The role of the carapace in the accumulation of metals from seawater in the green crab (Carcinus maenas): Studies with radio-labeled calcium, zinc, and nickel.

The role of the carapace in the uptake and storage of newly accumulated metals was investigated in the green crab exposed to environmentally relevant concentrations of calcium ([Ca] = 389 mg L-1 or 9.7 mmol L-1), zinc ([Zn] = 82 µg L-1 or 1.25 µmol L-1), and nickel ([Ni] = 8.2 µg L-1 or 0.14 µmol L-1) in 12 °C seawater, using radio-tracers (45Ca, 65Zn, 63Ni). After 24-h exposure, carapace exhibited the highest concentration of newly accumulated Ca, whereas carapace and gills exhibited the highest concentrations of both newly accumulated Zn and Ni relative to other tissues. For all three metals, the carapace accounted for >85 % of the total body burden. Acute temperature changes (to 2 °C and 22 °C) revealed the highest overall temperature coefficient Q10 (2.15) for Ca uptake into the carapace, intermediate Q10 for Ni (1.87) and lowest Q10 (1.45) for Zn. New Ca uptake into the carapace continued linearly with time for 24 h, new Zn uptake gradually deviated from linearity, whereas Ni uptake reached a plateau by 6 h. Attachment of a rubber membrane to the dorsal carapace, thereby shielding about 20 % of the total crab surface area from the external water, eliminated both new Zn and Ni incorporation into the shielded carapace, whereas 36 % of new Ca incorporation persisted. When recently euthanized crabs were exposed, new Zn uptake into the carapace remained unchanged, whereas Ca and Ni uptake were reduced by 89 % and 71 %, respectively. We conclude that the carapace is a very important uptake and storage site for all three metals. All of the uptake of new Zn and new Ni, and most of the uptake of new Ca into this tissue comes directly from the external water. For Zn, the mechanism involves only physicochemical processes, whereas for Ca and Ni, life-dependent processes make the major contribution.

Influence of a single locking throw on the in vitro holding security of five friction throws.

OBJECTIVE: To compare the in vitro security of four different friction throws (square, surgeon's, Miller's, and strangle), with and without a locking throw, and a slip knot, on a vascular ligation model. STUDY DESIGN: Experimental study. SAMPLE POPULATION: Nine groups with nine samples per group. METHODS: Four throws (square, surgeon's, Miller's, and strangle), with and without a locking throw, and a square throw locked in slip knot fashion, were tested to measure leakage pressure. RESULTS: The square and surgeon's throws leaked at a lower pressure than the Miller's and strangle throws (p < .0001). The leakage pressure was below physiologic arterial pressure for the square and the surgeon's throws. After the addition of a locking throw, the five knots leaked at a similar pressure (p = .5233) above physiologic arterial pressure. CONCLUSION: Following the addition of a locking throw, all the constructs tested in this study leaked at a similar pressure. The leakage pressure for all knots exceeded physiologic arterial pressures. CLINICAL SIGNIFICANCE: Any throw tested may be appropriate for secure vascular ligation if the initial throw is applied correctly.

Do extreme postprandial levels of oxygen, carbon dioxide, and ammonia in the digestive tract equilibrate with the bloodstream in the freshwater rainbow trout (Oncorhynchus mykiss)?

The gastrointestinal tract (GIT) lumen of teleosts harbors extreme conditions, especially after feeding: high PCO2 (20-115 Torr), total ammonia (415-3710 µM), PNH3 (79-1760 µTorr in the intestine), and virtual anoxia (PO2 < 1 Torr). These levels could be dangerous if they were to equilibrate with the bloodstream. Thus, we investigated the potential equilibration of O2, CO2, and ammonia across the GIT epithelia in freshwater rainbow trout by monitoring postprandial arterial and venous blood gases in vivo and in situ. In vivo blood was sampled from the indwelling catheters in the dorsal aorta (DA) and subintestinal vein (SIV) draining the posterior intestine in the fasting state and at 4 to 48 h following catheter-feeding. To investigate possible ammonia absorption in the anterior part of the GIT, blood was sampled from the DA, SIV and hepatic portal vein (HPV) from anaesthetized fish in situ following voluntary feeding. We found minimal equilibration of all three gases between the GIT lumen and the SIV blood, with the latter maintaining pre-feeding levels (PO2 = 25-49 Torr, PCO2 = 6-8 Torr, and total ammonia = 117-134 µM and PNH3 = 13-30 µTorr at 48 h post-feeding). In contrast to the SIV, we found that the HPV total ammonia more than doubled 24 h after feeding (128 to 297 µM), indicative of absorption in the anterior GIT. Overall, the GIT epithelia of trout, although specialized for absorption, prevent dangerous levels of PO2, PCO2 and ammonia from equilibrating with the blood circulation.

Lessons learnt from the global iodinated contrast media shortage in head and neck imaging.

A recent shortage in the global supply of iodinated contrast media (ICM) has required health service providers to review their contrast administration policies and implement strategies to conserve inventory. This article will review the current best practices in head and neck imaging for a variety of common presentations and provide examples where alternative imaging can be considered due to the recent ICM shortage. Ultrasound and MRI techniques can feature heavily in many diagnostic processes in head and neck pathology, and a variety of common presentations can be appropriately investigated through clinical evaluation or naso-endoscopy. In many instances, for the routine assessment of non-acute adult and paediatric head and neck presentations, the use of contrast-enhanced CT can be safely minimised to conserve ICM if required.

The roles of calcium and salinity in protecting against physiological symptoms of waterborne zinc toxicity in the euryhaline killifish (Fundulus heteroclitus).

In fresh water, environmental Ca ameliorates Zn toxicity because Ca2+ and Zn2+ compete for uptake at the gills. Zn toxicity is also lower in sea water, but it is unclear whether this is due to increased Ca2+ concentration, and/or to the other ions present at higher salinity. Using the euryhaline killifish, we evaluated the relative roles of Ca2+ (as CaNO3) versus the other ions contributing to salinity in protecting against physiological symptoms of Zn2+ toxicity. Killifish were exposed to a sublethal level of Zn (500 µg/L, as ZnSO4) for 96 h in either fresh water (0 % salinity) at low (1 mmol/L) and High Ca (10 mmol/L) or 35 ppt sea water (100 % salinity) at low (1 mmol/L) and High Ca (10 mmol/L). At 0 % salinity, High Ca partly or completely protected against the following effects of Zn seen at Low Ca: elevated plasma Zn, hypocalcaemia, inhibited unidirectional Ca2+ influx, inhibited branchial Na+/K+ATPase and Ca2+ATPase activities, and oxidative stress in gills, liver, intestine, and muscle. At 100 % salinity, in the presence of 1 mmol/L (Low Ca), Zn caused no disturbances in most of these same parameters, showing that the "non-Ca" components of sea water alone provided complete protection. However, for a few endpoints (inhibited intestinal Ca2+ATPase activity, oxidative stress in gill and liver), High Ca (10 mmol/L) was needed to provide full protection against Zn in 100 % salinity. There was no instance where the combination of 100 % salinity and High Ca failed to provide complete protection against Zn-induced disturbances in sea water.

Risk factors and outcome in dogs with recurrent massive hepatocellular carcinoma: A Veterinary Society of Surgical Oncology case-control study.

Local recurrence after surgical excision of canine massive hepatocellular carcinoma (HCC) has been poorly studied in veterinary medicine with scant information published regarding risk factors for and outcome following recurrence. The aim of this case-control study was to describe the time to recurrence, evaluate potential risk factors for recurrence, and report the outcome in dogs with massive HCC. Medical records for 75 dogs who developed recurrence and 113 dogs who did not develop recurrence were reviewed. Statistical analyses were performed to determine risk factors for recurrence as well as the median time to develop recurrence and overall survival time (OS). None of the risk factors evaluated were significant for the development of recurrence. The median time to develop recurrence was 367 days (range 32-2096 days). There was no significant difference in median OS for dogs who developed recurrence vs. those who did not (851 vs. 970 days). For dogs with recurrent HCC, treatment at recurrence trended toward prolonged OS but was not significantly different from dogs not undergoing treatment at recurrence. There was no significant difference in median OS for dogs with histologically complete vs. incomplete tumour excision (990 vs. 903 days). Although specific risk factors for recurrence were not identified, elevations in liver values were noted in patients with recurrent disease and could act as a noninvasive surveillance tool. Recurrence was noted earlier in dogs who had routine post-operative surveillance (228 vs. 367 days). Routine surveillance for recurrence is recommended especially in dogs where further intervention is possible and should extend beyond 1 year. Patients with massive HCC have a good long-term prognosis regardless of incomplete excision, pulmonary metastasis, or recurrent local disease.

Exosomal DEK removes chemoradiotherapy resistance by triggering quiescence exit of breast cancer stem cells.

Tumor therapeutics often target the primary tumor bulk but fail to eradicate therapy-resistant cancer stem cells (CSCs) in quiescent state. These can then become activated to initiate recurrence and/or metastasis beyond therapy. Here, we identified and isolated chemoradiotherapy-resistant CSCs in quiescent state with high capacity of tumor-initiation and tumorsphere formation from three types of breast tumors in mice. Experiments of knockdown and rescue revealed DEK, a nuclear protein, as essential for CSC activation. Exogenous DEK was then used to trigger quiescence exit of CSCs. ChIP-seq and ATAC-seq showed that DEK directly binds to chromatin, facilitating its genome-wide accessibility. The resulting epigenetic events upregulate the expression of cellular activation-related genes including MYC targets, whereas cellular quiescence-related genes including the p53 signaling pathway are silenced. However, twinned with DEK-induced activation, formerly resistant CSCs were then destroyed by chemotherapy in vitro. In mice, traditional chemoradiotherapy concurrent with the injection of DEK-containing exosomes resulted in eradication of primary tumors together with formerly resistant CSCs without recurrence or metastasis. Our findings advance knowledge of the mechanism of quiescent CSC activation and may provide novel clinical opportunities for removal of quiescence-linked therapy resistance.

Transepithelial potential remains indicative of major ion toxicity in rainbow trout (Oncorhynchus mykiss) after 4-day pre-exposure to major salts.

The Multi-Ion Toxicity (MIT) Model uses electrochemical theory to predict the transepithelial potential (TEP) across the gills as an index of major ion toxicity in freshwater animals. The goal is to determine environmental criteria that will be protective of aquatic organisms exposed to salt pollution. In recent studies, TEP disturbances above baseline (ΔTEP) during short-term exposures to major ions have been proven as indicative of their toxicity to fish, in accord with the MIT model. However, the acute 1-h exposures used in these previous studies might not be realistic relative to the 24 h or 96 h test periods used for toxicity assessment. To address this temporal inconsistency, the current study investigated both the TEP responses to serial concentrations of 10 major salts (NaCl, Na2SO4, NaHCO3, KCl, K2SO4, KHCO3, CaCl2, CaSO4, MgCl2, MgSO4) and plasma ion levels in juvenile rainbow trout after they had been pre-exposed to 50% of the 96h-LC50 levels of these same salts for 4 days. The pre-exposures caused no mortalities. In general, plasma ions (Na+, K+, Ca2+, Mg2+, Cl-) were well-regulated; however, pre-exposure to sulfate salts resulted in the greatest number of alterations in plasma ion levels. TEP responses remained largely similar to those of naïve trout (without salt pre-exposure). All salts caused hyperbolic concentration-dependent increases in TEP that were well-described by the Michaelis-Menten equation. In the pre-exposed trout, the variation of ∆TEP at the 96h-LC50 concentrations was only 2.2-fold, compared to nearly 28-fold variation among the molar concentrations of the various salts at the 96h-LC50s, identical to the conclusion for naïve trout. Overall, the results remove the temporal inconsistency of previous tests and remain supportive of the MIT model. In addition, the recorded alterations in certain plasma ions, baseline TEP, and Michaelis-Menten constants improve our knowledge on specific physiological responses after extended major ion exposure.

Arapaima gigas maintains gas exchange separation in severe aquatic hypoxia but does not suffer branchial oxygen loss.

One of the most air-reliant obligate air-breathing fish is the South American Arapaima gigas, with substantially reduced gills impeding gas diffusion, thought to be a result of recurring aquatic hypoxia in its habitat. In normoxic water, A. gigas is reported to satisfy 70-80% of its O2 requirement from the air while excreting 60-90% of its CO2 to the water. If this pattern of gas exchange were to continue in severely hypoxic water, O2 loss at the gills would be expected. We hypothesized therefore that partitioning of CO2 would shift to the air phase in severe aquatic hypoxia, eliminating the risk of branchial O2 loss. By adapting a respirometer designed to measure aquatic MO2/MCO2, we were able to run intermittent closed respirometry on both water and air phase for both of these gasses as well as sample water for N-waste measurements (ammonia-N, urea-N) so as to calculate metabolic fuel utilization. In contrast to our prediction, we found that partitioning of CO2 excretion changed little between normoxia and severe hypoxia (83% versus 77% aquatic excretion, respectively) and at the same time there was no evidence of branchial O2 loss in hypoxia. This indicates that A. gigas can utilize distinct transfer pathways for O2 and CO2. Routine and standard MO2, N-waste excretion and metabolic fuel utilization did not change with water oxygenation. Metabolism was fuelled mostly by protein oxidation (53%), while carbohydrates and lipids accounted for 27% and 20%, respectively.

Post-prandial respiratory gas and acid-base profiles in the gastrointestinal tract and its venous drainage in freshwater rainbow trout (Oncorhynchus mykiss) and seawater English sole (Parophrys vetulus).

The basic respiratory gas and acid-base conditions inside the lumen of the gastrointestinal tract (GIT) and blood draining the tract are largely unestablished in teleost fishes after feeding, though there have been some recent novel discoveries on freshwater rainbow trout (Oncorhynchus mykiss) and seawater English sole (Parophrys vetulus). The present study examined in greater detail the gas (PO2, PCO2, PNH3) and acid-base profiles (pH, [HCO3-], total [ammonia]) in the lumen of the stomach, the anterior, mid, and posterior intestine, as well as the venous drainage (subintestinal and/or hepatic portal vein) of the GIT in these two species 20 h post-feeding. Both species had high PCO2, PNH3, and total [ammonia], and low PO2 (virtual anoxia) in the lumens throughout all sections of the GIT, and high [HCO3-] in the intestine. Total [ammonia], PNH3, and [HCO3-] increased from anterior to posterior intestine in both species. P. vetulus had higher intestinal total [ammonia] and lower [HCO3-] than O. mykiss post feeding, but total [ammonia] was much higher in the stomach of O. mykiss. Despite the extreme conditions in the lumen, both arterial and venous blood showed relatively lower PCO2, total [ammonia] and higher PO2, implying limited equilibration between the two compartments. The higher [HCO3-] and lower total [ammonia] in the intestinal lumen of the freshwater O. mykiss than the seawater P. vetulus suggest the need for future comparative studies using conspecifics fed identical diets but acclimated to the two different salinities in order to understand the potential role of environmental salinity and associated osmoregulatory processes underlying these differences.

The effect of marine dissolved organic carbon on nickel accumulation in early life-stages of the sea urchin, Strongylocentrotus purpuratus.

Dissolved organic carbon (DOC) is known to ameliorate the toxicity of the trace metal nickel (Ni) to aquatic animals. In theory, this effect is mediated by the capacity of DOC to bind Ni, rendering it less bioavailable, with the resulting reduction in accumulation limiting toxicological effects. However, there is a lack of experimental data examining Ni accumulation in marine settings with natural sources of DOC. In the current study, radiolabelled Ni was used to examine the time- and concentration-dependence of Ni accumulation, using naturally sourced DOC, on developing larvae of the sea urchin Strongylocentrotus purpuratus. Contrary to prediction, the two tested natural DOC samples (collected from the eastern United States, DOC 2 (Seaview park, Rhode Island (SVP)) and DOC 7 (Aubudon Coastal Center, Connecticut)) which had previously been shown to protect against Ni toxicity, did not limit accumulation. The control (artificial seawater with no added DOC), and the DOC 2 sample could mostly be described as having saturable Ni uptake, whereas Ni uptake in the presence of DOC 7 was mostly linear. These data provide evidence that DOC modifies the bioavailability of Ni, through either indirect effects (e.g. membrane permeability) or by the absorption of DOC-Ni complexes. There was some evidence for regulation of Ni accumulation in later-stage embryos (96-h) where the bioconcentration factor for Ni declined with increasing Ni exposure concentration. These data have implications for predictive modelling approaches that rely on known relationships between Ni speciation, bioavailability and bioreactivity, by suggesting that these relationships may not hold for natural marine DOC samples in the developing sea urchin model system.

Trans-epithelial potential (TEP) response as an indicator of major ion toxicity in rainbow trout and goldfish exposed to 10 different salts in ion-poor water.

Freshwater ecosystems are facing increasing contamination by major ions. The Multi-Ion Toxicity (MIT) model, a new tool for risk assessment and regulation, predicts major ion toxicity to aquatic organisms by relating it to a critical disturbance of the trans-epithelial potential (TEP) across the gills, as predicted by electrochemical theory. The model is based on unproven assumptions. We tested some of these by directly measuring the acute TEP responses to a geometric series of 10 different single salts (NaCl, Na2SO4, KCl, K2SO4, CaCl2, CaSO4, MgCl2, MgSO4, NaHCO3, KHCO3) in the euryhaline rainbow trout (Oncorhynchus mykiss) and the stenohaline goldfish (Carassius auratus) acclimated to very soft, ion-poor water (hardness 10 mg CaCO3/L). Results were compared to 24-h and 96-h LC50 data from the literature, mainly from fathead minnow (Pimephales promelas). All salts caused concentration-dependent increases in TEP to less negative/more positive values, in patterns well-described by the Michaelis-Menten equation, or a modified version incorporating substrate inhibition. The ΔTEP above baseline became close to a maximum at the 96-h LC50, except for the HCO3- salts. Furthermore, the range of ΔTEP values at the LC50 within one species was much more consistent (1.6- to 2.1-fold variation) than the molar concentrations of the different salts at the LC50 (19- to 25-fold variation). ΔTEP responses were related to cation rather than anion concentrations. Overall patterns were qualitatively similar between trout and goldfish, with some quantitative differences, and also in general accord with recently published data on three other species in harder water where ΔTEP responses were much smaller. Blood plasma Na+ and K+ concentrations were minimally affected by the exposures. The results are in accord with most but not all of the assumptions of the MIT model and support its further development as a predictive tool.

The Central Dot Sign : A Specific Post-gadolinium Enhancement Feature of Intramedullary Spinal Cord Metastases.

PURPOSE: Peripheral enhancement characteristics on magnetic resonance imaging (MRI), namely the rim and flame signs, are specific for intramedullary spinal cord metastases (ISCM) compared to primary cord masses. The study compared the frequency of a novel finding-the central dot sign-in ISCMs versus primary intramedullary masses. METHODS: In this study 45 patients with 64 ISCMs and 64 control patients with 64 primary intramedullary cord masses were investigated and 2 radiologists blinded to lesion type independently evaluated MR images for the presence of a central dot sign: a punctate focus of enhancement in/near the center of an enhancing intramedullary mass. The frequency of this sign in the two patient groups was compared. RESULTS: A total of 63 enhancing ISCMs in 44 patients and 54 enhancing primary cord masses in 54 patients were included. The central dot sign was identified in 6% (4/63) of enhancing ISCMs in 9% (4/44) of patients and in none (0/54) of the enhancing primary cord masses (p = 0.038, per patient). The specificity for diagnosing ISCMs among spinal cord masses was 100%. The central dot sign was present in the axial plane only in two ISCMs and in the axial and sagittal planes in two ISCMs. The two ISCMs harboring the central dot sign also demonstrated both the previously described rim and flame signs, and two also demonstrated the rim sign alone. CONCLUSION: The central dot sign is not sensitive but highly specific for ISCMs compared to primary spinal cord masses. The rim and/or flame signs may or may not be concurrently present in ISCMs.

The potential for salt toxicity: Can the trans-epithelial potential (TEP) across the gills serve as a metric for major ion toxicity in fish?

An emerging Multi-Ion Toxicity (MIT) model for assessment of environmental salt pollution is based on the premise that major ion toxicity to aquatic organisms is related to a critical disturbance of the trans-epithelial potential across the gills (ΔTEP), which can be predicted by electrochemical theory. However, the model has never been evaluated physiologically. We directly tested key assumptions by examining the individual effects of eight different salts (NaCl, Na2SO4, MgCl2, MgSO4, KCl, K2SO4, CaCl2, and CaSO4) on measured TEP in three different fish species (fathead minnow, Pimephales promelas = FHM; channel catfish, Ictalurus punctatus = CC; bluegill, Lepomis macrochirus = BG). A geometric concentration series based on previously reported 96-h LC50 values for FHM was used. All salts caused concentration-dependent increases in TEP to less negative/more positive values in a pattern well-described by the Michaelis-Menten equation. The ΔTEP responses for different salts were similar to one another within each species when concentrations were expressed as a percentage of the FHM LC50. A plateau was reached at or before 100 % of the LC50 where the ΔTEP values were remarkably consistent, with only 1.4 to 2.2-fold variation. This relative uniformity in the ΔTEP responses contrasts with 28-fold variation in salt concentration (in mmol L-1), 9.6-fold in total dissolved solids, and 7.9-fold in conductivity at the LC50. The Michaelis-Menten Km values (salt concentrations causing 50 % of the ΔTEPmax) were positively related to the 96-h LC50 values. ΔTEP responses were not a direct effect of osmolarity in all species and were related to specific cation rather than specific anion concentrations in FHM. These responses were stable for up to 24 h in CC. The results provide strong physiological support for the assumptions of the MIT model, are coherent with electrochemical theory, and point to areas for future research.

Cellular oxygen consumption, ROS production and ROS defense in two different size-classes of an Amazonian obligate air-breathing fish (Arapaima gigas).

In air-breathing fish a reduction of gill surface area reduces the danger of losing oxygen taken up in the air-breathing organ (ABO) to hypoxic water, but it also reduces the surface area available for ion exchange, so that ion regulation may at least in part be transferred to other organs, like the kidney or the gut. In the air-breathing Arapaima gigas, gill lamellae regress as development proceeds, and starting as a water-breathing embryo Arapaima turns into an obligate air-breathing fish with proceeding development, suggesting that ion regulation is shifted away from the gills as the fish grows. In Arapaima the kidney projects medially into the ABO and thus, probably a unique situation among fishes, is in close contact to the gas of the ABO. We therefore hypothesized that the kidney would be predestined to adopt an increased importance for ion homeostasis, because the elevated ATP turnover connected to ion transport can easily be met by aerobic metabolism based on the excellent oxygen supply directly from the ABO. We also hypothesized that in gill tissue the reduced ion regulatory activity should result in a reduced metabolic activity. High metabolic activity and exposure to high oxygen tensions are connected to the production of reactive oxygen species (ROS), therefore the tissues exposed to these conditions should have a high ROS defense capacity. Using in vitro studies, we assessed metabolic activity and ROS production of gill, kidney and ABO tissue, and determined the activity of ROS degrading enzymes in small (~ 5g, 2-3 weeks old) and larger (~ 670 g, 3-4 months old) A. gigas. Comparing the three tissues revealed that kidney tissue oxygen uptake by far exceeded the uptake measured in gill tissue or ABO. ROS production was particularly high in gill tissue, and all three tissues had a high capacity to degrade ROS. Gill tissue was characterized by high activities of enzymes involved in the glutathione pathway to degrade ROS. By contrast, the tissues of the ABO and in particular the kidney were characterized by high catalase activities, revealing different, tissue-specific strategies in ROS defense in this species. Overall the differences in the activity of cells taken from small and larger fish were not as pronounced as expected, while at the tissue level the metabolic activity of kidney cells by far exceeded the activity of ABO and gill cells.

A Mystery Tale: Nickel Is Fickle When Snails Fail-Investigating the Variability in Ni Toxicity to the Great Pond Snail.

Dissolved Ni concentrations inhibiting the growth of juvenile great pond snails (Lymnaea stagnalis) have been documented to vary from about 1 to 200 µg L-1 Ni. This variability makes L. stagnalis either a moderately sensitive or the most sensitive freshwater species to chronic Ni exposure tested to date. Given the role of sensitive species in environmental risk assessment frameworks, it is particularly important to understand this variability, i.e., to characterize the factors that modulate Ni toxicity and that may confound toxicity test outcomes when uncontrolled. In the present study, we tested if this variability was due to analytical (growth calculation: biomass versus growth rate), environmental (water quality), lab-specific practices, and/or snail population differences among earlier studies. Specifically, we reanalyzed previously published Ni toxicity data and conducted additional measurements of Ni aqueous speciation, short-term Ni uptake, and chronic Ni toxicity with test waters and snail cultures used in previous studies. Corrections for Ni bioavailability and growth calculations explained a large degree of variability in the published literature. However, a residual 16-fold difference remained puzzling between 2 studies: Niyogi et al. (2014) (low ECxs) and Crémazy et al. (2018) (high ECxs). Indeed, differences in metal bioavailability due to water chemistry, lab-specific practices, and snail population sensitivity could not explain the large variation in Ni toxicity in these 2 very similar studies. Other potentially important toxicity-modifying factors were not directly evaluated in the present work: test duration, diet, snail holding conditions, and snail age at onset of testing. The present analysis highlights the need for further studies to elucidate 1) the mechanisms of growth inhibition in Ni-exposed L. stagnalis and 2) the important abiotic and biotic factors affecting this biological response. Until these processes are understood, substantial uncertainties will remain about inclusion of this species in Ni environmental risk assessment. Integr Environ Assess Manag 2020;16:983-997. © 2020 SETAC.

MTL-CEBPA, a Small Activating RNA Therapeutic Upregulating C/EBP-α, in Patients with Advanced Liver Cancer: A First-in-Human, Multicenter, Open-Label, Phase I Trial.

PURPOSE: Transcription factor C/EBP-α (CCAAT/enhancer-binding protein alpha) acts as a master regulator of hepatic and myeloid functions and multiple oncogenic processes. MTL-CEBPA is a first-in-class small activating RNA oligonucleotide drug that upregulates C/EBP-α. PATIENTS AND METHODS: We conducted a phase I, open-label, dose-escalation trial of MTL-CEBPA in adults with advanced hepatocellular carcinoma (HCC) with cirrhosis, or resulting from nonalcoholic steatohepatitis or with liver metastases. Patients received intravenous MTL-CEBPA once a week for 3 weeks followed by a rest period of 1 week per treatment cycle in the dose-escalation phase (3+3 design). RESULTS: Thirty-eight participants have been treated across six dose levels (28-160 mg/m2) and three dosing schedules. Thirty-four patients were evaluable for safety endpoints at 28 days. MTL-CEBPA treatment-related adverse events were not associated with dose, and no maximum dose was reached across the three schedules evaluated. Grade 3 treatment-related adverse events occurred in nine (24%) patients. In 24 patients with HCC evaluable for efficacy, an objective tumor response was achieved in one patient [4%; partial response (PR) for over 2 years] and stable disease (SD) in 12 (50%). After discontinuation of MTL-CEBPA, seven patients were treated with tyrosine kinase inhibitors (TKIs); three patients had a complete response with one further PR and two with SD. CONCLUSIONS: MTL-CEBPA is the first saRNA in clinical trials and demonstrates an acceptable safety profile and potential synergistic efficacy with TKIs in HCC. These encouraging phase I data validate targeting of C/EBP-α and have prompted MTL-CEBPA + sorafenib combination studies in HCC.

Effect of Augmented Antimicrobial Prophylaxis and Rectal Swab Culture-guided Targeted Prophylaxis on the Risk of Sepsis Following Transrectal Prostate Biopsy.

BACKGROUND: Sepsis is a severe complication following transrectal ultrasound-guided prostate biopsy (TRUSPBx). Ciprofloxacin is commonly used for prophylaxis; however, there is an increasing incidence of resistant enteric organisms worldwide. OBJECTIVE: To investigate the effect of a targeted prophylactic antimicrobial regimen based on rectal swab cultures in reducing the rate of sepsis. DESIGN, SETTING, AND PARTICIPANTS: A total of 1012 patients were included. Group A (609 patients) received an empirical prophylactic antimicrobial regimen of gentamicin, metronidazole, and ciprofloxacin. Targeted antimicrobial prophylaxis was introduced due to significant ciprofloxacin and gentamicin resistance in patients admitted with sepsis following TRUSPBx. The remaining 403 patients (Group B) had rectal swab cultures performed prior to biopsy. Patients with organisms resistant to ciprofloxacin or gentamicin received a targeted prophylaxis regimen of fosfomycin, amikacin, and metronidazole. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: We retrospectively collected and analysed data on sepsis and bacteraemia for all patients as well as data on rectal swab culture, recent foreign travel, and recent antibiotic use for patients in Group B. RESULTS AND LIMITATIONS: In group A, 12 (2.0%) patients developed sepsis following TRUSPBx, while in group B, 9 (2.2%) patients developed sepsis despite targeted prophylaxis (p=0.82). Patients with ciprofloxacin-resistant rectal flora had a significantly higher rate of sepsis (9.1% vs 1.1%; p=0.003). There was a reduction in patients admitted with bacteraemia and severe sepsis between group A (1.2%) and group B (0.3%) which did not reach statistical significance (p=0.16). In group B, 55 of 403 (13.6%) patients had ciprofloxacin-resistant rectal flora, while 66 (16.4%) had organisms resistant to both ciprofloxacin and gentamicin. A recent foreign travel history was associated with an increased incidence of ciprofloxacin-resistant rectal flora (23.6% vs 10.8%; p=0.007). The main limitations of our study include its retrospective nature and potential under-reporting of less severe infectious complications. CONCLUSIONS: Rectal swab cultures identify patients with ciprofloxacin-resistant rectal flora who have an eight-fold risk of sepsis. Targeted antimicrobial prophylaxis may not be beneficial in reducing the sepsis rate when compared with augmented empirical prophylaxis. In an era of increasing antimicrobial resistance, transperineal prostate biopsies should be considered to reduce the risk of infective complications. PATIENT SUMMARY: Performing rectal swab culture prior to transrectal prostate biopsy can help identify patients at risk of developing sepsis despite targeted prophylactic antibiotics.

Effects of copper on a reconstructed freshwater rainbow trout gill epithelium: Paracellular and intracellular aspects.

The barrier properties and intracellular responses of a primary cultured trout gill epithelium (containing both mitochondria-rich and pavement cells) were examined over 24 h of copper (Cu) exposure (0, 200 and 1000 µg/L) in apical fresh water. Transepithelial resistance (TER) and mRNA abundance of tight junction proteins zonula occludens-1, occludin, cingulin, claudin-8d and -28b were examined as endpoints of barrier function and the paracellular pathway. Intracellular endpoints analyzed were Cu accumulation, Na+ content, carbonic anhydrase activity and mRNA abundance of carbonic anhydrase (ca-II) and Na+/K+ ATPase (nka α1a and nka α1b isoforms). After a brief initial drop in TER in the 1000 µg Cu/L treatment, Cu at both levels increased TER over the first 6 h of exposure but there were no differences among groups from 12 h onwards. After 24 h of Cu exposure, there were no differences in mRNA abundance of any of the tight junction proteins examined. Cu accumulation occurred at 1000 µg Cu/L (5.5-fold increase), but no depletion of Na+ content. Carbonic anhydrase activity decreased significantly (by 76%), however Cu exposure did not alter the transcript abundance of ca-II, nka α1a, or nka α1b. This study provides a first report of carbonic anhydrase sensitivity to Cu exposure in a cultured model gill epithelium. We conclude that Cu impacts the permeability of this model during the early stages of exposure and that the use of carbonic anhydrase inhibition as an endpoint of metal toxicity in this model preparation may be useful for future mechanistic investigations and environmental monitoring.

Effects of sublethal Cd, Zn, and mixture exposures on antioxidant defense and oxidative stress parameters in early life stages of the purple sea urchin Strongylocentrotus purpuratus.

Oxidative stress parameters were evaluated during the first 72 h of embryonic development of purple sea urchin Strongylocentrotus purpuratus continuously exposed to control conditions, to cadmium alone (Cd, 30 µg/L), to zinc alone (Zn, 9 µg/L) or to a Cd (28 µg/L) plus Zn (9 µg/L) mixture. These sublethal concentrations represent ∼ 10% of the acute EC50. Bioaccumulation, antioxidant capacity against peroxyl radicals (ACAP), total glutathione (GSH) level, glutathione-S-transferase (GST), glucose-6-phosphate dehydrogenase (G6PDH) and superoxide dismutase (SOD) activity, and lipid peroxidation (LPO) were analyzed at 24 h (blastula), 48 h (gastrula), and 72 h (pluteus) stages of development. Zinc (an essential metal) was well-regulated, whereas Cd (non-essential) bioaccumulated and whole-body [Cd] increased from blastula to pluteus stage in sea urchin larvae. In controls, ACAP progressively declined from 24 h to 72 h, while LPO reciprocally increased, but other parameters did not change. Cd alone was more potent than Zn alone as a pro-oxidant, with the major effects being decreases in SOD activity and parallel increases in LPO throughout development; GST activity also increased at 24 h. Zn alone caused only biphasic disturbances of ACAP. In all cases, the simultaneous presence of the other metal prevented the effects, and there was no instance where the oxidative stress response in the presence of the Cd/Zn mixture was greater than in the presence of either Cd or Zn alone. Therefore the sublethal effects of joint exposures were always less than additive or even protective, in agreement with classical toxicity data. Furthermore, our results indicate that SOD and Zn can play important roles in protecting sea urchin embryos against Cd-induced lipid peroxidation.