Fernye, Csaba;Ancsin, Zsolt;Bocsai, Andrea;Balogh, Krisztian;Mezes, Miklos;Erdelyi, Marta
Toxicological Research
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제34권3호
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pp.249-257
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2018
The purpose of the present study was to evaluate the effects of different dietary concentrations of T-2 toxin on blood plasma protein content, lipid peroxidation and glutathione redox system of pheasant (Phasianus colchicus). A total of 320 one-day-old female pheasants were randomly assigned to four treatment groups fed with a diet contaminated with different concentrations of T-2 toxin (control, 4 mg/kg, 8 mg/kg and 16 mg/kg). Birds were sacrificed at early (12, 24 and 72 hr) and late (1, 2 and 3 weeks) stages of the experiment to demonstrate the effect of T-2 toxin on lipid peroxidation and glutathione redox status in different tissues. Feed refusal and impaired growth were observed with dose dependent manner. Lipid-peroxidation was not induced in the liver, while the glutathione redox system was activated partly in the liver, but primarily in the blood plasma. Glutathione peroxidase activity has changed parallel with reduced glutathione concentration in all tissues. Based on our results, pheasants seem to have higher tolerance to T-2 toxin than other avian species, and glutathione redox system might contribute in some extent to this higher tolerance, in particular against free-radical mediated oxidative damage of tissues, such as liver.
Long interspersed nuclear element-1 (LINE-1) is a retrotransposon that contains a CpG island in its 5'-untranslated region. The CpG island of LINE-1 is often heavily methylated in normal somatic cells, which is associated with poor prognosis in various cancers. DNA methylation can differ between formalin-fixed paraffin-embedded (FFPE) and frozen tissues. Therefore, this study aimed to compare the LINE-1 methylation status between the two tissue-storage conditions in gastric cancer (GC) clinical samples and to evaluate whether LINE-1 can be used as an independent prognostic marker for each tissue-storage type. We analyzed four CpG sites of LINE-1 and examined the methylation levels at these sites in 25 FFPE and 41 frozen GC tissues by quantitative bisulfite pyrosequencing. The LINE-1 methylation status was significantly different between the FFPE and frozen GC tissues (p < 0.001). We further analyzed the clinicopathological features in the two groups separately. In the frozen GC tissues, LINE-1 was significantly hypomethylated in GC tissues compared to their corresponding normal gastric mucosa tissues (p < 0.001), and its methylation status was associated with gender, differentiation state, and lymphatic and venous invasion of GC. In the FFPE GC tissues, the methylation levels of LINE-1 differed according to tumor location and venous invasion of GC. In conclusion, LINE-1 can be used as a useful methylation marker for venous invasion in both FFPE and frozen tumor tissues of GC.
This study was conducted to determine effects of different selenium (Se) sources on performance, carcass characteristics, blood measures (whole blood Se concentration and plasma glutathione peroxidase (GSH-Px) activity), and Se concentrations in tissues of finishing Hanwoo steers (Korean native steers). Twenty finishing Hanwoo steers (average body weight=536${\pm}$23.4 kg, average age=approximately 20 months) were allotted to treatments in four groups of five steers per pen for 16 weeks preceding slaughter. Treatments were control (CON), spent mushroom composts from Se-enriched mushrooms (Se-SMC), selenized yeast (Se-Y), and sodium selenite (SS). Dietary Se levels of all treatments except CON were 0.9 mg Se/kg on the dry matter basis. Body weight was measured at the first and final day of trial, and blood samples were collected to analyze whole blood Se concentration and plasma GSH-Px activity at 2, 4, 8, and 16 weeks. At the end of trial, steers were slaughtered to collect muscle and liver samples for their Se analyses, and carcass data were recorded. In terms of dry matter intake, body weight gain and carcass characteristics, no significant differences among treatments were observed. Whole blood Se concentrations were significantly higher (p<0.05) for Se-SMC and Se-Y treatments than for CON at each collection period, with no significant difference between SS and CON. For weeks 2 and 8, there was no significant difference for whole blood Se concentration between Se-SMC and Se-Y, but for weeks 4 and 16, Se-Y treatments were significantly higher (p<0.05) than Se-SMC. No differences were observed for plasma GSH-Px activity between Se-SMC and Se-Y. The Se concentrations in hind leg and liver were significantly different among treatments (p<0.05) and those in both tissues ranked the greatest in Se-Y, followed by Se-SMC, SS, and CON treatments. However, tissue Se concentration for SS was not different from that for CON. These results showed that feeding organic Se sources such as Se-SMC and Se-Y enhanced Se concentration in tissues, while SS, the most common supplement of inorganic Se, was inefficient in Se deposition. Even though Se-Y had a higher Se concentration in tissues than Se-SMC, replacing Se-Y with Se-SMC in diets of beef steers would be an inexpensive way to increase Se concentration in beef.
The change in fatty acid composition in brain tissue of the second generation rats(Sprague-Dawley strain) was studied using four different fat diets(Corn oil=CO, Soybean oil=SO, Perilla oil=PO, Fish oil=FO, 10% by Wt). The experimental diets were started from pregnancy in four different groups, each consisting of 9 rats. The seound generation rats were fed the same diet as their mothers. Animals were anesthetized with ether at 0, 3, 9 & 16 weeks of age. Whole brains were dissected out, brain tissues were, then, homogenized and lipids were extracted from brain tissues. The fatty acid compositions were measured after methylation by gas-liquid chromatography at 0, 3, 9 and 16 weeks of age of offspring. The changes in the relative concentrations of polyunsaturated fatty acids(PUFA) or more specifically docosahexaenoic acid(22 : 6, $\omega$3, DHA), the major $\omega$3 fatty acid component in rat brain at different age were similar to changes in the amount of DNA in brain tissue showing the maximum value during the lactation. The changes in saturated fatty acid(SFA) content showed a contrasting patten to those of PUFA, while monounsaturated fatty acid(MUFA) increased steadily throughout the experimental period. At birth, the relative concentrations of $\omega$3 series fatty acids the relative concentrations of PUFA, MUFA and SFA converged to very similar values respectively regardless of the dietary fatty acid compositions. In brain tissue, it is of value to note that while changes in relative concentrations of linoleic acid (18 : 2, $\omega$6, LA) and arachidonic acid(20 : 4, $\omega$6, AA) showed a precursor-product-like relationship, $\alpha$-linolenic acid(18 : 3, $\omega$3, $\alpha$-LnA) and DHA showed a different pattern. Even when the $\omega$3 fatty acid content in very low in maternal diet(CO), the second generation rat brain tissues appeared to secure DHA content, suggesting an essential role of this fatty acid in the brain. The fact that a large amount of $\alpha$-LnA in the maternal diet did not have a significant effect on the second generation rat brain $\alpha$-LnA content, indicated that DHA seemed essential component for the brain development in our experimental condition. In all groups, the relative content of $\alpha$-LnA in the brain tissues remained relatively constant throughout the experimental period at the very low level. The study of the specific concentrations and essential role(s) of DHA in each parts of brain tissue is needed in more details.
Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.
The synthesis of acyl-CoA catalyzed by acyl-CoA synthetase (ACS, EC 6.2.1.3) from fatty acid, ATP, and CoA is a crucial reaction in mammalian fatty acid metabolism. In arachidonate metabolism, acyl-CoA synthetase(ACS) plays a key role in the esterification of free arachidonate into membrane phospholipids. Following its release by the action of calcium dependent phospholipase, free arachidonate is believed to be rapidly converted to arachidonoyl-CoA and reesterified into phospholipids in order to prevent excessive synthesis of eicosanoids. In previous studies, we have characterized five ACSs (designated as ACS1-5) with different tissue distribution. ACS1, ACS2, and ACS5 are similar in structure and fatty acid preference, and completely different from ACS3 and ACS4. The latter are arachidonate-preferring enzymes closely related in structure but expressed in different tissues: ACS3 mRNA is highly expressed in the brain and the mRNA for ACS4 is expressed in steroidogenic tissues including adrenal gland, ovary, and testis. To learn more about the potential function of ACS4 in arachidonate metabolism, we have produced knock-out mice for ACS4 gene. ACS4+/- females become pregnant less frequently and produce small litters with extremely low transmission of the disrupted alleles. Striking morphological changes including extremely enlarged uterine filled with numerous proliferative cysts of various size were detected in ACS4+/- females. Furthermore, marked accumulation of prostaglandins were seen in the uterus of heterozygous females. These results indicate that ACS4 is critical for the uterine arachidonate metabolism and heterozygous disruption of its gene lead to impaired pregnancy.
MicroRNAs (miRNAs) are single-stranded, small RNAs (21-23 nucleotides) that function in gene silencing and translational inhibition via the RNA interference mechanism. Most miRNAs originate from host genomic regions, such as intergenic regions, introns, exons, and transposable elements (TEs). Here, we focused on the palindromic structure of medium reiteration frequencies (MERs), which are similar to precursor miRNAs. Five MER consensus sequences (MER5A1, MER53, MER81, MER91C, and MER117) were matched with paralogous transcripts predicted to be precursor miRNAs in the horse genome (equCab2) and located in either intergenic regions or introns. The MER5A1, MER53, and MER91C sequences obtained from RepeatMasker were matched with the eca-miR-544b, eca-miR-1302, and eca-miR-652 precursor sequences derived from Ensembl transcript database, respectively. Each precursor form was anticipated to yield two mature forms, and we confirmed miRNA expression in six different tissues (cerebrum, cerebellum, lung, spleen, adrenal gland, and duodenum) of one thoroughbred horse. MER5A1-derived miRNAs generally showed significantly higher expression in the lung than in other tissues. MER91C-derived miRNA-5p also showed significantly higher expression in the duodenum than in other tissues (cerebellum, lung, spleen, and adrenal gland). The MER117-overlapped expressed sequence tag generated polycistronic miRNAs, which showed higher expression in the duodenum than other tissues. These data indicate that horse MER transposons encode miRNAs that are expressed in several tissues and are thought to have biological functions.
Gene expression profiling is a useful tool for identifying critical genes and pathways in metabolism. The objective of this study was to determine the major differences in the expression of genes associated with metabolism and metabolic regulation in liver and mammary tissues of lactating cows. We used the Michigan State University bovine metabolism (BMET) microarray; previously, we have designed a bovine metabolism-focused microarray containing known genes of metabolic interest using publicly available genomic internet database resources. This is a high-density array of 70mer oligonucleotides representing 2,349 bovine genes. The expression of 922 genes was different at p<0.05, and 398 genes (17%) were differentially expressed by two-fold or more with 222 higher in liver and 176 higher in mammary tissue. Gene ontology categories with a high percentage of genes more highly expressed in liver than mammary tissues included carbohydrate metabolism (glycolysis, glucoenogenesis, propanoate metabolism, butanoate metabolism, electron carrier and donor activity), lipid metabolism (fatty acid oxidation, chylomicron/lipid transport, bile acid metabolism, cholesterol metabolism, steroid metabolism, ketone body formation), and amino acid/nitrogen metabolism (amino acid biosynthetic process, amino acid catabolic process, urea cycle, and glutathione metabolic process). Categories with more genes highly expressed in mammary than liver tissue included amino acid and sugar transporters and MAPK, Wnt, and JAK-STAT signaling pathways. Real-time PCR analysis showed consistent results with those of microarray analysis for all 12 genes tested. In conclusion, microarray analyses clearly identified differential gene expression profiles between hepatic and mammary tissues that are consistent with the differences in metabolism of these two tissues. This study enables understanding of the molecular basis of metabolic adaptation of the liver and mammary gland during lactation in bovine species.
A patient complaining of chronic dull pain in the right maxillary area showed slight haziness and small ovoid radiopacity in the right antrum, which was not extended into the choanal area in radiographic views. At operation, lots of mucoid fluid admixed myxoid soft tissues was discharged and the polypoid mucosal tissues were removed. In histological examination, the removed tissues showed a polyp by the overgrowth of dermal connective tissues exhibiting severe myxoid degeneration. Throughout the entire specimen, the inflammatory reaction was diffuse but not so remarkable to produce the mucosal thickening and necrosis. The polypoid tissues were diffusely infiltrated with neutrophiles and plasma cells, but few eosinophils, resulted in the extensive myxoid degeneration together with severe vascular degeneration. Therefore, we suggest that the antral polyp is basically different in its pathogenesis and prognosis from the common maxillary sinusitis of odontogenic origin, thus the antral polyp should be carefully diagnosed when the inflamed antral lesion is recurred and diffusely degenerative with myxoid changes.
Human activities have enhanced the influx of heavy metals to aquatic ecosystem and hanged the abiotic environment such as the sediments supporting benthic organisms. The levels of heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) in the benthic gastropod Oxyloma hirasei and the sediments around their habitat were investigated to examine heavy metal levels and the potential of the gastropod as a bioindicator. We detected different levels of heavy metals in the sediments of two wetland areas, Upo and Mokpo, in the relatively well-conserved Upo wetland, Ramsar Convention Area. Oxyloma hirasei had higher concentrations of heavy metals except Cr and Ni in their soft tissues than in their shells (Cd: $2.10{\sim}3.16$, Cu: $19.73{\sim}28.66$, Pb: $0.67{\sim}1.17$, Zn: $216.1{\sim}285.7\;{\mu}g/g$ dry weight in the soft tissues; Cr: $1.19{\sim}2.58$, Ni: $0.47{\sim}1.16\;{\mu}g/g$ dry weight in the shells). Differences in the Cd, Cu, Ni and Pb concentrations in O. hirasei soft tissues reflected differences in heavy metal concentrations in the sediments at the sampling sites. The coefficients of variation for Cd, Cu and Pb were lower than those for other metals in the soft tissues. Levels of Cd in the tissues of O. hirasei were the highest among the metals examined in this study. Therefore, the soft tissue of O. hirasei appears to be a promising bioindicator particularly for Cd.
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