Toxicology

Verification of soman-related nerve agents via detection of phosphonylated adducts from rabbit albumin in vitro and in vivo Abstract A major challenge in organophosphate compound (OP) and OP nerve agent (OPNA) research has been in the identification and utilization of reliable biomarkers for rapid, sensitive, and efficient detection of OP exposure. Albumin has been widely studied as a biomarker for retrospective verification of exposure to OPNAs, including soman (GD), by detecting the phosphonylation of specific amino acid residues. The aim of the present study was to identify binding sites between GD and rabbit serum albumin in vitro and in vivo. A nano-liquid chromatography coupled with a quadrupole-orbitrap mass spectrometry (nLC-Q-Orbitrap-MS) was used to examine the GD-modified adducts of rabbit albumin. A total of 11 GD-modified sites were found in rabbit serum albumin across three experimental models. The following five GD-modified rabbit albumin sites, which were all lysine residues, were established in vivo: K188, K329, K162, K233, and K525. Two of these five lysine residues, K188 in peptide EK*ALISAAQER and K162 in peptide YK*AILTECCEAADK, were stable for at least 7 days in vivo. Molecular simulation of the GD–albumin interaction provided theoretical evidence for reactivity of the identified lysine residues. The findings suggest that these modifiable lysine residues are potential biomarkers of GD exposure for retrospective analysis by Q-Orbitrap-MS.

Comment on 'Kim, S.-J., Choi, E.-J., Choi, G.-W., Lee, Y.-B., and Cho, H.-Y. (2019). Exploring sex differences in human health risk assessment for PFNA and PFDA using a PBPK model, Arch Toxicol 93:311–330'

Uptake and effects of orally ingested polystyrene microplastic particles in vitro and in vivo Abstract Evidence exists that humans are exposed to plastic microparticles via diet. Data on intestinal particle uptake and health-related effects resulting from microplastic exposure are scarce. Aim of the study was to analyze the uptake and effects of microplastic particles in human in vitro systems and in rodents in vivo. The gastrointestinal uptake of microplastics was studied in vitro using the human intestinal epithelial cell line Caco-2 and thereof-derived co-cultures mimicking intestinal M-cells and goblet cells. Different sizes of spherical fluorescent polystyrene (PS) particles (1, 4 and 10 µm) were used to study particle uptake and transport. A 28-days in vivo feeding study was conducted to analyze transport at the intestinal epithelium and oxidative stress response as a potential consequence of microplastic exposure. Male reporter gene mice were treated three times per week by oral gavage with a mixture of 1 µm (4.55 × 107 particles), 4 µm (4.55 × 107 particles) and 10 µm (1.49 × 106 particles) microplastics at a volume of 10 mL/kg/bw. Effects of particles on macrophage polarization were investigated using the human cell line THP-1 to detect a possible impact on intestinal immune cells. Altogether, the results of the study demonstrate the cellular uptake of a minor fraction of particles. In vivo data show the absence of histologically detectable lesions and inflammatory responses. The particles did not interfere with the differentiation and activation of the human macrophage model. The present results suggest that oral exposure to PS microplastic particles under the chosen experimental conditions does not pose relevant acute health risks to mammals.

Proteomic analysis of hippocampal proteins in acrylamide-exposed Wistar rats Abstract Acrylamide has been used industrially and also found in certain foods cooked at high temperatures. Previous reports described acrylamide-related human intoxication who presented with ataxia, memory impairment, and/or illusion. The aim of this study was to characterize the molecular mechanisms of neurotoxicity of acrylamide by analyzing the expression levels of various proteins in the hippocampus of rats exposed to acrylamide. Male Wistar rats were administered acrylamide by gavage at 0, 2, and 20 mg/kg for 1 week or 0, 0.2, 2, and 20 mg/kg for 5 weeks. At the end of the experiment, the hippocampus was dissected out and proteins were extracted for two-dimensional difference gel electrophoresis combined with matrix-assisted laser-desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF/MS). MALDI-TOF/TOF/MS identified significant changes in two proteins in the 1-week and 22 proteins in the 5-week exposure groups. These changes were up-regulation in 9 and down-regulation in 13 proteins in the hippocampus of rats exposed to acrylamide at 20 mg/kg for 5 weeks. PANTHER overrepresentation test based on the GO of biological process showed significant overrepresentation in proteins annotated to nicotinamide nucleotide metabolic process, coenzyme biosynthetic process, pyruvate metabolic process, and carbohydrate metabolic process. The test also showed significant overrepresentation in proteins annotated to creatinine kinase activity for the GO of molecular function as well as myelin sheath, cytoplasmic part, and cell body for the GO of cellular component. Comparison with a previous proteomic study on hippocampal proteins in rats exposed to 1-bromopropane identified triosephosphate isomerase, mitochondrial creatine kinase U-type, creatine kinase β-type and proteasome subunit α type-1 as proteins affected by exposure to acrylamide and 1-bromopropane, suggesting a common mechanism of neurotoxicity for soft electrophiles.

Ecdysteroids as non-conventional anabolic agent: performance enhancement by ecdysterone supplementation in humans Abstract Recent studies suggest that the anabolic effect of ecdysterone, a naturally occurring steroid hormone claimed to enhance physical performance, is mediated by estrogen receptor (ER) binding. In comparison with the prohibited anabolic agents (e.g., metandienone and others), ecdysterone revealed to be even more effective in a recent study performed in rats. However, scientific studies in humans are very rarely accessible. Thus, our project aimed at investigating the effects of ecdysterone-containing products on human sport exercise. A 10-week intervention study of strength training of young men (n = 46) was carried out. Different doses of ecdysterone-containing supplements have been administered during the study to evaluate the performance-enhancing effect. Analysis of blood and urine samples for ecdysterone and potential biomarkers of performance enhancement has been conducted. To ensure the specificity of the effects measured, a comprehensive screening for prohibited performance-enhancing substances was also carried out. Furthermore, the administered supplement has been tested for the absence of anabolic steroid contaminations prior to administration. Significantly higher increases in muscle mass were observed in those participants that were dosed with ecdysterone. The same hypertrophic effects were also detected in vitro in C2C12 myotubes. Even more relevant with respect to sports performance, significantly more pronounced increases in one-repetition bench press performance were observed. No increase in biomarkers for liver or kidney toxicity was noticed. These data underline the effectivity of an ecdysterone supplementation with respect to sports performance. Our results strongly suggest the inclusion of ecdysterone in the list of prohibited substances and methods in sports in class S1.2 "other anabolic agents".

Neurotoxicity of Micrurus lemniscatus lemniscatus (South American coralsnake) venom in vertebrate neuromuscular preparations in vitro and neutralization by antivenom Abstract We investigated the effect of South American coralsnake (Micrurus lemniscatus lemniscatus) venom on neurotransmission in vertebrate nerve–muscle preparations in vitro. The venom (0.1–30 µg/ml) showed calcium-dependent PLA2 activity and caused irreversible neuromuscular blockade in chick biventer cervicis (BC) and mouse phrenic nerve–diaphragm (PND) preparations. In BC preparations, contractures to exogenous acetylcholine and carbachol (CCh), but not KCl, were abolished by venom concentrations ≥ 0.3 µg/ml; in PND preparations, the amplitude of the tetanic response was progressively attenuated, but with little tetanic fade. In low Ca2+ physiological solution, venom (10 µg/ml) caused neuromuscular blockade in PND preparations within ~ 10 min that was reversible by washing; the addition of Ca2+ immediately after the blockade temporarily restored the twitch responses, but did not prevent the progression to irreversible blockade. Venom (10 µg/ml) did not depolarize diaphragm muscle, prevent depolarization by CCh, or cause muscle contracture or histological damage. Venom (3 µg/ml) had a biphasic effect on the frequency of miniature end-plate potentials, but did not affect their amplitude; there was a progressive decrease in the amplitude of evoked end-plate potentials. The amplitude of compound action potentials in mouse sciatic nerve was unaffected by venom (10 µg/ml). Pre-incubation of venom with coralsnake antivenom (Instituto Butantan) at the recommended antivenom:venom ratio did not neutralize the neuromuscular blockade in PND preparations, but total neutralization was achieved with a tenfold greater volume of antivenom. The addition of antivenom after 50% and 80% blockade restored the twitch responses. These results show that M. lemniscatus lemniscatus venom causes potent, irreversible neuromuscular blockade, without myonecrosis. This blockade is apparently mediated by pre- and postsynaptic neurotoxins and can be reversed by coralsnake antivenom.

The in vivo developmental toxicity of diethylstilbestrol (DES) in rat evaluated by an alternative testing strategy Abstract In the present study, we evaluated an alternative testing strategy to quantitatively predict the in vivo developmental toxicity of the synthetic hormone diethylstilbestrol (DES). To this end, a physiologically based kinetic (PBK) model was defined that was subsequently used to translate concentration–response data for the in vitro developmental toxicity of DES, obtained in the ES-D3 cell differentiation assay, into predicted in vivo dose–response data for developmental toxicity. The previous studies showed that the PBK model-facilitated reverse dosimetry approach is a useful approach to quantitatively predict the developmental toxicity of several developmental toxins. The results obtained in the present study show that the PBK model adequately predicted DES blood concentrations in rats. Further studies revealed that DES tested positive in the ES-D3 differentiation assay and that DES-induced inhibition of the ES-D3 cell differentiation could be counteracted by the estrogen receptor alpha (ERα) antagonist fulvestrant, indicating that the in vitro ES-D3 cell differentiation assay was able to mimic the role of ERα reported in the mode of action underlying the developmental toxicity of DES in vivo. In spite of this, combining these in vitro data with the PBK model did not adequately predict the in vivo developmental toxicity of DES in a quantitative way. It is concluded that although the EST qualifies DES as a developmental toxin and detects the role of ERα in this process, the ES-D3 cell differentiation assay of the EST apparently does not adequately capture the processes underlying DES-induced developmental toxicity in vivo.

Novel insight in estrogen homeostasis and bioactivity in the ACI rat model of estrogen-induced mammary gland carcinogenesis Abstract Despite being widely used to investigate 17β-estradiol (E2)-induced mammary gland (MG) carcinogenesis and prevention thereof, estrogen homeostasis and its significance in the female August Copenhagen Irish (ACI) rat model is unknown. Thus, levels of 12 estrogens including metabolites and conjugates were determined mass spectrometrically in 38 plasmas and 52 tissues exhibiting phenotypes ranging from normal to palpable tumor derived from a representative ACI study using two different diets. In tissues, 40 transcripts encoding proteins involved in estrogen (biotrans)formation, ESR1-mediated signaling, proliferation and oxidative stress were analyzed (TaqMan PCR). Influence of histo(patho)logic phenotypes and diet on estrogen and transcript levels was analyzed by 2-way ANOVA and explanatory variables influencing levels and bioactivity of estrogens in tissues were identified by multiple linear regression models. Estrogen profiles in tissue and plasma and the influence of Hsd17b1 levels on intra-tissue levels of E2 and E1 conclusively indicated intra-mammary formation of E2 in ACI tumors by HSD17B1-mediated conversion of E1. Proliferation in ACI tumors was influenced by Egfr, Igf1r, Hgf and Met levels. 2-MeO-E1, the only oxidative estrogen metabolite detected above 28–42 fmol/g, was predominately observed in hyperplastic tissues and intra-tissue conversion of E1 seemed to contribute to its levels. The association of the occurrence of 2-MeO-E1 with higher levels of oxidative stress observed in hyperplastic and tumor tissues remained equivocal. Thus, the present study provides mechanistic explanation for previous and future results observed in the ACI model.

Okadaic acid activates Wnt/β-catenin-signaling in human HepaRG cells Abstract The lipophilic phycotoxin okadaic acid (OA) occurs in the fatty tissue and hepatopancreas of filter-feeding shellfish. The compound provokes the diarrhetic shellfish poisoning (DSP) syndrome after intake of seafood contaminated with high levels of the DSP toxin. In animal experiments, long-term exposure to OA is associated with an elevated risk for tumor formation in different organs including the liver. Although OA is a known inhibitor of the serine/threonine protein phosphatase 2A, the mechanisms behind OA-induced carcinogenesis are not fully understood. Here, we investigated the influence of OA on the β-catenin-dependent Wnt-signaling pathway, addressing a major oncogenic pathway relevant for tumor development. We analyzed OA-mediated effects on β-catenin and its biological function, cellular localization, post-translational modifications, and target gene expression in human HepaRG hepatocarcinoma cells treated with non-cytotoxic concentrations up to 50 nM. We detected concentration- and time-dependent effects of OA on the phosphorylation state, cellular redistribution as well as on the amount of transcriptionally active β-catenin. These findings were confirmed by quantitative live-cell imaging of U2OS cells stably expressing a green fluorescent chromobody which specifically recognize hypophosphorylated β-catenin. Finally, we demonstrated that nuclear translocation of β-catenin mediated by non-cytotoxic OA concentrations results in an upregulation of Wnt-target genes. In conclusion, our results show a significant induction of the canonical Wnt/β-catenin-signaling pathway by OA in human liver cells. Our data contribute to a better understanding of the molecular mechanisms underlying OA-induced carcinogenesis.

The Toll-like receptor agonist imiquimod is metabolized by aryl hydrocarbon receptor-regulated cytochrome P450 enzymes in human keratinocytes and mouse liver Abstract The Toll-like receptor 7 agonist imiquimod (IMQ) is an approved drug for the topical treatment of various skin diseases that, in addition, is currently tested in multiple clinical trials for the immunotherapy of various types of cancers. As all of these trials include application of IMQ to the skin and evidence exists that exposure to environmental pollutants, i.e., tobacco smoke, affects its therapeutic efficacy, the current study aims to elucidate the cutaneous metabolism of the drug. Treatment of human keratinocytes with 2.5 µM benzo[a]pyrene (BaP), a tobacco smoke constituent and aryl hydrocarbon receptor (AHR) agonist, for 24 h induced cytochrome P450 (CYP) 1A enzyme activity. The addition of IMQ 30 min prior measurement resulted in a dose-dependent inhibition of CYP1A activity, indicating that IMQ is either a substrate or inhibitor of CYP1A isoforms. Incubation of 21 recombinant human CYP enzymes with 0.5 µM IMQ and subsequent LC–MS analyses, in fact, identified CYP1A1 and CYP1A2 as being predominantly responsible for IMQ metabolism. Accordingly, treatment of keratinocytes with BaP accelerated IMQ clearance and the associated formation of monohydroxylated IMQ metabolites. A co-incubation with 5 µM 7-hydroxyflavone, a potent inhibitor of human CYP1A isoforms, abolished basal as well as BaP-induced IMQ metabolism. Further studies with hepatic microsomes from CD-1 as well as solvent- and β-naphthoflavone-treated CYP1A1/CYP1A2 double knock-out and respective control mice confirmed the critical contribution of CYP1A isoforms to IMQ metabolism. Hence, an exposure to life style-related, dietary, and environmental AHR ligands may affect the pharmacokinetics and, thus, treatment efficacy of IMQ.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Radiological Physics and Technology

Early-age Ndufs4 knockout mice are an inappropriate animal model of Leigh syndrome

Early detection of elevated lactate levels in a mitochondrial disease model using chemical exchange saturation transfer (CEST) and magnetic resonance spectroscopy (MRS) with 7T MR imaging

Proposal of a new method to prove that unnecessary information is not drawn on the image using statistical analysis Abstract The purpose of this study is to propose a new method of image evaluation using statistical analysis. We used the Sign test and the Wilcoxon test to analyze the statistical significance of image differences. Using this method, we evaluated whether the small electrode of the DAP meter appears in the X-ray image. Two observed values, which were obtained by averaging all values under all exposure conditions, were compared. All the observation tests showed the same sign. Thus, the results proved that the small electrode of the DAP meter is not present on the image. Using this method, it became possible to prove that the electrode was not depicted, which was impossible to determine using conventional methods. The method combining both the Sign test and the Wilcoxon test can be useful in image evaluation.

Development of a new image manipulation system based on detection of electroencephalogram signals from the operator's brain: a feasibility study Abstract Physicians require an adequate display system with a console within arm's reach to view images during surgical operations and interventional radiological examinations. However, manipulation of the console by physicians themselves may not be possible because their hands may be otherwise engaged. In this study, an image manipulation system using an electroencephalogram (EEG) sensor mounted on the operator's head was developed. In this system, data acquired by the device is used to manipulate images, and the output can be converted to commands for various actions such as paging, which can be controlled by the operator's eye-blink, and zooming of a region indicated by the cursor, which can be controlled by the operator's mental concentration. In this study, the MindWave Mobile headset was used as EEG sensor, and AZEWIN for the display system. Ten observers were enrolled and fitted with EEG device to determine the threshold values of blink strength and attention; threshold value of 100 for blink strength and 65 for attention were determined. Thirty-one observers were enrolled and fitted with EEG device to investigate average response-time; the average response time for detecting paging was 0.43 ± 0.02 s, and that for zooming was 5.85 ± 0.56 s. Thus, the proposed image manipulation system using the operator's EEG signals enabled physicians to assess and manipulate images without using their hands.

Voxel-based morphometry analysis of double inversion-recovery magnetic resonance imaging for detecting microscopic lesions: a simulation study Abstract Double inversion-recovery (DIR) imaging has the potential to improve the detection of subcortical lesions through the use of voxel-based morphometry (VBM) analysis. The aim of this study was to clarify the characteristics of detectable lesions by performing a VBM analysis on DIR images of simulated lesions. Twenty healthy volunteers underwent magnetic resonance imaging using a head three-dimensional DIR sequence. The images were processed using SPM12; then, the selected images with simulated lesions were analyzed via VBM. The VBM results were evaluated using free-response receiver-operating characteristic curves and a receiver-operating characteristic analysis. The sensitivity was 100% (5/5), with 5.6 false-positive objects per case, in simulated lesions with a contrast of 0.6 and a size of 2.4 mm. The sensitivity was 80% (4/5), with 5.4 false-positive objects per case, in simulated lesions with a contrast of 0.5 and a size of 2.4 mm. The mean area under the curve value was increased from 0.783 to 0.883 using VBM, with a statistically significant difference (p < 0.01). The VBM analysis of the DIR images using SPM alone showed the potential to detect subcortical microscopic lesions. Early detection of Alzheimer's disease may be possible by adapting VBM in the clinical setting.

Estimation and validation of the frequency responses of a scanner system and an image reconstruction system in X-ray computed tomography Abstract In computed tomography, factors that theoretically affect the modulation transfer function (MTF) in the region near the isocenter are the frequency responses of the scanner system (MTFS) and reconstruction processing (MTFA). Although MTFS and MTFA are performance indices that are not disclosed to the users, both can be estimated by the measured MTF with the use of theoretical formulas. In this study, we proposed two methods to obtain the MTFS and MTFA, and confirm their validity. The first method to obtain the MTFS and MTFA uses a theoretical formula and the measured MTF. Another method uses the measured MTF and the noise power spectrum. In both the methods, the MTFS and MTFA were obtained separately. By our proposed methods, performance indices that are not usually disclosed to the users can be known.

Appropriate echo time selection for quantitative susceptibility mapping Abstract The purpose of our study was to clarify the dependence of quantitative susceptibility mapping (QSM) on echo time (TE). We constructed a phantom consisting of six tubes; three tubes were filled with different concentrations (0.5, 1.0, and 2.5 mM) of gadopentetate dimeglumine (Gd-DTPA), and three were filled with different concentrations (100, 200, and 350 mg/mL) of calcium hydroxyapatite. Real and imaginary images from multi-echo spoiled gradient-echo data (12 echoes) were acquired. We then used four datasets with three serial echoes. The QSM procedure consists of four steps: field map estimation, phase unwrapping, background removal, and dipole inversion. For each sample, we compared the measured mean susceptibility value with the theoretical susceptibility value and conducted a linear regression analysis. Accordingly, the relationship between the measured susceptibility and concentration of Gd-DTPA was shown to agree well with the theoretical values (TEs = 16.4, 20.8, and 25.2 ms; slope = 0.24, R2 = 1.00). Furthermore, the relationship between the measured susceptibility and concentration of hydroxyapatite also showed good linearity (TEs = 16.4, 20.8, and 25.2 ms; slope = − 0.00121, R2 = 1.00). In conclusion, the optimization of the TE in QSM makes it possible to obtain more detailed information regarding the susceptibility of biomaterials.

Clinical application of biological fingerprints extracted from averaged chest radiographs and template-matching technique for preventing left–right flipping mistakes in chest radiography Abstract We aimed to evaluate the identification performance achieved using biological fingerprints extracted from averaged chest radiographs and template-matching techniques for the prevention of left–right flipping mistakes. We produced averaged chest radiographs for each sex by averaging 100 posteroanterior chest radiographs. Further, 400 and 566 chest radiographs were used in consistency and validation tests, respectively, and they were flipped horizontally to produce flipped chest radiographs under the assumption that the left–right flipping mistake occurred. The correlation values obtained with chest radiographs and those obtained with flipped chest radiographs were calculated. When we used correlation indices calculated from the correlation values from four biological fingerprints except for the lung apex, 96.5% (386/400) and 95.8% (542/566) of the left or right sides were identified correctly in the consistency and validation tests, respectively. This result indicates that our proposed method would be promising for the prevention of left–right flipping mistakes.

A new approach for detecting abnormalities in mammograms using a computer-aided windowing system based on Otsu's method Abstract Breast cancer is the most common cancer and the leading cause of cancer deaths in women worldwide. This study aimed to provide an automatic windowing method in mammograms, based on the principles of Otsu's thresholding function, to help radiologists more easily detect abnormalities on mammograms. A total of 322 mammographic images from the Mammographic Image Analysis Society (MIAS) database were used in the present study. The image background was removed based on Otsu's method. After selecting the threshold in the computer-aided windowing (CAW) system, the pixel values were kept larger than the threshold and displayed on a grayscale. A radiologist evaluated images randomly before and after CAW. Using CAW, the radiologist correctly diagnosed all healthy images (207 images). A total of 115 mammograms were evaluated to differentiate malignancy from benign masses. All 63 benign images were accurately diagnosed after using CAW. Moreover, of 52 malignant images, all were accurately recognized as malignant except one, which was recognized as benign. Therefore, specificity and sensitivity were significantly improved to 98% and 99.6%, respectively, and the area under the receiver operating characteristic (ROC) curve was calculated to be 0.99. The study showed that the use of CAW can potentially lead to quicker image assessment and improve the diagnostic accuracy of radiologists in differentiating between benign and malignant masses on mammograms.

Comparison of volumetric-modulated arc therapy and intensity-modulated radiation therapy prostate cancer plans accounting for cold spots Abstract This study compared dosimetric indices of volumetric-modulated arc therapy (VMAT) with intensity-modulated radiation therapy (IMRT) accounting for cold spots in prostate cancer plans. IMRT plans were retrospectively generated from 30 prostate cancer patients with ten cases for each risk group, who received VMAT plans. The mean, maximum, and minimum doses, and conformity and homogeneity indexes were evaluated for planning target volume (PTV) and the mean dose and V20–V70 for organs at risk (OAR) including the rectum, bladder, right and left femoral heads, and rectum overlapped with PTV (ROP) regions. The numbers and volume percentages of cold spots within PTVs and ROP regions were measured using in-house software. Three-dimensional probabilistic distributions of the probability and distributions of cold spots were generated using a centroid matching technique for visualization and analysis. There was a statistically better dose conformity in the PTV, rectum, and bladder dose-sparing in VMAT compared to that in the IMRT plans, whereas VMAT had statistically worse target dose homogeneity, and right and left femoral head dose-sparing than those of the IMRT plans. The average volume percentage of cold spots per PTV for the VMAT was 4.37 ± 2.68%, which was smaller than the 5.72 ± 1.84% observed for IMRT plans (P = 0.007). The volume percentage of cold spots per ROP for the VMAT did not significantly differ from those for the IMRT plans. Compared with IMRT, the VMAT plans achieved better PTV dose conformity, OAR dose-sparing, and smaller cold spots in the treatment of prostate cancer.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Radiology

Change in hepatic hemodynamics assessed by hepatic arterial blood pressure and computed tomography during hepatic angiography with the double balloon technique Abstract Purpose To assess the change in hepatic arterial blood pressure (HABP) and computed tomography during hepatic arteriography (CTHA) using the double balloon technique. Materials and methods Nine patients with hepatocellular carcinoma (HCC) were enrolled. We inserted a 5.2-Fr balloon catheter into the common or proper hepatic artery and a 1.8-Fr microballoon catheter into the lobar or segmental artery feeding the HCC. HABPs were measured with the 1.8-Fr microballoon catheter (usual-HABP), with the 1.8-Fr balloon inflated (B-HABP), and with both the 5.2-Fr and 1.8-Fr balloons inflated (BB-HABP). CTHAs were performed via a 1.8-Fr microcatheter (usual-CTHA), with the 1.8-Fr balloon inflated (B-CTHA selective), with both the 5.2-Fr and 1.8-Fr balloons inflated (BB-CTHA selective), and via the 5.2-Fr catheter with the 1.8-Fr balloon inflated (B-CTHA whole) and with both the 5.2-Fr and 1.8-Fr balloons inflated (BB-CTHA whole). Results In all cases, B-HABP was lower than usual-HABP. There was a decrease in BB-HABP in comparison with B-HABP in cases with occlusion of the proper hepatic artery. The contrast effect of B-CTHA selective increased in four cases. The contrast effect on B-CTHA whole remained in all cases. Conclusion This technique can be useful in decreasing HABP and collateral blood flow from the adjacent hepatic segment.

T2*-weighted MR imaging findings of giant cell tumors of bone: radiological–pathological correlation Abstract Purpose To assess the correlation between T2*-weighted MR imaging and pathological findings of giant cell tumors (GCT) of bone. Methods Of the 33 patients with histopathologically proven GCT of bone, 12 were examined using 1.5-T MR imaging, including T2*-weighted imaging, and were included in this study. The imaging and pathological findings of GCTs were compared between GCTs with and without hypointensity on T2*-weighted images (T2* hypointensity). Results T2* hypointensity was observed in 6 out of 12 (50%) GCTs. Septal formation (83% vs. 17%; p < 0.05) and cystic formation (67% vs. 0%; p < 0.05) on T2-weighted images was significantly more frequent in the GCTs with T2* hypointensity compared with those without T2* hypointensity. Among the six GCTs with T2* hypointensity, a large amount of hemosiderin deposition was pathologically observed in five (83%) cases, whereas small amounts of hemosiderin deposition was seen in one (17%) case. In contrast, among the six GCTs without T2* hypointensity, a small amount of hemosiderin deposition was pathologically observed in all six (100%). Conclusion Half of the GCTs showed T2* hypointensity, which is characteristic of hemosiderin deposition; whereas, the other half did not show T2* hypointensity due to a small amount of hemosiderin deposition.

Distinction between benign and malignant breast masses at breast ultrasound using deep learning method with convolutional neural network Abstract Purpose We aimed to use deep learning with convolutional neural network (CNN) to discriminate between benign and malignant breast mass images from ultrasound. Materials and Methods We retrospectively gathered 480 images of 96 benign masses and 467 images of 144 malignant masses for training data. Deep learning model was constructed using CNN architecture GoogLeNet and analyzed test data: 48 benign masses, 72 malignant masses. Three radiologists interpreted these test data. Sensitivity, specificity, accuracy, and area under the receiver operating characteristic curve (AUC) were calculated. Results The CNN model and radiologists had a sensitivity of 0.958 and 0.583–0.917, specificity of 0.925 and 0.604–0.771, and accuracy of 0.925 and 0.658–0.792, respectively. The CNN model had equal or better diagnostic performance compared to radiologists (AUC = 0.913 and 0.728–0.845, p = 0.01–0.14). Conclusion Deep learning with CNN shows high diagnostic performance to discriminate between benign and malignant breast masses on ultrasound.

Radiation with concomitant superselective intra-arterial cisplatin infusion for maxillary sinus squamous cell carcinoma Abstract Purpose To assess the efficacy and prognostic factors after superselective intra-arterial chemoradiation (RADPLAT) for maxillary sinus squamous cell carcinoma (MS-SCC). Materials and methods Prognostic significance of age, gender, T and N factors, gross tumor volume of the primary-site (GTV), total cisplatin dosage, and total cisplatin dosage per GTV (CDDP/GTV) for primary-site recurrence-free survival rate (PRFS) were analyzed. RADPLAT was administered to 27 patients. The median follow-up period was 42.1 months. Results The 3-year rates of overall survival and PRFS were 59.2% and 53.9%, respectively. In univariate analysis, age, male, and total cisplatin dosage were significant factors for PRFS. In multivariate analysis, lymph node metastasis was significant factors for PRFS, and gender and total cisplatin dosage weakly influenced PRFS. In acute phase, no patient showed ≥ grade 3 hematologic toxicity, and grade 3 mucositis developed in 5 patients. Late toxicities were recognized in 3 patients (grade 2 phlegmon of the face, grade 3 maxillofacial osteonecrosis, and retinopathy). Twelve patients (44%) experienced recurrences. Of them, 8 patients showed recurrence at the primarysite. Conclusion RADPLAT was effective for MS-SCC, with acceptable toxicity. Total cisplatin dosage is suggested to be important for primary tumor control.

Detectability of the choroid plexus of the third ventricle with magnetic resonance ventriculography Abstract Purpose To clarify the detectability of the choroid plexus of the third ventricle (ChPl3V) with magnetic resonance ventriculography (MRVn) employing a steady-state free precession (SSFP) sequence in comparison to surgical endoscopic movies as a golden standard, as we encountered some clinical cases of total agenesis of corpus callosum (ACC) where we could not recognize the choroid plexus of the third ventricle and found no previous article addressing this problem. Materials and methods This retrospective study included consecutive patients from 2010 to 2016 for whom endoscopic evaluation of the third ventricle was conducted. The anterior portion of the right and left streaks of ChPl3V was evaluated in 8 patients on 16 sites, while the posterior portion of both streaks of ChPl3V was evaluated in 13 patients on 26 sites. Sensitivity of MRVn to visualize ChPl3V with endoscopic movies as the golden standard was calculated. Results Sensitivity of MRVn in visualizing the anterior portion of ChPl3V was 0.813, and that for the posterior portion 0.692. The anterior portion of ChPl3V was visualized in all cases where no tumor contacted the foramen of Monro. Conclusion MRVn visualizes the anterior portion of ChPl3V with significant sensitivity and the posterior portion with lower one.

Changes in tissue gadolinium biodistribution measured in an animal model exposed to four chelating agents Abstract Purpose This study investigated the potential to reduce gadolinium levels in rodents after repetitive IV Gadodiamide administration using several chelating agents. Materials and methods The following six groups of rats were studied. Group 1: Control; Group 2: Gadodiamide only; Group 3: Meso-2,3-Dimercaptosuccinic acid (DMSA) + Gadodiamide; Group 4: N-Acetyl-l-cysteine (NAC) + Gadodiamide; Group 5: Coriandrum sativum extract + Gadodiamide; and Group 6: Deferoxamine + Gadodiamide. Brain, kidney, and blood samples were evaluated via inductively coupled plasma mass spectrometry. The brain was also evaluated histologically. Results Kidney gadolinium levels in Groups 4 and 5 were approximately double that of Group 2 (p = 0.033 for each). There was almost no calcification in rat hippocampus for Group 4 rodents when compared with Groups 2, 3, 5 and 6. Conclusion Our preliminary study shows that excretion to the kidney has a higher propensity in NAC and Coriandrum sativum groups. It may be possible to change the distribution of gadolinium by administrating several agents. NAC may lower Gadodiamide-induced mineralization in rat hippocampus.

Ethanol fixation method for heart and lung imaging in micro-CT Abstract Purpose The soft tissue imaging in micro-CT remains challenging due to its low intrinsic contrast. The aim of this study was to create a simple staining method omitting the usage of contrast agents for ex vivo soft tissue imaging in micro-CT. Materials and methods Hearts and lungs from 30 mice were used. Twenty-seven organs were either fixed in 97% or 50% ethanol solution or in a series of ascending ethanol concentrations. Images were acquired after 72, 168 and 336 h on a custom-built micro-CT machine and compared to scans of three native samples. Results Ethanol provided contrast enhancement in all evaluated fixations. Fixation in 97% ethanol resulted in contrast enhancement after 72 h; however, it caused hardening of the samples. Fixation in 50% ethanol provided contrast enhancement after 336 h, with milder hardening, compared to the 97% ethanol fixation, but the visualization of details was worse. The fixation in a series of ascending ethanol concentrations provided the most satisfactory results; all organs were visualized in great detail without tissue damage. Conclusions Simple ethanol fixation improves the tissue contrast enhancement in micro-CT. The best results can be obtained with fixation of the soft tissue samples in a series of ascending ethanol concentrations.

Monitoring of fatigue in radiologists during prolonged image interpretation using fNIRS Abstract Purpose To determine whether functional near-infrared spectroscopy (fNIRS) allows monitoring fatigue in radiologists during prolonged image interpretation. Materials and methods Nine radiologists participated as subjects in the present study and continuously interpreted medical images and generated reports for cases for more than 4 h under real clinical work conditions. We measured changes in oxygenated hemoglobin concentrations [oxy-Hb] in the prefrontal cortex using 16-channel fNIRS (OEG16ME, Spectratech) every hour during the Stroop task to evaluate fatigue of radiologists and recorded fatigue scale (FS) as a behavior data. Results Two subjects showed a subjective feeling of fatigue and an apparent decrease in brain activity after 4 h, so the experiment was completed in 4 h. The remaining seven subjects continued the experiment up to 5 h. FS decreased with time, and a significant reduction was observed between before and the end of image interpretation. Seven out of nine subjects showed a minimum [oxy-Hb] change at the end of prolonged image interpretation. The mean change of [oxy-Hb] at the end of all nine subjects was significantly less than the maximum during image interpretation. Conclusion fNIRS using the change of [oxy-Hb] may be useful for monitoring fatigue in radiologists during image interpretation.

Cryoablation of renal cell carcinoma for patients with stage 4 or 5 non-dialysis chronic kidney disease Abstract Purpose To evaluate the safety and efficacy of cryoablation for renal cell carcinoma (RCC) in patients with stage 4 or 5 non-dialysis chronic kidney disease (CKD). Materials and methods This retrospective multicenter study included patients with maximum tumor diameter ≤ 4 cm, estimated glomerular filtration rate (eGFR) < 30 ml/min/1.73 m2, in whom cryoablation was performed percutaneously with curative intent between July 2011 and May 2016. Results Of 541 patients who underwent renal tumor cryoablation, 17 (3.1%; 4 women, 13 men; mean age 70.1 ± 10.6 years) with stage 4 or 5 non-dialysis CKD were included in this study. The pre-cryoablation eGFR was 22.5 ± 6.3 ml/min/1.73 m2. The mean tumor diameter was 2.8 ± 0.7 cm. No Grade 3 or higher adverse events occurred post-cryoablation. The eGFR at each time point was significantly lower than that before treatment. One patient required hemodialysis initiation at 21 months post-procedure. None of the patients showed residual RCC at their last follow-up. Conclusion Cryoablation of RCC is safe in patients with stage 4 or 5 non-dialysis CKD and yields treatment results comparable to those in patients without CKD. This treatment could be completed without the early initiation of hemodialysis after the procedure.

Cinematic rendering: a new imaging approach for ulcerative colitis Abstract Purpose Cinematic rendering (CR) is a new technique for visualizing volumetric three-dimensional data. The purpose of this study was to investigate the added value of CR to conventional computed tomography (CT) in the diagnosis and evaluation of ulcerative colitis (UC). Materials and methods We retrospectively evaluated the CT data of 48 patients (33 men, 15 women; mean age, 44.35 years) with a definitive diagnosis of UC. All patients underwent conventional CT and CR, and had colonoscopy results. Two radiologists independently reviewed the conventional CT images first without and then with CR. Then, the imaging value of CR was evaluated by both radiologists together. The readers were blinded to the disease extent. The diagnostic performance of CR for both readers was assessed by receiver-operating characteristic (ROC) curve analysis. Results There were 23 cases of mild to moderate UC and 25 cases of severe UC, which were divided into two groups. Both readers showed improved diagnostic performance with the addition of CR (the area under the ROC curve improved from 0.676 to 0.804, P = 0.0255, and from 0.679 to 0.826, P = 0.0049, for readers 1 and 2, respectively). Full view of the lesion and contrast enhancement was not significantly different between the two groups (P > 0.05). Increased mesenteric vascularity and the comb sign on CR were more clearly observed in the severe group (P < 0.05). Conclusion Adding CR to conventional CT improved the diagnostic performance of evaluating the extent of UC.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Radiation Protection and Environment

Radon in dwellings and workplaces: An update on current regulations Pushparaja  Radiation Protection and Environment 2019 42(1):1-4

A comparative study for 235U radioactivity concentration calculation methods in phosphate samples A Salman, Z Ahmed, Kh A Allam, S El-Sharkawy Radiation Protection and Environment 2019 42(1):5-9 In this study, a comparison is made between the selection of the most proper and accurate methods for 235U activity determination according to the available conditions with the smallest error possible. The detection of 235U activity is used to determine the mass of the 235U in the samples and consecutively, the uranium enrichment which is important for the nuclear safeguards' needs and the monitoring of the radionuclides in the environment. Various methods have been studied to distinguish between counting rate contributions from both235U and 226Ra to the 186 keV energy region in samples of uranium in natural form.

Measurement of radon gas activity concentrations in drinking water in the city center of Adıyaman, Turkey Mehmet Fatih Aydin, Ömer Söǧüt Radiation Protection and Environment 2019 42(1):10-14 In this study, 16 water samples collected from homes and the water tanks in various districts from two different natural water sources, supplying Adıyaman's domestic drinking water. Measurements of radon gas activity concentrations in collected samples have been performed by AlphaGUARD PQ2000 PRO radon detector. The average radon (222Rn) activity concentrations of samples taken from the natural water resources and homes were found to be 0.39 ± 0.11 and 0.51 ± 0.14 Bq/L, respectively. In addition, the annual effective dose equivalent that exposed due to radon gas in drinking water is also calculated and found to be 1.30 ± 0.36 μSv/y. The upper limit for radon gas activity concentrations in drinking waters has been defined by the United States Environmental Protection Agency (USEPA), United Nations Scientific Committee on the Effects of Atomic Radiation, and World Health Organization to be 11, 40, and 100 Bq/L, respectively. The measured values of radon gas activity concentrations are lower than that of limit value defined by the USEPA.

A new look at blind test procedures in personnel monitoring Sneha Chandrasekhar, SM Pradhan, Madhumita Bhattacharya, AK Bakshi, D Datta Radiation Protection and Environment 2019 42(1):15-21 In India, a single type of dosimeter is used all over the country for estimation of occupational doses due to X, beta, and gamma radiations. The quality assurance program of external personnel monitoring in India includes an annual announced test and periodic blind tests. The announced test carried out by a central agency includes various energies and angles of irradiation. Blind tests are carried out by specified users of personnel monitoring service who are distributed throughout the country. Due to the lack of easy availability of various energy sources, blind tests are mostly carried out for a single energy. Analysis of the results is carried out by means of ANSI criteria or trumpet curve analysis which may not be sufficiently stringent given that a single energy and angle are used. In view of this and also to meet the specific requirements of the Indian monitoring scenario, keeping in mind existing practices, an attempt is made to have a new look at the analysis of blind test results. The criteria which have been considered are zeroing of dosimeters, coefficient of variation, and bias. The results of a blind test based on harmonized procedures can be utilized to understand the actual quality of service and also identify deviations in procedures, if any.

Effect of natural radioactivity content in the beach sands along the east coast of Tamil Nadu, India, due to tsunami KS Lakshmi, V Meenakshisundaram, J Punniyakotti Radiation Protection and Environment 2019 42(1):22-27 In the aftermath of devastating tsunami, studies were undertaken to assess its impact on the environmental radioactivity profile in the beach sands along the entire east coastal region of Tamil Nadu state (~1000 km) and compared with previously established pretsunami distribution profile. The results of the present study clearly indicate that the radioactivity content of 232Th and 238U in the beach sands registered a steep fall (52% and 41%, respectively), whereas the 40K activity has increased by 26%. This is attributed to the hydrodynamics of tsunami waves, which removed huge quantities of beach minerals that have been built-up over a few decades. The individual activity content of the three primordial radionuclides, total activity content, comparison between pre- and post-tsunami data for all the 32 sites, and their statistical analyses are covered in the paper.

Measurement of patient skin dose and establishment of local diagnostic reference levels for interventional cardiology procedures Arti R Kulkarni, Philomina Akhilesh, Sunil Dutt Sharma Radiation Protection and Environment 2019 42(1):28-33 Use of diagnostic reference levels (DRLs) is an important tool for patient dose optimization in interventional radiology. DRLs are normally expressed in terms of kerma-area product (Pka). However, no direct method is available to estimate the probability of skin reactions using system displayed quantities. The aim of this work was to measure skin entrance doses (SEDs) in patients undergoing coronary angiography (CA) and percutaneous coronary interventions (PCI) using Gafchromic films and establish DRLs. The details of patients, exposure parameters, and dose quantities were recorded for 572 patients. Out of these, skin doses were measured for selected 64 patients using Gafchromic film. Measured SEDs are in the range of 48.2–740 mGy and 84–1242 mGy for CA and PCI, respectively. The data of 572 patients were analyzed, and 75th percentile of Pkawas calculated. DRLs for CA and PCI in terms of Pkaare found to be 34 and 134 Gy.cm2, respectively. DRLs for CA and PCI in terms of cumulative air kerma (Ka) at reference point are found to be 590 mGy and 1930 mGy, respectively. SED has a good correlation with Ka, however, it does not correlate well with Pka.

Naturally occurring radioactive material and naturally occurring mercury assessment of black powder in sales gas pipelines Michael Ian Cowie, AM El-Sherik Radiation Protection and Environment 2019 42(1):34-39 Black powder solids are formed inside the sales gas pipelines as a result of internal corrosion due to condensed moisture and the presence of corrosive gases, namely, H2S, CO2, and O2. Naturally occurring radioactive material, principally the radionuclides lead-210 and polonium-210, and naturally occurring mercury (NOM) (Hg) have been identified in black powder. A detailed investigation into both the radiological components and the presence of NOM associated with black powder was carried out by Saudi Aramco. The aim of the investigation was to determine if waste produced during pipeline scraping operations presented a workers protection or environmental control problem due to the radioactivity and mercury present. The investigation looked at Saudi Aramco's entire sales gas pipeline network and spanned scraping operations over a 3-year period. This article details the sampling and analysis methods used to assess black powder samples, the results of sample analysis and advice provided to ensure workers' protection, and environmental control of the waste produced during pipeline scraping activities.

Assessment of radiological safety of abandoned tantalite mining sites in Oke-Ogun, Oyo State, Nigeria AE Ajetunmobi, AO Mustapha, IC Okeyode, AM Gbadebo, D Al-Azmi Radiation Protection and Environment 2019 42(1):40-46 Introduction: Tantalite and other solid mineral deposits in Oke-Ogun occupy a very large landscape, and the possibilities of further exploration in new landscape and abandoning unproductive sites cannot be ruled out. These landscapes when abandoned become hideout for criminals and ritualistic activities which pose a great threat to the wellness of the people in the area and Nigerians at large. Objectives: This study attempts to investigate the radiological safety of these abandoned mines so as to convert them to economical valuable sites that can serve as a means of employment for population of the unemployed youth in the area and Nigeria at large. Materials and Methods: A total of seventeen soil samples were randomly selected. These samples were processed and analyzed for activity concentrations of 226Ra, 232Th, and 40K using Hyper Pure Germanium Gamma Spectrometer. RESidual RADioactivity software version (6.5), a computer program developed by the Environmental Assessment Division of Argonne National Laboratory, was used for dose prediction for 70 years using the activity concentrations of soil samples from all the selected sites as its input parameter for the software. Predicted doses were estimated with respect to the three exposure pathways, namely external gamma irradiation from radionuclides in the soil samples, inhalation of dust particles, and inadvertent ingestion of soil. Results: The measured activity concentrations of radionuclides: 226Ra for all the sites is in the range of (17–177) Bq/kg, 232Th in the range of (2–107) Bq/kg, and 40K in the range of (650–1667) Bq/kg. The predicted dose for all the sites is within the range of 0.05–0.35 mSv/y with Eluku mining site having values greater than the permissible limit of 0.25 mSv/y. The health implication of these values is that three of the sites (Gbedu, Sepenteri, and Komu) are safe for other usages. Conclusion: Any of these three mining sites may be used for re-creational purposes, fishing and building of estates to mention but a few.

Lead nitrate loaded, novel clay-based bricks as radiation shielding materials for building applications Madhuri Dumpala Radiation Protection and Environment 2019 42(1):47-56 A new class of clay bricks designed, developed, and tested for their radiation shielding efficiency. The bricks were prepared using natural clay, lake clay, and lightweight clay and loaded with different concentrations of Pb (NO3)2. The gamma-ray shielding parameters were measured for these bricks with a transmission type of good geometry setup and operated at 511 and 662 keV energies. The gamma-ray spectrometer consists of a 2“×2” NaI (Tl) detector, 8K multichannel analyzer. The 22Na and 137Cs point isotropic gamma-ray sources were used in this study. To evaluate the efficiency of these bricks, various shielding parameters, such as linear attenuation coefficient, mass attenuation coefficient, mean-free path, half-value layer and tenth-value layer were calculated. The X-COM software was used to calculate these parameters theoretically, and both the experimental and theoretical values were compared, and they are in good agreement. These results suggest that the clay bricks prepared with the natural and lake clay attenuate more radiation than the lightweight clay bricks. It is also observed that the lightweight bricks prepared with the solutions with higher loadings of lead nitrate showing increased attenuation, this may be attributed to the reason that at higher loadings of LN, the porous gaps of lightweight bricks are being filled by the LN particles and contributed to the attenuation of radiation.

Assessment of ambient gamma dose rate in different locations of Amritsar city, Punjab, India Sumit Sharma, Ajay Kumar Radiation Protection and Environment 2019 42(1):57-62 Preliminary results of ambient indoor and outdoor gamma dose rates estimated in 20 distinct areas of Amritsar city, Punjab, are exhibited by utilizing Dosimeter-Radiometer MKS-03 (SARAD). This study was planned in such a way to get a uniform and representative distribution of estimations. The indoor to outdoor dose rate ratio was ascertained which showed that the indoor gamma dose rate as contrasted with outdoor gamma dose rate has elevated levels of exposure due to confined space and poor ventilation. The indoor and outdoor annual effective dose rate was likewise estimated from the exposure point of view, and it varied from 0.35 to 1.61 mSv/y and from 0.11 to 0.44 mSv/y. The average values of outdoor and indoor effective dose rate levels in all of these locations were well below the world average value prescribed by the United Nations Scientific Committee on Effects of Atomic Radiations.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Natural Medicines

Rapid discovery and identification of the anti-inflammatory constituents in Zhi-Shi-Zhi-Zi-Chi-Tang Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Hai-Qiang WANG, Yun-Xiang ZHU, Yi-Ning LIU, Ruo-Liu WANG, Shu-Fang WANG Abstract The anti-inflammatory active ingredients of Zhi-Shi-Zhi-Zi-Chi-Tang (ZSZZCT), a traditional Chinese medicine formula, were predicted and identified using an approach based on activity index, LC-MS, semi-preparative LC and NMR. Firstly, the whole extract of ZSZZCT was analyzed using liquid chromatography-quadrupole time of flight-mass spectrometry (LC-Q-TOF-MS) and liquid chromatography - ion trap mass spectrometry (LC-IT-MS), 79 constituents were detected and 39 constituents were identified unambiguously or tentatively. Subsequently, the whole extract of the formula was separated into multiple components and the activity index method was used to calculate index values of the 79 constituents by integrating the chemical and pharmacological information of multiple components. Four polymethoxyl flavones were predicted as the major active constituents according to the activity index values. Furthermore, three polymethoxyl flavones were prepared using the strategy with semi-preparative LC guided by LC-MS, and their anti-inflammatory activities were validated. The results show that three polymethoxyl flavones with higher positive index values, i.e., 3, 5, 6, 7, 8, 3′, 4′-heptamethoxyflavone, 3-hydroxynobiletein and tangeretin had significant anti-inflammatory effects. In conclusion, the predicted results indicated that the activity index method is feasible for the accurate prediction of active constituents in TCM formulae.

New octadecanoid derivatives from the seeds of Ipomoea nil Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Xiu-Qing SONG, Jun-Sheng ZHANG, Shu-Juan YU, Jin-Hai YU, Hua ZHANG Abstract Four new octadecanoid derivatives (1–4) including a pair of enantiomers (1/2), along with 12 known analogues (5–16), were isolatedfrom the seeds of Ipomoea nil. Their structures were determined by detailed spectroscopic analyses and comparison with reported data of structurally related compounds, with the absolute configurations of 1 and 2 being assigned by an in situ dimolybdenum ECD method. Our bioassays revealed that these isolates did not show ABTS radical scavenging activity while 10 and 13 displayed better α-glucosidase inhibitory activity than the positive control acarbose (IC50 167.7 ± 1.55 μmol·L−1), with IC50 of 92.73 ± 3.12 and 11.39 ± 2.18μmol·L−1, respectively.

Jatrogricaine A: a new diterpenoid with a 5/6/6/4 carbon ring system from the stems of Jatropha podagrica Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Lin CHEN, Jia-Luo HUANG, Lei ZHANG, Hai-Yan TIAN, Sheng YIN Abstract Jatrogricaine A (1), a new diterpenoid possessing a 5/6/6/4 carbon ring system, together with eight known diterpenoids (2–9) were isolated from the stems of Jatropha podagrica. Their structures were elucidated by extensive spectroscopic methods and the absolute configuration of 1 was determined by single crystal X-ray diffraction analysis. All compounds were evaluated for their anti-inflammatory activities in vitro, and compound 3 showed significant inhibitory effects against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW264.7 macrophage cells with an IC50 of 13.44 ± 0.28 μmol·L−1, being comparable to the positive control, quercetin (IC50 17.00 ± 2.10 μmol·L−1).

Eight new cytotoxic annonaceous acetogenins from the seeds of Annona squamosa Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Cheng-Yao MA, Jia-Hui LU, Xiang LI, Xiao LIU, Jian-Wei CHEN Abstract Eight new annonaceous acetogenins, squamotin A-D (1–4), annosquatin IV-V (5 and 6), muricin O (7) and squamosten B (8), together with four known ones (9–12) were isolated from the seeds of Annona squamosa. Their structures were elucidated by chemical methods and spectral data. The inhibitory activities of compound 1–9 against three multidrug resistance cell lines were evaluated. All tested compounds showed strong cytotoxicity.

A novel approach based on metabolomics coupled with network pharmacology to explain the effect mechanisms of Danggui Buxue Tang in anaemia Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Yong-Li HUA, Qi MA, Zi-Wen YUAN, Xiao-Song ZHANG, Wan-Ling YAO, Peng JI, Jun-Jie HU, Yan-Ming WEI Abstract Danggui Buxue Tang (DBT) is a famous Chinese medicinal decoction. Mechanism of DBT action is wide ranging and unclear. Exploring new ways of treatment with DBT is useful. Sprague-Dawley(SD) rats were randomly divided into 3 groups including control (NC, Saline), the DBT (at a dose of 8.10 g−1), and blood deficiency(BD) (Cyclophosphamide (APH)-andCyclophosphamide(CTX)-induced anaemia). A metabolomics approach using Liquid Chromatography-Quadrupole-Time-of-Flight/Mass Spectrometry (LC/Q-TOFMS) was developed to perform the plasma metabolic profiling analysis and differential metaboliteswerescreened according to the multivariate statistical analysiscomparing the NC and BD groups, andthe hub metabolites were outliers with high scores of the centrality indices. Anaemia disease-related protein target and compound of DBT databases were constructed. The TCMSP, ChemMapper and STITCH databases were used to predict the protein targets of DBT. Using the Cytoscape 3.2.1 to establish a phytochemical component-target protein interaction network and establish a component, protein and hub metabolite protein-protein interaction (PPI) network and merging the three PPI networks basing on BisoGenet. The gene enrichment analysis was used to analyse the relationship between proteins based on the relevant genetic similarity by ClueGO. The results shown DBT effectively treated anaemia in vivo. 11 metabolic pathways are involved in the therapeutic effect of DBT in vivo; S-adenosyl-l-methionine, glycine, l-cysteine, arachidonic acid (AA) and phosphatidylcholine(PC) were screened as hub metabolites in APH-and CTX-induced anaemia. A total of 288 targets were identified as major candidates for anaemia progression. The gene-set enrichment analysis revealed that the targets are involved in iron ion binding, haemopoiesis, reactive oxygen species production, inflammation and apoptosis. The results also showed that these targets were associated with iron ion binding, haemopoiesis, ROS production, apoptosis, inflammation and related signalling pathways. DBT can promote iron ion binding and haemopoiesis activities, restrain inflammation, production of reactive oxygen, block apoptosis, and contribute significantly to the DBT treat anaemia.

Involvement of mitochondrial apoptotic pathway and MAPKs/NF-κ B inflammatory pathway in the neuroprotective effect of atractylenolide III in corticosterone-induced PC12 cells Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Wen-Xia GONG, Yu-Zhi ZHOU, Xue-Mei QIN, Guan-Hua DU Abstract Atractylenolide III (ATL-III), a sesquiterpene compound isolated from Rhizoma Atractylodis Macrocephalae, has revealed a number of pharmacological properties including anti-inflammatory, anti-cancer activity, and neuroprotective effect. This study aimed to evaluate the cytoprotective efficiency and potential mechanisms of ATL-III on corticosterone injured rat phaeochromocytoma (PC12) cells. Our results demonstrate that ATL-III increases cell viability and reduces the release of lactate dehydrogenase (LDH). The results suggest that ATL-III protects PC12 cells from corticosterone-induced injury by inhibiting the intracellular Ca2+ overloading, inhibiting the mitochondrial apoptotic pathway and modulating the MAPK/NF-ΚB inflammatory pathways. These findings provide a novel insight into the molecular mechanism by which ATL-III protected the PC12 cells against corticosterone-induced injury for the first time. Our results provide the evidence that ATL-III may serve as a therapeutic agent in the treatment of depression.

Astragalus membranaceus improves therapeutic efficacy of asthmatic children by regulating the balance of Treg/Th17 cells Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Wei WANG, Qing-Bin LIU, Wei JING Abstract Astragalus membranaceus may be a potential therapy for childhood asthma but its driving mechanism remains elusive. The main components of A. membranaceus were identified by HPLC. The children with asthma remission were divided into two combination group (control group, the combination of budesonide and terbutaline) and A. membranaceus group (treatment group, the combination of budesonide, terbutaline and A. membranaceus). The therapeutic results were compared between two groups after 3-month therapy. Porcine peripheral blood mononuclear cells (PBMCs) were isolated from venous blood by using density gradient centrifugation on percoll. The levels of FoxP3, EGF-β, IL-17 and IL-23 from PBMCs and serum IgE were measured. The relative percentage of Treg/Th17 cells was determined using flow cytometry. The main components of A. membranaceus were calycosin-7-O-glucoside, isoquercitrin, ononin, calycosin, quercetin, genistein, kaempferol, isorhamnetin and formononetin, all of which may contribute to asthma therapy. Lung function was significantly improved in the treatment group when compared with a control group (P < 0.05). The efficacy in preventing the occurrence of childhood asthma was higher in the treatment group than the control group (P < 0.05). The levels of IgE, IL-17 and IL-23 were reduced significantly in the treatment group when compared with the control group, while the levels of FoxP3 and TGF-β were increased in the treatment group when compared with the control group (P < 0.05). A. membranaceus increased the percentage of Treg cells and reduced the percentage of Th17 cells. A. membranaceus is potential natural product for improving the therapeutic efficacy of combination therapy of budesonide and terbutaline for the children with asthma remission by modulating the balance of Treg/Th17 cells.

Roles of integrin in tumor development and the target inhibitors Publication date: April 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 4 Author(s): Zhao-He LI, You ZHOU, You-Xiang DING, Qing-Long GUO, Li ZHAO Abstract Integrin is a large family of cell adhesion molecules (CAMs) which involves in the interaction of cells/cells and cells/ extracellular matrix (ECM) to mediate cell proliferation, differentiation, adhesion, migration, etc. In recent years, aberrant expression of integrin has been clearly found in many tumor studies, indicating that integrin is closely related to tumor formation and development. Meanwhile, it has effects on tumor cell differentiation, cell migration, proliferation and tumor neovascularization. The study of drugs targeting integrins is of great significance for the clinical treatment of tumors. Because of its important role in tumorigenesis and development, integrin has become a promising target for the treatment of cancer. This review summarizes the role of integrin in tumor development and the current state of integrin inhibitors to provide a valuable reference for subsequent research.

Quantification of Panax notoginseng saponins metabolites in rat plasma with in vivo gut microbiota-mediated biotransformation by HPLC-MS/MS Publication date: March 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 3 Author(s): Yin-Ping GUO, Man-Yun CHEN, Li SHAO, Wei ZHANG, Tai RAO, Hong-Hao ZHOU, Wei-Hua HUANG Abstract Panax notoginseng saponins (PNS) are the major components of Panax notoginseng, with multiple pharmacological activities but poor oral bioavailability. PNS could be metabolized by gut microbiota in vitro, while the exact role of gut microbiota of PNS metabolism in vivo remains poorly understood. In this study, pseudo germ-free rat models were constructed by using broad-spectrum antibiotics to validate the gut microbiota-mediated transformation of PNS in vivo. Moreover, a high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) was developed for quantitative analysis of four metabolites of PNS, including ginsenoside F1 (GF1), ginsenoside Rh2 (GRh2), ginsenoside compound K (GCK) and protopanaxatriol (PPT). The results showed that the four metabolites could be detected in the control rat plasma, while they could not be determined in pseudo germ-free rat plasma. The results implied that PNS could not be biotransformed effectively when gut microbiota was disrupted. In conclusion, gut microbiota plays an important role in biotransformation of PNS into metabolites in vivo.

Two new nimbolinin- and trichilin-class limonoids isolated from the fruits of Melia azedarach Publication date: March 2019 Source: Chinese Journal of Natural Medicines, Volume 17, Issue 3 Author(s): Lu QIU, Li HENG, Rong XU, Jun LUO, Yi LI Abstract Two new furan fragment isomerized limonoids, meliazedalides A and B (compounds 1 and 2), were isolated from the fruits of Melia azedarach Linn.. Their chemical structures were elucidated on the basis of HR-ESI-MS and 1D and 2D NMR data, which belonged to nimbolinin- and trichilin-class, respectively. Compound 2 exhibited weak inhibitory effect on NO production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages with IC50 being 37.41 μmol·L−1.

Alexandros Sfakianakis
Anapafseos 5 . Agios Nikolaos
Crete.Greece.72100
2841026182

6948891480

alsfakia@gmail.com

Acta Histochemica

Prognostic significance of resident CD103+CD8+T cells in human colorectal cancer tissues Publication date: Available online 30 May 2019 Source: Acta Histochemica Author(s): Wenwei Hu, Runzi Sun, Lujun Chen, Xiao Zheng, Jingting Jiang Abstract The prognostic significance and clinical implications of resident CD103+CD8+T cells in human colorectal cancer tissues still remains largely unexplored. In our present study, we aimed to characterize the resident CD8+T cells in human colorectal cancer tissues by using double staining of CD103 and CD8, and further evaluated the prognostic significance of resident CD8+T cells in colorectal cancer. We found that the OS rate of the colorectal cancer patients with higher infiltration of CD8+T cells, or with higher numbers of resident CD103+CD8+T cells, or with higher ratio of CD103+CD8+T cells over total CD8+T cells in cancer tissues was significantly better than that of the patients with lower infiltration of CD8+T cells, or with lower numbers of resident CD103+CD8+T cells, or with higher ratio of CD103+CD8+T cells over total CD8+T cells in cancer tissues, respectively. Moreover, higher infiltration of CD8+T cells in colorectal cancer tissues was significantly and inversely correlated with advanced TNM stage. Higher numbers of resident CD103+CD8+T cells in colorectal cancer tissues were significantly and inversely correlated with distant metastasis status. Higher ratio of CD103+CD8+T cells over total CD8+T cells in colorectal cancer tissues was significantly and inversely correlated with age status. The COX model analysis demonstrated that higher infiltration of CD8+T cells, higher numbers of resident CD103+CD8+T cells, or higher ratio of CD103+CD8+T cells over total CD8+T cells in colorectal cancer tissues, could serve as independent prognostic predictors for colorectal cancer patients. Taken together, our present study demonstrated the density of tumor infiltrating CD8+T cells or the numbers of resident CD103+CD8+T cells in colorectal tissues could be used as an important prognostic predictor for this malignancy.

A histomorphometric study on the hepatoprotective effects of a green rooibos extract in a diet-induced obese rat model Publication date: Available online 29 May 2019 Source: Acta Histochemica Author(s): J.I. Layman, D.L. Pereira, N. Chellan, B. Huisamen, S.H. Kotzé Abstract Obesity, type two diabetes mellitus and insulin resistance are associated with increased oxidative stress and inflammation. Unfermented green rooibos is an aspalathin rich variant of traditional fermented rooibos (Aspalathus linearis) and has a high polyphenol content. The present study aimed to determine the histologically observable effects of a commercially produced, aspalathin-rich green rooibos extract, Afriplex GRT™ (GRE) in a diet-induced obese rat model. Male Wistar rats (N = 28) were randomly assigned to four study groups (n = 7): control (C), green rooibos (GRT), high-fat diet (HFD) and experimental (HFD-GRT) group. Body mass was determined prior to euthanasia and liver mass was determined after death. The left lateral lobe of the liver was processed to wax and stained using haematoxylin and eosin (H & E), Masson's trichrome stain, Gordons and Sweet's reticulin impregnation and periodic acid-Schiff stain. Frozen liver tissue sections were used for Oil red O staining. Morphometric quantification of steatosis, semiquantitative pathology grading and scoring were performed and verified by a veterinary histopathologist. A significant increase in body and liver mass was observed in the HFD groups while co-treatment with green rooibos significantly reduced both. The volume and area of steatosis were significantly increased in the HFD groups while the area of steatosis significantly reduced with green rooibos co-treatment. The percentage, location and type of steatosis as well as presence of inflammation and hepatocellular injury were reduced in the HFD group co-treated with GRE. These findings suggest that a GRE has potential as an anti-steatotic, anti-inflammatory and weight reducing agent in vivo. Graphical abstract

Pancreatic endocrine cell arrangement during human ontogeny Publication date: Available online 27 May 2019 Source: Acta Histochemica Author(s): Alexandra E. Proshchina, Yuliya S. Krivova, Valeriy M. Barabanov, Sergey V. Saveliev Abstract In the human pancreas, various forms of endocrine cell arrangement are found: single endocrine cells, endocrine cell clusters, and mantel, bipolar and mosaic cell (mixed) islets. Our aim was to analyse the distribution and dynamics of insulin-, glucagon- and somatostatin-containing cells within the various forms of endocrine pancreas arrangement during human prenatal development and in adults and to suggest a mechanism of change in the endocrine cell ratio in adult islets. Pancreatic autopsies derived from human foetuses from the 10th to the 40th weeks of development and from adults were examined using histological, immunohistochemical and morphometric methods. During development, the human endocrine pancreas undergoes not only de novo differentiation of endocrine cells and islet formation, but morphogenetic restructuring, which is revealed as a change of the α-, β- and δ-cell ratio in the islets. In particular, increased proportion of glucagon- and somatostatin-containing cells and decreased proportion of β-cells were shown in the largest mosaic islets in adults. Our results indicate that the distribution and proportion of α-, β- and δ-cells depend on the islets size and vascularisation. Studying of the mechanism of such restructuring may contribute to the development of new approaches in the treatment of diabetes mellitus.

High expression of meningioma 1 is correlated with reduced survival rates in colorectal cancer patients Publication date: Available online 24 May 2019 Source: Acta Histochemica Author(s): Yi-Jung Ho, Feng-Cheng Liu, Jungshan Chang, Bingyang Shi, Kun-Tu Yeh, Yueh-Min Lin, Jeng-Wei Lu Abstract The identification of prognostic markers for colorectal cancer (CRC) has important clinical implications. However, the association between meningioma 1 (MN1) expression and clinical outcomes of CRC has not been fully investigated. The aim of this study was to investigate the expression of MN1 in the clinical context of CRC. We first used immunohistochemistry (IHC) staining to examine and compare MN1 expression between multiple human cancer tissues and normal tissues. Initial screening revealed that the expression of MN1 proteins was significantly higher in tumor tissues of the breast, colon, and liver than in normal tissues. In further testing conducted on 59 paired CRC samples, we observed that the expression of MN1 in CRC tissue samples was significantly higher than in adjacent normal tissues. Moreover, high MN1 expression was not significantly associated with clinicopathological characteristics. Kaplan-Meier survival analysis revealed that high expression of MN1 mRNA or MN1 protein was significantly associated with poor CRC prognosis. Furthermore, univariate Cox analysis revealed that a high MN1 score was significantly associated with prognostic factors. Multivariate Cox analysis further indicated that gender, histologic grade, tumor-node-metastasis (TNM) stage, and a high MN1 score were independent factors of overall CRC survival rates. Finally, MN1 and PCNA protein levels were positively correlated, which suggests that MN1 may be involved in the cell proliferation process during CRC formation. Our results, which confirm those of other studies, indicate that (1) high levels of MN1 expression contribute to poor CRC prognosis and (2) MN1 can serve as a novel potential biomarker in predicting the prognosis of CRC patients.

Age related changes in the dermal mast cells and the associated changes in the dermal collagen and cells: A histological and electron microscopy study Publication date: Available online 22 May 2019 Source: Acta Histochemica Author(s): Sara Mohamed Naguib Abdel Hafez Abstract Mast cells are widely distributed bone marrow cells. They have a crucial role in the dermal aging process. The aim of the present study was to describe the biochemical and the histological changes that occur in the aged dermal mast cells and to demonstrate the associated changes in the dermal cells and fibers as well. Sixteen male albino rats were used and divided into two groups; the control group (8–10 weeks) and the aged group (20–22 weeks). The rats were decapitated then processed for further biochemical and histological studies. The mean area fraction for collagen fibers was measured. In the aged group, there was a significant increase in the skin histamine and heparin levels if compared with the control one. Furthermore, there was an apparent increase in intact and degranulated dermal mast cells if compared with the control one. The dermal collagen bundles were apparently decreased and appeared distorted with wide spacing. Additionally, there were apparently large sized eosinophils with more cytoplasmic granules. Direct contact between mast, fibroblast, and macrophage cells was noticed. The average area fraction of collagen fibers was significantly increased in the aged group if compared with the control one. It could be concluded that the secretory activity of dermal mast cells was significantly increased in the aged skin group. Also, this study demonstrated the implicated role of mast cell in aged skin changes. Further long-term studies are needed to validate the prophylactic or therapeutic potential by intentional hindering of mast cell degranulation in aged skin.

Do estrogens regulate lipid status in testicular steroidogenic Leydig cell? Publication date: Available online 21 May 2019 Source: Acta Histochemica Author(s): A. Milon, M. Kaczmarczyk, P. Pawlicki, B. Bilinska, M. Duliban, E. Gorowska-Wojtowicz, W. Tworzydlo, M. Kotula-Balak Abstract In this study mouse Leydig cell (MA-10) were treated with G-protein coupled membrane estrogen receptor antagonist (G-15; 10 nM). Cells were analyzed by Western blotting for expression of estrogen-related receptors (ERRα, β and γ), steroidogenic markers (lutropin receptor; LHR and 3β-hydroxysteroid dehydrogenase; 3β-HSD) and lipid droplet markers (perilipin; PLIN and microtubule-associated protein 1 A/1B-light chain 3; LC3). Concomitantly, microscopic analyses by light microscope (immunofluorescent staining for lipid droplets, PLIN and LC3) as well as by electron microscope (for lipid droplet ultrastructure) were utilized. For analysis of cholesterol content, cAMP level and progesterone secretion, G-15, estrogen receptor (ER) antagonist (ICI 182,780; 10 μM), 17β-estradiol (10 mM) and, bisphenol A (BPA; 10 nM) were used alone or in combinations. We revealed no changes in ERRs expression but alterations in ERRβ and γ localization in G-15-treated cells when compared to control. Partial translocation of ERRβ and γ from the cell nucleus to cytoplasm was observed. Decreased expression of LHR, 3β-HSD, PLIN and LC3 was detected. Moreover, in treated cells large lipid droplets and differences in their distribution were found. Very strong signal of co-localization for PLIN and LC3 was found in treated cells when compared to control. In ultrastructure of treated cells, degenerating lipid droplets and double membrane indicating on presence of lipophagosome were observed. We found, that only (i) BPA and G-15 did not effect on cholesterol content, (ii) BPA, G-15 and ICI did not effect on cAMP level and (iii) BPA, ICI alone and in combination, and BPA with G-15 did not modulate progesterone secretion. These findings showed complex and diverse estrogen effects on mouse Leydig cells at various steps of steroid hormone production (cholesterol storage, release and processing). Lipid homeostasis and metabolism in these cells were affected by endogenous and exogenous estrogen, interactions of receptors (GPER, ER and ERR) and GPER and ER antagonists.

Effect of acrylamide on BEAS-2B normal human lung cells: Cytotoxic, oxidative, apoptotic and morphometric analysis Publication date: Available online 18 May 2019 Source: Acta Histochemica Author(s): Sedat Kacar, Varol Sahinturk, Hatice Mehtap Kutlu Abstract Due to the broad toxic relevance of acrylamide, many measures have been taken since the 1900s. These measures increased day by day when acrylamide was discovered in foods in 2002, and its toxic spectrum was found to be wider than expected. Therefore, in some countries, the products with higher acrylamide content were restricted. On the other hand, the effects of acrylamide on the respiratory system cells have yet to be well understood. In this study, we aimed at investigating the effect of acrylamide on lung epithelial BEAS-2B cells. Initially, the cytotoxic effect of acrylamide on BEAS-2B was determined by MTT assay. Then, cellular oxidative stress was measured. Flow cytometry analysis was conducted for Annexin-V and caspase 3/7. Furthermore, Bax, Bcl-2 and Nrf-2 proteins were evaluated by immunocytochemistry. Finally, acrylamide-induced cellular morphological changes were observed under confocal and TEM microscopes. According to MTT results, the IC50 concentration of acrylamide was 2.00 mM. After acrylamide treatment, oxidative stress increased dose-dependently. Annexin V-labelled apoptotic cells and caspase 3/7 activity were higher than untreated cells in acrylamide-treated cells. Immunocytochemical examination revealed a marked decrease in Bcl-2, an increase in Bax and Nrf-2 protein staining upon acrylamide treatment. Furthermore, in confocal and TEM microscopy, apoptotic hallmarks were pronounced. In the present study, acrylamide was suggested to display anti-proliferative activity, decrease viability, induce apoptosis and oxidative stress and cause morphological changes in BEAS-2B cells.

Gaseous transmitters in human retinogenesis Publication date: Available online 18 May 2019 Source: Acta Histochemica Author(s): Sergei G. Kalinichenko, Natalya Yu. Matveeva, Igor I. Pushchin Abstract Endogenous gaseous transmitters (nitric oxide, carbon monoxide, and hydrogen sulphide) form a special neuromodulation system mediating the development and modification of nerve centers. Here, we examined the localization of key gaseous transmitter enzymes: cystathionine β-synthetase (CBS), cystathionine γ-lyase (CSE), heme oxygenase 2 (HO-2), and constitutive NO synthase (nNOS) in the fetal human retina at different stages of development. The number of CBS- and CSE-positive photoreceptors and intermediate retinal neurons was high in trimester I and gradually decreased to the end of trimester III. The number of HO-2-positive cells followed the same trend. The number of nNOS-positive intermediate retinal neurons and neurons within the ganglion cell layer showed the opposite dynamics with the peak in trimester III. The results are interpreted in terms of the role of gaseous transmitters in retinogenesis and cytoprotection.

Cryoembedder, automatic processor/stainer, liquid nitrogen freezing, and manual staining for frozen section examination: A comparative study Publication date: Available online 9 May 2019 Source: Acta Histochemica Author(s): Salvatore Lorenzo Renne, Silvia Redaelli, Biagio Paolini Abstract Frozen section examination (FSE) reshaped surgical pathology and is characterized by a high accuracy. Nonetheless pathologists can experience artefacts that can compromise or defer the diagnosis. We compared a commercial system, composed by a cryoembedder and a processor/stainer, to our FSE protocol. Feasibility of diagnosis as well as overall architecture, cytology, and staining, were scored under the following conditions: Traditional (liquid nitrogen freezing and manual staining), Only-Presto (liquid nitrogen freezing and commercial processor/stainer), Only-PrestoCHILL (cryoembedder and manual staining), and PrestoSystem (cryoembedder and processor/stainer). Scores were compared across the different experimental conditions. PrestoSystem had significantly higher scores than Traditional, Only-Presto or Only-PrestoCHILL in all categories (Wilcoxon test; all P-value <.001); similarly, also Only-Presto and Only-PrestoChill had significantly higher scores than Traditional in all categories. Only-PrestoCHILL had significantly higher scores than Only-Presto in Cytology and Architecture. In conclusion the control of pre-analytical variables provided reproducible results, of a higher quality.

Impact of renal ischemia/reperfusion injury on the rat Kupffer cell as a remote cell: A biochemical, histological, immunohistochemical, and electron microscopic study Publication date: Available online 9 May 2019 Source: Acta Histochemica Author(s): Sara Mohamed Naguib Abdel Hafez, Rehab Ahmed Rifaai, Asmaa M.A. Bayoumi Abstract Almost all transplanted solid organs are exposed to some degree of ischemia-reperfusion (IR) damage. It is interesting to know that this IR damage affects various remote tissues including the liver and resulted in serious adverse effects. Liver injury triggers different responses of liver tissue especially Kupffer cells (KCs). The goal of this current study is to assess the biochemical and morphological changes of hepatic KCs after the induction of renal ischemia-reperfusion (RIR) and point out their role in remote liver injury after RIR. Sixteen male Sprague-Dawley rats were randomly divided into two equal groups: Group I; sham group. Group II; renal ischemia reperfusion (IR) group in which rats were exposed to renal ischemia for 45 min followed by renal reperfusion for 48 h. Three rats from each group were subjected to charcoal injection to evaluate KCs activity. Specimens of rat liver from each group were obtained and processed for biochemical, light microscopic and ultramicroscopic examination. The current results showed elevated serum levels of AST and ALT. The liver HGF-α protein expression increased in IR group compared to the sham group. In IR group, numerous charcoal labeled KCs were observed mainly localized around the central vein. Scanning electron micrographs showed complex primary and secondary foot process of the KCs. Ultrastructural study showed KCs with multiple cytoplasmic vacuoles, lysosomes and mitochondria, rough endoplasmic reticulum and ribosomes. Immuno-histochemical study showed more tumor necrosis factor-α (TNF-α) expression in KCs than the sham group. These results collectively demonstrated that renal IR produced biochemical and morphological changes in the liver KCs and theses cells might have a role in the remote liver injury after renal IR. This might be one of the mechanisms through which RIR affects the liver.

Alexandros Sfakianakis
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