Fatty acid composition of lipids in erythrocytes and blood plasma in cadmium intoxication and its correction with unitiol

The effect of cadmium chloride on the fatty acid composition of lipids of the erythrocytes and blood plasma has been studied. It has been established that in the process of cadmium intoxication the decrease of the level of unsaturated fatty acids takes place and the most considerable changes in the content of oleic and arachidonic acids. It makes possible to assume the disturbance of both the erythrocytes membrane structure and metabolism in the erythrocyte. Unitiol injection promotes the normalisation of the indices under investigation.     

Nature and store of fatty acid lipids in the blood of newborn calves with dyspepsia

The content of fatty acids of lipids extracted from the whole blood as well as from erythrocytes, leucocytes, plasma and serum samples of newborn dyspeptic calves were investigated for the first time. Twenty three (23) saturated, mono-non-saturated and poly-non-saturated fatty acids were detected. Native, palmitoleic, stearic, oleic, linoleic and arachidonic fatty acids, are the main components of the whole blood lipid fraction and its components. The fatty acids found in lipids and other components of the whole blood in the samples of dyspeptic and healthy calves are the same, their ratios, however, are different. The decrease in the nonsaturated fatty acids content and its increase in saturated fatty acids are considerable. The saturation coefficient is different in native blood lipids, erythrocytes, leukocytes, serum and plasma.     

Fatty acid values in the plasma of neonates, umbilical cord and maternal blood

The proportion of a wide range of fatty acids was studied in the plasma of 15 healthy newborn infants following a physiological pregnancy and delivery. The same measurements were done in seven healthy mothers (immediately after parturition) and the proportion of fatty acids was analysed in mixed umbilical cord blood (n = 7). The fatty acids were identified by gas chromatography as their methyl esters (FAME). In newborn infants the proportion of saturated fatty acids was found to be 42.3%, of monoene fatty acids 31.3% and of polyene fatty acids 25.4%; type n-6 fatty acids formed 13.9% and n-3 11.1%. The proportion of the various fatty acids in healthy maternal plasma was 37.9% (saturated), 34.4% (monoene) and 25.0% (polyene) respectively; type n-6 fatty acids formed 21.6% and type n-3 only 3.6%. The values in mixed cord blood were 44% (saturated FA), 34.8% (monoene FA) and 20% (polyene FA); group n-6 FA accounted for 17.4% and n-3 for only 2.1%. In the above three series we also described the sequence of the fatty acids present in the largest amounts. This is part of an extensive study, in a large series, of the commonest perinatal risks. We particularly draw attention to the high proportion of long-chain fatty acids (C 22 - C 26) in the plasma of healthy newborn infants.     

Acid-base state of blood in calves with dyspepsia treated using different therapeutic approaches

As a result of our investigations it has been found, that the state of sub-compensated respiratory-metabolic acidosis with gradual recovery of electrolytic composition of blood serum was observed in calves which have had dyspepsia by the 30th day of life. At the same time in calves, which obtained milk phospholipids in a form of biologically active additive FLP-MD, more rapid recovery of acid-base balance of blood was diagnosed. The mentioned above was proved by the presence of compensated metabolic acidosis with simultaneous stabilization of blood serum electrolytic composition in calves which obtained biologically active additive FLP-MD.     

Fatty acid compositions of Taenia solium metacestode and its surrounding tissues

Fatty acids (FAs) are the main energy sources of living organisms and are the major components of cellular and organelle membranes. Their compositions also affect the flexibility/rigidity of cells and cell vitality. The Taenia solium metacestode (TsM) causes neurocysticercosis (NC), which is one of the most common helminthic infections of the central nerve system. We investigated the FA composition of the cyst fluid (CF) and parenchyma of the TsM, together with those of the granuloma and swine tissue surrounding the granuloma. The FA fractions of the TsM CF and swine tissue showed a composition and proportional contents comparable to each other, in which C18:0 (stearic acid), C18:1n9c (oleic acid), C20:4 (arachidonic acid) and C16:0 (palmitic acid) constituted the major fractions. However, the relative amount of individual FAs of the TsM parenchyma and granuloma differed from those of TsM CF and swine tissue, which contained enriched C16:0 and a lower amount of C20:4. Saturated FAs were the major constituents in parenchyma and granuloma, 50.4% and 46.1%, respectively. Conversely, monounsaturated FAs were the major constituents of CF and swine tissue, 38.7% and 40.3%, respectively. Our results strongly suggest that host-derived FAs might translocate across the parasite syncytial membrane and be stored in the CF.     

Fatty acid composition of plasma, blood cells and whole blood: relevance for the assessment of the fatty acid status in humans

The composition and incorporation of fatty acids (FA) in plasma and blood cells is the result of distinct processes: intake, metabolism and peripheral utilization. Aim of the study: was to compare the FA profile in plasma, lipoproteins and blood cells with that in whole blood (WB) from healthy volunteers; to assess the quantitative distribution of selected FA in triacylglycerols, cholesteryl esters and phospholipids. Lipid FA profiles are comparable in plasma and lipoproteins but differ from those in blood cells. In WB, the FA profile results from the balanced proportion of FA pools in plasma and cells. The contribution of each lipid class to the total amount of FA differs among blood specimens. Phospholipids of plasma and red blood cell are the major contributors to the FA amount and profile in WB. In conclusion, the FA profile of WB reflects the FA status and WB could be an adequate specimen for the assessment of FA intakes.     

Proteins from bovine tissues and biological fluids: defining a reference electrophoresis map for liver,kidney, muscle,plasma and red blood cells

A number of high resolution two-dimensional electrophoresis (2-DE) reference maps for bovine tissues and biological fluids have been determined for animals in basal state. Among the 1863 distinct protein features detected in samples of liver, kidney, muscle, plasma and red blood cells, 509 species were identified and associated to 209 different genes. Difficulties in the identification were related to the poorly characterized Bos taurus genome and were solved by a combined matrix-assisted laser desorption/ionisation-mass spectrometry and liquid chromatography-electrospray ionization tandem mass spectrometry approach. The experimental output allowed us to establish a 2-DE database accessible through the World Wide Web network at the URL address (http://www.iabbam.na.cnr.it/Biochem). These reference maps may serve as a tool in future veterinary medical studies aimed at the evaluation of changes in protein repertoire for altered animal physiological conditions and infectious diseases, to the definition of molecular markers for novel diagnostic kits and vaccines, as well as the characterization of protein modifications in bovine materials following technological processes used in the food industry.     

Formation of bile acid glucosides and dolichyl phosphoglucose by microsomal glucosyltransferases in liver, kidney and intestine of man

Formation of glucosides of the bile acids chenodeoxycholic, ursodeoxycholic, deoxycholic and hyodeoxycholic acids has been detected in microsomes from human liver, kidney and intestinal mucosa. Hepatic and extrahepatic bile acid glucosyltransferase activities were characterized with respect to kinetic parameters and other catalytic properties. Whereas no marked organ-specific differences in the affinities of individual bile acids toward hepatic and extrahepatic glucosyltransferases were observed, microsomes from extrahepatic sources showed twice to 5-times the maximal rates of bile acid glucosidation estimated with microsomes from liver. In addition to bile acid glucoside formation, microsomes from human liver, kidney and intestinal mucosa catalyzed the synthesis of dolichyl phosphoglucose acting as natural glucosyl donor in bile acid glucosidation.     

Glucuronidation of bile acids in human liver, intestine and kidney. An in vitro study on hyodeoxycholic acid

The activities of UDP-glucuronyl transferase(s) in homogenates and microsomal preparations of human liver, kidney and intestine were tested with hyodeoxycholic acid (HDC). The various kinetic parameters of the UDC-glucuronidation were determined from time course experiments. In both liver and kidney preparations, HDC underwent a very active metabolic transformation: liver Km = 78 microM, Vmax = 3.3 nmol . min-1 . mg-1 protein; kidney Km = 186 microM, Vmax = 9.9 nmol . min-1 . mg-1 protein. To our knowledge this is the first observation of both an extensive and comparable bile acid glucuronidation occurring in renal and hepatic tissues.     

小鼠成长过程中肾、肝、小肠碱性磷酸酶活性变化及组织细胞定位

利用比色、酶的原位复性电泳、组织化学和电镜酶化学等方法,观察了小鼠出生后的成长过程中肝、肾和小肠内碱性磷酸酶(AKP)的活性变化及组织细胞定位.结果表明,在小鼠成长过程中,3种器官内的AKP活性依次为小肠肾肝,小肠和肝AKP活性先上升后下降,肾内酶活性呈上升趋势.AKP活性在小肠主要分布在小肠绒毛上皮细胞的细胞膜、细胞浆、微绒毛和微绒毛表面的糖衣上,在肝主要分布住胆小管,在肾主要分布在皮质肾小管,尤以近曲小管的刷状缘、近曲小管上皮细胞膜和符种管状结构之间的腔隙内分布较多.小鼠成长过程中肝内共出现3条AKP同工酶带,小肠内出现2条,肾内出现3条,各同工酶活性随着小鼠的生长发育发生不同的变化.     

Determination of 5-aminolevulinic acid in blood plasma, tissues and cell cultures by high-performance liquid chromatography with electrochemical detection

A simple and sensitive method for determining 5-aminolevulinic acid (ALA) in biological samples is described. ALA is derivatized with o-phthaldehyde to give a compound with favorable properties for high-performance liquid chromatography with electrochemical detection. The method does not require extensive pretreatment of the samples and its detection limit is in the range of 1 pmol/20 μl injection. This method was applied to the determination of plasma ALA from normal and lead-exposed subjects, where 0.26±0.08 μM (n=30) and 2.6±0.75 μM (n=30), respectively were found. We also determined ALA in rat tissues, namely liver and brain, and the uptake of ALA by cultured fibroblasts and hepatocytes to illustrate the diversified applicability of the method.     

Increase in DPP-IV in the intestine, liver and kidney of the rat treated with high fat diet and streptozotocin

High fat diet or insulin deficiency is commonly seen in Type II diabetes, while the mechanism remains unclear. To test our hypothesis that DPP-IV contributes to Type II diabetes, we examined the expression and activity of DPP-IV in rats (n=8 to each group) treated for 12 weeks with 3 separate diets: a) normal control; b) a high fat diet; and c) a high fat diet plus streptozotocin, a chemical for induction of insulin-deficient diabetes. Compared to rats on the normal diet, the rats with a high fat diet significantly increased DPP-IV's expression and activity about 142-152% in the intestine (P<0.05) and 153-240% in kidneys (P<0.05), but there was no change in the liver. Administration of streptozotocin to the rats treated with the high fat diet showed an insufficient insulin secretion and higher blood glucose in response to glucose/insulin tolerance test, and an increase in expression of DPP-IV and activity by 188-242% in the intestine (P<0.01); 191-225% in liver (P<0.01); and 211-321% in the kidneys (P<0.01). Immunohistochemistry studies confirmed the above results, showing increased DPP-IV immunostaining localized primarily in intestinal epithelium, hepatocytes and renal tubular cells. This study, for the first time reports an increase in DPP-IV associated with a high fat diet, as well as in the combination of a high fat diet with an insulin deficiency. Since both high fat diet and insulin deficiency are closely linked with etiology of Type II diabetes, the evidence in this study suggests a role of DPP-IV in development of Type II diabetes.     

WGEF is a novel RhoGEF expressed in intestine, liver, heart, and kidney

Rho family GTPases regulate multiple cellular processes through their downstream effectors, where their activities are stimulated by the guanine nucleotide exchange factors. Here, we report a new member of RhoGEF, WGEF, which has the classical structure of DH-PH domain and a C-terminal SH3 domain. WGEF was shown to activate RhoA, Cdc42, and Rac1 by pulldown assay, and forced expression of WGEF resulted in marked rearrangement of the actin cytoskeleton, which is typically seen by the activation of RhoA, Cdc42, and Rac1. WGEF was highly expressed in intestine and also in liver, heart and kidney, which may suggest the involvement of WGEF in the development and functions of these organs. The expression pattern may also suggest the possible importance of WGEF in the understanding of diseases based on metabolic disorder.     

Regulation of transporter expression in mouse liver, kidney, and intestine during extrahepatic cholestasis

It is hypothesized that during cholestasis, the liver, kidney, and intestine alter gene expression to prevent BA accumulation; enhance urinary excretion of BA; and decrease BA absorption, respectively. To test this hypothesis, mice were subjected to either sham or bile-duct ligation (BDL) surgery and liver, kidney, duodenum, ileum, and serum samples were collected at 1, 3, 7, and 14 days after surgery. Serum total BA concentrations were 1-5 mumol/l in sham-operated mice and were elevated at 1, 3, 7, and 14 days after BDL, respectively. BDL decreased liver Ntcp, Oatp1a1, 1a5, and 1b2 mRNA expression and increased Bsep, Oatp1a4, and Mrp1-5 mRNA levels. In kidney, BDL decreased Oatp1a1 and increased Mrp1-5 mRNA levels. In intestine, BDL increased Mrp3 and Ibat mRNA levels in ileum. BDL increased Mrp1, 3, 4, and 5 protein expression in mouse liver. These data indicate that the compensatory regulation of transporters in liver, kidney, and intestine is unable to fully compensate for the loss of hepatic BA excretion because serum BA concentration remained elevated after 14 days of BDL. Additionally, hepatic and renal Oatp and Mrp genes are regulated similarly during extrahepatic cholestasis, and may suggest that transporter expression is regulated not to remove bile constituents from the body, but instead to remove bile constituents from tissues.     

Regulation of transporter expression in mouse liver, kidney, and intestine during extrahepatic cholestasis

It is hypothesized that during cholestasis, the liver, kidney, and intestine alter gene expression to prevent BA accumulation; enhance urinary excretion of BA; and decrease BA absorption, respectively. To test this hypothesis, mice were subjected to either sham or bile-duct ligation (BDL) surgery and liver, kidney, duodenum, ileum, and serum samples were collected at 1, 3, 7, and 14 days after surgery. Serum total BA concentrations were 1-5 μmol/l in sham-operated mice and were elevated at 1, 3, 7, and 14 days after BDL, respectively. BDL decreased liver Ntcp, Oatp1a1, 1a5, and 1b2 mRNA expression and increased Bsep, Oatp1a4, and Mrp1-5 mRNA levels. In kidney, BDL decreased Oatp1a1 and increased Mrp1-5 mRNA levels. In intestine, BDL increased Mrp3 and Ibat mRNA levels in ileum. BDL increased Mrp1, 3, 4, and 5 protein expression in mouse liver. These data indicate that the compensatory regulation of transporters in liver, kidney, and intestine is unable to fully compensate for the loss of hepatic BA excretion because serum BA concentration remained elevated after 14 days of BDL. Additionally, hepatic and renal Oatp and Mrp genes are regulated similarly during extrahepatic cholestasis, and may suggest that transporter expression is regulated not to remove bile constituents from the body, but instead to remove bile constituents from tissues.     

Heat-killed bacteria induce genome instability in mouse small intestine,liver and spleen tissues

Bacterial infection has been associated with several malignancies, yet the exact mechanism of infection-associated carcinogenesis remains obscure. Furthermore, it is still not clear whether oncontransformation requires an active infection process, or merely the presence of inactivated bacteria remnants is enough to cause deleterious effects. Here, we analyzed whether or not consumption of non-pathogenic and pathogenic heat-killed Escherichia coli leads to changes in genome stability in somatic tissues of exposed animals. For one week, mice were given to drink filtered or not-filtered water contaminated with heat-killed non-pathogenic E. coli DH5α or heat-killed pathogenic E. coli O157:H7 Sakai. Control animals received tap water. One week after exposure, molecular changes were analyzed in the small intestine, an organ that is in immediate contact with contaminated water. Additionally, we studied the effect in the distant spleen and liver, the organs that are involved in an immune response and detoxification, respectively. Finally, muscles were chosen as neutral tissues that were not supposed to be affected. Intestinal, liver and spleen but not muscle cells responded to all bacterial treatments with an increased level of DNA damage monitored by the induction of γH2AX foci. In the intestine, elevated levels of DNA damage were in parallel with an increase in Ku70 and p53 expression. We have also found an elevated level of cellular proliferation in the intestine, liver and spleen but not in muscle tissues of all exposed animals as measured by increase in PCNA levels. Our data suggest that exposure to heat-killed filtered bacteria can trigger substantial molecular responses and cause genomic instability in target and distant organs. Even though bacteria were non-pathogenic and unable to cause infection, their remnants still caused a profound effect on exposed animals.