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PubMed2024

Genetic and Microbial Analysis of Invasiveness for Escherichia coli Strains Associated With Inflammatory Bowel Disease.

BACKGROUND & AIMS: The adherent-invasive Escherichia coli (AIEC) pathotype is implicated in inflammatory bowel disease (IBD) pathogenesis. AIEC strains are currently defined by phenotypic measurement of their pathogenicity, including invasion of epithelial cells. This broad definition, combined with the genetic diversity of AIEC across patients with IBD, has complicated the identification of virulence determinants. We sought to quantify the invasion phenotype of clinical isolates from patients with IBD and identify the genetic basis for their invasion into epithelial cells. METHODS: A pangenome with core and accessory genes (genotype) was assembled using whole genome sequencing of 168 E coli samples isolated from 13 patients with IBD. A modified assay for invasion of epithelial cells (phenotype) was established with consideration of antibiotic resistance phenotypes. Isolate genotype was correlated to invasiveness phenotype to identify genetic factors that cosegregate with invasion. RESULTS: Pangenome-wide comparisons of E coli clinical isolates identified accessory genes that can cosegregate with invasion phenotype. These correlations found the acquisition of antibiotic resistance genes in clinical isolates compromised the traditional gentamicin protection assays used to quantify invasion. Therefore, an alternate assay, based on amikacin resistance, identified genes cosegregating with invasion. These genes encode an arylsulfatase, a glycoside hydrolase, and genetic islands carrying propanediol utilization and sulfoquinovose metabolism pathways. CONCLUSIONS: This study highlights the importance of incorporating antibiotic resistance screening for invasion assays used in AIEC identification. Accurately screened invasion phenotypes identified accessory genome elements among E coli IBD isolates that correlate with their ability to invade epithelial cells. These results help explain why single genetic markers for the AIEC phylotype are challenging to identify.

Humans↗
PubMed2006

Prostaglandin inhibitory and antioxidant components of Cistus laurifolius, a Turkish medicinal plant.

As Cistus laurifolius has been used traditionally to treat inflammatory and rheumatic disorders, its leaves were tested for prostaglandin (PG) inhibitory and antioxidant activities. The leaf extract showed both activities, i.e., inhibitory effect at 300 microg/ml on PGE1- and E2-induced contractions in guinea pig ileum and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effect. The separation guided by the activities shown by these dual assays provided sixteen compounds, 1-16. Known compounds 1-12 and 15 were identified as 3-O-methyl quercetin (1), 3,7-O-dimethyl quercetin (2), genkwanin (3), 3,7-O-dimethyl kaempferol (4), 3,4'-O-dimethyl quercetin (5), apigenin (6), 3,4'-O-dimethyl kaempferol (7), ellagic acid (8), beta-sitosterol-3-O-beta-glucoside (9), quercetin 3-O-alpha-rhamnoside (10), 5-O-p-coumaroyl quinic acid methyl ester (11), 1-(4-hydroxy-3-methoxyphenyl)-2-[4-(3-alpha-l-rhamnopyranoxypropyl)-2-methoxyphenoxy]-1,3-propanediol (12) and 2,3-dihydro-2-(4'-alpha-l-rhamnopyranosyloxy-3'-methoxyphenyl)-3-hydroxymethyl-7-methoxy-5-benzofuranpropanol (15). New lignan glycosides 13 and 14 were determined to be olivil 9-O-beta-D-xyloside and berchemol 9-O-rhamnoside, respectively. Compound 16 was isolated as a 2:1 mixture of two diastereomers, the major one of which was determined to be (7S,8R)-dihydrodehydrodiconiferyl alcohol 9'-O-alpha-L-rhamnoside. The structures were determined by detailed 2D NMR analysis together with NOEDF and CD. PG inhibitory effect was observed in 1, 5, 10, 12 and 16 at 30 microg/ml and antioxidant activity, in 1, 2, 8, 10, 12-14 and 16.

Alprostadil↗
PubMed2005

Molecular cloning and structural analysis of quinohemoprotein alcohol dehydrogenase ADH-IIG from Pseudomonas putida HK5.

Depending on the alcohols used as growth substrates, Pseudomonas putida HK5 produces two distinct quinohemoprotein alcohol dehydrogenases, ADH-IIB and ADH-IIG, both of which contain pyrroloquinoline quinone (PQQ) and heme c as the prosthetic groups but show different substrate specificities, especially for diol substrates. Molecular cloning of the gene of ADH-IIB and its crystal structure are already reported. Here, molecular cloning of the gene, qgdA, and solution of the three-dimensional structure of ADH-IIG are reported. The enzyme consists of 718 amino acid residues including a signal sequence of 29 amino acid residues. The PQQ domain is highly homologous to other quinoproteins, especially to quinohemoproteins. The crystal structure of ADH-IIG, determined at 2.2A resolution, shows that the overall structure and the amino acid residues involved in PQQ binding are quite similar to ADH-IIB and to another quinohemoprotein ADH, qhEDH from Comamonas testosteroni. However, the lengths of the linker regions connecting the PQQ and the cytochrome domains are different from each other, leading to a significant difference in orientation of the cytochrome domain with respect to the PQQ domain. Apart from ADH-IIB and qhEDH, ADH-IIG has an extra 12-residue helix within loop 3 in the PQQ domain and an extra 3(10) helix in the C terminus of the cytochrome domain, and both helices appear parallel and linked by a hydrogen bond. The amino acid residues contacting substrate/product in the crystal structures are also different among them. In the crystal structure of ADH-IIG with 1,2-propanediol, one of the hydroxyl groups of the substrate forms a hydrogen bond with O5 of PQQ and OD1 of Asp300, and the other interacts with a water molecule and with NE2 of Trp386, the corresponding residue of which is not found in ADH-IIB and qhEDH, and might be the residue responsible for making ADH-IIG prefer diol substrates.

Alcohol Oxidoreductases↗
PubMed2006

The structural and cellular viability in cryopreserved rabbit carotid arteries.

OBJECTIVE: We investigated the histological and mechanical changes in addition to viable cellular recovery in cryopreserved blood vessels. MATERIALS AND METHODS: Rabbit carotids were cryopreserved in a cryoprotective medium containing 1.5 M of 1,2-propanediol (PD) and then were thawed slowly in an ice bag that had been precooled in liquid nitrogen. Fresh carotids were used as the control. The fresh and freeze-thawed arteries were cultured for the growth of vascular smooth muscle cells (VSMCs). The freeze-thawed arterial tissues were perfused in vitro for 6, 12, or 24 h, respectively, to assess the integrity of carotid walls and the mechanical properties. RESULTS: The results showed that it took almost the same time (24 approximately 36 h) for the VSMCs of the PD-cryopreserved arteries to regenerate as those from the fresh arteries. Their growing speeds also were similar. On the contrary, Me2SO-cryopreserved (1.5 M) arteries were unable to regenerate VSMCs in culture. After freeze-thawing, the mechanical properties decreased significantly (P < 0.003 for elastic modulus and P < 0.001 for fracture strength). After in vitro perfusion of the freeze-thawed carotid arteries, all of the survived endothelial cells fell off, and some of the VSMCs denaturalized or necrosed. The internal elastic fibers and collagen showed various degrees of cracking. The mechanical properties were decreased (P < 0.05). CONCLUSION: Our findings demonstrate that the PD-containing cryoprotective medium can preserve regenerative capacity of VSMCs, which makes it a useful technique for viable VSMC recovery. However, the freeze-thawing process and the in vitro perfusion caused serious disruption in the arterial mechanical properties, rendering the cryopreserved blood vessels less useful for vessel reconstruction.

Animals↗
PubMed2004

3-monochloropropane-1,2-diol does not cause neurotoxicity in vitro or neurobehavioral deficits in rats.

3-monochloro-1,2-propanediol (3-MCPD) is a contaminant of acid-hydrolyzed vegetable protein. Several reports have suggested that chronic exposure to 3-MCPD could produce neurotoxicity in vitro or neurobehavioral effects in experimental animals. The present study further explored the in vitro neurotoxic effects of 0.1-100 microM 3-MCPD on PC12 and N18D3 cell lines. In addition, to investigate the effects of repeated ingestion of 3-MCPD on neurobehavioral impairments parameters in rats, motor activity, landing foot splay, and grip strength tests were performed, following treatment with 3-MCPD at dose levels of 10, 20, and 30 mg/kg per day for 11 weeks. We demonstrated that no significant neurotoxic effects were present in 3-MCPD-treated rats compared to saline-treated control rats, whereas, acrylamide, used as a positive control, induced significant deficits in all neurobehavioral parameters in both male and female rats. On the other hand, body weight gain was significantly decreased in high dose 3-MCPD-treated male rats as well as in acrylamide-treated rats. Taken together, these results suggest that 3-MCPD, at the dose levels used for this study, does not produce in vitro neurotoxicity or neuromotor deficits in vivo.

Animals↗
PubMed2005

Mechanism responsible for the salutary effects of flutamide on cardiac performance after trauma-hemorrhagic shock: Upregulation of cardiomyocyte estrogen receptors.

BACKGROUND: Although flutamide (FTM), an androgen-receptor antagonist, normalizes the depressed immune and cardiac function in males after trauma hemorrhage (T-H), the mechanism responsible for its salutary effects remains unknown. We hypothesized that the salutary effects of FTM are mediated via upregulation of estrogen receptors (ERs). METHODS: Male Sprague-Dawley rats underwent T-H (laparotomy and 90 minutes of hemorrhage (35-40 mm Hg) and then resuscitated with 4x the volume of shed blood in the form of Ringer's lactate). FTM (25 mg/kg) or vehicle (propanediol) was injected subcutaneously 30 minutes before the end of resuscitation. At 2 hours after T-H or sham operation, cardiac output, stroke volume, heart rate, mean arterial pressure, +/- dp/dt, and total peripheral resistance were measured (n = 6 rats per group). Immediately after the measurement of cardiac function, cardiomyocytes were isolated, RNA was extracted, and expression of ER-alpha, ER-beta, and androgen-receptor (AR) mRNA in cardiomyocytes was determined by quantitative real-time polymerase chain reaction. ER-alpha, ER-beta, and AR protein levels in cardiomyocytes were also measured by Western blot analysis. RESULTS: The depressed cardiac output, stroke volume, and +/- dp/dt after T-H were significantly improved in the FTM-treated T-H group. Moreover, the decrease in expression of ER-alpha and ER-beta mRNA and protein in cardiomyocytes in the T-H group was prevented with FTM treatment after T-H. However, expression of cardiomyocytes AR mRNA and protein were not significantly different between the T-H or sham group with or without FTM treatment. CONCLUSIONS: These findings collectively suggest that, in addition to blockade of androgen receptors, flutamide-mediated ER upregulation is likely to play a role in mediating the salutary effect of flutamide on cardiac function after trauma hemorrhage.

Androgen Antagonists↗
PubMed2004

Effect of cryoprotectants and their concentration on in vitro development of vitrified-warmed immature oocytes in buffalo (Bubalus bubalis).

Experiments were conducted to study the effect of cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), 1,2-propanediol (PROH), and glycerol at different concentrations (3.5, 4, 5, 6, and 7 M each with 0.5 M sucrose and 0.4% BSA in DPBS) on survival, in vitro maturation, in vitro fertilization, and post-fertilization development of vitrified-thawed immature buffalo oocytes. The COCs were harvested from the ovaries by aspirating the visible follicles. The recovery of post-thaw morphologically normal oocytes was lower in 3.5 and 4 M DMSO, EG, and PROH compared to 5, 6, and 7 M. In all the concentrations of glycerol, an overall lower numbers of oocytes recovered were normal compared to other cryoprotectants. Less number of oocytes reached metaphase-II (M-II) stage from the oocytes cryopreserved in any of the concentrations of DMSO, EG, PROH, and glycerol compared to fresh oocytes. Among the vitrified groups, highest maturation was obtained in 7 M solutions of all the cryoprotectants. The cleavage rates of oocytes vitrified in different concentrations of DMSO, EG, PROH, and glycerol were lower than that of the fresh oocytes. The cleavage rates were higher in oocytes cryopreserved in 6 and 7 M DMSO, EG, PROH, and glycerol compared with oocytes cryopreserved in other concentrations. However, the percentage of morula and blastocyst formation from the cleaved embryos did not vary in fresh oocytes and vitrified oocytes. In conclusion, this report describes the first successful production of buffalo blastocysts from immature oocytes cryopreserved by vitrification.

Animals↗
PubMed2004

Effect of different cryoprotectants on the structural preservation of follicles in frozen zebu bovine (Bos indicus) ovarian tissue.

Cryopreservation of ovarian tissue is a new and promising technique for germ-line storage. The objective of this study was to evaluate the effect of four cryoprotectants (at two concentrations each) on the preservation of zebu bovine preantral follicles after ovarian cryostorage. Strips of ovarian cortex were cryopreserved using glycerol (GLY; 10 or 20%), ethylene glycol (EG), propanediol (PROH) or dimethylsulphoxide (DMSO; 1.5 or 3M). In addition, a toxicity test was performed for each cryoprotectant by exposing the ovarian tissue to them without freezing. Tissues were analyzed by histology and transmission electron microscopy. Ovarian tissue frozen in either concentration of DMSO or PROH or in 10% GLY retained a higher percentage of morphologically normal follicles (73-88%) than tissue frozen in 20% GLY or in either concentration of EG (16-52%). In the toxicity test, exposure of tissues to DMSO, PROH or GLY resulted in higher percentages of normal follicles (80-97%) than exposure to EG (49%). Electron microscopy revealed damage to the ultrastructure of follicles frozen in 10% GLY, while follicles cryopreserved in DMSO and PROH at either concentration exhibited normal ultrastructure. In conclusion, DMSO and PROH were the most effective cryoprotectants for zebu ovarian tissue, preserving the structural integrity of somatic and reproductive cells within the ovary.

Animals↗
PubMed2005

Effects of removal of necrotic blastomeres from mouse cryopreserved embryos on blastocyst formation and hatching.

To evaluate whether the developmental potential of embryos that were partially damaged after freezing and thawing can be improved by removal of necrotic blastomeres. Eight-cell mouse embryos were cryopreserved using 1,2-propanediol and sucrose as cryoprotectant with slow cooling procedure. After thawing, blastocyst formation and hatching of fully intact embryos were compared between no treatment and with laser-assisted hatching. For partially intact embryos, the effects of removal of necrotic blastomeres with micromanipulation were evaluated. Laser-assisted hatching of mouse cryopreserved fully intact embryos significantly increased blastocyst hatching (63.4% versus 48.3%, P<0.05), but had little effect on blastocyst formation (72.0% versus 70.1%, P>0.05). The removal of necrotic blastomeres from partially damaged mouse cryopreserved embryos with micromanipulation significantly increased blastocyst formation (52.9% versus 32.0%, P<0.05) and blastocyst hatching (41.2% versus 22.0%, P>0.05) compared with the control group. The developmental potential of partially damaged cryopreserved embryos can be improved by removal of necrotic blastomeres with micromanipulation.

Animals↗
PubMed2006

Analysis of oocyte physiology to improve cryopreservation procedures.

In contrast to the preimplantation mammalian embryo, it has been notoriously difficult to cryopreserve the metaphase II oocyte. The ability to store oocytes successfully at -196 degrees C has numerous practical and financial advantages, together with ethical considerations, and will positively impact animal breeding programs and assisted conception in the human. Differences in membrane permeability and in physiology are two main reasons why successful oocyte cryopreservation has remained elusive. It is proposed, therefore, that rather than relying on technologies already established for the preimplantation embryo, the development of cryopreservation techniques suitable for the mammalian oocyte needs to take into account the idiosyncratic physiology of this cell. Analysis of intracellular calcium, for example, has revealed that exposure to conventional permeating cryoprotectants, such as propanediol, ethylene glycol and DMSO, all independently result in an increase in calcium, which in turn has the potential to initiate oocyte activation, culminating in zona hardening. Quantification of the metabolome and proteome of the oocyte has revealed that whereas slow freezing has a dramatic effect on cell physiology, vitrification appears to have limited effect. This is plausibly achieved by the limited exposure to cryoprotectants. Analysis of meiotic spindle dynamics and embryo development following IVF, also indicate that vitrification is less traumatic than slow freezing, and therefore has the greatest potential for successful oocyte cryopreservation.

Animals↗
PubMed2005

Immunotoxic effect of beta-chlorolactic acid on murine splenocyte and peritoneal macrophage function in vitro.

Beta-chlorolactic acid is a major intermediate of 3-monochloro-1,2-propanediol (MCPD) in mammalian species, which a well-known by-product of acid-hydrolyzed soy sauce during its manufacturing process. beta-Chlorolactic acid has not been studied on immunotoxicity. To evaluate the immunomodulatory effect of beta-chlorolactic acid on murine splenocyte and macrophage in vitro, we investigated splenocyte blastogenesis by concanavalin A (Con A), anti-CD3 and lipopolysaccharide (LPS), the production of cytokines from splenocyte, and the activity of mouse peritoneal macrophages. beta-Chlorolactic acid suppressed significantly splenic blastogenesis to Con A or anti-CD3 from 8.5 to 54.7% at doses comprised between 200 and 800 microM. beta-Chlorolactic acid also suppressed significantly splenic blastogeneis to LPS from 8.5 to 71.5% at doses comprised between 200 and 800 microM. The production level of interferon (IFN)-g on splenocyte culture with Con A was significantly reduced from 21.5 to 51.4% at the higher concentration than 100 microM of beta-chlorolactic acid. The levels of interleukin (IL)-2 and IL-4 were also decreased 22.6-58.4 and 10.2-36.6%, respectively, at high concentrations of beta-chlorolactic acid. There was a significant decrease from 6.1 to 40.8% in the production of nitric oxide (NO) by peritoneal macrophages treated with 400-1000 microuM beta-chlorolactic acid. These results indicate that beta-chlorolactic acid might be able to induce immunotoxic effect on immune response of lymphocytes and peritoneal macrophages in vitro.

Animals↗
PubMed2005

Endocytosis and intracellular trafficking of fatty acid esters of phenylaminopropanediol, the putative etiologic agents of the toxic oil syndrome (TOS).

The toxic oil syndrome (TOS) caused by ingestion of rapeseed oil adulterated with aniline is characterized by symptoms of an allergic and/or autoimmune illness associated with vessel wall lesions similar to those of atherosclerosis. Fatty acid esters of 3-(N-phenylamino)-1,2-propanediol (PAP) have been incriminated as the etiologic agents of TOS. However, the pathogenesis of TOS is yet unknown. Here, we addressed whether PAP fatty acid esters are incorporated into lipoproteins, which after transport to vascular endothelial cells are taken up to initiate TOS vasculopathy. After loading (14)C-dioleyl-ester of PAP into LDL labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindolcarbocyanine (DiI) we assessed receptor mediated endocytosis and intracellular localization of these lipopoproteins in vascular endothelial cells. Our data suggest that these lipoprotein-derivatives are internalized into endothelial cells by LDL receptor mediated endocytosis. Confocal microscopy revealed that DiI-LDL loaded with dioleyl-ester of PAP and incubated for 60 min with endothelial cells colocalizes with the lysosomotropic compound LysoTracker Green, indicating that internalized PAP-loaded LDL are targetted to the endolysosomal compartment for further processing. Subcellular fractionation of endothelial-like ECV-304 cells after incubation with LDL loaded with the (14)C-dioleyl-ester of PAP for 6h showed that the radioactive label accumulated in fractions containing endosomes, the Golgi apparatus and the endoplasmic reticulum.

Animals↗
PubMed1992

Identification of type 1 and type 2 light-harvesting chlorophyll a/b-binding proteins using monospecific antibodies.

The amino acid sequences of more than 40 apoproteins of the light-harvesting complex associated with Photosystem II (LHC II) of various plants have been deduced by sequencing their corresponding genes. These highly conserved sequences fall into two major categories, type 1 and type 2, that differ mainly in a small number of domains close to the N-terminus. We have made polyclonal, monospecific antibodies against synthetic peptides corresponding to the most unique sequence domains of the N-terminal regions of type 1 and type 2 LHC II apoproteins, using sequences derived from petunia genes. On Western blots our anti-type 1 and 2 antibodies crossreact with light-harvesting proteins of petunia, tomato, spinach and several other plants. By using a new gel-system based on ammediol (2-amino-2-methyl-1,3-propanediol), we are able to resolve up to eight LHC II apoproteins. On petunia, tomato and spinach blots the anti type 1 antibodies bind to two or more of the higher molecular weight LHC II polypeptides, whereas the anti type 2 antibodies recognize very specifically only one or two of the lower molecular weight LHC-proteins. In all plants studied, the type 1 LHC II apoproteins are more numerous and span a greater size range than the type 2 apoproteins. This is consistent with the smaller number of type 2 LHC II CAB genes that have been discovered to date.

Amino Acid Sequence↗
PubMed1984

Inhibition of peptidyltransferase and possible mode of action of a dipeptidyl chloramphenicol analog.

A dipeptidyl chloramphenicol analog, D-threo-2-(L-phenylalanylglycyl)amino-3-p-nitrophenyl-1,3- propanediol, has been prepared and examined as an inhibitor of ribosomal peptidyltransferase. The analog is a more effective inhibitor of poly (U,C) directed protein biosynthesis in an Escherichia coli cell-free system than chloramphenicol and shows inhibitory activity equal to the parent antibiotic in the transpeptidation reaction. These results and the common structural features of puromycin and this compound suggest a model for the binding modes of chloramphenicol and chloramphenicol analogs. This proposal invokes four major binding pockets at the A-site of the peptidyltransferase center.

Acyltransferases↗
PubMed1998

Stimulation of biosynthetic activity by novel succinate esters in rat pancreatic islets.

Selected esters of succinic acid are currently under investigation as possible insulinotropic agents for the treatment of noninsulin-dependent diabetes mellitus. The aim of the present study was to investigate the effects of ten novel esters of succinic acid upon biosynthetic activity in rat pancreatic islets. In the absence of any other exogenous nutrient, glycerol-3-hydroxy-1,2-dimethyl succinate (0.5 mM), D-arabitol-5-hydroxy-1,2,3,4-tetramethylsuccinate (0.5 mM), and 4-tert-butylsuccinate (2.5 mM) exerted little or no effect upon L-[4-3H]phenylalanine incorporation into trichloroacetic acid-precipitable material. A modest but significant increase in biosynthetic activity to approximately 150% of basal value was found in the presence of L-threitol-1,2,4-trimethylsuccinate (2.0 mM) and ethanediol-1,2-diethylsuccinate (2.5 mM). A two- to five-fold increase in protein biosynthesis was observed in islets exposed to propanediol-1,2-dimethylsuccinate, glycerol-1,2-dimethylsuccinate-3-hydrogenosuccinate, L-threitol-3-succinoyl-1,2,4-trimethylsuccinate, glycerol-1,2-dimethylsuccinate or ethanediol-1,2-dimethylsuccinate (2.5 mM each), these esters being mentioned in order of increasing biological efficiency. There was a significant correlation between these results and the insulinotropic action of the same esters. The present findings thus reinforce the view that such esters act as nutrients in islet cells and, therefore, offer the advantage over pharmacological agents currently used for the treatment of type-2 diabetes in stimulating both the biosynthetic and secretory activity of insulin-producing B-cells.

Animals↗
PubMed1975

Ethanolamine ammonia-lyase: inactivation of the holoenzyme by N2O and the mechanism of action of Coenzyme B12.

Functional ethanolamine ammonia-lyase is inactivated by N2O as well as by O2, indicating that the active form of coenzyme B12 is an enzyme-bound corrin derivative in which the Co-C bond of the coenzyme is broken and the cobalt ion is in the +1 state of oxidation. The nucleoside fragment formed in the process of coenzyme activation is tentatively identified as 4',5'-didehydro-5'-deoxyadenosine. A mechanism of action of ethanolamine ammonia-lyase is formulated in analogy to that of DL-1,2-Propanediol dehydrase and compared to proposed alternative reaction schemes.

Ammonia-Lyases↗
PubMed1998

Structure of the acidic polysaccharide chain of the lipopolysaccharide of Shewanella alga 48055.

A lipopolysaccharide (LPS) with an acidic polysaccharide chain was isolated from the bacterium Shewanella alga strain 48055 and cleaved selectively at the glycosidic linkage of N-acetylneuraminic acid to give a tetrasaccharide. Studies of the tetrasaccharide and the O-deacylated LPS by 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, rotating-frame NOE spectroscopy (ROESY), and H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC) experiments, revealed the following structure of the polysaccharide repeating unit: -->3)-beta-D-GalpA6GroN-(1-->3)-beta-D-GlcpNAc-(1-->3)-alpha-D- GalpA6GroN- (1-->4)-alpha-Neup5Ac-(2--> where GroN is an amidically linked residue of 2-amino-1,3-propanediol (2-amino-2-deoxyglycerol). A similar structure, but with 2-acetamido-2,6-dideoxy-D-glucose instead of 2-acetamido-2-deoxy-D-glucose, has been reported previously for the polysaccharide chain of a non-O1 Vibrio cholerae H11 LPS [E. V. Vinogradov, O. Holst, J.E. Thomas-Oates, K.W. Broady, and H. Brade, Eur. J. Biochem., 210 (1992) 491-498].

Carbohydrate Conformation↗
PubMed1980

Utilization of a glycol-stabilized liquid NAD for the measurement of three enzymes on the GEMSAEC.

Some analytical parameters have been investigated for a recently described stabilized liquid coenzyme technology in which water-free NAD is dissolved in 1,2 propanediol. Correlation for 108 specimens assayed for AST, ALT and LD with a reference method in which glycol-based NAD was absent was greater than or equal to 0.998 with near identical reproducibility over a period of at least 107 days. Mean recovery of exogenous serum enzymes in this linear kinetic assay is 103%. With the option of mixing only the volume of reagent needed for the enzymatic assay, waste can be eliminated as compared to more costly preparations stabilized by lyophilization. Hazards from an impure water supply are avoided since no reconstituting volume is required.

Alanine Transaminase↗