Quantitative prediction of CYP3A induction-mediated drug-drug interactions in clinical practice.

There have been no reports on the quantitative prediction of CYP3A induction-mediated decreases in AUC and Cmax for drug candidates identified as a "victims" of CYP3A induction. Our previous study separately evaluated the fold-induction of hepatic and intestinal CYP3A by known inducers using clinical induction data and revealed that we were able to quantitatively predict the AUC ratio (AUCR) of a few CYP3A substrates in the presence and absence of CYP3A inducers. In the present study, we investigate the predictability of AUCR and also Cmax ratio (CmaxR) in additional 54 clinical studies. The fraction metabolized by CYP3A (fm), the intestinal bioavailability (Fg), and the hepatic intrinsic clearance (CLint) of substrates were determined by the in vitro experiments as well as clinical data used for calculating AUCR and CmaxR. The result showed that 65-69% and 65-67% of predictions were within 2-fold of observed AUCR and CmaxR, respectively. A simulation using multiple parameter combinations suggested that the variability of fm and Fg within a certain range might have a minimal impact on the calculation output. These findings suggest that clinical AUCR and CmaxR of CYP3A substrates can be quantitatively predicted from the preclinical stage.

Changes in spinal motoneuron excitability during the improvement of fingertip dexterity by actual execution combined with motor imagery practice.

Since there is an upper limit to skill improvement through the repetition of actual execution, we examined whether motor imagery could be used in combination with actual execution to maximize motor skill improvement. Fingertip dexterity was evaluated in 25 healthy participants performing a force adjustment task using a pinch movement with the left thumb and index finger. In the intervention condition, six sets of repetitions of combined actual execution and motor imagery were performed, while in the control condition, the same flow was performed, but with motor imagery replaced by rest. Changes in the excitability of spinal motoneurons during motor imagery compared to rest were compared in terms of the F/M amplitude ratio. Motor skill changes were compared before and after repeated practice and between the conditions, respectively, using the absolute amount of adjustment error between the target pinch force value and the delivered pinch force value (absolute error) as an index. The results showed that the repetition of exercise practice and motor imagery decreased the absolute error, which was greater than that of exercise practice alone in terms of motor skill improvement. The F/M amplitude ratio for motor imagery compared to rest did not increase. This suggests that motor imagery is involved in the degree of the increase of spinal motoneuron excitability based on the real-time prediction of motor execution and that there may be no need for an increase in excitability during motor skill control.

Improving the Continuous Multiple Tube Reactor: an Innovative Bioreactor Configuration with Great Potential for Dark Fermentation Processes.

The continuous multiple tube reactor (CMTR) has been developed as a promising technology to maximize biohydrogen production (BHP) by dark fermentation (DF) by preventing excess biomass accumulation, leading to suboptimum values of specific organic loading rates (SOLR). However, previous experiences failed to achieve stable and continuous BHP in this reactor, as the low biomass retention capacity in the tube region limited controlling the SOLR. This study goes beyond the evaluation of the CMTR for DF by inserting grooves in the inner wall of the tubes to ensure better cell attachment. The CMTR was monitored in 4 assays at 25ºC using sucrose-based synthetic effluent. The hydraulic retention time (HRT) was fixed at 2 h, while the COD varied between 2-8 g L-1 to obtain organic loading rates in the 24 - 96 g COD L-1 d-1. Long-term (90 d) BHP was successfully attained in all conditions due to the improved biomass retention capacity. Optimal values for the SOLR (4.9 g COD g-1 VSS d-1) were observed when applying up to 48 g COD L-1 d-1, in which BHP was maximized. These patterns indicate a favorable balance between biomass retention and washout was naturally achieved. The CMTR looks promising for continuous BHP and is exempt from additional biomass discharge strategies.

In hot water: Interactions of temperature, nitrogen form and availability and photosynthetic and nitrogen uptake responses in natural Karenia brevis populations.

During 2020-2021, an unusually prolonged bloom of the toxigenic dinoflagellate Karenia brevis persisted for more than 12 months along the Gulf coast of Florida, resulting in severe environmental effects. Motivated by the possibility that unusual nutrient conditions existed during summer 2021, the short-term interactions of temperature, nitrogen (N) forms (ammonium (NH4+), nitrate (NO3-), and urea) and availability on photosynthesis-irradiance responses and N uptake rates were examined in summer 2021 and compared to such responses from the earlier winter. Winter samples were exposed to temperatures of 15, 20, 25, 30 °C while summer samples were incubated at 15, 25, 30, 33 °C, representing the maximum range the cells might experience throughout the water column due to daytime surface heating or extreme weather events. Depending on thermal history of the cells, photosynthetic performance differed when cells were exposed to the same temperature, showing a capacity for thermal acclimation in this species. Although blooms generally do not persist throughout the summer, bloom biomass was remarkably higher in summer than during the winter. However, most of the photosynthetic parameters and N uptake rates, as well as total carbon (C) and N cell-1 were significantly lower in the summer populations, showing that the summer populations were photosynthetically and nutritionally stressed. When the summer cells were treated with urea, however, uptake rates and total C and N cell-1 were higher than with the other N substrates, especially in warmer waters, showing differential thermal responses depending on N forms.

Deciphering the molecular basis for photosynthetic parameters in Bambara groundnut (Vigna subterranea L. Verdc) under drought stress.

BACKGROUND: Assessment of segregating populations for their ability to withstand drought stress conditions is one of the best approaches to develop breeding lines and drought tolerant varieties. Bambara groundnut (Vigna subterranea L. Verdc.) is a leguminous crop, capable of growing in low-input agricultural systems in semi-arid areas. An F4 bi-parental segregating population obtained from S19-3 × DodR was developed to evaluate the effect of drought stress on photosynthetic parameters and identify QTLs associated with these traits under drought-stressed and well-watered conditions in a rainout shelter. RESULTS: Stomatal conductance (gs), photosynthesis rate (A), transpiration rate (E) and intracellular CO2 (Ci) were significantly reduced (p < 0.05) while water use efficiency (WUE) was significantly increased (p < 0.05) under drought-stressed conditions. A strong linear correlation was observed between gs, WUE, A, E and Ci under both water regimes. The variability between different water treatment, among individual lines and the interaction between lines and environment for photosynthetic parameters provides resources for superior lines selection and drought resistant variety improvement. Significant QTL for gs and FV/FM under well-watered conditions were mapped on LG5 and LG3, respectively, with more than 20% of the PVE, which could be considered as the major QTL to control these traits. Five clustered QTLs for photosynthetic traits under drought-stressed and well-watered conditions were mapped on LG5, LG6A, LG10 and LG11, respectively. CONCLUSIONS: Significant and putative QTLs associated with photosynthetic parameters and the effect of drought stress on these traits have been revealed by QTL linkage mapping and field experiment in the F4 segregating population derived from S19-3 × DodR in bambara groundnut. The study provides fundamental knowledge of how photosynthetic traits response to drought stress and how genetic features control these traits under drought-stressed and well-watered conditions in bambara groundnut.

Respiratory function monitoring during neonatal resuscitation: A systematic review.

Aim: Positive pressure ventilation via a facemask is critical in neonatal resuscitation, but frequently results in mask leak, obstruction, and inadequate respiratory support. This systematic review aimed to determine whether the display of respiratory function monitoring improved resuscitation or clinical outcomes. Methods: Randomized controlled trials comparing outcomes when respiratory function monitoring was displayed versus not displayed for newborns requiring positive pressure ventilation at birth were selected and from databases (last search August 2022), and assessed for risk of bias using Cochrane Risk of Bias Tools for randomized control trials. The study was registered in the Prospective Register of Systematic Reviews. Grading of Recommendations, Assessment, Development and Evaluations was used to assess the certainty of evidence. Treatment recommendations were approved by the Neonatal Life Support Task Force of the International Liaison Committee on Resuscitation. Results reported primary and secondary outcomes and included resuscitation and clinical outcomes. Results: Of 2294 unique articles assessed for eligibility, three randomized controlled trials were included (observational studies excluded) (n = 443 patients). For predefined resuscitation and clinical outcomes, these studies either did not report the primary outcome (time to heart rate ≥ 100 bpm from birth), had differing reporting methods (achieving desired tidal volumes, significant mask leak) or did not find significant differences (intubation rate, air leaks, death before hospital discharge, severe intraventricular hemorrhage, chronic lung disease). Limitations included limited sample size for critical outcomes, inconsistent definitions amongst studies and unreported long-term outcomes. Conclusion: Although respiratory function monitoring has been utilized in clinical care, there is currently insufficient evidence to suggest its benefit for newborn infants receiving respiratory support for resuscitation at birth. Registration: PROSPERO CRD42021278169 (registered November 27, 2021). Funding: The International Liaison Committee on Resuscitation provided support that included access to software platforms and teleconferencing.

Formulation of a protocol to evaluate the aerobic granulation potential (AGP) of an inoculum.

This paper proposes and develops a protocol for measuring the aerobic granulation potential of sludge, aiming to provide an affordable and simple alternative that can facilitate the development of aerobic granulation technology. In this sense, the protocol comprises a set of parameters and considerations that interact to create a controlled environment and stimulate cell population clustering. All of this is done in the context of procedural simplicity, low cost, and the speed at which results are obtained. The protocol is essentially a three-stage method: preparation of the substrate, adaptation of the inoculum, and implementation of the protocol. Simple parameters were measured to evaluate the granulation process: SVI, settling velocity, and morphological parameters. The protocol was validated according to optimal ranges and criteria previously established in the literature. For this purpose, an activated sludge inoculum from a domestic wastewater treatment plant was submitted to the protocol, obtaining an optimal response of the biomass (SVI5 =13.90 mL g-1, settling velocity= 25,79 m h-1, Diameter > 0.2 mm) in a relatively short time (7 d). The results show that this protocol can constitute a tool for evaluation and decision-making using traditional laboratory equipment and is applicable at different scales.

[Impacts of F/M Ratio on Microbial Networks in Activated Sludge].

The performance of wastewater treatment plants (WWTPs) mainly relies on the microbial community in activated sludge (AS). The food to microorganisms (F/M) ratio is an important operational parameter, the recommended value of which is between 0.2 and 0.5 kg·(kg·d)-1. The F/M directly influences microbial growth and metabolism, but how the F/M ratio affects AS microbial networks and the related mechanisms remain unclear. In this study, 63 AS samples with different F/M ratios were analyzed by utilizing a random-matrix-theory based network pipeline. Firstly, all 63 samples were divided into three groups based on their F/M ratios (lower F/M group, normal F/M group, and higher F/M group). The results indicated that the lower F/M decreased microbial diversity and evenness significantly, but the higher F/M had no significant effects on the diversity of the microbial community. All three constructed networks were scale-free, small world, and modular, but network size and complexity were decreased in the lower and higher F/M groups. The network of the normal F/M ratio group was detected with the most nodes, the highest average clustering coefficient, and the shortest geodesic distance. The proportion of positive links was 76.8% when the F/M was within the normal range. On the contrary, 71.1% and 60.0% of positive links were identified when the F/M was below 0.2 and above 0.5 kg·(kg·d)-1. Moreover, 24 keystones were detected in the normal F/M network, whereas only 4 and 7 keystones were detected in the lower and higher F/M networks. Overall, our results provided clear evidence that the AS microbial community could be more stable and resilient to environmental disturbance when the F/M ratio was between 0.2 and 0.5 kg·(kg·d)-1.

My time with Franz Enzinger at the Armed Forces Institute of Pathology.

By a set of curious chances, Dr. F.M.Enzinger went from being captured as a prisoner of war by the American Army in Normandy in 1944 to become Head of the Soft Tissue Department of the Armed Forces Institute of Pathology (AFIP) in Washington DC and a friend of one of the directors, Major General Joe M. Blumberg. The author relates how he worked under Enzinger from 1967 to 1970. Enzinger would review all the slides of referred consultation cases on alternate days and record his diagnoses on filing cards kept in his personal file which supplemented the computerised AFIP central file. Periodic reviews of the files allowed him to retrieve large numbers of recognised tumors and to study potential cases of previously undescribed entities. During his 31 years at the AFIP, Enzinger lectured widely, wrote numerous papers on recognised entities, co-authored with Dr. Sharon W. Weiss their famous text book and discovered the following 26 previously undefined soft tissue tumors: clear cell sarcoma, intramuscular myxoma, epithelioid sarcoma, extra-skeletal myxoid chondrosarcoma, fetal rhabdomyoma, extra-skeletal Ewing's sarcoma, spindle cell lipoma, fibroma of tendon sheath, angiomatoid (malignant) fibrous histiocytoma, cranial fasciitis of childhood, intravascular fasciitis, pleomorphic lipoma, neuromuscular hamartoma, epithelioid hemangioendothelioma, spindle cell hemangioma, fibro-osseous pseudotumor of the digits, plexiform fibrohistiocytic tumor, giant cell fibroblastoma as a juvenile form of dermatofibrosarcoma protuberans, ossifying fibromyxoid tumor, myolipoma of soft tissues, inflammatory fibrosarcoma, juxta-articular myxoma, atypical decubital fibroplasia, chondroid lipoma, sclerosing epithelioid fibrosarcoma, and lipofibromatosis. His last discovery, co-authored by some of his AFIP followers, was published in 2000. He died in 2006. The AFIP itself was "disestablished" only five years later, which will make it very difficult for any future pathologist to exceed Enzinger's astonishing record of 26 "new" tumor discoveries.

De novo chromosome-level genome of a semi-dwarf cultivar of Prunus persica identifies the aquaporin PpTIP2 as responsible for temperature-sensitive semi-dwarf trait and PpB3-1 for flower type and size.

Peach (Prunus persica) is one of the most important fruit crops globally, but its cultivation can be hindered by large tree size. 'Zhongyoutao 14' (CN14) is a temperature-sensitive semi-dwarf (TSSD) cultivar which might be useful as breeding stock. The genome of CN14 was sequenced and assembled de novo using single-molecule real-time sequencing and chromosome conformation capture assembly. A high-quality genome was assembled and annotated, with 228.82 Mb mapped to eight chromosomes. Eighty-six re-sequenced F1 individuals and 334 previously re-sequenced accessions were used to identify candidate genes controlling TSSD and flower type and size. An aquaporin tonoplast intrinsic protein (PpTIP2) was a strong candidate gene for control of TSSD. Sequence variations in the upstream regulatory region of PpTIP2 correlated with different transcriptional activity at different temperatures. PpB3-1, a candidate gene for flower type (SH) and flower size, contributed to petal development and promoted petal enlargement. The locus of another 12 agronomic traits was identified through genome-wide association study. Most of these loci exhibited consistent and precise association signals, except for flesh texture and flesh adhesion. A 6015-bp insertion in exon 3 and a 26-bp insertion upstream of PpMYB25 were associated with fruit hairless. Along with a 70.5-Kb gap at the F-M locus in CN14, another two new alleles were identified in peach accessions. Our findings will not only promote genomic research and agronomic breeding in peach but also provide a foundation for the peach pan-genome.

Application of Positron Annihilation Spectroscopy in Accelerator-Based Irradiation Experiments.

Positron annihilation spectroscopy (PAS) is widely recognized as a powerful characterization technique in all types of radiation damage studies in nuclear materials. In the past, fission reactor irradiation of reactor pressure vessel (RPV) steels was a primary aim in most studies, while today's applications of PAS in this field are centered around ion implantation experiments in advanced structural materials. These experiments use hydrogen, helium, heavy ions, and their combination to simulate various radiation environments of future nuclear reactors or nuclear research facilities. The spectrum of ion energies used ranges from a few tens of keV to tens or even hundreds of MeV in proton irradiation or spallation neutron source irradiation experiments. The variety of ion energies, irradiation temperatures, and other experimental conditions poses a major challenge to researchers, who often fail to successfully incorporate the lessons learned from their research. In this paper, we review and supplement recent PAS studies in which structural materials irradiated under a variety of irradiation conditions were investigated using positron annihilation spectroscopy. It summarizes the most important conclusions and lessons learned from the application of PAS in accelerator-based irradiation experiments.

Direct measurement of neuronal ensemble activity using photoacoustic imaging in the stimulated Fos-LacZ transgenic rat brain: A proof-of-principle study.

Measuring neuroactivity underlying complex behaviors facilitates understanding the microcircuitry that supports these behaviors. We have developed a functional and molecular photoacoustic tomography (F/M-PAT) system which allows direct imaging of Fos-expressing neuronal ensembles in Fos-LacZ transgenic rats with a large field-of-view and high spatial resolution. F/M-PAT measures the beta-galactosidase catalyzed enzymatic product of exogenous chromophore X-gal within ensemble neurons. We used an ex vivo imaging method in the Wistar Fos-LacZ transgenic rat, to detect neuronal ensembles in medial prefrontal cortex (mPFC) following cocaine administration or a shock-tone paired stimulus. Robust and selective F/M-PAT signal was detected in mPFC neurons after both conditions (compare to naive controls) demonstrating successful and direct detection of Fos-expressing neuronal ensembles using this approach. The results of this study indicate that F/M-PAT can be used in conjunction with Fos-LacZ rats to monitor neuronal ensembles that underlie a range of behavioral processes, such as fear learning or addiction.

Grafting in Hylocereus (Cactaceae) as a tool for strengthening tolerance to high temperature stress.

The Hylocereus species that are grown as exotic fruit crops are very often farmed under marginal agronomic conditions, which may include exposure to high temperatures. Here we present a pioneering investigation of grafting as an agro-technique to improve heat tolerance in Hylocereus. To this end, we studied the diploid species H. undatus, the tetraploid H. megalanthus and its di-haploid gamete-derived line 2719, and the interspecific-interploid tetraploid Z-10, all grafted onto H. undatus as the rootstock. Self-grafted, grafted and non-grafted plants were acclimated for one week (to obtain baseline values) and then exposed to heat stress (45/35 °C day/night) for three days, followed by a one-week recovery period under optimal temperatures (30/22 °C). A comparison of the physiological, biochemical and molecular performances of the grafted and self-grafted plants under heat stress and during the recovery period vs those of non-stressed plants (control; 30/22 °C) showed that the grafted and self-grafted plants performed better in most of the assessments: grafted and self-grafted plants recovered more rapidly from the heat stress and suffered far less stem damage. An unexpected - but important - finding that may have implications for other crop was that the self-grafted plants showed better performance than non-grafted plants throughout the trial. Our findings provide support for grafting as a strategy for coping with the stress induced by extremely high temperatures. This study thus paves the way for further investigations of grafting in Hylocereus as a valuable technique that will maintain crop productivity in the face of increasing worldwide temperatures.

Current condition of Bathypogon types (Diptera: Asilidae) described by Frank M. Hull and housed in the South Australian Museum of Natural History with notes on other Bathypogon types.

The current condition of the types of Bathypogon, described by Frank M. Hull 1956-1959, are listed. By his own admission, a portion of the material borrowed from the South Australian Museum was destroyed by "pests". Note is made that the holotype of Bathypogon microdonturus is missing and presumed destroyed. Additions are made to the published data based on data on the Type specimen labels. Photos of the hypandrium of some Bathypogon holotypes that reside in the SA Museum collection are provided.

Clinical and Physiological Significance of F-Wave in Spinocerebellar Ataxia Type 3.

Objective: To evaluate the characteristics of F-wave in spinocerebellar ataxia type 3 (SCA3) patients and preclinical carriers of SCA3 gene mutation (PreSCA3), and explore the relationship between disease severity and F-wave parameters and evaluate F-wave parameters as potential biomarkers for monitoring of disease progression in SCA3. Methods: We performed F-wave recordings in median, ulnar and tibial nerves of 39 SCA3 patients, 20 PreSCA3, and 27 healthy controls, and compared F-wave parameters between them. Results: In all nerves studied, the mean F-wave amplitude, maximum F-wave amplitude, and F/M amplitude ratio were significantly increased in the SCA3 patients in comparison with the normal controls. And the minimal F-wave latency of SCA3 patients was significantly prolonged and the F-wave persistence (%) was significantly decreased in the median nerve. For the PreSCA3, the maximum F-wave amplitude was significantly higher than normal controls for both median, ulnar, and tibial nerves. The mean F-wave amplitude and F/M amplitude ratio in all nerves were comparable between PreSCA3 and normal controls. The frequency of giant F-wave and frequency of patients with giant F-wave were similar between PreSCA3 and SCA3. The values of F/M amplitude ratio in both median, ulnar, and tibial nerves were correlated positively with disease severity and disease duration. Conclusion: Significant F-wave abnormalities occur in patients with SCA3, even in PreSCA3. F-wave may therefore reveal subclinical alterations and provide objective parameters for evaluating the progression of SCA3.

Cell-by-cell estimation of PAH sorption and subsequent toxicity in marine phytoplankton.

Polycyclic Aromatic Hydrocarbons (PAHs) have elicited increasing concern due to their ubiquitous occurrence in coastal marine environments and resultant toxicity in organisms. Due to their lipophilic nature, PAHs tend to accumulate in phytoplankton cells and thus subsequently transfer to other compartments of the marine ecosystem. The intrinsic fluorescence properties of PAHs in the ultraviolet (UV)/blue spectral range have recently been exploited to investigate their uptake modes, localization, and aggregation in various biological tissues. Here, we quantitatively evaluate the sorption of two model PAHs (phenanthrene and pyrene) in three marine phytoplankton species (Chaetoceros tenuissimus, Thalassiosira sp. and Proteomonas sp.) using a combined approach of UV excitation flow cytometry and fluorescence microscopy. Over a 48-h exposure to a gradient of PAHs, Thalassiosira sp. showed the highest proportion of PAH-sorbed cells (29% and 97% of total abundance for phenanthrene and pyrene, respectively), which may be attributed to its relatively high total lipid content (33.87 percent dry weight). Moreover, cell-specific pulse amplitude modulation (PAM) microscope fluorometry revealed that PAH sorption significantly reduced the photosynthetic quantum efficiency (Fv/Fm) of individual phytoplankton cells. We describe a rapid and precise hybrid method for the detection of sorption of PAHs on phytoplankton cells. Our results emphasize the ecologically relevant sub-lethal effects of PAHs in phytoplankton at the cellular level, even at concentrations where no growth inhibition was apparent. This work is the first study to address the cell-specific impacts of fluorescent toxicants in a more relevant toxicant-sorbed subpopulation; these cell-specific impacts have to date been unidentified in traditional population-based phytoplankton toxicity assays.

Identification and quantitation of enzyme and transporter contributions to hepatic clearance for the assessment of potential drug-drug interactions.

Drug-drug interactions (DDIs) involving drug-metabolizing enzymes and membrane transporters can lead to alteration in substrate drug (victim) exposure, and can influence the pharmacological and toxicological effects. In order to predict DDI potential, it is important to quantitatively characterize the major enzyme(s) and/or transporter(s) involved in the clearance of drugs, in terms of fraction metabolized (fm) and fraction transported (ft). In this review, we discuss a strategy using Extended Clearance Classification System (ECCS) to identify the clearance mechanism(s) early in drug discovery, and subsequently rational staging of in vitro characterization to determine fm and ft. In addition, the examples of complex DDIs due to involvement of transporter-enzyme interplay in the hepatic clearance are discussed.

Anaerobic co-digestion of industrial landfill leachate and glycerin: methanogenic potential, organic matter removal and process optimization.

The objective of this study was to evaluate the performance of the anaerobic co-digestion of different concentrations of industrial landfill leachate associated with crude residual glycerin, in relation to the methanogenic potential, COD removal, accumulated methane production, the effects of the factors (food/microorganism ratio and percentage of glycerin added to the leachate) and their interactions on kinetic parameters of methane production (CH4) using the modified Gompertz model. Co-digestion tests were carried out in bench scale (400 mL of useful volume) under batch mode at 30 ± 1°C during 30-day incubation of anaerobic sewage sludge as inoculum. The parameters glycerin addition to the leachate (v/v) (0%, 1.5%, 5%, 8.5% and 10%) and F/M ratio (0.3, 0.5, 1, 1.5 and 1.7) were investigated using Central Composite Rotational Design method (CCRD). The results indicated significant effect to the response variables: methanogenic potential, COD removal, accumulated production of CH4 and maximum estimated production of CH4, considering a confidence interval of 95% (p < .05). The ideal mixture of 95.13% of leachate with 4.87% of raw glycerin was obtained by desirability test to F/M of 1.61 gCOD of substrate per gVSS (volatile suspended solids) of sludge. Methanogenic potential was 0.19 LNCH4 gTVSrem -1, and the average removal of COD was 92%, resulting in accumulated production and maximum estimated production of CH4 of 74 and 80 mL, respectively. It was noted that the process of co-digestion of the industrial landfill leachate with the crude residual glycerin is promising, due to is potential of complementing and balancing organic materials, nutrients and other components that influence the biological process. Abbreviations: AN: ammoniacal nitrogen; BMP: biochemical methane potential; CCRD: central composite rotational design; COD: chemical oxygen demand; F/M ratio: food/microorganism ratio; FSS: fixed suspended solids; NTP: normal temperature and pressure; TSS: total suspended solids; TA: total alkalinity; TFS: total fixed solids; TKN: total Kjeldahl nitrogen; TP: total phosphorus; TS: total solids; TVA: total volatile acids; TVS: total volatile solids; VSS: volatile suspended solids; WWTP: wastewater treatment plant.

Is Karenia brevis really a low-light-adapted species?

Despite nearly annual blooms of the neurotoxic dinoflagellate Karenia brevis (Davis) G. Hansen and Moestrup in the Gulf of Mexico, defining the suite of biological traits that explain its proliferation has remained challenging. Studies have described K. brevis as a low-light-adapted species, incapable of sustaining growth under high light, which is at odds with observed surface aggregations sometimes within centimeters of the sea surface and also with short-term experiments showing photosynthetic machinery accommodating high irradiances. Here, growth and photophysiology of three K. brevis isolates were evaluated under a range of environmentally relevant irradiances (10-1500 µmol photons m-2 s-1) in the laboratory. No differences in growth-irradiance curves were observed among isolates; all sustained maximum growth rates at the highest irradiances examined, even in exposures as long as three weeks. The growth efficiency α of K. brevis under light-limiting conditions appeared mediocre among dinoflagellates, and poorer than that of other phytoplankton (e.g., diatoms, cyanobacteria), implying that K. brevis is not a low-light specialist. This finding substantially alters earlier parameterizations of K. brevis growth-irradiance curves. Therefore, a model was developed to contextualize how these new growth-irradiance curves might affect bottom growth rates. This model was subsequently applied to a case study comparing seasonal light forcing offshore of Pinellas County, FL, USA, with a single empirical value for light attenuation, and seasonal bottom water temperatures. Predictions suggested that light may limit bottom growth as close as 1 km from shore in winter, but would only begin limiting growth 20 km from shore in summer. Population maintenance (no net growth) was possible as far offshore as 90 km in summer and 68 km in winter. These ranges intercept areas thought to be important for bloom initiation.

Cell cycle regulation of the mixotrophic dinoflagellate Dinophysis acuminata: Growth, photosynthetic efficiency and toxin production.

The mixotrophic dinoflagellate Dinophysis acuminata is a widely distributed diarrhetic shellfish poisoning (DSP) producer. Toxin variability of Dinophysis spp. has been well studied, but little is known of the manner in which toxin production is regulated throughout the cell cycle in these species, in part due to their mixotrophic characteristics. Therefore, an experiment was conducted to investigate cell cycle regulation of growth, photosynthetic efficiency, and toxin production in D. acuminata. First, a three-step synchronization approach, termed "starvation-feeding-dark", was used to achieve a high degree of synchrony of Dinophysis cells by starving the cells for 2 weeks, feeding them once, and then placing them in darkness for 58 h. The synchronized cells started DNA synthesis (S phase) 10 h after being released into the light, initiated G2 growth stage eight hours later, and completed mitosis (M phase) 2 h before lights were turned on. The toxin content of three dominant toxins, okadaic acid (OA), dinophysistoxin-1 (DTX1) and pectenotoxin-2 (PTX2), followed a common pattern of increasing in G1 phase, decreasing on entry into the S phase, then increasing again in S phase and decreasing in M phase during the diel cell cycle. Specific toxin production rates were positive throughout the G1 and S phases, but negative during the transition from G1 to S phase and late in M phase, the latter reflecting cell division. All toxins were initially induced by the light and positively correlated with the percentage of cells in S phase, indicating that biosynthesis of Dinophysis toxins might be under circadian regulation and be most active during DNA synthesis.