Incorporation of radioactive polyunsaturated fatty acids into liver and brain of developing rat

The incorporation of radioactivity from orally administered linoleic acid-1-¹⁴C, linolenic acid-1-¹⁴C, arachidonic acid-³Hg, and docosahexaenoic acid-¹⁴C into the liver and brain lipids of suckling rats was studied. In both tissues, 22 hr after dosing, 2 distinct levels of incorporation were observed: a low uptake (from 18∶2-1-¹⁴C and 18∶3-1-¹⁴C) and a high uptake (from 20∶4-³H₈ and 22∶6-¹⁴C). In adult rats, the incorporation of radioactivity into brain lipids from 18∶2-1-¹⁴C and 20∶4-³H was considerably lower than the incorporation into the brains of the young rats. In the livers of the suckling rats, the activity from the 18 carbon acids was associated mostly with the triglyceride fraction, whereas the activity from the 20∶4-³H₈ and 22∶6-¹⁴C was concentrated in the phospholipid fraction. In the brain lipids, the activity from the different fatty acids was associated predominantly with the phospholipids. In the liver and brain phospholipid fatty acids, some of the activity in the 18∶2-1-¹⁴C and 18∶3-1-¹⁴C experiments was associated with 20 and 22 carbon polyunsaturated fatty acids; however, radioactivity from orally administered 20∶4-³H₈ and 22∶6-¹⁴C was incorporated intact into the tissue phospholipid to a much greater extent compared with the incorporation of radioactivity into 20∶4 and 22∶6 in the experiments where 18∶2-1-¹⁴C and 18∶3-1-¹⁴C, respectively, were administered. Possible reasons for these differences are discussed. Rat milk contains a wide spectrum of polyunsaturated fatty acids, including linoleate, linolenate, arachidonate, and docosahexaenoate. During the suckling period in the rat, there is a rapid deposition of 20∶4 and 22∶6 in the brain. The results of the present experiments suggested that dietary 20∶4 and 22∶6 were important sources of brain 20∶4 and 22∶6 in the developing rat.     

Phosphatidylinositol 4,5-bisphosphate phospholipase C activity in particulate preparations from rat brain

We describe the hydrolysis of phosphatidylinositol 4,5-bisphosphate by a particulate rat brain preparation in the presence of the cationic detergent, cetrimide. The optimum cetrimide concentration was in the range 0.2–0.4 mg/ml; at higher or lower concentrations, the reaction rate diminished abruptly, suggesting that the electrical charge density of the micelle is critical for enzymatic attack. In other respects, such as its partial requirement for Ca⁺⁺ and its pH optimum of about 7.0, the particulate enzyme seems similar to soluble preparations which have been reported previously. Interestingly, the particulate preparation could be stimulated about fourfold by a soluble brain extract in the presence of 1 mM guanosine triphosphate, confirming that the enzyme is the catalytic subunit in a membrane-bound signal-transduction system.     

Incorporation oftrans long-chain n−3 polyunsaturated fatty acids in rat brain structures and retina

During heat treatment, polyunsaturated fatty acids and specifically 18∶3n−3 can undergo geometrical isomerization. In rat tissues, 18∶3 Δ9c, 12c, 15t, one of thetrans isomers of linolenic acid, can be desaturated and elongated to givetrans isomers of eicosapentaenoic and docosahexaenoic acids. The present study was undertaken to determine whether such compounds are incorporated into brain structures that are rich in n−3 long-chain polyunsaturated fatty acids. Two fractions enriched intrans isomers of α-linolenic acid were prepared and fed to female adult rats during gestation and lactation. The pups were killed at weaning. Synaptosomes, brain microvessees and retina were shown to contain the highest levels (about 0.5% of total fatty acids) of thetrans isomer of docosahexaenoic acid (22∶6 Δ4c, 7c, 10c, 13c, 16c, 19t). This compound was also observed in myelin and sciatic nerve, but to a lesser extent (0.1% of total fatty acids). However, the ratios of 22∶6trans to 22∶6cis were similar in all the tissues studied. When the diet was deficient in α-linolenic acid, the incorporation oftrans isomers was apparently doubled. However, comparison of the ratios oftrans 18∶3n−3 tocis 18∶3n−3 in the diet revealed that thecis n−3 fatty acids were more easily desaturated and elongated to 22∶6n−3 than the correspondingtrans n−3 fatty acids. An increase in 22∶5n−6 was thus observed, as has previously been described in n−3 fatty acid deficiency. These results encourage further studies to determine whether or not incorporations of suchtrans isomers into tissues may have physiological implications. Presented in part at the 32nd International Conference on the Biochemistry of Lipids, 1991, Granada, Spain. Delta nomenclature (Δ) is used fortrans polyunsaturated fatty acids to specify the position and geometry of ethylenic bonds. Polyunsaturated fatty acids containingtrans double bonds are abbreviated giving the locations of thetrans double bonds only; e.g., 20∶5 Δ17t 20∶5 Δ5c,8c,11c,14c,17t; 22∶5 Δ19t, 22∶5 Δ7c,10c,13c,16c,19t; 22∶6 Δ19t 22∶6 Δ4c,7c,10c,13c,16c,19t.     

Identification of Genes Essential for Sulfamate and Fluorine Incorporation During Nucleocidin Biosynthesis

Nucleocidin is an adenosine derivative containing 4’-fluoro and 5’-O-sulfamoyl substituents. In this study, nucleocidin biosynthesis is examined in two newly discovered producers, Streptomyces virens B-24331 and Streptomyces aureorectus B-24301, which produce nucleocidin and related derivatives at titers 30-fold greater than S. calvus. This enabled the identification of two new O-acetylated nucleocidin derivatives, and a potential glycosyl-O-acetyltransferase. Disruption of nucJ, nucG, and nucI, within S. virens B-24331, specifying a radical SAM/Fe−S dependent enzyme, sulfatase, and arylsulfatase, respectively, led to loss of 5’-O-sulfamoyl biosynthesis, but not fluoronucleoside production. Disruption of nucN, nucK, and nucO specifying an amidinotransferase, and two sulfotransferases respectively, led to loss of fluoronucleoside production. Identification of S. virens B-24331 as a genetically tractable and high producing strain sets the stage for understanding nucleocidin biosynthesis and highlights the utility of using 16S-RNA sequences to identify alternative producers of valuable compounds in the absence of genome sequence data.     

Sphingolipid biosynthesis by particulate fractions of normal and “Quaking” mouse brain

No differences were detected in the ability of brain particulate fractions isolated from “Quaking” and normal mice for the incorporation of:¹⁴C-L-serine into 3-keto-dihydrosphingosine; 1-¹⁴C-stearoyl acid and 1-¹⁴C-stearoyl CoA into ceramide and¹⁴C-choline from CDP¹⁴C-choline into sphingomyelin.     

Incorporation and metabolism of fatty acids by cultured dissociated cells from rat cerebrum

Dissociated brain cells in culture incorporate a variety of saturated and unsaturated fatty acids into their cellular lipids. Of the various fatty acids studied, uptake of radioactivity was greatest for stearic acid and decreased progressively with decreasing chain length. Incorporation of radioactivity from linoliec and linolenic acids was more extensive than from oleic acid. Cellular phospholipids and triacylglycerols were labeled preferentially from all fatty acid precursors, with the relative amount of label in phospholipids being greatest when cells were incubated with linolenic acid. Fatty acids underwent desaturation and chain elongation. Changes in the labeling pattern of phospholipid fatty acids in the course of incubation demonstrated precursor-product relationships for laurate (12∶0), myristate (14∶0), palmitate (16∶0), and stearate (18∶0) and for linolenate (18∶3), eicosapentaenoate (20∶5), docosapentaenoate (22∶5), and docosahexaenoate (22∶6). The appearance of label in 22∶5 and 22∶6 paralleled the entrance of label into the ethanolamine phosphoglyceride fraction. Conversion of linoleate (18∶2 ω6) to arachidonate (20∶4 ω6) could be demonstrated but did not proceed via 18∶3 ω6.     

Superhydrophobic graphene/ceramic templates for the preparation of particulate drugs

This paper proposes the use of superhydrophobic graphene/ceramic templates fabricated through laser texturing and patterning for the preparation of particulate drugs. A nanosecond pulse fiber laser was used to texture a graphene film coated on ceramic substrates for obtaining a superhydrophobic surface. Then, laser patterning was conducted on the laser-textured surface to define the diameter of the prepared particulate drugs. Laser-textured graphene/ceramic substrates with a laser areal fluence of 17.51 J/cm² and a hatch distance of 0.01 mm exhibited a maximum water contact angle of 151.5°. This result was obtained because the laser-textured graphene films contained coral reef structures with nanoscale pores. Raman analyses indicated that the intensities of the G and 2D bands gradually decreased after the laser texturing and patterning processes. Moreover, the sheet resistance of the laser-textured and laser-patterned graphene films was larger than that of untextured ones because the thickness of the graphene films was reduced through laser thinning. The maximum heating temperature of the graphene-based heater was 140 °C for an input direct current voltage of 36 V. In addition, graphene-based self-heating devices were developed and successfully used to dry liquid roflumilast.     

Enzymatic synthesis of glucosylsphingosine by rat brain microsomes

Labeled glucosylceramide was formed when rat brain microsomes were incubated with¹⁴C-UDP-glucose. When sphingosine was added to the incubation mixture, labeled glucosylsphingosine was synthesized, and the formation of glucosylceramide increased.     

贵金属纳米颗粒及其催化剂的生物还原制备技术

利用自行筛选的具有强还原能力的菌株制备得到银纳米颗粒,所得颗粒的粒径基本在10 nm以下,主要集中在2～8nm.将生物还原过程进一步引入催化剂制备过程.得到负载犁银催化剂和负载型钯催化剂,并将催化剂分别应用于1,2一丙二醇空气氧化合成丙酮醛、CO氧化生产CO2以及蒽醌加氢反应中.     

Functionalization of dextran: Incorporation of carboxy groups by O-succinoylation

The modification reaction of dextran with succinic anhydride using pyridine as catalyst and N,N-dimethylformamide/lithiumchloride system as solvent was studied. The structure of the resulting polymers was determined by means of ¹H and ¹³C NMR spectroscopy. A linear dependence of the reaction rate on succinic anhydride concentration was found. The orders with respect to pyridine and polymer concentrations were 1 and 2, respectively. The activation energy was found to be 69.0 kJ/mol. ¹³C NMR spectra at 75.4 MHz of partially modified dextran with monosuccinate groups were studied in order to evaluate the selectivity of the reaction of dextran with succinic anhydride in the homogeneous phase. Analysis of the spectra of ring carbons in the anhydroglucose units shows that the reactivity of individual hydroxy groups decreases in the order C-2 ∽ C-3 > C-4. For samples with higher degree of substitution the order is C-4 > C-3 > C-2. The results were explained by considering the formation of intramolecular hydrogen bonds as well as by steric considerations.     

Biosynthesis of brain sphingolipids and myelin accumulation in the mouse

Microsomal fractions were prepared from the brains of mice sacrificed at 13 are points between 5 and 48 days of age. Several enzymatic activities implicated in sphingolipid biosynthesis were assayed. The developmental pattern for the terminal step in galactosylceramide synthesis, transfer of galactose from UDP-galactose to ceramide, peaked sharply at 17–19 days of age. Thus maximal activity for biosynthesis of this relatively myelin specific lipid appears slightly before the time period of maximal rate of accumulation of myelin (21–23 days). These latter data were obtained by isolating myelin from mice at 10 age points from 9 to 45 days of age. A control enzymatic activity, transfer of glucose from UDP-glucose to ceramide to form glucosylceramide, was high at all age points with a broad peak between 20 and 35 days of age. Condensation of palmitoyl-CoA and serine to form ketodihydrosphingosine, a common precursor to both glucosyl- and galactosylceramide also followed a developmental pattern of high activity at all age points, but peaked slightly at ca. 10–12 days of age.     

Uptake of fatty acids by the developing rat brain

Polyunsaturated fatty acids are avidly taken up by the developing rat brain. To explore the specificity of this process, [1-¹⁴C]labeled 16∶0, 18∶2n−6, 18∶3n−3, and 22∶6n−3 each were co-injected with [³H]18∶1n−9 into the jugular vein of two-wk-old functionally hepatectomized and shamoperated control rats. The radioactivities present in the brain, liver and serum were assessed 30 min after injection. Uptake of labeled fatty acids into brain lipids steadily increased with increasing degree of unsaturation, with more than twice as much uptake of 22∶6n−3 compared to 16∶0. Phosphatidylcholine was the principal radioactive species in the brain except for animals injected with [1-¹⁴C]22∶6n−3, in which more of the label was incorporated into phosphatidylethanolamine. Determination of watersoluble oxidation products in the brain and serum revealed that the greater uptake of the more unsatrated fatty acids did not result from differences in rates of degradation.     

Lipid and fatty acyl composition of rat brain capillary endothelia isolated by a new technique

A method is described for the isolation of pure capillary endothelia from rat brain and the phospholipid composition of these cells is reported. This method is rapid and requires only a small amount of starting material. It involves: (a) tissue disruption by high speed homogenization, (b) separation of the capillary endothelia from other brain structures using sucrose gradients, and (c) a final purification using a glass bead column. Choline and ethanolamine phosphoglycerides were found to be the predominant lipid classes of these cells amounting to 31.9% and 24.4%, respectively, of total phospholipids. The choline phosphoglycerides consisted almost exclusively of 1,2-diacyl glycerophosphorylcholine, whereas the ethanolamine phosphoglycerides consisted of approximately equal amounts of 1,2-diacyl and 1-alk-l’-enyl-2-acyl glycerophosphorylethanolamine. The composition of the constituent fatty acids of both choline and ethanolamine phosphoglycerides and the alk-1-enyl composition of ethanolamine phosphoglycerides is reported. Saturated fatty acids accounted for 45% of the total fatty acids in choline phosphoglycerides and for 53% in ethanolamine phosphoglycerides. Arachidonic acid accounted for approximately 48% of the total fatty acids in alk-1-enyl ethanolamine phosphoglyceride.     

Incorporation ofcis- andtrans-octadecenoic acids into the membranes of rat liver mitochondria

The incorporation of dietary isomeric fatty acids into the membranes of liver mitochondria was investigated. Three groups of rats were fed diets containing 3% sunflower seed oil plus 15%, 20%, or 25% partially hydrogenated arachis oil. A fourth group was fed 25% partially hydrogenated arachis oil, but no sunflower seed oil. All diets were given for 3, 6, or 10 weeks. After 10 weeks, the content oftrans fatty acids in the lipids of the mitochondrial membranes was 15–19% of the total fatty acids. The composition of thetrans- and thecis-octadecenoic acids in the lipids of the mitochondrial membranes was similar for all groups supplemented with sunflower seed oil (SO), irrespective of time and dietary level of partially hydrogenated arachis oil (HAO). Thecis 18∶1 (n−8), which was a major isomer of the partially hydrogenated arachis oil, was almost excluded from the mitochondrial fatty acids. Likewise, the content oftrans 18∶1 (n−8) was considerably lower in the mitochondrial lipids than in the diet. On the contrary, the content oftrans 18∶1 (n−6) was higher in the mitochondrial lipids than in the diet. In the group fed without sunflower seed oil, isomers of linoleic acid and arachidonic acid were observed in the lipids of mitochondrial membranes. Presented in part at the ISF Congress, Marseille, September 1976.     

Incorporation of Cesium by Hydrating Calcium Aluminosilicates

The leach behavior of cesium from a series of cement-based waste forms has been modeled in terms of pore solution concentration and phase formation data. Cement-based Materials enriched in alumina and silica, and compositionally related glasses, were hydrated in the presence of cesium hydroxide to gauge the degree of partitioning of cesium by the developing hydrates. The compositions of the pore solutions extracted from the cement-based waste forms were compared both pore solution and leachates, the retention of cesium was found to be directly related to the bulk composition of the waste form; the degree of partitioning of cesium by the hydrated phases increased as more alumina and/or silica were added to the formulation, suggesting that cesium-containing phases were forming. Additionally, the leachate concentration of cesium for any given composition was also found to be related to the pore solution concentration. This observation led to the development of a leaching model for cement-based waste forms in which the concentration of cesium in the leachate could be described in terms of pore solution concentration, diffusion, dissolution, and development of cesium host phases. Using this model, effective diffusion coefficients for cesium were calculated to be ∼2 × 10⁻⁷ cm²· s⁻¹. Pollucite and cesium-substituted herschelite were identified as two of the hydration products of the studied glasses. The ability of these zeolites to strongly partition cesium suggests that small quantities of zeolitic phases may also be forming in the cement-based waste forms, thus explaining their compositionally related leach behavior.     

Electrochemical study of rat brain acetylcholinesterase inhibition by chlorofos: Kinetic aspects and analytical applications

An electrochemical sensor was applied for investigating the immobilized rat brain acetylcholinesterase inhibition by chlorofos. Two alternative routes were explored as response-generating reactions: (i) direct electrochemical oxidation of thiocholine produced upon acetylthiocholine enzymatic hydrolysis and (ii) reduction of the produced thiocholine with hexacyanoferrate (III), followed by hexacyanoferrate (II) electrochemical detection. The advantages of the direct way are simplicity and higher sensitivity compared to the indirect one, which however avoids the interferences because of the lower potential applied.Enzyme inhibition was identified as competitive, the increasing from 1.31 to 1.43 mmol L⁻¹ with chlorofos concentration in the range 0.2-1.0 mmol L⁻¹ and the maximal rate of the enzyme reaction remaining constant (I_max = 579.30 ± 5.71 μA) in the presence of chlorofos. The inhibition constant was calculated using the Dixon method (K_I = 10.07 mmol L⁻¹).The suppression of the acetylcholinesterase activity by the inhibitor, expressed as current decrease at a constant substrate concentration, was exploited for chlorofos quantification optimized by the design of experiments methodology. Optimal response was obtained for an acetylthiocholine concentration of 0.2 mmol L⁻¹, at 26 °C and pH 7.     

Role of particulate fillers in elastomer reinforcement: a review

In a treatise of this size it is impossible to discuss reinforcement in all its ramifications. The author has attempted to put the emphasis on an understanding of the observed phenomena wherever possible and has gone into detail only when necessary. Naturally the judgement of the importance of details is a subjective matter and not everybody may agree with the author's treatment. An attempt is made to present a unified picture in which differences between concepts and theories are given secondary importance and the agreements between them are emphasized. Much of the subject will deal with carbon black, the most universally used reinforcing filler, but other fillers such as the silicas with their coupling agents are also discussed and compared with carbon black. Similarities and differences between fillers are related to the resulting differences in the properties of the final elastomer—filler composite. Some attention is also given to the growing importance of the fibrous filler. Several review articles and books have dealt with reinforcement. See references 1 to 5.     

Metabolism of linolenic acid in developing brain: I. Incorporation of radioactivity from 1-14C linolenic acid into brain fatty acids

Twelve-thirteen day old rats were given 1-¹⁴C linolenic acid by intraperitoneal injection. Fatty acids were isolated from the brains of animals sacrificed at the end of 8 and 48 hr and 15 and 45 days. Eight hr after the tracer, radioactivity was found neither in 18∶3 nor its endproduct, 22∶6, and palmitate was the most highly radioactive component. At longer intervals, 22∶6 seemed to retain much of the radioactivity, whereas palmitate showed a precipitous decline in radioactivity. Initial oxidation of linolenate and sparing of the linolenate complexed with polar lipids are discussed.     

Sphingolipid metabolism II. The biosynthesis of 3-keto-dihydrosphingosine by a partially-purified enzyme from rat brain

A condensation reaction between serine and palmitoyl CoA by a partially-purified enzyme from rat brain is described. The product of the reaction, 3-keto-dihydrosphingosine is characterized by the conversion to several derivatives. The addition of EDTA to the incubation mixture results in inhibition of the conversion of serine to phospholipid, with the result that 3-keto-dihydro-sphingosine is the sole product.