where exchange is a problem; if, as in small and medium-sized molecules, T2 is not too short, they can be significantly more sensitive than their STE counterparts, because the SE retains the full signal while the STE discards half. However, as there is signal loss due to T2 during the SE delay, for signals with a short T2 a compromise between degree of diffusion weighting and signal-to-noise ratio may be required. Other important examples of applications for PROJECTED include T2-filtered DOSY and convection compensation. T2-filtration is most commonly used where broad signals, for example from polymers or proteins, obscure signals of interest, and is typically implemented in STE-based DOSY pulse sequences by adding CPMG sequences either before [34], or after [35] the STE element. With PROJECTED, diffusion encoding and T2-filtering can be performed simultaneously, minimising signal losses, sample heating and J-modulation. Convection compensation is a particularly attractive application. In STE-based DOSY experiments, convection compensation is normally achieved using a double stimulated echo (DSTE) [22], with a fourfold loss in signal compared to a SE experiment.

Most participants (88%) had sustained moderate or severe TBI and

Most participants (88%) had sustained moderate or severe TBI and over half were more than 1-year postinjury. Standard neurorehabilitation consisted primarily of individual, discipline-specific therapies (physical therapy, occupational therapy, and speech therapy) along with 1 hour KU-60019 price of individual cognitive rehabilitation. The holistic neuropsychologic intervention included individual and group therapies that emphasized metacognitive and emotional regulation for cognitive deficits, emotional difficulties, interpersonal behaviors, and functional skills. Neuropsychologic functioning improved in both conditions, but the holistic neuropsychologic rehabilitation produced greater improvements in community functioning

and productivity, self-efficacy, and life satisfaction. An earlier (class II) study compared these interventions for clinical referrals.119 The study found that participants, despite being more severely disabled and further postinjury, receiving comprehensive-holistic rehabilitation were twice as likely to make clinically significant gains in community functioning than those receiving conventional rehabilitation. Several class II studies of comprehensive-holistic rehabilitation demonstrated reductions in symptoms, improvements in community functioning, and better quality of life compared with conventional treatment120 or no treatment.121 and 122 Results from a class I study,118 several class II studies,119,

120, 121 and 122 and class III studies,123, 124, 125, 128 and 129 are consistent with prior findings suggesting that comprehensive-holistic neuropsychologic rehabilitation Palbociclib price can improve community integration, functional independence, and productivity, even for patients who are many years postinjury.118, 119 and 124 The task force recommends that postacute, comprehensive-holistic neuropsychologic rehabilitation should be provided to reduce cognitive and functional disability after moderate or severe TBI (Practice Standard) ( table 7). Within this context, interventions should address the cognitive, emotional, and interpersonal difficulties of

people with acquired brain injury. Comprehensive-holistic programs typically incorporate a combination in individual and group therapies. There is also evidence Reverse transcriptase for the effectiveness of group treatment for memory deficits, 79 and 91 social communication skills, 38 and 41 aphasia, 131 and executive functioning and problem solving. 109 and 110 Based on this evidence, the task force recommends that group interventions be considered for treating cognitive and communication deficits after TBI and left hemisphere stroke (Practice Option) (see Table 4, Table 5, Table 6 and Table 7). In this systematic review, we evaluated 112 studies of cognitive rehabilitation after TBI or stroke. Based on our current review, we recommend 2 new Practice Standards and the strengthening or refinement of several Practice Standards previously advanced.

The highest DNA binding by 3-NBA in ES cells was observed at 10 μ

The highest DNA binding by 3-NBA in ES cells was observed at 10 μM after 24 h with 863 ± 74 adducts per 108 nucleotides (Fig. 3C). Interestingly, and in contrast to BaP, adduct levels for 3-NBA in MEFs were only 1.5-fold higher

(1266 ± 188 adduct per 108 nucleotides) under the same experimental conditions (Fig. 3D). DNA binding Selleckchem BMS-907351 was highest in MEFs at 10 μM after 48 h with 2478 ± 455 adducts per 108 nucleotides. Previously, in primary HUFs previously treated with 10 μM 3-NBA for 48 h, adduct levels were 680 ± 147 adducts per 108 nucleotides (Kucab et al., 2012). As 3-NBA is predominantly activated by NQO1 (Arlt et al., 2005), the expression of Nqo1 was studied in ES cells and MEFs by RT-PCR and revealed that Nqo1 mRNA expression increased in both cell types up to ∼60-fold; the induction was higher in MEFs than in ES cells ( Fig. 6C and D). This is in line with a previous study showing that Nqo1 protein levels were inducible in primary and immortal HUFs upon treatment with nitro-PAHs such as 1,8-dinitropyrene and 3-NBA ( Kucab et al., 2012). However, that study also showed that there was not a clear relationship between nitro-PAH-induced DNA adduct formation and the expression of Nqo1, suggesting

that other cytosolic nitroreductases such as xanthine oxidase might also contribute to the activation of nitro-PAHs like 3-NBA in HUFs ( Kucab et al., 2012). As shown in Fig. 5C and D, 3-NBA also induced Cyp1a1 mRNA expression, the induction in MEFs being manifoldly higher than in very ES cells. Other studies have Dasatinib price demonstrated the induction of Cyp1a1 protein levels in mouse Hepa1c1c7 cells after exposure to 3-NBA treatment ( Landvik et al., 2010) and in vivo in rats treated with 3-NBA ( Mizerovska et al., 2011, Stiborova et al., 2006 and Stiborova et al., 2008). The major activation pathway of AAI is

nitroreduction, cytosolic NQO1 being the most efficient activating enzyme while CYP1A-mediated demethylation contributes to AAI detoxification (Fig. 1C) (Stiborova et al., 2014a and Stiborova et al., 2013). Exposure to AAI resulted in loss of cell viability of both ES cells and MEFs (Fig. 2E and F). However, in contrast to 3-NBA which showed strong cytotoxicity in ES cells, AAI cytotoxicity was higher in MEFs. We therefore chose 20 μM and 50 μM AAI in MEFs while ES cells were treated with up to 100 μM for DNA adduct analysis by 32P-postlabelling (Fig. 3E and F). The AAI-induced adduct patterns in ES cells and MEFs were the same and identical to the patterns observed in kidney and ureter tissue of AAN patients (Gokmen et al., 2013 and Nortier et al., 2000). These adducts have previously been identified as 7-(deoxyadenosine-N6-yl)aristololactam I (dA-AAI; spot A1), 7-(deoxyguanosin-N2-yl)aristolactam I (dG-AAI; spot A2) and 7-(deoxyadenosin-N6-yl)aristolactam II (dA-AAII; spot A3) ( Bieler et al., 1997 and Schmeiser et al., 2014).

At fixed intervals of 10, 30, 60, 90, 120, 180 and 240 min after

stylifera females (N = 20) into its central part (0). At fixed intervals of 10, 30, 60, 90, 120, 180 and 240 min after the start, percentages of copepods in (+), (−) and (0) were assessed by counting the number of females in each area and dividing it by the total number of copepods

actually counted in the vessel at that time. Three replicate experiments were performed, every time using freshly prepared agarose gels and changing the orientation of the vessel with respect to the experimenter and to the light conditions in the room. In two replicates, the vessel was placed vertically with (+) located at the same side or at the opposite side of the observer, whereas in the third replicate, the vessel was placed horizontally with (+) located on the left side of the observer. Filtration and ingestion rates of T. stylifera females on P. minimum were higher in DD treatments ( Fig. 1A, B). On average, Talazoparib chemical structure filtration rates selleck chemicals increased from 0.19 ± 0.12 mL ind−1 h−1 for controls to 0.40 ± 0.14 and 0.47 ± 0.04 mL ind−1 h−1 for 2.0 μg mL−1 and 0.5 μg mL−1 DD, respectively ( Fig. 1A). Ingestion rates increased from 0.20 ± 0.11 μg C ind−1 h−1 for controls to 0.40 ± 0.13 and 0.44 ± 0.03 μg C ind−1 h−1 for 1.0 μg mL−1 and 0.5 μg mL−1 DD, respectively ( Fig. 1B). Although the differences between the control (DD 0), 0.5 μg mL−1 and 2.0 μg mL−1 DD were only significant for filtration rate (1-way ANOVA, df = 2, F = 5.368, p = 0.0461),

but not ingestion rate (1-way ANOVA, df = 2, F = 4.997, p = 0.0532), ingestion and filtration rates almost doubled between controls and 0.5 μg mL−1 (Student-t test p < 0.05, for both rates). Egg production rate (EPR) increased with increasing DD concentration, with values ranging from 23.5 eggs female−1 day−1 (0.0 μg mL−1 DD) in controls to 33.8 eggs female−1 day−1 at 2 μg mL−1 DD (Fig. 2A). Egg hatching time (EHT) increased in DD treatments, ranging on average from 19.4 h in controls to 20.7 h at 1.0 μg mL−1 DD (Fig. 2B). Egg hatching success (EHS) decreased in DD treatments with values ranging on average from 97% in controls to 54% at 2 μg mL−1 DD (Fig. 2C).

There was no significant difference between treatments for fecundity (1-way ANOVA, df = 3, F = 1.846, p = 0.161) and EHS (1-way ANOVA, df = 3, F = 2.482, p = 0.081), but a ID-8 significant difference for EHT (1-way ANOVA, df = 3, F = 4.603, p = 0.010). Survivorship was high for both females and males (on average 75–100%) for controls (0.0 DD) and DD concentrations between 0.5 and 2.0 μg mL−1 (Fig. 3). Survivorship decreased drastically above 3.0 μg mL−1 DD, with values ranging from 0 to 42% and 0 to 17% for females and males, respectively. The percentage of apoptotic nauplii increased from 25% in controls to a maximum of 64% at 1.0 μg mL−1.