Chemical Studies on Ambergris

So called ambreinolal (IV),* a component of ambergris, was first synthesized by CrO₃ oxidation of ambreinolol (III)* which was obtained from ambreinolide (II) by reduction with LiAlH₄. Ambreinolol (III) was converted to the C₁₇-saturated oxide (VII) in a good yield through the monotosylate (VIII) by treatment with p-toluenesulfonyl chloride in pyridine.

Ambrein (I), a major constituent of ambergris, was easily converted to ambrein-tetrahydropyranylether (II), of which thermal decomposition gave back ambrein (I). The tetrahydropyranylether (II) was oxidized to ambreinolal-tetrahydropyranylether (V) in two steps. Ambreinolal-tetrahydropyranylether (V) was synthesized from ambreinolol (VII) in four steps and converted to the C₁₇-unsaturated oxide (VI) on heating.     

Regulation of homoserine dehydrogenase in developing organs of soybean seedlings

The regulation of homoserine dehydrogenase (HSD) activity (EC 1.1.1.3) by L-threonine, L-cysteine and K⁺ was examined using extracts of organs of soybean seedlings harvested 3, 6, 11, and 19 days after germination. K⁺ stimulated HSD activity from each source at least 2-fold. HSD activity was completely inhibited by 10 mM L-cysteine while 10 mM D-cysteine was not inhibitory. A progressive decrease in sensitivity of NAD-dependent HSD to inhibition by 10 mM L-threonine occurred in all organs except the leaf during the sampling period. This progressive decrease in sensitivity of the HSD to threonine inhibition was detected only when K⁺ was present in the assay mixtures. Four major molecular forms, including one rapidly migrating form (form I) and three more slowly migrating forms (forms II, III, IV) of HSD, were identified in extracts of soybean organs by polyacrylamide electrophoresis. Chromatographic and electrophoretic data indicate that form I, which was not inhibited by threonine or stimulated by K⁺, was of lower MW than forms II, III and IV which were of similar MW. These latter 3 forms were inhibited by threonine and stimulated by K⁺. During soybean seedling development form II increased in amount and forms I and IV decreased in amount. This alteration in the amounts of the forms of HSD occurred during the same period as the decrease in the amount of threonine inhibition. Since K⁺ stimulation of HSD decreased during soybean organ development and K⁺ enhanced threonine inhibition, this might account for the observed decrease in threonine inhibition.     

Isolation, characterization and protein and polar lipid compositions of Paracoccus denitrificans outer membrane

Sucrose density gradient centrifugation of Paracoccus denitrificans strains ATCC 13543 and ATCC 17741 cell envelopes plus poly-β-hydroxybutyrate, isolated from organisms broken using a French pressure cell, revealed three bands of densities: I, 1.16 g/ml; II, 1.19 g/ml; III, 1.24 g/ml. On the basis of chemical and enzymatic assays and sodium dodecyl sulfate-polyacrylamide gel electrophoresis the bands were identified as: I, cytoplasmic membrane; II, poly-β-hydroxybutyrate; III, outer membrane plus poly-β-hydroxybutyrate. Poly-β-hydroxybutyrate was removed by increased low-speed centrifugation before deposition of cell envelopes. Density gradient centrifugation of cell envelopes gave a simple pattern of two bands, cytoplasmic and outer membranes. In both strains outer membranes showed a broad protein band at M_r 70 000–83 000 upon SDS-polyacrylamide gel electrophoresis of samples solubilized at 25°C, which was not present in samples solubilized at 100°C, where a single major band was present of M_r 32 000 in strain ATCC 13543 and 35 000 in strain ATCC 17741. The major outer membrane protein stained positively for lipid in both strains, as did an M_r 70 000 protein, which was the second major protein in strain ATCC 17741. The second major outer membrane protein of stain ATCC 13543 had an M_r of 20 000 in unheated samples but 23 000 in heated samples. This protein was not present in strain ATCC 17741. Quantitative data on the polar lipid compositions of cell envelope fractions are presented.     

Formation of Glucose Isomerase by Streptomyces sp.

Sophoradin (I) [2′,4,4′-trihydroxy-3,3′,5-tris(3-methyl-2-butenyl)chalcone] which had been isolated from “Guang-Dou-Gen” (the root of Sophora subprostrata Chun et T. Chen) was synthesized through Claisen rearrangement. The reaction of p-hydroxybenzaldehyde and 3-chloro-3-methyl-1-butyne (III) gave 4-(1,1-dimethylpropargyloxy)benzaldehyde (VIII), which was catalytically hydrogenated over Lindlar catalyst to afford 4-(1,1-dimethylallyloxy)benzaldehyde (IX). IX was converted to 4-hydroxy-3-(3-methyl-2-butenyl)benzaldehyde (X) by Claisen rearrangement. The reaction of X and III gave 3-(3-methyl-2-butenyl)-4-(1,1-dimethylpropargyloxy)benzaldehyde (XI). Condensation of 2-hydroxy-4-(1,1-dimethylpropargyloxy)acetophenone (IV) and XI in alkaline solution gave a chalcone (XIII), which was catalytically hydrogenated over Lindlar catalyst to give 2′-hydroxy-4,4′-bis(1,-dimethylallyloxy)-3-(3-methyl-2-butenyl)chalcone (XIV). XIV was converted to I by Claisen rearrangement.     

Effects of repeated desflurane and sevoflurane anesthesia on enzymatic free radical scavanger system

Background: To investigate the possible effects of repeated sevoflurane and desflurane anesthesia on hepatocellular system by evaluating the free radical metabolism, hepatocellular enzymes and histopatholgical changes in rats. Methods: Four groups of animals were studied. Sevoflurane 2% (v/v) and desflurane 6% (v/v) in air/O₂ were administered to animals in group II (n = 9) and III (n = 9) respectively. 100% (v/v) O₂ was administered in group IV (n = 9). Administration was done for 60 minutes over 3 days. Nine animals were allocated to control group (group I), superoxide dismutase (SOD), catalase (CAT), glutathion peroxidase (GSH-Px), glutathione-s-transferase (GST) and thiobarbituric acid reactive substances (TBARS) were studied. Also electron microscopy was performed. Results: Catalase, SOD, GSH-Px, GST activities and TBARS levels were significantly higher in groups II and III than in group I (p < 0.05). All parameters were significantly higher in groups II versus group IV (p < 0.05). On the other hand, SOD, GSH-Px and GST activities were significantly elevated in group III than IV, but CAT activity and TBARS levels were not significantly. Catalase, SOD, GSH-Px, GST but not TBARS levels were significantly higher in groups II and III than in group IV (p < 0.05). TBARS levels were higher in group III than in group IV, but this elevation was not statistically significant. CAT, SOD and GSH-Px activities were significantly higher in groups II than in group III (p < 0.05). Conclusion: Although electron microscopy findings were similar for group II and III, we can conclude that sevoflurane might cause more cellular damage than desflurane by causing higher activation of free radical metabolising enzymes.     

Low Resolution Mass Spectra of Some Unsaturated δ-Lactones

Mass spectra of the δ-lactones of the following 5-hydroxy-2-enoic acids were determined: 5-hydroxyhex-2-enoic acid (I), 5-hydroxyoct-2-enoic acid (II), 5-hydroxydec-2-enoic acid (III), 5-hydroxydodec-2-enoic acid (IV), 5-hydroxy-8-methylnon-2-enoic acid (V), 5-hydroxy-6-ethyloct-2-enoic acid (VI), 5-hydroxy-5, 6, 6-trimethylhept-2-enoic acid (VII), and 5-hydroxy-5-methylnon-2-enoic acid (VIII). The following modes of fragmentation are consistent with observed m/e values, metastable peaks, and established modes of breakdown in compounds containing similar atomic groupings:—1. Loss of side chain, resulting in ions at m/e 97 for I-VI and at m/e 111 and 153 for VII and VIII (diagnostic peaks); 2. 1,4-Rupture of the ring giving an ion at m/e 68 (diagnostic peak) which loses CO to give m/e 40; 3. Loss of CO from m/e 97 fragment to give m/e 69 which breaks down further to m/e 41→m/e 39; 4. 1, 4-Rupture of m/e 111 and m/e 153 fragments to give m/e 43 and 85, further breakdown of m/e 85→57→41→39; 5. Loss of H₂O from the molecular ion providing there is a hydrogen atom on C₅ and the side chain is at least 3 carbon atoms in length, further loss of H₂O when the side chain is equal to C5 or C7; 6. Loss of CO₂ from the molecular ion of I, IV-VIII; 7. Loss of CO from all molecular ions; 8. Loss of 2×28 from the molecular ions of III, IV, V, VI; 9. Loss of (18 + 28) from the molecular ion of III, IV, V, VI; 10. Loss of 60 from the molecular ion of II, III, IV, V, VI; 11. Formation of M + 1 ion (169) of VII and VIII; 12. Formation of M + 1 ion (143) of saturated δ-octalactone and loss of H₂O from this M + 1 ion.     

Transformation of Schizandrin into Gomisin A by Use of Microbial O-Demethylation and Chemical Reactions

The transformation of schizandrin (I) into gomisin A (II) was accomplished by use of a combination of biotransformation and chemical reactions. The biotransformation, microbial O-demethylation of I by Cuntiinghamella echinulata var. elegans (ATCC 9245) produced two novel metabolites [3-norschizandrin (IV) and 2-norschizandrin (VI)] and two known metabolites [gomisin T (III) and 13-norschizandrin (V)]. Among those metabolites, compound III was derived to II by the O-demethylation with a Lewis acid in the presence of an acid scavenger, followed by methylenation.     

Metabolism of purine bases, nucleosides and alkaloids in theobromine-forming Theobroma cacao leaves

We examined the purine alkaloid content and purine metabolism in cacao (Theobroma cacao L.) plant leaves at various ages: young small leaves (stage I), developing intermediate size leaves (stage II), fully developed leaves (stage III) from flush shoots, and aged leaves (stage IV) from 1-year-old shoots. The major purine alkaloid in stage I leaves was theobromine (4.5 μmol g^–1 fresh weight), followed by caffeine (0.75 μmol g^–1 fresh weight). More than 75% of purine alkaloids disappeared with subsequent leaf development (stages II–IV). In stage I leaves, ¹⁴C-labelled adenine, adenosine, guanine, guanosine, hypoxanthine and inosine were converted to salvage products (nucleotides and nucleic acids), to degradation products (ureides and CO₂) and to purine alkaloids (3- and 7-methylxanthine, 7-methylxanthosine and theobromine). In contrast, ¹⁴C-labelled xanthine and xanthosine were not used for nucleotide synthesis. They were completely degraded, but nearly 20% of [8-¹⁴C]Xanthosine was converted in stage I leaves to purine alkaloids. These observations are consistent with the following biosynthetic pathways for theobromine: (a) AMP → IMP → 5′-xanthosine monophosphate → xanthosine → 7-methylxanthosine → 7-methylxanthine → theobromine; (b) GMP → guanosine → xanthosine → 7-methylxanthosine → 7-methylxanthine → theobromine; (c) xanthine → 3-methylxanthine → theobromine. Although no caffeine biosynthesis from ¹⁴C-labelled purine bases and nucleosides was observed during 18 h incubations, exogenously supplied [8-¹⁴C]Theobromine was converted to caffeine in young leaves. Conversion of theobromine to caffeine may, therefore, be slow in cacao leaves. No purine alkaloid synthesis was observed in the subsequent growth stages (stages II–IV). Significant degradation of purine alkaloids was found in leaves of stages II and III, in which [8-¹⁴C]Theobromine was degraded to CO₂ via 3-methylxanthine, xanthine and allantoic acid. [8-¹⁴C]Caffeine was catabolised to CO₂ via theophylline (1,3-dimethylxanthine) or theobromine.     

MORPHOLOGY AND CYTOLOGY OF SYNTHETIC HYBRIDS OF AGROPYRON TRICHOPHORUM X AGROPYRON CRISTATUM

Dewey, Douglas R. (Utah State U., Logan.) Morphology and (cytology of synthetic hybrids of Agropyron trichophorum X Agropyron cristatum. Amer. Jour. Bot. 50(10): 1028–1034. Illus 1963.—Three hybrids were obtained from controlled crosses of pubescent wheatgrass, A. trichophorum (2n = 42), and hexaploid crested wheatgrass, A. cristatum (211 = 42). The hybrids were intermediate between the parent plants for all vegetative and spike characteristics observed. Under open pollination, 2 of the hybrids set 2 seeds each, and the other hybrid produced 60 seeds. Meiosis in the parent plants was basically regular. Average motaphase-I chromosome associations were 0.09 I, 20.56 II, 0.05 III, and 0.16 IV per cell in the A. trichophorum parent, which was described as a segmental autoallohexaploid. The hexaploid A. cristatum parent averaged 0.18 I, 7.44 II, 0.81 III, 2.86 IV, 0.08 V, and 2.11 VI per cell at diakinesis and was described as an autohexaploid. Chromosome pairing in the hexaploid hybrid averaged 5.08 I, 8.94 II, 4.33 III, 1.11 IV, 0.27 V, and 0.05 VI per cell. On the basis of chromosome pairing in the parent species and their hybrids, it was concluded that 1 of the A. trichophorum genomes was partially homologous with the 3 genomes of hexaploid A. cristatum. Genome formulae for hexaploid A. cristatum, A. trichophorum, and their hybrids were represented as AAAAAA, A₁A₁B₁B₁B₂B₂, and AAAA₁B₁B₂ respectively.     

Initial reactions in the fungal degradation of guaiacylglycerol-β-coniferyl ether,a lignin substructure model

Fusarium solani M-13-1 was shake-cultured in a medium containing guaiacylglycerol-β-coniferyl ether (I), a model compound representing the arylglycerol-β-aryl ether linkage in lignin, as sole carbon source. From the culture filtrate guaiacylglycerol-β-coniferyl aldehyde ether (II) and guaiacylglycerol-β-ferulic acid ether (III) were isolated as metabolic products. Incubation with (III) resulted in formation of guaiacylglycerol-β-vanillin ether (IV), which was further metabolized to guaiacyglycerol-β-vanillic acid ether (V). The results indicate that the cinnamyl alcohol group of (I) is initially oxidized to an aldehyde group, which is further oxidized to a carboxyl group, yielding (II) and (III). Compound (III) is converted to (IV) by the release of a C₂ fragment, and the aldehyde group of (IV) is further oxidized to a carboxyl group, giving (V). In the pathway from (I) to (V), neither oxidation of the benzylic secondary alcohol to ketone nor cleavage of the arylglycerol-β-aryl ether linkage was observed. The fungus was found to attack both erythro and threo form without distinction.     

Bacteriophages of l-Glutamic Acid-Producing Bacteria

Deoxyribonucleic acids were isolated from Brevibacterium lacto fermentum No. 2256 and its four representative phages belonging to different serological groups, i.e., P465 (group I), P 468 II (group II), Ap 85 III (group III) and P4 (group IV), by phenol extraction.

DNA’s isolated from the four phages contained only usual four bases, i.e., guanine, adenine, cytosine and thymine. The G-C content of the phage DNA was determined by thermal denaturation method (T_m); the values of P465, P 468 II, Ap 85 III and P4 were 54.0, 54.6, 56.6 and 55.3%, respectively. Sedimentation coefficient was measured by ultracentrifugal analysis using ultraviolet optics; s_20,w of the phage DNA’s were 15.5 to 31.8 s.

Unusual bases were not detected also in the host bacterial DNA. The G-C content of the bacterial DNA determined by paperchromatography was 55.1% which came very close to the G-C content derived from the T_m.

Morphological properties of the P- and Ap-series phages described in previous papers were examined by means of electron microscopy, analytical centrifugation, CsCl density-gradient centrifugation and ultrafiltration.

In view of the buoyant density of phage particles, fourteen Brevibacterium phages were classified into four groups, i.e., the phages under group I had buoyant density of 1.511 to 1.514g cm^?3, and those under groups II, III and IV, densities of 1.482, 1.492 and 1.508 g cm^?3, respectively; and above grouping corresponded to that by the serological chracteristics of the phages.

Electron microscopic observation by means of shadow-casting or negative staining recognized all the phages as tadpole-shaped; their particles having polyhedral head (40 ~ 70 mμ in diameter) and long tail (80 ~ 275 mμ in length).

In relation to particle sizes of the phages as estimated by all of above-mentioned methods, no significant differences were observed between the sizes calculated from ultrafiltration and those obtained directly from electron micrographs by negative staining.     

Bacteriophages of l-Glutamic Acid-Producing Bacteria

The growth characteristics of phages were investigated with the four phages, active on Brevibacterium lactofermentum, which were selected from the respective serological groups, namely, P465 (group I), P468II (group II), Ap85III (group III) and P4 (group IV).

The adsorption rate of the phages, P465 and P468II, on the host bacteria was low, whereas that of the phages, Ap85III and P4, was higher. The adsorption rate constants for the four phages were respectively calculated at 2.02 × 10^?10, 1.87 × 10^?10, 4.32 × 10^?10 and 3.15 × 10^?10 cm³ per minute, at 30°C in G₅B₂ medium. With reference to the ionic environment for adsorption, the phages, P465 and Ap85III, specifically required either for Ca⁺⁺ or Mg⁺⁺; the phage P468II, for both; and the phage P4, for neither.

The growth characteristics of these phages were examined by the one-step growth experiment. The latent periods of the phages were 50, 53, 57 and 47 minutes, respectively; and the corresponding average burst sizes were about 98, 31, 145 and 126. The growth of the phage P4 was completely suppressed at above 34°C, although the host bacteria and the other three phages were capable of the full growth at that temperature.     

The endo-(1→4)-β- -glucanase system of Penicillium pinophilum cellulase: Isolation, purification, and characterization of five major endoglucanase components

Five major endo-(1→4)-β- -glucanases (I–V) have been isolated from a cellulase preparation of P. pinophilum. The pI values for I–V were 7.4, 4.8, 4.1, 3.7, and 4.0, respectively, and the respective molecular weights were 25,000, 39,000, 62,500, 54,000, and 44,500, when determined by SDS-gel electrophoresis. Endoglucanase V was optimally active at 65–70° and I–IV were most active at 50–60°. The pH optima of I and III–V were in the range 4.0–5.0. Antiserum prepared to I reacted only with I; II antiserum reacted only with II. Endoglucanases I and V were more random in their attack on CM-cellulose and H₃PO₄-swollen cotton cellulose, and showed no activity against cello-oligosaccharides containing less than five -glucose residues, whereas III and IV were active against all the cello-oligosaccharides tested and acted in a less random manner, and II was intermediate in its catalytic action. III was adsorbed completely on both Avicel PH101 and H₃PO₄-swollen cellulose, whereas IV was not adsorbed. The endoglucanases I–V have distinct roles in the digestion of cellulose.     

The phenoloxidases of the ascomycete Podospora anserina: The three forms of the major laccase activity

In Podospora anserina three laccase activities (I, II and III) were identified. Present results show the existence of an additional lacaase (an anodic protein; MW 80,000; R_f 0.07). Laccase IV derived from the dissociation at acid pH (4.5) of a protein which showed identical molecular weight (390,000) and R_f (0.1) to the oligomeric laccase I. The recovery of laccase I activity when starting from laccase IV (purified by means of isoelectric focusing) suggests that laccase I itself was the source of laccase IV. In turn, laccase IV gave rise to the laccase III after electrophoresis or dialysis at basic pH (8.5).     

Pyruvate kinase isozymes in adult tissues and eggs of Rana pipiens. Comparative studies on the properties of the different isozymes

The properties of the isozymes of pyruvate kinase (ATP: pyruvate phosphotransferase, EC 2.7.1.40) found in unfertilized frog egg have been compared to those found in adult tissues of Rana pipiens. Chromatographic, kinetic, and electrophoretic data indicate that, of the five electrophoretic forms found in egg, the isozyme with the least anodic mobility (isozyme I) is the same molecular species as the only isozyme found in heart, and the egg isozyme with the greatest anodic mobility (isozyme V) is identical to the major isozyme found in liver.The activity of egg isozyme I was markedly inhibited by the antibody to the skeletal muscle enzyme, which has been shown previously to cross-react with the cardiac enzyme, but was unaffected by the antibody to liver isozyme V; the opposite effects were observed with egg isozyme V. The antibody to the skeletal muscle enzyme inhibited egg isozymes II > III > IV whereas the antibody to the liver enzyme gave the reverse inhibitory pattern, e.g., isozyme IV > III > II.In vitro dissociation-reassociation of mixtures of isozyme I and V led to the formation of the other three isozymes. Similar experiments performed individually with either egg isozyme III or IV resulted in the production of predominantly isozymes III, II, and I due to the instability of isozyme V during the hybridization procedure.The above results indicate that isozymes I and V are tetramers of the respective parental subunits and that isozymes II, III, and IV are hybrid molecules with subunit assignments of (I₃V₁), I₂V₂), and (I₁V₃), respectively.     

PLOIDAL CHANGES IN CLONAL CULTURES OF SPIROGYRA COMMUNIS AND IMPLICATIONS FOR SPECIES DEFINITION

A clonal culture of Spirogyra filaments of initially uniform width produced filaments of three additional significantly different widths. Group I filaments of the original clone were 30.9 ± 0.7 μm wide (mean ± SD, N = 50). Group I filaments produced Group II filaments (22.0 ± 1.1 μm) through vegetative growth and sexual reproduction. Zygospores from homothallic Group I filaments produced germlings representative of Groups I and II; zygospores from homothallic Group II filaments produced germlings representative of Group II only. Germlings of Groups III (27.7 ± 1.0 μm) and IV (44.9 ± 0.8 μm) were produced in the cross of I × II. Viable zygospores from homothallic Group III filaments were obtained. Cells of Group IV filaments were initially binucleate and did not conjugate. Of the six intergroup crosses possible, four resulted in conjugation-tube formation only; two crosses yielded zygospores (I × II and III × IV). Germlings from the successful cross of Groups III and IV produced filaments of all four groups. Chromosome counts were: Group I (24), Group II (12), Group III (18), and Group IV (24, one nucleus). Relative nuclear fluorescence values of mithramycin-stained DNA were (mean ± SD, N ≥ 30): Group I (11.1 ± 1.4), Group II (5.7 ± 0.7), Group III (8.8 ± 1.3), and Group IV (10.0 ± 0.9, one nucleus). Cytologically, Group II appears to be a diploid (2x), Group I a tetraploid (4x), and Group III a triploid (3x). Systematically, Groups I, II, and III key out to Spirogyra singularis, S. communis, and S. fragilis, respectively, using Transeau's mongraph of the family Zygnemataceae. These species are interpreted to represent a species complex of S. communis (whose name has priority) with the ancestral haploid (x = 6) missing.     

Lignin degradation byPhanerochaete chrysosporium: Metabolism of a phenolic phenylcoumaran substructure model compound

The degradation of a lignin substructure model compound, 5-formyl-3-hydroxymethyl-2-(4-hydroxy-3,5-dimethoxyphenyl)-7-methoxycoumaran (I), in ligninolytic culture of a white-rot wood decay fungus,Phanerochaete chrysosporium, was investigated. It was found that I was hydroxylated or dehydrogenated in its coumaran ring to give 2-(5-formyl-2-hydroxy-3-methoxyphenyl)-3-hydroxypropiosyringone (II) and two coumarones, 5-formyl-3-hydroxymethyl-2-(4-hydroxy-3,5-dimethyoxyphenyl)-7-methoxycoumarone (V) and 3,5-diformyl-2-(4-hydroxy-3,5-dimethoxyphenyl)-7-methoxycoumarone (VI), II was further converted to 2,6-dimethoxy-p-benzoquinone (IV), syringic acid (III), and 5-carboxyvanillic acid (VIII). These metabolic products were identified by mass spectrometric comparison with the authentic compounds. A proposed pathway for the degradation of I is presented on the basis of these metabolic products. The degradation could be catalyzed mainly by phenol-oxidizing enzymes.Non-Standard Abbreviations TLC thin layer chromatography     

Improving the endothelial dysfunction in type 2 diabetes with chromium and vitamin D3 byreducing homocysteine and oxidative stress: A randomized placebo-controlled trial

BackgroundChromium picolinate (CrPic) and vitamin D3 are known as two antioxidant micronutrients. Through inducing endothelial dysfunction, oxidants such as homocysteine (Hct) and malondialdehyde (MDA) lead to cardiovascular disease in type 2 diabetes mellitus (T2DM). No published data has directly examined the effects of these two antioxidants on improving the endothelial dysfunction in T2DM throughreducing homocysteine and oxidative stress.MethodsSubjects (n = 92) in this randomized, double blind, placebo-control study were randomly assigned to receive oral placebo (group I), D₃ (group II: 50,000 IU/ week), chromium picolinate (CrPic) (group III: 500 μg/day), and both vitamin D₃ and CrPic (group IV) for four months. Fasting blood samples were drawn at study baseline and following intervention to determine Hct, MDA, total antioxidant capacity (TAC), total thiol groups (SHs), vascular cell adhesion molecule- 1 (VCAM-1), and plasminogen activator inhibitor-1 (PAI-1).ResultsAfter intervention, MDA significantly decreased in groups II and IV; TAC significantly increased in group IV, and SHs significantly augmented in group III; Hct was significantly reduced in groups II, III, and IV; and VCAM-1 significantly decreased in groups III and IV and PAI-1 was significantly reduced in groups II, III, and IV.ConclusionOur findings suggest that through reducing homocysteine and oxidative stress and improving endothelial dysfunction, chromium and vitamin D₃ co-supplementation might be predictive and preventive of cardiovascular diseasesassociated with T2DM.IRCT, IRCT20190610043852N1, registered 21 October 2019, https://fa.irct.ir/user/trial/42293/view     

Protective effect of omega-3 fatty acid against mercury chloride intoxication in mice

The aim of this study was to investigate the protective effect of omega-3 fatty acid in HgCI₂ toxicity in mice. Levels of malondialdehyde (MDA), reduced glutathione (GSH), nitric oxide (NO) and total sialic acid (TSA), and histopathological changes in selected organs were evaluated. Twenty-eight mice were equally divided into 4 groups, namely Groups I–IV. Group I animals received intraperitoneal (ip) injection of physiological saline solution; Group II animals received ip injection of 0.4 mg/kg/day HgCI₂; Group III animals received ip injection of 0.4 mg/kg/day HgCI₂ in addition to subcutaneous (sc) injection of 0.5 g/kg/day omega-3 fatty acid; and Group IV animals received sc injection of 0.5 g/kg/day omega-3 fatty acid. All treatments lasted 7 days. The levels of MDA, NO and TSA were significantly higher in Group II and lower in Groups III and IV as compared to the Group I. GSH level was the highest in Group IV. In histopathology, severe degeneration in liver and kidney was observed in Group II animals. These degrading changes were seen to be reduced greatly in Group III animals. The results suggested that omega-3 fatty acid might attenuate HgCI₂-induced toxicity by improving antioxidant status and acute phase response in mice.     

Biochemical and immunological characterization and the synthesis of rat intestinal glycoproteins following the induction of rapid synchronous vitamin A deficiency

Glycoproteins and proteins were extracted from segments or scrapings of the intestine in tube-fed, vitamin-A-deficient and control rats on the eight day after withdrawal of retinoic acid from the diet by using either 1% sodium dodecyl sulfate (SDS) or aqueous 5 mM EDTA (pH 7.4). They were then fractionated on columns of Sepharose 4B. Water-soluble peak I material contained large (M_r > 10⁶; S₂₀ = 11.7) glycoprotein aggregates which were rich in hexose, fucose and sialic acid. These aggregates dissociated into several non-identical glycoprotein and protein subunits upon treatment with dithiothreitol. The protein matrix was rich in threonine, valine, proline, serine, glutamate and aspartate. Peak II consisted of smaller proteins and glycoproteins, the latter with much lower carbohydrate content. Some peak II glycoproteins also dissociated into subunits in the presence of dithiothreitol. Peak III consisted mainly of a heterogenous assortment of proteins, including some glycoproteins of low carbohydrate content. Antibodies either to peak II or to peak III reacted both with peaks II and III but not with peak I.The total weight, carbohydrate composition of glycoproteins and the ratio of carbohydrate to protein in the total extract or in each of the three fractions were not significantly affected in vitamin A deficiency despite decreased incorporation of all labeled precursors. Rather, the relatively lower incorporation (approx. 0.8) of radioactive sulfate, D-glucosamine and L-fucose into total SDS-soluble duodenal glycoproteins of vitamin-A-deficient rats could be explained on the basis of a reduced prevalence of goblet cells alone. In contrast, the relative incorporation rate of L-fucose into peak I, but not into peaks II and III, ranged from 0.25 to 0.45, less than expected on the basis of fewer goblet cells alone. The incorporation of radioactive threonine into all protein fractions was reduced to 60% of normal in vitamin A deficiency. Thus, the well established observation that intestinal tissue of vitamin-A-deficient rats synthesizes high molecular weight glycoproteins poorly might be due to several interacting factors: (1) a reduced prevalence of goblet cells, (2) a lower rate of protein synthesis, (3) a lack of retinyl phosphate for the formation of mannosyl or other carbohydrate derivatives, and (4) secondary, and as yet undefined, cellular changes which preferentially reduce the rate of synthesis of high molecular weight fucose- and sialic-acid-enriched glycoproteins.