Protective effect of cannabidiol against cadmium hepatotoxicity in rats

The protective effect of cannabidiol, the non-psychoactive component of Cannabis sativa, against liver toxicity induced by a single dose of cadmium chloride (6.5 mg kg^?1 i.p.) was investigated in rats. Cannabidiol treatment (5 mg kg^?1/day, i.p.) was applied for five days starting three days before cadmium administration. Cannabidiol significantly reduced serum alanine aminotransferase, and suppressed hepatic lipid peroxidation, prevented the depletion of reduced glutathione and nitric oxide, and catalase activity, and attenuated the elevation of cadmium level in the liver tissue resulted from cadmium administration. Histopathological examination showed that cadmium-induced liver tissue injury was ameliorated by cannabidiol treatment. Immunohistochemical analysis revealed that cannabidiol significantly decreased the cadmium-induced expression of tumor necrosis factor-α, cyclooxygenase-2, nuclear factor-κB, caspase-3, and caspase-9, and increased the expression of endothelial nitric oxide synthase in liver tissue. It was concluded that cannabidiol may represent a potential option to protect the liver tissue from the detrimental effects of cadmium toxicity.     

δ15N and δ13C diet–tissue discrimination factors for large sharks under semi-controlled conditions

Stable isotopes (δ¹⁵N and δ¹³C) are being widely applied in ecological research but there has been a call for ecologists to determine species- and tissue-specific diet discrimination factors (?¹³C and ?¹⁵N) for their study animals. For large sharks stable isotopes may provide an important tool to elucidate aspects of their ecological roles in marine systems, but laboratory based controlled feeding experiments are impractical. By utilizing commercial aquaria, we estimated ?¹⁵N and ?¹³C of muscle, liver, vertebral cartilage and a number of organs of three large sand tiger (Carcharias taurus) and one large lemon shark (Negaprion brevirostris) under a controlled feeding regime. For all sharks mean ± SD for ?¹⁵N and ?¹³C in lipid extracted muscle using lipid extracted prey data were 2.29‰ ± 0.22 and 0.90‰ ± 0.33, respectively. The use of non-lipid extracted muscle and prey resulted in very similar ?¹⁵N and ?¹³C values but mixing of lipid and non-lipid extracted data produced variable estimates. Values of ?¹⁵N and ?¹³C in lipid extracted liver and prey were 1.50‰ ± 0.54 and 0.22‰ ± 1.18, respectively. Non-lipid extracted diet discrimination factors in liver were highly influenced by lipid content and studies that examine stable isotopes in shark liver, and likely any high lipid tissue, should strive to remove lipid effects through standardising C:N ratios, prior to isotope analysis. Mean vertebral cartilage ?¹⁵N and ?¹³C values were 1.45‰ ± 0.61 and 3.75‰ ± 0.44, respectively. Organ ?¹⁵N and ?¹³C values were more variable among individual sharks but heart tissue was consistently enriched by ~ 1–2.5‰. Minimal variability in muscle and liver δ¹⁵N and δ¹³C sampled at different intervals along the length of individual sharks and between liver lobes suggests that stable isotope values are consistent within tissues of individual animals. To our knowledge, these are the first reported diet–tissue discrimination factors for large sharks under semi-controlled conditions, and are lower than those reported for teleost fish.     

Quantification of titanium from TiO2 particles in biological tissue

This study presents the development of a strategy for the quantification of titanium from titanium dioxide polydisperse particles (TiO₂) in dry biological tissue. Calf liver was chosen as laboratory testing material. The challenge was to (i) obtain a complete mineralization of the solid material (biological tissue and TiO₂) and (ii) ensure the accuracy of the determined concentrations with a sufficient sensitivity. Mineralization was performed using a mixture of concentrated nitric and hydrofluoric acids. Atomic mass spectrometry associated with light-scattering technique was used to control the physical state (dissolved and particle forms) of titanium and reliably estimate the total titanium concentration in calf liver. The monitoring of ⁴⁶Ti and ⁴⁹Ti, operating in helium collision/reaction cell mode, and using external calibration with internal standard addition, allowed the quantification of Ti while removing isobaric interferences. The limit of detection and quantification were 0.7 and 2.3 μg (Ti) g⁻¹ (tissue) respectively. The mean analytical recovery over the whole procedure was (103 ± 6)% in a range of concentrations from LOD to 200 μg(Ti) g⁻¹ (tissue).     

Potential of using cerium oxide nanoparticles for protecting healthy tissue during accelerated partial breast irradiation (APBI)

The purpose of this study is to investigate the feasibility of using cerium oxide nanoparticles (CONPs) as radical scavengers during accelerated partial breast irradiation (APBI) to protect normal tissue. We hypothesize that CONPs can be slowly released from the routinely used APBI balloon applicators—via a degradable coating—and protect the normal tissue on the border of the lumpectomy cavity over the duration of APBI. To assess the feasibility of this approach, we analytically calculated the initial concentration of CONPs required to protect normal breast tissue from reactive oxygen species (ROS) and the time required for the particles to diffuse to various distances from the lumpectomy wall. Given that cerium has a high atomic number, we took into account the possible inadvertent dose enhancement that could occur due to the photoelectric interactions with radiotherapy photons. To protect against a typical MammoSite treatment fraction of 3.4 Gy, 5 ng·g⁻¹ of CONPs is required to scavenge hydroxyl radicals and hydrogen peroxide. Using 2 nm sized NPs, with an initial concentration of 1 mg·g⁻¹, we found that 2–10 days of diffusion is required to obtain desired concentrations of CONPs in regions 1–2 cm away from the lumpectomy wall. The resultant dose enhancement factor (DEF) is less than 1.01 under such conditions. Our results predict that CONPs can be employed for radioprotection during APBI using a new design in which balloon applicators are coated with the NPs for sustained/controlled in-situ release from within the lumpectomy cavity.     

Modelling biomass of mountainous grasslands by including a species composition map

High mountain grasslands offer multiple goods and services to society but are severely threatened by improper land use practices such as abandonment or rapid intensification. In order to reduce abandonment and strengthen the common extensive agricultural practice a sustainable land use management of high mountain grasslands is needed. A spatially detailed yield assessment helps to identify possible meadows or, on the contrary, areas with a low carrying capacity in a region, making it easier to manage these sites. Such assessments are rarely available for remote and inaccessible areas. Remotely sensed vegetation indices are able to provide valuable information on grassland properties. These indices tend, however, to saturate for high biomass. This affects their applicability to assessments of high-yield grasslands.The main aim of this study was to model a spatially explicit grassland yield map and to test whether saturation issues can be tackled by consideration of plant species composition in the modelling process. The high mountain grassland of the subalpine belt (1800 – 2500 m a.s.l.) in the Kazbegi region, Greater Caucasus, Georgia, was chosen as test site for its strong species composition and yield gradients.We first modelled the species composition of the grassland described as metrically scaled gradients in the form of ordination axes by random forest regression. We then derived vegetation indices from Rapid Eye imagery, and topographic variables from a digital elevation model, which we used together with the multispectral bands as predictive variables. For comparison, we performed two yield models, one excluding the species composition maps and one including the species composition map as predictors. Moreover, we performed a third individual model, with species composition as predictors and a split dataset, to produce the final yield map.Three main grassland types were found in the vegetation analysis: Hordeum violaceum-meadows, Gentianella caucasea-grassland and Astragalus captiosus-grassland. The three random forest regression models for the ordination axes explained 64%, 33% and 46% of the variance in species composition. Independent validation of modelled ordination scores against a validation data set resulted in an R² of 0.64, 0.32 and 0.46 for the first, second and third axes, respectively. The model based on species composition resulted in a R² = 0.55, whereas the benchmark model showed weaker relationships between yield and the multispectral reflectance, vegetation indices, and topographical parameters (R² = 0.42). The final random forest yield model used to derive the yield map resulted in 62% variance explained and an R² = 0.64 between predicted and observed biomass. The results further indicate that high yields are generally difficult to predict with both models.The benefit of including a species composition map as a predictor variable for grassland yield lies in the preservation of ecologically meaningful features, especially the occurrence of high yielding vegetation type of Hordeum violaceum meadows is depicted accurately in the map. Even though we used a gradient based design, sharp boundaries or immediate changes in productivity were visible, especially in small structures such as arable fields or roads (Fig. 6b), making it a valuable tool for sustainable land use management. The saturation effect however, was mitigated by using species composition as predictor variables but is still present at high yields.     

High performance liquid chromatography determination of sulphachloropyrazine residues in broiler and turkey edible tissues

A HPLC method to determine and quantify sulphachloropyrazine residues from broilers and turkeys is reported. This procedure permitted sulphachloropyrazine to be separated from muscle tissue, liver, kidneys and fat with skin after extraction with dichloromethane under slightly acidic conditions. The analytical methodology showed a high specificity and sensitivity and an adequate precision and accuracy with a limit of quantification of 56 ng mL^?1. The peak area showed a linear relationship with a concentration over the range 50–750 ng mL^?1 for sulphachloropyrazine standard solutions. Recovery dates were also satisfactory with values between 69.7 and 77.5%.     

A colorimetric assay method to measure acetyl-CoA synthetase activity: Application to woodchuck model of hepatitis virus-induced hepatocellular carcinoma

A new spectrophotometric method for quantitation of acetyl-CoA synthetase (ACAS) activity is developed. It has been applied for ACAS assay in the liver tissues of a woodchuck model of hepatitis virus-induced hepatocellular carcinoma (HCC). The assay is based on the established pyrophosphate (PPi) detection system. ACAS activity is indexed by the amount of PPi, the product of ACAS reaction system of activated form of acetate (acetyl-CoA) with ACAS catalysis. PPi is determined quantitatively as the amount of chromophore formed with molybdate reagent, 1-amino-2-naphthol-4-sulfonic acid in bisulfite and 2-mercaptoethanol. PPi reacts with molybdate reagent to produce phosphomolybdate and PPi-molybdate complexes. 2-mercaptoethanol is responsible for color formation which has the peak absorbance at 580 nm. This method was sensitive from 1 to 20 nmol of PPi in a 380-μl sample (1-cm cuvette). A ten-fold excess of Pi did not interfere with the determination of PPi. To study the major metabolic pathways of imaging tracer [1-¹¹C]-acetate in tumors for detection of HCC by Positron Emission Tomography (PET), the activity of one of the key enzymes involved in acetate or [1-¹¹C]-acetate metabolism, ACAS was assayed by this newly developed assay in the tissue samples of woodchuck HCCs. A significant increase of ACAS activity was observed in the liver tissues of woodchuck HCCs as compared with neighboring regions surrounding the tumors (P < 0.05). The respective ACAS activities in the subcellular locations were also significantly higher in HCCs than in the surrounding tissues (P < 0.05) (total soluble fraction: 876.61 ± 34.64 vs. 361.62 ± 49.97 mU/g tissue; cytoplasmic fraction: 1122.02 ± 112.39 vs. 732.32 ± 84.44 mU/g tissue; organelle content: 815.79 ± 100.77 vs. 547.91 ± 97.05 mU/ g tissue; sedimentable fragment: 251.92 ± 51.56 vs. 90.94 ± 18.98 mU/ g tissue). The finding suggests an increase in ACAS activity in the liver cancer of woodchuck models of HCC as compared to that in the normal woodchuck liver. The developed assay is rapid, simple and accurate and is suitable for the investigation of ACAS activity under physiologic and pathophysiologic conditions.     

End-Permian stratigraphic timeline applied to the timing of marine and non-marine extinctions

A latest Permian timeline (251.9 Ma) can be constructed from the perspectives of: a global nickel spike attributed to emissions from the coeval Siberian flood-basalt eruptions, the correlative end-Permian marine mass extinction (EPME), a transition from reversed to normal paleomagnetism, and a negative anomaly in δ¹³C_carb and δ¹³C_org. In a number of marine and non-marine localities, this timeline is also correlated (to within ≤30 ky) with palynological evidence for the latest Permian destruction of terrestrial vegetation and the accompanying short-lived global fungal (Reduviasporonites) event. This correlation suggests that devastation in marine and non-marine environments was essentially coeval at a time marked by hyperthermal conditions and anoxic oceans.We utilized this proposed timeline to estimate the relative timing of the extinction of latest Permian vertebrates in the Karoo Basin of South Africa. In several sections in the Karoo, the LAD of the therapsid Dicynodon, is correlated with the proposed timeline. In the Carlton Heights section in the Karoo we estimate that the palynological changes and the fungal event occurred within ≤30 ky of the LAD of Dicynodon. Further sampling in the Karoo and other Permian–Triassic non-marine basins would help to clarify the relative timing of the global marine extinctions, plant devastation and the disappearance of non-marine vertebrates.     

Is seed availability enough to ensure colonization success?: An experimental study in road embankments

We tested the hypothesis that seed availability is a limiting factor for plant colonization of road embankments under Mediterranean climate conditions. Experimental sowing on 10 road embankments was carried out to compare the colonization success of plants that successfully colonize the road embankment and species that appear only occasionally in the road embankments. After sowing, we measured plant establishment, biomass production, and reproductive capacity of the species.The species that appear only occasionally in the road embankments had lower emergence rates (l.l ± 0.3%) than species that were successful colonizers (18.8 ± 2.9%). None of the species of the former group survived or reproduced. The results did not support the hypothesis that seed availability was the main factor limiting plant colonization in the road embankments. We concluded that the arrival of seeds to road embankments under Mediterranean climate conditions was not enough to ensure colonization success of plants. Other factors, like hydric stress, appeared to affect seedling establishment and plant growth. Reclamation measures such as species selection should be taken in account to ensure revegetation success of road embankments.     

Stereotactic body radiation therapy in hepatocellular carcinoma: Optimal treatment strategies based on liver segmentation and functional hepatic reserve

AimTo discuss current dosage for stereotactic body radiation therapy (SBRT) in hepatocellular carcinoma (HCC) patients and suggest alternative treatment strategies according to liver segmentation as defined by the Couinaud classification.BackgroundSBRT is a safe and effective alternative treatment for HCC patients who are unable to undergo liver ablation/resection. However, the SBRT fractionation schemes and treatment planning strategies are not well established.Materials and methodsIn this article, the latest developments and key findings from research studies exploring the efficacy of SBRT fractionation schemes for treatment of HCC are reviewed. Patients’ characteristics, fractionation schemes, treatment outcomes and toxicities were compiled. Special attention was focused on SBRT fractionation approaches that take into consideration liver segmentation according to the Couinaud classification and functional hepatic reserve based on Child–Pugh (CP) liver cirrhosis classification.ResultsThe most common SBRT fractionation schemes for HCC were 3 × 10–20 Gy, 4–6 × 8–10 Gy, and 10 × 5–5.5 Gy. Based on previous SBRT studies, and in consideration of tumor size and CP classification, we proposed 3 × 15–25 Gy for patients with tumor size <3 cm and adequate liver reserve (CP-A score 5), 5 × 10–12 Gy for patients with tumor sizes between 3 and 5 cm or inadequate liver reserve (CP-A score 6), and 10 × 5–5.5 Gy for patients with tumor size >5 cm or CP-B score.ConclusionsTreatment schemes in SBRT for HCC vary according to liver segmentation and functional hepatic reserve. Further prospective studies may be necessary to identify the optimal dose of SBRT for HCC.     

Regulatory effects on central carbon metabolism from poly-3-hydroxybutryate synthesis

Poly-3-hydroxybutyrate (PHB) synthesis in Escherichia coli elicits regulatory responses that affect product yield and productivity. We used controlled, steady-state cultures (chemostats) of a genetically stable strain to determine growth-independent metabolic flux regulation. We measured flux and steady-state intracellular metabolite concentrations across different dilution rates (0.05, 0.15, 0.3 h⁻¹), limitations (glucose, gluconate and nitrogen), and operon copy counts of the PHB pathway (0, 6, 17, and 29). As PHB flux increases, specific substrate consumption and lactate secretion increase while formate and acetate secretion decreases in N-limited, glucose-fed conditions.To understand the regulatory mechanisms that resulted in these macroscopic changes, we used a flux balance analysis model to analyze intracellular redox conditions. Our model shows that under N-limited conditions, synthesis of PHB creates excess reducing equivalents. Cells, under these conditions, secrete more reduced metabolites in order to recycle reducing equivalents. By switching to a more oxidized substrate (gluconate) that decreased excess reducing equivalents, PHB flux yield increased 1.6 fold compared to glucose-fed fermentations. High flux of PHB (~1.2 mmol/g DCW h) was maintained under these steady-state, oxidized conditions. These results imply redox imbalance is a driving force in industrial production of PHB, and substrates that are more oxidized than glucose can increase productivity.     

Feasibility study of detecting surface electromyograms in severely obese patients

The aims of this study were to examine if surface EMG signals can be detected from the quadriceps femoris muscle of severely obese patients and to investigate if differences exist in quadriceps force and myoelectric manifestations of fatigue between obese patients and lean controls.Fourteen severely obese patients (body mass index, BMI, mean ± SD: 44.9 ± 6.3 kg/m²) and fourteen healthy controls (BMI: 23.7 ± 2.5 kg/m²) were studied. The vastus medialis and lateralis of the dominant thigh were concurrently investigated during voluntary isometric contractions (10-s long at submaximal and maximal intensities and intermittent submaximal contractions until exhaustion) and sustained (120-s long) electrically elicited contractions.We found that the detection of surface EMG signals from the quadriceps is feasible also in severely obese subjects presenting increased thickness of the subcutaneous fat tissue. In addition, we confirmed and extended previous findings showing that the volume conductor properties determine the amplitude and spectral features of the detected surface EMG signals: the lower the subcutaneous tissue thickness, the higher the amplitude and mean frequency estimates. Further, we found no differences in the mechanical and myoelectric manifestations of fatigue during intermittent voluntary and sustained electrically elicited contractions between obese patients and lean controls.     

Statins inhibit expression of thioredoxin reductase 1 in rat and human liver and reduce tumour development

BackgroundStatins have been reported to have anti-carcinogenic properties in addition to their cholesterol-lowering effects, but the mechanism is unknown. Thioredoxin reductases (TrxR) are selenium-containing enzymes of great importance for carcinogenesis and their levels are increased in neoplastic cells. The aim of the present study was to investigate if statin treatment is associated with alterations in the hepatic expression of TrxR.MethodsHuman liver biopsies from a study where patients had been randomised to statin treatment or placebo were analysed. In addition we used liver tissue from a human liver bank where statin treated subjects were compared with non-treated. We also used tissue from a rat liver cancer model in which we have previously shown anti-carcinogenic effects of statins. Real-time PCR and activity assay were used to determine TrxR-levels and activity in tissue extracts.ResultsIn humans 80 mg atorvastatin treatment for 4 weeks (n = 6) was associated with 85% lower levels of TrxR1 and TrxR2 compared to placebo-treated patients (n = 8) (p = 0.03). In liver biopsies from a human donor liver bank 3 statin treated subjects had 90% lower expression of TrxR1 than 15 non-treated subjects (p = 0.04). Statin treatment was associated with 45% lower expression and activity of TrxR1 in a rat model for liver cancer (p = 0.03). There was a clear correlation between inhibition of carcinogenesis and decreased TrxR1-levels (p = 0.003).ConclusionStatin treatment decreases the hepatic expression of TrxR1 in humans and rats. Suppression of TrxR1 expression could explain possible anti-carcinogenic effects of statins. In addition, decreased levels of TrxR1 during statin treatment may shed light on the mechanism of other side-effects of statins.     

Ecological effects of forest roads on plant species diversity in Caspian forests of Iran

Current research includes the effects of asphalt forest roads on changes of plant cover and tree regeneration from asphalt forest roads edges towards its inner parts in two compartments of Nave Asalem forests located in the north of Iran. For this reason, in each side of road, 6 sample plots (20 m × 20 m) were established for measuring plant species diversity. In each sample plot, ground vegetation and tree regeneration were assessed within nine 2 × 2 m micro plots. In total, 12 sample plots and 108 μ plots were established. Results indicated that the road positions were effective on plant species diversity. The highest diversity and evenness indices value were observed down of the road compared to the up of the road position for herbal and tree regeneration layers. The same results were found also for herbal richness indices. Up of road position had the greatest value of richness indices in comparison to the other road position for tree regeneration layer. Also, the results showed that diversity, richness, and evenness indices were decreased with the increasing of distance from the road side for herbs and tree regeneration layers. This study indicated that roads can increase plant biodiversity; that is, tree regeneration density.     

Fluorescence kinetics of PSII crystals containing Ca2 + or Sr2 + in the oxygen evolving complex

Photosystem II (PSII) is the pigment–protein complex which converts sunlight energy into chemical energy by catalysing the process of light-driven oxidation of water into reducing equivalents in the form of protons and electrons. Three-dimensional structures from x-ray crystallography have been used extensively to model these processes. However, the crystal structures are not necessarily identical to those of the solubilised complexes. Here we compared picosecond fluorescence of solubilised and crystallised PSII core particles isolated from the thermophilic cyanobacterium Thermosynechococcus elongatus. The fluorescence of the crystals is sensitive to the presence of artificial electron acceptors (K₃Fe(CN)₃) and electron transport inhibitors (DCMU). In PSII with reaction centres in the open state, the picosecond fluorescence of PSII crystals and solubilised PSII is indistinguishable. Additionally we compared picosecond fluorescence of native PSII with PSII in which Ca² in the oxygen evolving complex (OEC) is biosynthetically replaced by Sr^2 +. With the Sr^2 + replaced OEC the average fluorescence decay slows down slightly (81 ps to 85 ps), and reaction centres are less readily closed, indicating that both energy transfer/trapping and electron transfer are affected by the replacement.     

Three new players in energy regulation: Preptin,adropin and irisin

Homeostasis of energy is regulated by genetic factors, food intake, and energy expenditure. When energy input is greater than expenditure, the balance is positive, which can lead to weight gain and obesity. When the balance is negative, weight is lost. Regulation of this homeostasis is multi-factorial, involving many orexigenic (appetite-stimulating) and anorexigenic (appetite-suppressing) peptide hormones. Peripheral tissues are now known to be involved in weight regulation and research on its endocrine characteristics proceeds apace. Preptin with 34 amino acids (MW 3948 Da), adropin with 43 amino acids and a molecular weight of (4999 Da), and irisin with 112 amino acids (12587 Da), are three newly discovered peptides critical for regulating energy metabolism. Preptin is synthesized primarily in pancreatic beta cells, and adropin mainly in the liver and brain, and many peripheral tissues. Irisin, however, is synthesized principally in the heart muscle, along with peripheral tissues, including salivary glands, kidney and liver. The prime functions of preptin and adropin include regulating carbohydrate, lipid and protein metabolisms by moderating glucose-mediated insulin release. Irisin is an anti-obesitic and anti-diabetic hormone regulating adipose tissue metabolism and glucose homeostasis by converting white to brown adipose tissue. This review offers a historical account of these discovery and function of these peptides, including their structure, and physiological and biochemical properties. Their roles in energy regulation will be discussed. Their measurement in biological fluids will be considered, which will lead to further discussion of their possible clinical value.     

Neutron shielding for a new projected proton therapy facility: A Geant4 simulation study

In this work, we used the Monte Carlo-based Geant4 simulation toolkit to calculate the ambient dose equivalents due to the secondary neutron field produced in a new projected proton therapy facility. In particular the facility geometry was modeled in Geant4 based on the CAD design. Proton beams were originated with an energy of 250 MeV in the gantry rooms with different angles with respect to the patient; a fixed 250 MeV proton beam was also modeled. The ambient dose equivalent was calculated in several locations of interest inside and outside the facility, for different scenarios. The simulation results were compared qualitatively to previous work on an existing facility bearing some similarities with the design under study, showing that the ambient dose equivalent ranges obtained are reasonable. The ambient dose equivalents, calculated by means of the Geant4 simulation, were compared to the Australian regulatory limits and showed that the new facility will not pose health risks for the public or staff, with a maximum equivalent dose rate equal to 7.9 mSv/y in the control rooms and maze exit areas and 1.3·10⁻¹ mSv/y close to the walls, outside the facility, under very conservative assumptions. This work represents the first neutron shielding verification analysis of a new projected proton therapy facility and, as such, it may serve as a new source of comparison and validation for the international community, besides confirming the viability of the project from a radioprotection point of view.