Search

Abstract We have reported that an environmental pollutant, cadmium, promotes cell death in the human renal tubular cells (RTCs) through hyperactivation of a serine/threonine kinase Akt. However, the molecular mechanisms downstream of Akt in this process have not been elucidated. Cadmium has a potential to accumulate misfolded proteins, and proteotoxicity is involved in cadmium toxicity. To clear the roles of Akt in cadmium exposure-induced RTCs death, we investigated the possibility that Akt could regulate proteotoxicity through autophagy in cadmium chloride (CdCl2)-exposed HK-2 human renal proximal tubular cells. CdCl2 exposure promoted the accumulation of misfolded or damaged proteins, the formation of aggresomes (pericentriolar cytoplasmic inclusions), and aggrephagy (selective autophagy to degrade aggresome). Pharmacological inhibition of Akt using MK2206 or Akti-1/2 enhanced aggrephagy by promoting dephosphorylation and nuclear translocation of transcription factor EB (TFEB)/transcription factor E3 (TFE3), lysosomal transcription factors. TFEB or TFE3 knockdown by siRNAs attenuated the protective effects of MK2206 against cadmium toxicity. These results suggested that aberrant activation of Akt attenuates aggrephagy via TFEB or TFE3 to facilitate CdCl2-induced cell death. Furthermore, these roles of Akt/TFEB/TFE3 were conserved in CdCl2-exposed primary human RTCs. The present study shows the molecular mechanisms underlying Akt activation that promotes cadmium-induced RTCs death.

Objective: Previous studies have demonstrated synergistic antitumor effects of angiotensin system inhibition (ASI) combined with cisplatin therapy in pancreatic cancer. This study examines whether or not synergistic antitumor effects occur with combination ASI and cisplatin treatment in lung cancer, and whether or not ASI-induced changes in epithelial-mesenchymal transition play a role in the mechanism of this antitumor phenomenon. Methods: A set of lung cancer cell lines representing a spectrum of epithelial to mesenchymal phenotypes were identified and characterized. Response of epithelial-mesenchymal transition markers to losartan was characterized. Cell culture models of lung cancer were next treated with losartan, cisplatin, or combination of both. Markers of epithelial-mesenchymal transition or surrogates of other signaling pathways (AKT, Stat3, and programmed death-ligand), and cell viability were quantified. Findings were confirmed in both allogenic and syngeneic in vivo murine flank tumor models. Results: Losartan treatment significantly increased E-cadherin and reduced vimentin in human lung cancer cell lines. Combination treatment with losartan and cisplatin enhanced epithelial markers, reduced mesenchymal markers, inhibited promesenchymal signaling mediators, and reduced cell viability. Findings were confirmed in vivo in a murine flank tumor model with transition from mesenchymal to epithelial phenotype and reduced tumor size following combination losartan and cisplatin treatment. Conclusions: Combination losartan and cisplatin treatment attenuates the epithelial-mesenchymal transition pathway and enhances the cytotoxic effect of chemotherapy with in vitro and in vivo models of non–small cell lung cancer. This study suggests an important role for ASI therapy in the treatment of lung cancer.

Background and Aims: Effective approaches for prevention of hepatocellular carcinoma (HCC) will have a significant impact on HCC-related mortality. There are strong preclinical data and rationale to support targeting epidermal growth factor receptor (EGFR) for HCC chemoprevention. Small molecule inhibitors of EGFR have been Food and Drug Administration–approved for cancer therapy, which provides an opportunity to repurpose one of these drugs for chemoprevention of HCC. Unfortunately, the frequency of side effects associated with administration of these drugs at oncology doses renders them ineffective for chemoprevention. This clinical trial assesses whether lower doses of one of these inhibitors, erlotinib, still engages EGFR in the liver to block signaling (eg, EGFR phosphorylation). The objective of this clinical trial was determination of a safe and minimum effective dose of erlorinib for which ≥ 50% reduction phospho-EGFR immunohistochemical staining in the liver was observed. Methods: Forty six participants were preregistered and 25 participants were registered in this multicenter trial. By dose de-escalation trial design, cohorts of participants received a 7-day course of erlotinib 75 mg/day, 50 mg/day or 25 mg/day with liver tissue acquisition prior to and after erlotinib. Results: A ≥50% reduction phospho-EGFR immunohistochemical staining in the liver was observed in a minimum of 40% of participants (predetermined threshhold) at each of the dose levels. Erlotinib was very well tolerated with few side effects observed, particularly at the dose of 25 mg/day. Favorable modulation of the Prognostic Liver Signature was observed in participants who received erlotinib. Conclusion: These data support the selection of erlotinib doses as low as 25 mg/day of for a longer intervention to assess for evidence of efficacy as an HCC chemoprevention drug (ClinicalTrials.gov NCT02273362).

Abstract Sea turtles are migratory with nesting and foraging areas in distinct and often widely separated habitats. Telemetry has been a vital tool for tracking sea turtle migrations between these areas, but tagging efforts are often focused on only a few large rookeries in a given region. For instance, turtle tagging in the Red Sea has been focused in the north of the basin. We tagged five green turtles (Chelonia mydas) at a nesting site in the central-southern Red Sea and tracked them for 72–243 days. During the inter-nesting period, the turtles showed high site-fidelity, with a maximum home range of 161 km2. After the nesting season, the turtles migrated up to 1100 km to five distinct foraging locations in three countries (Saudi Arabia, Sudan, and Eritrea). Movements within foraging habitats were more wide-ranging compared to inter-nesting movements, with home ranges varying between 1.19 and 931 km2. The tracking data revealed that the creation of a relatively small marine reserve could protect the critical inter-nesting habitat in the Farasan Banks. The results also highlight the need for multinational collaboration to protect migratory corridors and foraging sites of this endangered species.

Abstract Semi‐enclosed seas are often associated with elevated local threats and distinct biogeographic patterns among marine fishes, but our understanding of how fish assemblage dynamics vary in relation to relatively small semi‐enclosed seas (e.g., the Gulf of Aqaba) remains limited. Baited remote underwater video surveys (n = 111) were conducted across ~300 km of coral reef habitats in the Gulf of Aqaba and the northern Red Sea. A total of 55 predatory fish species were detected, with less than half of all species (n = 23) observed in both basins. Relative abundance patterns between the Gulf of Aqaba and the northern Red Sea were variable among taxa, but nearly twice as many predatory fish were observed per unit of effort in the northern Red Sea. In general, assemblages in both basins were dominated by three taxa (Epinephelinae, Carangidae, and Lethrinidae). Large‐bodied and threatened species were recorded at very low abundances. Multivariate analysis revealed distinct assemblage structuring of coral reef predators between the Gulf of Aqaba and the northern Red Sea. Most of the species driving these differences were recorded in both basins, but occurred at varying levels of abundance. Environmental factors were largely unsuccessful in explaining variation in assemblage structuring. These findings indicate that biological assemblages in the Gulf of Aqaba are more distinct than previously reported and that reef fish assemblage structuring can occur even within a relatively small semi‐enclosed sea. Despite inter‐basin assemblage structuring, the overall low abundance of vulnerable fish species is suggestive of overexploitation in both the Gulf of Aqaba and the northern Red Sea of Saudi Arabia. As the region surveyed is currently undergoing large‐scale coastal development, the results presented herein aim to guide spatial management and recovery plans for these coral reef systems in relation to this development.

Background & Aims: Many liver diseases are driven by inflammation, but imaging to non-invasively diagnose and quantify liver inflammation has been underdeveloped. The inflammatory liver microenvironment is aberrantly oxidising owing in part to reactive oxygen species generated by myeloid leucocytes. We hypothesised that magnetic resonance imaging using the oxidatively activated probe Fe-PyC3A will provide a non-invasive biomarker of liver inflammation. Methods: A mouse model of drug-induced liver injury was generated through intraperitoneal injection of a hepatoxic dose of acetaminophen. A mouse model of steatohepatitis was generated via a choline-deficient, l-amino acid defined high-fat diet (CDAHFD). Images were acquired dynamically before and after intravenous injection of Fe-PyC3A. The contrast agent gadoterate meglumine was used as a non-oxidatively activated negative control probe in mice fed CDAHFD. The (post-pre) Fe-PyC3A injection change in liver vs. muscle contrast-to-noise ratio (ΔCNR) recorded 2 min post-injection was correlated with liver function test values, histologic scoring assigned using the NASH Clinical Research Network criteria, and intrahepatic myeloid leucocyte composition determined by flow cytometry. Results: For mice receiving i.p. injections of acetaminophen, intrahepatic neutrophil composition correlated poorly with liver test values but positively and significantly with ΔCNR (r = 0.64, p

Abstract Background Hawksbill turtles (Eretmochelys imbricata) are Critically Endangered throughout their global range, and concerningly little is known about this species in the Red Sea. With large-scale coastal development projects underway in the northern Red Sea, it is critical to understand the movement and habitat use patterns of hawksbill turtles in this environmentally unique region, so that effective conservation strategies can be implemented. We satellite tagged three hawksbill turtles, one 63 cm curved carapace length adult male captured near Wahlei Island, one 55 cm turtle captured in the Gulf of Aqaba, and one 56 cm turtle suffering from a floating syndrome which was captured at Waqqadi Island, rehabilitated, and released at Waqqadi Island. Turtles were tracked for 156, 199, and 372 days between October 2020 and November 2021. Results We calculated the home ranges and core use areas of hawksbill turtles using kernel-density estimations and found that each turtle showed high fidelity to their foraging sites. Home ranges calculated with GPS-derived locations ranged between 13.6 and 2.86 km2, whereas home ranges calculated with Argos-derived locations ranged from 38.98 to 286.45 km2. GPS-derived locations also revealed a higher proportion of time spent in coral and rock habitats compared to Argos, based on location overlap with the Allen Coral Reef Atlas. We also found that turtles were making shallow dives, usually remaining between 0 and 5 m. Conclusions While the number of tracked turtles in this study was small, it represents an important contribution to the current understanding of spatial ecology among foraging hawksbill turtles globally, and provides the first-ever reported hawksbill turtle tracking data from the Red Sea. Our results suggest that protecting coral reef habitats and implementing boating speed limits near reefs could be effective conservation measures for foraging hawksbill turtles in the face of rapid coastal development.

Background & Aims: During liver fibrosis, tissue repair mechanisms replace necrotic tissue with highly stabilized extracellular matrix proteins. Extracellular matrix stabilization influences the speed of tissue recovery. Here, we studied the expression and function of peroxidasin (PXDN), a peroxidase that uses hydrogen peroxide to cross-link collagen IV during liver fibrosis progression and regression. Methods: Mouse models of liver fibrosis and cirrhosis patients were analyzed for the expression of PXDN in liver and serum. Pxdn-/- and Pxdn+/+ mice were either treated with carbon tetrachloride for 6 weeks to generate toxin-induced fibrosis or fed with a choline-deficient L-amino acid-defined high-fat diet for 16 weeks to create nonalcoholic fatty liver disease fibrosis. Liver histology, quantitative real-time polymerase chain reaction, collagen content, flowcytometry and immunostaining of immune cells, RNA-sequencing, and liver function tests were analyzed. In vivo imaging of liver reactive oxygen species (ROS) was performed using a redox-active iron complex, Fe-PyC3A. Results: In human and mouse cirrhotic tissue, PXDN is expressed by stellate cells and is secreted into fibrotic areas. In patients with nonalcoholic fatty liver disease, serum levels of PXDN increased significantly. In both mouse models of liver fibrosis, PXDN deficiency resulted in elevated monocyte and pro-fibrolysis macrophage recruitment into fibrotic bands and caused decreased accumulation of cross-linked collagens. In Pxdn-/- mice, collagen fibers were loosely organized, an atypical phenotype that is reversible upon macrophage depletion. Elevated ROS in Pxdn-/- livers was observed, which can result in activation of hypoxic signaling cascades and may affect signaling pathways involved in macrophage polarization such as TNF-a via NF-kB. Fibrosis resolution in Pxdn-/- mice was associated with significant decrease in collagen content and improved liver function. Conclusion: PXDN deficiency is associated with increased ROS levels and a hypoxic liver microenvironment that can regulate recruitment and programming of pro-resolution macrophages. Our data implicate the importance of the liver microenvironment in macrophage programming during liver fibrosis and suggest a novel pathway that is involved in the resolution of scar tissue.

Drug and target discovery for advanced liver disease are hampered by a lack of suitable models for clinical translation. Here the authors present a human liver cell-based system modeling a clinical prognostic signature allowing to propose nizatidine for treatment of advanced liver fibrosis and hepatocellular carcinoma prevention.

Architectures based on multi-bit magnetic domain walls (DWs) take advantage of the fast speed, high density, nonvolatility, and flexible design of DWs to process and store data bits. However, controlling multi-bit DWs driven by electric current at an ideal position remains a significant challenge for developing integrated spintronic applications with high reliability and low power consumption. We exhibit the possibility of driving fast and stable multi-bit DWs at low current density without an in-plane external magnetic field in Fe-rich GdFeCo magnetic wires. When an in-plane magnetic field is applied in the wire direction, the front edge accelerates, although the rear edge decelerates, and the recorded data are destroyed. Hence, this method is not practical. Here, the DW speed of the multi-bit DWs is 1500 m/s under a low current density of 29 × 1010 (A/m2). A straight DW shape is required to accurately read the bits of information by the tunneling magnetoresistance head in real DW memory devices. Moreover, we demonstrate that the DW position is related to the DW shape after injecting a pulse current into the magnetic wire. A straight DW shape is exhibited for 3 ns pulse duration width, while the DW shape became rounded for 30 and 50 ns pulse duration widths. Our finding provides a practical concept for multiple-bit-per-cell memory and presents a viable platform for DW memory applications.

Background Anthropogenic sources can lead to the accumulation of heavy metals in marine organisms through ingestion, absorption, or inhalation. For sea turtle embryos, heavy metals can be absorbed into the egg from the incubation environment or be maternally transferred to the offspring causing neurological, reproductive, and developmental problems. Here, we report heavy metal concentrations in green turtle hatchlings from the largest rookery on the Red Sea, Ras Baridi. Methods Deceased hatchlings were collected from two beaches near a cement factory at Ras Baridi, from which heavy metal concentrations (chromium (Cr), manganese (Mn), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), selenium (Se), cadmium (Cd), and lead (Pb)) were measured from the liver, muscle, and residual yolk of the hatchlings. Results Although based on a small sample of hatchlings, the data presented here provides the first measurements of heavy metals from sea turtles in the Red Sea and highlights the link between human activity and its impact on the ecology of sea turtles. In general, the heavy metal concentrations of heavy metals were not significantly different between the beach next to the cement factory and the beach downwind from the factory. However, the concentrations of heavy metals were significantly different between sampled tissues (liver, muscle, and residual yolk). Discussion This study provides insight into current heavy metal levels in green turtle hatchlings, which can be used as bio-indicators for environmental contaminants as coastal development increases in the Red Sea. Moreover, we found a lack of standardized methodology to evaluate heavy metals in hatchling sea turtles. Future efforts should work toward creating comparable techniques for long-term heavy metal monitoring, as this is a useful determinant of anthropogenic pollution.

Background There is relatively little published information about sea turtle nesting distribution and seasonality in the Saudi Arabian Red Sea. Upcoming large-scale developments occurring along the Saudi Arabian Red Sea coast could negatively affect many sea turtle nesting beaches with potential impacts on the survival of local populations. Methods In 2019, two coastal beaches and three near-shore islands were surveyed for turtle nesting in the central Red Sea. We recorded all emergences, examined beach morphology, and collected sand samples to determine grain size, moisture content and colour. Results Sea turtle nesting was found at all surveyed sites, though emergence counts were often low. The limited occurrence of nesting at several previously undocumented sites suggests that nesting activity may be widespread, but sparsely distributed, in the central Red Sea region. In addition, nesting at novel sites appeared to favour the seaward side of islands, a pattern that was not observed in previously documented areas. The substrate of most surveyed sites was composed of calcium carbonate with Ras Baridi as the only exception; it was composed of dark quartz-rich sediment. This study highlights several important sea turtle rookeries while also demonstrating that low levels of nesting occur throughout the region, although inter-annual nesting patterns still need to be determined. Future developments should be steered away from key nesting areas and the seaward bias in marginal rookeries should be taken into account where possible.

Hepatocellular carcinoma (HCC) is a leading cause of death among cirrhotic patients, for which chemopreventive strategies are lacking. Recently, we developed a simple human cell-based system modeling a clinical prognostic liver signature (PLS) predicting liver disease progression and HCC risk. In a previous study, we applied our cell-based system for drug discovery and identified captopril, an approved angiotensin converting enzyme (ACE) inhibitor, as a candidate compound for HCC chemoprevention. Here, we explored ACE as a therapeutic target for HCC chemoprevention. Captopril reduced liver fibrosis and effectively prevented liver disease progression toward HCC development in a diethylnitrosamine (DEN) rat cirrhosis model and a diet-based rat model for nonalcoholic steatohepatitis–induced (NASH-induced) hepatocarcinogenesis. RNA-Seq analysis of cirrhotic rat liver tissues uncovered that captopril suppressed the expression of pathways mediating fibrogenesis, inflammation, and carcinogenesis, including epidermal growth factor receptor (EGFR) signaling. Mechanistic data in liver disease models uncovered a cross-activation of the EGFR pathway by angiotensin. Corroborating the clinical translatability of the approach, captopril significantly reversed the HCC high-risk status of the PLS in liver tissues of patients with advanced fibrosis. Captopril effectively prevents fibrotic liver disease progression toward HCC development in preclinical models and is a generic and safe candidate drug for HCC chemoprevention.

BackgroundSorafenib, a kinase inhibitor, is a standard treatment for advanced hepatocellular carcinoma (HCC) but provides only a limited survival benefit. Disulfiram (DSF), a drug for treating alcoholism and a chelator of copper (Cu), forms a complex with Cu (DSF/Cu). DSF/Cu is a potent inducer of autophagic apoptosis of cancer stem cells, which can demonstrate drug resistance. Thus, we hypothesized that DSF/Cu could increase the sensitivity of HCC cells to sorafenib by targeting hepatic cancer stem cells.MethodsThe synergistic effect of DSF/Cu and sorafenib on human HCC cell lines was assessed by cell viability MTT assay. Changes in stemness gene expression in HCC cells were investigated by assessing the presence of hepatic cancer stem cells (HCSCs) (defined as ALDH+ cells) using flow cytometry, sphere formation ability as an index of in vitro tumorigenicity, and expression of stemness gene-encoded proteins by western blot. Autophagic apoptosis and the ERK signaling pathway were also assessed by western blot. Most importantly, the in vivo anti-tumor efficacy of DSF/Cu and sorafenib was tested using orthotopic HCC xenografts in mice.ResultsCompared with sorafenib alone, DSF/Cu + sorafenib synergistically inhibited proliferation of all HCC cell lines, decreased the stemness of HCC cells, and increased the autophagy and apoptosis of HCC cells. The mechanism by which DSF/Cu mediated these phenomena with sorafenib was sustained activation of the ERK pathway. The combination of DSF/Cu (formed with endogenous Cu2+) and sorafenib was significantly more effective than sorafenib alone in inhibiting the growth of orthotopic HCC xenografts in mice. This in vivo anti-tumor efficacy was associated with decreased stemness in treated HCC tumors.ConclusionsDSF/Cu and sorafenib can synergistically and effectively treat HCC by targeting HCSCs in vitro and in vivo. Our data provide a foundation for clinical translation.

Abstract Findings of ammonoid soft tissues are extremely rare compared to the rich fossil record of ammonoid conchs ranging from the Late Devonian to the Cretaceous/Paleogene boundary. Here, we apply the computed-tomography approach to detect ammonoid soft tissue remains in well-preserved fossils from the Early Cretaceous (early Albian) of NE-Germany of Proleymeriella. The ammonites were found in glauconitic–phosphatic sandstone boulders. Analyses of the high-resolution Ct-data revealed the presence of cameral sheets, the siphuncular tube wall, and the siphuncle itself. The siphuncle is a long, segmented soft tissue that begins at the rear end of the body chamber and comprises blood vessels. Chemical analyses using energy-dispersive spectroscopy (EDS) showed that all preserved soft tissues were phosphatized and are now composed of fluorapatite. The same holds true for preserved shell remains that locally show the nacreous microstructure. We provide a short description of these soft tissue remains and briefly discuss the taphonomic pathway.

Abstract Non-alcoholic steatohepatitis (NASH) is an increasing cause of chronic liver disease characterized by steatosis, inflammation, and fibrosis which can lead to cirrhosis, hepatocellular carcinoma, and mortality. Quantitative, noninvasive methods for characterizing the pathophysiology of NASH at both the preclinical and clinical level are sorely needed. We report here a multiparametric magnetic resonance imaging (MRI) protocol with the fibrogenesis probe Gd-Hyd to characterize fibrotic disease activity and steatosis in a common mouse model of NASH. Mice were fed a choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) to induce NASH with advanced fibrosis. Mice fed normal chow and CDAHFD underwent MRI after 2, 6, 10 and 14 weeks to measure liver T1, T2*, fat fraction, and dynamic T1-weighted Gd-Hyd enhanced imaging of the liver. Steatosis, inflammation, and fibrosis were then quantified by histology. NASH and fibrosis developed quickly in CDAHFD fed mice with strong correlation between morphometric steatosis quantification and liver fat estimated by MRI (r = 0.90). Sirius red histology and collagen quantification confirmed increasing fibrosis over time (r = 0.82). Though baseline T1 and T2* measurements did not correlate with fibrosis, Gd-Hyd signal enhancement provided a measure of the extent of active fibrotic disease progression and correlated strongly with lysyl oxidase expression. Gd-Hyd MRI accurately detects fibrogenesis in a mouse model of NASH with advanced fibrosis and can be combined with other MR measures, like fat imaging, to more accurately assess disease burden.

The Red Sea is particularly biodiverse, hosting high levels of endemism and numerous populations whose extinction risk is heightened by their relative isolation. Elasmobranchs and sea turtles have likely suffered recent declines in this region, although data on their distribution and biology are severely lacking, especially on the eastern side of the basin in Saudi Arabian waters. Here, we present sightings of elasmobranchs and sea turtles across the north-eastern Red Sea and Gulf of Aqaba collected through a combination of survey methods. Over 455 survey hours, we recorded 407 sightings belonging to 26 elasmobranch species and two sea turtle species, more than 75% of which are of conservation concern. We identified 4 species of rays and 9 species of sharks not previously recorded in Saudi Arabia and report a range extension for the pink whipray (Himantura fai) and the round ribbontail ray (Taeniurops meyeni) into the Gulf of Aqaba. High density of sightings of conservation significance, including green and hawksbill sea turtles and halavi guitarfish were recorded in bay systems along the eastern Gulf of Aqaba and the Saudi Arabian coastline bordering the north-eastern Red Sea, and many carcharhinid species were encountered at offshore seamounts in the region. Our findings provide new insights into the distribution patterns of megafaunal assemblages over smaller spatial scales in the region, and facilitate future research and conservation efforts, amidst ongoing, large-scale coastal developments in the north-eastern Red Sea and Gulf of Aqaba.

(1) Background: Plastic pollution is a major environmental concern confronting marine animals. Sea turtles are considered a bio-indicator of plastic pollution, but there is little information regarding plastic ingestion by turtles in the Red Sea. With large-scale development projects being built along the Saudi Arabian coast, it is important to have a baseline for plastic ingestion before construction is complete. (2) Methods: Ten deceased sea turtles (four hawksbill and six green turtles) were collected along the Saudi Arabian coastline. Necropsies were conducted, and the entire gastrointestinal tracts were extracted and the contents were passed through a 1 mm mesh sieve. (3) Results: We found that 40% of the turtles in this study had ingested plastics. Thread-like plastics were the most common plastic category, and multi-colored was the most prevalent color category. (4) Conclusions: Monitoring of the plastic ingestion by marine megafauna should be conducted as a long-term assessment of the developments’ impacts. Additionally, conservation efforts should be focused on removing plastics (namely ghost nests and fishing lines) from the reefs and reducing the amount of plastic entering the sea.

Major aggregations of nesting green turtles (Chelonia mydas) occur in the northern Red Sea, although little is known about the reproductive ecology of this endangered species in the region. To address this issue, we satellite-tracked 30 female green turtles to document their movements and to identify factors driving habitat use at two major rookeries in the Red Sea, Jazirat Mashabah (Mashabah Island) and Ras Al Baridi in Saudi Arabia. Between successive nesting events, turtles displayed high fidelity to nesting beaches and adjacent in-water habitats (inter-nesting habitats). Using generalized linear mixed models, we estimated the mean probability of nesting per beach emergence (nesting success rate) to be 0.628, and the mean duration between a successful nesting event and the successive emergence onto the beach (re-nesting interval) to be 10.8 days at each site. The nesting success rate was relatively high (>0.8) when the preceding daytime land surface temperature (LST) was lower than 37°C but decreased with elevated daytime LST (47°C). Re-nesting interval was longer at lower water temperatures and towards the end of the nesting season of individuals. Our study improves the robustness of abundance estimates from census data (e.g., track counts) and shows that the protection of nesting and inter-nesting habitats during a breeding season would be an effective conservation strategy for the species. We discuss how global warming could increase energy expenditure due to lowered nesting success, ultimately compromising the reproductive fitness of these populations.

Studies of the volatility distribution of secondary organic aerosol (SOA) from aromatic compounds are limited compared with SOA from biogenic monoterpenes. In this study, the volatility distribution was investigated by composition, heating, and dilution measurements for SOA formed from the photooxidation of 1,3,5-trimethylbenzene in the presence of NOx. Composition studies revealed that highly oxygenated monomers (C9H14Ox, x = 4–7) and dimers (C18H26Ox, x = 8–12) are the major products in SOA particles. Highly oxygenated molecules (HOMs) with five or more oxygens were formed during photochemical aging, whereas dimers degraded during photochemical aging. HOMs with five or more oxygens may be produced from the photooxidation of phenol-type gaseous products, whereas dimers in the particle phase may be photolyzed to smaller molecules during photochemical aging. The results of composition, heating, and dilution measurements showed that fresh SOA that formed from 1,3,5-trimethylbenzene (TMB) photooxidation includes low-volatility compounds with  µg m−3 saturation concentrations, which are attributed to dimers. Similar results were reported for α-pinene SOA in previous studies. Low-volatility compounds with  µg m−3 saturation concentrations are not included in the volatility distributions employed in the standard volatility basis-set (VBS) approach. Improvements in the organic aerosol model will be necessary for the study of anthropogenic SOA as well as biogenic SOA.

Abstract Climate change poses a serious threat to species that demonstrate temperature‐dependent sex determination, including marine turtles. Increased temperatures can result in highly female‐skewed sex ratios and decreased hatching success. The pivotal temperature that delineates hatchling sex ratios is commonly considered to be 29.2°C, but whether this threshold applies to turtles in the Red Sea region has not been tested in situ. For all species of marine turtles, there is a supposed thermal range of 25–33°C in which egg incubation is successful, with prolonged temperatures above 33°C resulting in morphological abnormalities and hatchling mortality. Sand temperature data were collected from May–September 2018 from the average nesting depth of hawksbill (Eretmochelys imbricata) and green turtles (Chelonia mydas) at five study sites. We calculated the expected sex ratio based on a maximum likelihood model. The sand temperature profile at four of the sites exceeded the pivotal temperature (29.2°C) throughout the study duration, which suggests feminization of turtles could be occurring; however, the pivotal temperature in this region still needs to be empirically confirmed. The percentage of days with sand temperature exceeding the maximum thermal threshold between June 3, and September 16, 2018, was site‐specific rather than determined by latitudinal temperature gradients, and ranged between 0 and 100% of days. Maximum temperature recordings were as high as 36.0 and 35.3°C at 30 and 50 cm depth, respectively. Nesting sites in the Red Sea region could already be exceeding the thermal limits and may be particularly vulnerable to rising temperatures. Sites with lower sand temperatures, such as Small Gobal Island, may represent priority areas for conservation efforts. Alternatively, local adaptation may be a reality under extremely warm conditions, thus, further research into the thermal tolerance of hatchlings in the region could provide insight on how they might adapt to future climate change.

We examined a novel farnesoid X receptor agonist, EDP‐305, for its antifibrotic effect in bile duct ligation (BDL) and choline‐deficient, L‐amino acid‐defined, high‐fat diet (CDAHFD) models of hepatic injury. We used molecular magnetic resonance imaging with the type 1 collagen‐binding probe EP‐3533 and the oxidized collagen‐specific probe gadolinium hydrazide to noninvasively measure treatment response. BDL rats (n = 8 for each group) were treated with either low or high doses of EDP‐305 starting on day 4 after BDL and were imaged on day 18. CDAHFD mice (n = 8 for each group) were treated starting at 6 weeks after the diet and were imaged at 12 weeks. Liver tissue was subjected to pathologic and morphometric scoring of fibrosis, hydroxyproline quantitation, and determination of fibrogenic messenger RNA expression. High‐dose EDP‐305 (30 mg/kg) reduced liver fibrosis in both the BDL and CDAHFD models as measured by collagen proportional area, hydroxyproline analysis, and fibrogenic gene expression (all P < 0.05). Magnetic resonance signal intensity with both EP‐3533 in the BDL model and gadolinium hydrazide in the CDAHFD model was reduced with EDP‐305 30 mg/kg treatment (P < 0.01). Histologically, EDP‐305 30 mg/kg halted fibrosis progression in the CDAHFD model. Conclusion: EDP‐305 reduced fibrosis progression in rat BDL and mouse CDAHFD models. Molecular imaging of collagen and oxidized collagen is sensitive to changes in fibrosis and could be used to noninvasively measure treatment response in clinical trials. (Hepatology Communications 2018;2:821‐835)

Traditional yield curve analysis shows that semi-volatile organic compounds are a major component of secondary organic aerosols (SOAs). We investigated the volatility distribution of SOAs from α-pinene ozonolysis using positive electrospray ionization mass analysis and dilution- and heat-induced evaporation measurements. Laboratory chamber experiments were conducted on α-pinene ozonolysis, in the presence and absence of OH scavengers. Among these, we identified not only semi-volatile products, but also less volatile highly oxygenated molecules (HOMs) and dimers. Ozonolysis products were further exposed to OH radicals to check the effects of photochemical aging. HOMs were also formed during OH-initiated photochemical aging. Most HOMs that formed from ozonolysis and photochemical aging had 10 or fewer carbons. SOA particle evaporation after instantaneous dilution was measured at

Sangiovese is the most cultivated red grape variety in Italy where it is certified for the production of several Protected Designation of Origin (PDO) wines, and it is one of the most cultivated Italian red grape varieties in California. Despite the global distribution of this variety, there is a lack of international studies on Sangiovese grapes and wines. For this reason, the present study aimed to compare 20 commercial Sangiovese wines from 2017 harvest, 9 produced in Italy (Tuscany) and 11 in California, in order to evaluate the intrinsic and perceived quality. The eligibility, identity, and style properties (the intrinsic quality) of the wines were evaluated. A group of 11 Italian experts evaluated the perceived quality by rating the typicality of the wines. The experimental data showed that the intrinsic quality of Sangiovese wine samples was affected by the growing area; in particular, the wine resulted very different for the color indices and polyphenol composition. The above differences in intrinsic quality levels did not lead to a different evaluation of the perceived quality (typicality) by the wine experts. The results evidenced that Sangiovese variety is recognizable also if grown outside its original terroir, and fresh and fruity wines were considered more typical. This study expands our current knowledge of Sangiovese wines and the contribution of regional characteristics to the composition of wine.

The compositional authentication of wine is of great interest, as the geographic origin of the grapes is often associated with quality, uniqueness, and authenticity. Previous elemental fingerprinting studies mainly discriminated wines from different countries or regions within a country. Here, we report the use of element profiles to distinguish commercial Pinot noir wines from five sub-regional appellations or neighborhoods within one American viticultural area (AVA). Fifty-three single cultivar wines were collected over two harvests and analyzed using microwave plasma-atomic emission spectroscopy (MP-AES) and inductively coupled plasma-mass spectrometry (ICP-MS). Of 62 monitored elements that were quantified with fully validated methods, 24 and 32 elements differed significantly across the neighborhoods and vintages, respectively (p < 0.05). Targeted canonical variate analysis (CVA) explained 85%-90% of the variance ratio across the two vintages, indicating persistent and stable elemental fingerprints of wines at a sub-regional level. A sixth, newly founded neighborhood was correctly grouped separately from the others using a Soft Independent Modeling of Class Analogy (SIMCA), indicating the potential of elemental fingerprints for wine authenticity.

Syngeneic, immunocompetent allograft tumor models recapitulate important aspects of the tumor microenvironment and have short tumor latency with predictable growth kinetics, making them useful for trialing novel therapeutics. Here, we describe surgical techniques for orthotopic and heterotopic pancreatic ductal adenocarcinoma (PDAC) tumor implantation and characterize phenotypes based on implantation site. Mice (n=8 per group) were implanted with 104 cells in the pancreas or flank. Hy15549 and Han4.13 cell lines were derived from primary murine PDAC (Ptf1-Cre; LSL-KRAS-G12D; Trp53 Lox/+) on C57BL/6 and FVB strains, respectively. Single-cell suspension and solid tumor implants were compared. Tumors were treated with two intravenous doses of FOLFIRINOX and responses evaluated. All mice developed pancreatic tumors within 7 days. Orthotopic tumors grew faster and larger than heterotopic tumors. By 3 weeks, orthotopic mice began losing weight, and showed declines in body condition requiring euthanasia starting at 4 weeks. Single-cell injection into the pancreas had near 100% engraftment, but solid tumor implant engraftment was ∼50% and was associated with growth restriction. Orthotopic tumors were significantly more responsive to intravenous FOLFIRINOX compared with heterotopic tumors, with greater reductions in size and increased apoptosis. Heterotopic tumors were more desmoplastic and hypovascular. However, drug uptake into tumor tissue was equivalent regardless of tumor location or degree of fibrosis, indicating that microenvironment differences between heterotopic and orthotopic tumors influenced response to therapy. Our results show that orthotopic and heterotopic allograft locations confer unique microenvironments that influence growth kinetics, desmoplastic response and angiogenesis. Tumor location influences chemosensitivity to FOLFIRINOX and should inform future preclinical trials. This article has an associated First Person interview with the first author of the paper.