Differential mobilization of newly synthesized cholesterol and biosynthetic sterol precursors from cells

Previous work demonstrates that the biosynthetic precursor of cholesterol, desmosterol, is released from cells and that its efflux to high density lipoprotein or phosphatidylcholine vesicles is greater than that of newly synthesized cholesterol (Johnson, W. J., Fischer, R. T., Phillips, M. C., and Rothblat, G. H. (1995) J. Biol. Chem. 270, 25037-25046). Here we report that the release of individual precursor sterols varies with the efflux of newly synthesized zymosterol being greater than that of lathosterol and both exceeding that of newly synthesized cholesterol when using either methyl-beta-cyclodextrin or complete serum as acceptors. The transfer of newly synthesized lathosterol to methyl-beta-cyclodextrin was inhibited by actin polymerization but not by Golgi disassembly whereas that of newly synthesized cholesterol was inhibited by both conditions. Newly synthesized lathosterol associated with cellular detergent-resistant membranes more rapidly than newly synthesized cholesterol. Upon efflux to serum, newly synthesized cholesterol precursors associated with both high and low density lipoproteins. Stimulation of the formation of direct endoplasmic reticulum-plasma membrane contacts was accompanied by enhanced efflux of newly synthesized lathosterol but not of newly synthesized cholesterol to serum acceptors. The data indicate that the efflux of cholesterol precursors differs not only from that of cholesterol but also from each other, with the more polar zymosterol being more avidly effluxed. Moreover, the results suggest that the intracellular routing of cholesterol precursors differs from that of newly synthesized cholesterol and implicates a potential role for the actin cytoskeleton and endoplasmic reticulum-plasma membrane contacts in the efflux of lathosterol.     

Isolation and characterization of proteoglycans synthesized in ovo by embryonic chick cartilage and new bone

Cartilage proteoglycans have been well characterized in a number of developing systems, both in vitro and in vivo, but the newly synthesized molecules have been analyzed only from culture material. Because of potential culture artifacts, an attempt was made to characterize the proteoglycans newly synthesized in ovo in chick embryo sternum, tibial epiphysis, and tibial shaft. These in ovo synthesized proteoglycans share many structural features with chick proteoglycans synthesized by chondrocytes in culture including average monomer size, chondroitin sulfate chain size, keratan sulfate chain size, and the ability to aggregate with hyaluronic acid. Moreover, the newly synthesized in ovo proteoglycans, notably those of the tibial epiphysis, display reproducible changes in their structure as a function of embryonic age. These changes correlate with similar changes documented for chick cartilage proteoglycans synthesized in culture. Finally, the proteoglycans synthesized in ovo in the day 17 tibial shaft include, in addition to cartilage proteoglycans, one proteoglycan which seems to be characteristic of bone.     

Differential rates of processing and transport of herpes simplex virus type 1 glycoproteins gB and gC.

The kinetics of processing and transport of herpes simplex virus type 1 (HSV-1) glycoproteins gB and gC was investigated. The conversion of precursor to mature forms and the appearance of the glycoproteins at the infected-cell surface at different times postinfection (p.i.) were studied. gB, synthesized at 4 h p.i., was converted to the mature form with a half-time (t1/2) of 120 min and appeared at the plasma membrane with a t1/2 of 270 min. The gB synthesized at later times p.i. (6, 8, and 10.5 h) was transported less efficiently. Less than 50% of gB synthesized at later times p.i. was processed and transported to the cell surface. gB synthesized in transfected cells was transported to the plasma membrane with kinetics similar to that for gB synthesized at early times p.i. gC was processed efficiently when synthesized at both 8 and 10.5 h p.i., with t1/2 of conversion of pgC to gC of 40 and 60 min, respectively. Approximately 90 to 95% of the gC synthesized was converted to the mature form. The gC synthesized at 8 h p.i. was also transported rapidly to the cell surface, compared with the transport of gB synthesized at the same time, with a t1/2 of 240 min. Greater than 70% of the gC synthesized at 8 h p.i. appeared at the cell surface. The gC synthesized at 10.5 h was transported less efficiently to the cells surface during a 6-h chase.     

Green synthesis of silver nanoparticles from aqueous extract of Scutellaria barbata and coating on the cotton fabric for antimicrobial applications and wound healing activity in fibroblast cells (L929)

Scutellaria barbata is a perennial herb which was vastly prescribed in Chinese medicine to treat inflammations, infections and it is also used a detoxifying agent. We synthesized silver nanoparticles with Scutellaria barbata extract and characterized the nanoparticles with UV–Vis spectroscopic analysis, TEM, AFM, FTIR and XRD. The biofilm inhibiting property of synthesized silver nanoparticles were examined with XTT reduction assay and the antimicrobial property was examined with well diffusion method. The silver nanoparticles were also coated with cotton fabrics and their efficacy against antimicrobials was analyzed to prove its application. The cytotoxic property of synthesized silver nanoparticles was examined with L929 fibroblast cells using MTT assay. Finally we analyzed the wound healing property of synthesized silver nanoparticles with wound scratch assay. The result of our UV–Vis spectroscopic analysis confirms Scutellaria barbata aqueous extract reduced silver ions and synthesized silver nanoparticles. The characterization studies TEM, AFM, FTIR and XRD confirms the synthesized silver nanoparticles are in ideal shape and size to be utilized as a drug. The XTT reduction assay proves silver nanoparticles effectively inhibits the biofilm formation in both resistant and sensitive strains. Antimicrobial sensitivity tests confirms synthesized silver nanoparticles and cotton coated synthesized silver nanoparticles both are effective against gram positive, gram negative and fungal species. Further the results of MTT assay confirms the synthesized silver nanoparticles are non toxic and finally the wound healing potency of the nanoparticles was confirmed with wound scratch assay. Over all our results authentically confirms the silver nanoparticles synthesized with Scutellaria barbata aqueous extract is potent wound healing drug.     

Synthesis of various kinds of esters by four microbial lipases

Ester synthesis by microbial lipases, using homogeneous enzyme preparations, were investigated. The amount of synthesized ester was estimated by alkalimetry, and products were identified by thin-layer chromatography and infrared spectroscopy. Lipases from Aspergillus niger, Rhizopus delemar, Geotrichum candidum and Penicillium cyclopium synthesized esters from oleic acid and various primary alcohols. Only Geotrichum candidum lipase synthesized esters of secondary alcohols. Esters of tertiary alcohols, phenols or sugar alcohols were not synthesized by any lipase. Rather high concentrations of alcohol were required to synthesize the esters of ethylene glycol, propylene glycol or trimethylene glycol. Lipases from Aspergillus niger and Rhizopus delemar synthesized oleyl esters of various fatty acids and some dibasic acids. In contrast, lipases from Geotrichum candidum and Penicillium cyclopium synthesized oleyl esters only from medium or long chain fatty acids.     

Synthesis and pharmacological evaluation of a novel series of 3-aryl-2-(2-substituted-4-methylthiazole-5-yl)thiazolidin-4-one as possible anti-inflammatory and antimicrobial agents

A new series of 3-aryl-2-(2-aryl/benzyl-4-methylthiazole-5-yl)thiazolidin-4-one was synthesized by condensation of 2-aryl/benzyl-4-methylthiazole-5-carbaldehyde, aromatic amines and thioglycolic acid in toluene. All the synthesized compounds are characterized by IR, NMR and elemental or mass analysis. Sixteen out of the newly synthesized compounds were screened for in vivo anti-inflammatory activity using carrageenan-induced rat paw edema method. Some of the synthesized compounds exhibited good anti-inflammatory activity compared with indomethacin. The synthesized compounds were also evaluated for their in vitro antimicrobial activity. Some of the compounds showed mild antibacterial activity while most of the compounds showed good antifungal activity.     

Thiazolidin-4-one and hydrazone derivatives of capric acid as possible anti-inflammatory,analgesic and hydrogen peroxide-scavenging agents

Starting from capric acid, hydrazone and thiazolidin-4-one derivatives have been synthesized in the present investigation. Decanoic acid hydrazide was reacted appropriately to yield hydrazones, which were then cyclized to yield the corresponding thiazolidin-4-ones. The structures of the newly synthesized compounds were confirmed by analytical and spectral methods. Anti-inflammatory, analgesic, and hydrogen peroxide-scavenging activity of the title compounds were evaluated. Among synthesized compounds, 2-hydroxyphenyl thiazolidinone with 44.90% inhibition of inflammation was the most potent anti-inflammatory agent. Similarly, 4-methoxybenzylidine hydrazide with 64.90% inhibition of writhing was observed to be the most potent analgesic agent of the synthesized compounds. All the synthesized compounds exhibited potent hydrogen peroxide-scavenging activity.     

Thiazolidin-4-one and hydrazone derivatives of capric acid as possible anti-inflammatory, analgesic and hydrogen peroxide-scavenging agents

Starting from capric acid, hydrazone and thiazolidin-4-one derivatives have been synthesized in the present investigation. Decanoic acid hydrazide was reacted appropriately to yield hydrazones, which were then cyclized to yield the corresponding thiazolidin-4-ones. The structures of the newly synthesized compounds were confirmed by analytical and spectral methods. Anti-inflammatory, analgesic, and hydrogen peroxide-scavenging activity of the title compounds were evaluated. Among synthesized compounds, 2-hydroxyphenyl thiazolidinone with 44.90% inhibition of inflammation was the most potent anti-inflammatory agent. Similarly, 4-methoxybenzylidine hydrazide with 64.90% inhibition of writhing was observed to be the most potent analgesic agent of the synthesized compounds. All the synthesized compounds exhibited potent hydrogen peroxide-scavenging activity.     

Synthesis of the metabolites of a free radical scavenger edaravone (MCI-186, Radicut)

Two metabolites of a free radical scavenger, edaravone, were synthesized. Edaravone glucuronate was synthesized by glycosylation of a glucuronic acid precursor using silver (I) trifluoromethane-sulfonate with edaravone. Edaravone sulfate was synthesized by sulfonylation of edaravone using a sulfur trioxide-pyridine complex. The two synthesized metabolites were identical to isolated metabolites. X-ray analysis identified edaravone glucuronate as beta-O-glucuronate, although there were three possible edaravone glucuronate tautomers.     

Classes of Proteins Synthesized in Oocytes, Eggs, Embryos, and Differentiated Tissues of Xenopus Zaevis

Two-dimensional gel electrophoresis has been used to analyse protein synthesis in embryonic stages and in three differentiated tissues of Xenopus laevis. The patterns found in oocyte, unfertilized eggs, embryos shortly after fertilization and at progressively later stages of development have been characterized and compared with the patterns found in the brain, heart and liver of tadpoles. The results suggest that at least four classes of proteins can be recognized among the proteins synthesized, although other categories may exist. They also suggest that some proteins synthesized rapidly in the oocyte are likely to be synthesized in differentiated tissues as well, while proteins synthesized for the first time only after fertilization are much less likely to be synthesized in differentiated tissues.     

Dual action spirobicycloimidazolidine-2,4-diones: Antidiabetic agents and inhibitors of aldose reductase-an enzyme involved in diabetic complications

The desired 3-(arylsulfonyl)spiroimidazolidine-2,4-diones were synthesized by reacting spiroiminoimidazolidine-2,4-dione with arylsulfonyl chlorides. Spiroimidazolidine-2,4-dione was in turn synthesized from norcamphor. Structures of the synthesized molecules were established by modern spectroscopic techniques. The synthesized compounds were screened for in vivo antidiabetic activity and aldose reductase inhibition. Compounds 2a, 2b and 2g exhibited excellent dual activity, compound 2a being most prominent. These results reveal that the synthesized compounds may serve as the molecule of choice to treat diabetes and diabetic complications using a single medication.     

Polyglucose particles histochemically synthesized by glycogen synthetase activity in the living chick retina

Summary Electron histochemical techniques for glycogen synthetase has been applied to the living retina of the chick and the polyglucose particles synthesized from UDPG in the paraboloid of the accessory cone were compared with those synthesized by the conventional histochemical techniques.In the retinal incubated in the medium for glycogen synthetase in vivo, synthesized polyglucose particles were located in the cytoplasmic matrices and most of the particles were less than 200 Å in diameter. These particles were rather well stainable with lead citrate and filled the cytoplasmic matrices. However, the tubular structures were not flattened, but slightly dilated. Compared with polyglucose particles synthesized in vitro by glycogen synthetase, those demonstrated by the in vivo histochemical techniques showed closer resemblance to native glycogen particles in size and stainability with lead citrate.The polyglucose particles synthesized from UDPG by glycogen synthetase were apparently different from those synthesized from glucose-1-phosphate by phosphorylase and branching glycosyltransferase.     

The response of foetal sheep to the somatic and flagellar antigens of Salmonella typhimurium.

Following the injection of polymeric flagellin (POL), foetal sheep older than 70 days gestation produced haemagglutinating antibody and synthesized IgM. The maximum titre of antibody in the blood increased with the age at which the foetus was injected. All foetuses synthesized 2-mercapto-ethanol-sensitive antibodies, while older foetuses (approximately 120 days gestation) also produced 2-mercaptoethanol-resistant antibodies and synthesized IgG1. During the primary immune response, there was a poor correlation between the antibody titre and the amount of immunoglobulin synthesized. The majority of IgM synthesized and almost all IgG1 had no demonstrable specificity for POL. During the secondary response to POL, the majority of IgG1 synthesized was specific and in one case appeared to be monoclona. There was no detectable primary antibody response in foetal sheep to the somatic antigens of Salmonella typhimurium, although all foetuses synthesized IgM. Only one of six foetuses receiving a second injection of antigen produced antibody. There was an increase in the numbers of blood lymphocytes following the injection of both POL and S. typhimurium, but only POL induced a rapid increase in the numbers of neutrophils in the blood and produced histological changes in the draining lymph nodes and spleen.     

Rapid biosynthesis of PVP coated silver nanoparticles by <Emphasis Type="Italic">Kocuria rosea</Emphasis> and their antimicrobial activity

The need for more effective antimicrobial agent and propitious application of nanotechnology in therapeutics and diagnostics has prompted the research on ecofriendly synthesis of silver nanoparticles. The objective of present study was to investigate the antibacterial and antifungal activity of biologically synthesized silver nanoparticles. The silver nanoparticles were synthesized by extracellular method, using soil bacteria Kocuria rosea. The synthesized silver nanoparticles were characterized by UV-Visible spectroscopy, X-ray diffractometry (XRD), transmission electron microscopy (TEM) and fourier transformation infrared spectroscopy (FTIR). On the basis of TEM analysis, the synthesized nanoparticles were found to be spherical with an average size of 30–50 nm. The biologically synthesized silver nanoparticles showed significant antimicrobial activity against pathogens.     

Molecular size distribution of proteoglycan subunits and glycosaminoglycans synthesized by chondrocytes under conditions of reduced proteoglycan synthesis

The rate of proteoglycan synthesis by chondrocytes in vitro was depressed by either omitting l-glutamine from the incubation medium or by addition of proteoglycan subunit to the medium. The molecular size distribution on Sepharose 2B of the proteoglycan subunits synthesized by the chondrocytes under these conditions of reduced proteoglycan synthesis was found to be the same as those synthesized by the control cells. Likewise, the molecular size distribution on Sepharose 6B CL of the glycosaminoglycan chains synthesized by the depressed cells was found to be similar to that observed in untreated chondrocytes. This work demonstrates that, under conditions of reduced proteoglycan synthesis, fewer proteoglycan subunits are synthesized by chondrocytes and that the molecular size distribution of these macromolecules is similar to those synthesized by untreated cells.     

Rheology of native dextrans in relation to their primary structure

High molecular weight dextrans were synthesized at five temperatures (3, 10, 20, 25 and 30°C) using an in-vitro enzymatic method. The rheological properties of these dextrans in aqueous solution were assessed through their flow behaviour and their viscoelastic characteristics. The results were interpreted in relation to their primary structure and particularly to their branching.

It was shown that the relatively expanded conformation of the dextrans synthesized at 3, 10 and 20°C gives to these dextrans comparable properties which are not too different from those described in literature for random-coil linear polysaccharides. Dextran synthesized at 30°C exhibited flow properties which are typical of particle suspensions in dilute and semi-dilute solution. In the concentrated domain, this dextran yielded structured systems with properties typical of weak gels. This unexpected behaviour could be related to the highly-ramified structure of this dextran in comparison with the dextrans synthesized between 3 and 20°C. On the other hand, the dextran synthesized at 25°C displayed rheological behaviour which could also be related to an intermediate primary structure between those of dextran synthesized at 20°C and dextran synthesized at 30°C.