scholarly journals Polar Flagella Glycosylation in Aeromonas: Genomic Characterization and Involvement of a Specific Glycosyltransferase (Fgi-1) in Heterogeneous Flagella Glycosylation

2021 ◽  
Vol 11 ◽  
Author(s):  
Gabriel Forn-Cuní ◽  
Kelly M. Fulton ◽  
Jeffrey C. Smith ◽  
Susan M. Twine ◽  
Elena Mendoza-Barberà ◽  
...  

Polar flagella from mesophilic Aeromonas strains have previously been shown to be modified with a range of glycans. Mass spectrometry studies of purified polar flagellins suggested the glycan typically includes a putative pseudaminic acid like derivative; while some strains are modified with this single monosaccharide, others modified with a heterologous glycan. In the current study, we demonstrate that genes involved in polar flagella glycosylation are clustered in highly polymorphic genomic islands flanked by pseudaminic acid biosynthetic genes (pse). Bioinformatic analysis of mesophilic Aeromonas genomes identified three types of polar flagella glycosylation islands (FGIs), denoted Group I, II and III. FGI Groups I and III are small genomic islands present in Aeromonas strains with flagellins modified with a single monosaccharide pseudaminic acid derivative. Group II were large genomic islands, present in strains found to modify polar flagellins with heterogeneous glycan moieties. Group II, in addition to pse genes, contained numerous glycosyltransferases and other biosynthetic enzymes. All Group II strains shared a common glycosyltransferase downstream of luxC that we named flagella glycosylation island 1, fgi-1, in A. piscicola AH-3. We demonstrate that Fgi-1 transfers the first sugar of the heterogeneous glycan to the pseudaminic acid derivative linked to polar flagellins and could be used as marker for polysaccharidic glycosylation of Aeromonas polar flagella.

1999 ◽  
Vol 45 (11) ◽  
pp. 959-969 ◽  
Author(s):  
Eric Kurt Hani ◽  
David Ng ◽  
Voon-Loong Chan

Arginine biosynthetic genes from Campylobacter jejuni TGH9011 were cloned by functional complementation of the respective Escherichia coli arginine biosynthetic mutants. Complementation of argA, argB, argC, argD, argE, argF, and argH auxotrophs was accomplished using a pBR322-based C. jejuni TGH9011 plasmid library. By cross-complementation analyses, the first four steps of arginine biosynthesis were shown to be closely linked on the genome. Two additional clones complementing the first (ArgA) and fifth (ArgE) steps in arginine biosynthesis were obtained. Neither recombinant showed linkage to the arg cluster, to each other, nor to other arginine biosynthetic functions by cross-complementation. Genes argF and argH were not linked to other arginine biosynthetic genes by cross-complementation analysis. Restriction enzyme patterns of recombinant plasmids fell into five groups. Group I contained the arg(ABCD) complementing locus. Group II and Group III were the two genetic loci corresponding to the argA and argE complementing genes. Group II contains the hipO gene encoding N-benzoylglycine-amino-acid amidohydrolase, also known as hippurate hydrolase. Group III contains the hipO homolog of C. jejuni. Group IV represents the argF gene. GroupV is the argH gene. Functional complementation of mutations in the first four steps of the arginine biosynthetic pathway was obtained on recombinant plasmid pARGC2. The predicted order of gene complementation was argCargA(argBargD). The sequence of the insert in plasmid pARGC2 revealed direct homologs for argC, argB, and argD. However, sequence analysis of the gene complementing ArgA function in two separate E. coli argA mutants determined that the C. jejuni gene was not a canonical argA gene. The gene complementing the argA defect, which we call argO, showed limited homology to the streptothricin acetyltransferase gene (sat) of Escherichia coli. The flanking open reading frames in pARGC2 showed no homologies to arginine biosynthetic genes. The structure of the argCOBD gene arrangement is discussed with reference to the presence and location of other arginine biosynthetic genes on the genome of C. jejuni and other bacterial organisms.Key words: arginine synthesis, Campylobacter jejuni, arginine biosynthetic genes, gene sequence, gene arrangement.


Author(s):  
M Ashraf-Khorassani ◽  
N Nazem ◽  
LT Taylor ◽  
WM Coleman

AbstractIsolation, fractionation, and identification of sucrose esters from aged oriental tobacco employing supercritical fluids have been completed. Underivatized sucrose ester-rich extracts were obtained using supercritical CO2 at densities greater than 0.73 g/mL. Lower density CO2 provided extracts with notable amounts of tobacco derived material; yet, no detectable sucrose ester content. Preparative supercritical fluid chromatography (SFC) provided for an additional purification of the sucrose ester-enriched fraction after column optimization. Structural assignments of the SFC fractions were facilitated using gas chromatography/mass spectrometry (GC/MS) accompanied by N, O-bis(trimethylsilyl)trifluoroacetamide-dimethylformamide (BSTFA-DMF) derivatization of the free hydroxyl groups and high performance-liquid chromatography/mass spectrometry (HPLC/MS). From a relative quantitative perspective regardless of tobacco type, sucrose esters having an acetyl group on C6 of the glucose function (Group III) were in higher concentration compared to both the concentration observed for sucrose ester of Group I (acetyl group on C3 of fructose) and sucrose ester of Group II (no acetyl group on either glucose or fructose). Saturated fatty acid constituents were found to range from a maximum total of 18 carbons to a minimum total of 13 carbons. Unsaturated and isomeric fatty acid homologues were detected within the Group II sucrose ester.


2019 ◽  
Vol 21 (2) ◽  
pp. 341-350 ◽  
Author(s):  
N. B. Zakharova ◽  
L. Kh. Pastushkova ◽  
R. V. Lakh ◽  
I. M. Larina ◽  
E. N. Nikolaev ◽  
...  

We performed clinical observations and laboratory examination of 22 patients with chronic pyelonephritis (chronic renal failure, CRF) and 30 healthy individuals. The patients with CRF were examined twice. The first group (Group I) included patients with exacerbation of the disease. The comparison series (Group II) was represented by the same patients who were examined 1.5-3 months after completion of treatment, without clinical exacerbation of chronic pyelonephritis (CPN). Laboratory signs of acute renal damage were not detectable in all the patients examined. Concentrations of VEGF, MCP-1, IL-8 and IL-18 were determined in urine samples of all examined persons by ELISA technique. Protein spectrum of urine was assessed in six patients from Group I, and in six cases of Group II by means of mass spectrometry, using Agilent 1100 chromatographic device, and LTQ-FT Ultra hybrid mass spectrometer. The results of parallel determination of urine proteins by the two methods have shown that the evolving CPN exacerbation is associated with local secondary immune deficiency at the level of renal tubular urothelium. Determination of urine proteome by means of mass spectrometry in exacerbating disease allows identify the proteins associated with damage to epithelial lining of renal tubules and development of local immune response.


Author(s):  
K.K. SEKHRI ◽  
C.S. ALEXANDER ◽  
H.T. NAGASAWA

C57BL male mice (Jackson Lab., Bar Harbor, Maine) weighing about 18 gms were randomly divided into three groups: group I was fed sweetened liquid alcohol diet (modified Schenkl) in which 36% of the calories were derived from alcohol; group II was maintained on a similar diet but alcohol was isocalorically substituted by sucrose; group III was fed regular mouse chow ad lib for five months. Liver and heart tissues were fixed in 2.5% cacodylate buffered glutaraldehyde, post-fixed in 2% osmium tetroxide and embedded in Epon-araldite.


1998 ◽  
Vol 80 (09) ◽  
pp. 393-398 ◽  
Author(s):  
V. Regnault ◽  
E. Hachulla ◽  
L. Darnige ◽  
B. Roussel ◽  
J. C. Bensa ◽  
...  

SummaryMost anticardiolipin antibodies (ACA) associated with antiphospholipid syndrome (APS) are directed against epitopes expressed on β2-glycoprotein I (β2GPI). Despite a good correlation between standard ACA assays and those using purified human β2GPI as the sole antigen, some sera from APS patients only react in the latter. This is indicative of heterogeneity in anti-β2GPI antibodies. To characterize their reactivity profiles, human and bovine β2GPI were immobilized on γ-irradiated plates (β2GPI-ELISA), plain polystyrene precoated with increasing cardiolipin concentrations (CL/β2GPI-ELISA), and affinity columns. Fluid-phase inhibition experiments were also carried out with both proteins. Of 56 selected sera, restricted recognition of bovine or human β2GPI occurred respectively in 10/29 IgA-positive and 9/22 IgM-positive samples, and most of the latter (8/9) were missed by the standard ACA assay, as expected from a previous study. Based on species specificity and ACA results, IgG-positive samples (53/56) were categorized into three groups: antibodies reactive to bovine β2GPI only (group I) or to bovine and human β2GPI, group II being ACA-negative, and group III being ACA-positive. The most important group, group III (n = 33) was characterized by (i) binding when β2GPI was immobilized on γ-irradiated polystyrene or cardiolipin at sufficient concentration (regardless of β2GPI density, as assessed using 125I-β2GPI); (ii) and low avidity binding to fluid-phase β2GPI (Kd in the range 10–5 M). In contrast, all six group II samples showed (i) ability to bind human and bovine β2GPI immobilized on non-irradiated plates; (ii) concentration-dependent blockade of binding by cardiolipin, suggesting epitope location in the vicinity of the phospholipid binding site on native β2GPI; (iii) and relative avidities approximately 100-fold higher than in group III. Group I patients were heterogeneous with respect to CL/β2GPI-ELISA and ACA results (6/14 scored negative), possibly reflecting antibody differences in terms of avidity and epitope specificity. Affinity fractionation of 23 sera showed the existence, in individual patients, of various combinations of antibody subsets solely reactive to human or bovine β2GPI, together with cross-species reactive subsets present in all samples with dual reactivity namely groups III and II, although the latter antibodies were poorly purified on either column. Therefore, the mode of presentation of β2GPI greatly influences its recognition by anti-β2GPI antibodies with marked inter-individual heterogeneity, in relation to ACA quantitation and, possibly, disease presentation and pathogenesis.


Phlebologie ◽  
2003 ◽  
Vol 32 (05) ◽  
pp. 115-120 ◽  
Author(s):  
A. Franek ◽  
H. Koziolek ◽  
M. Kucharzewski

SummaryAim: The study of the influence of sulodexide in the treatment of venous leg ulcers. Patients and method: 44 patients with chronic venous ulceration were randomly divided into two groups. Group I: 21 patients (ulceration area: 12.7-18.9 cm2), Group II: 23 patients (ulceration size: 12.1-20.3 cm2). Both groups were treated by using Unna’s boot. This dressing was changed every seven days until the ulcer had healed. Additionally, the patients in group II received the systemic pharmacological treatment with sulodexide. Results: After 7 weeks of treatment ulcers of seven patients (35%) from group I had healed, and 3 weeks later the ulceration of two more patients had healed completely. After further 7 weeks the ulcers of 12 patients had healed completely. Whereas in group II after 7 weeks of treatment ulceration of 16 (70%, p <0.05) patient had healed completely and after further 3 weeks the ulcers of the remaining 7 patients had healed, too. Conclusion: The use of sulodexide in patients with chronic venous leg ulcers accelerates the healing process.


1997 ◽  
Vol 36 (08) ◽  
pp. 259-264
Author(s):  
N. Topuzović

Summary Aim: The purpose of this study was to investigate the changes in blood activity during rest, exercise and recovery, and to assess its influence on left ventricular (LV) volume determination using the count-based method requiring blood sampling. Methods: Forty-four patients underwent rest-stress radionuclide ventriculography; Tc-99m-human serum albumin was used in 13 patients (Group I), red blood cells was labeled using Tc-99m in 17 patients (Group II) in vivo, and in 14 patients (Group III) by modified in vivo/in vitro method. LV volumes were determined by a count-based method using corrected count rate in blood samples obtained during rest, peak exercise and after recovery. Results: In group I at stress, the blood activity decreased by 12.6 ± 5.4%, p <0.05, as compared to the rest level, and increased by 25.1 ± 6.4%, p <0.001, and 12.8 ± 4.5%, p <0.05, above the resting level in group II and III, respectively. This had profound effects on LV volume determinations if only one rest blood aliquot was used: during exercise, the LV volumes significantly decreased by 22.1 ± 9.6%, p <0.05, in group I, whereas in groups II and III it was significantly overestimated by 32.1 ± 10.3%, p <0.001, and 10.7 ± 6.4%, p <0.05, respectively. The changes in blood activity between stress and recovery were not significantly different for any of the groups. Conclusion: The use of only a single blood sample as volume aliquot at rest in rest-stress studies leads to erroneous estimation of cardiac volumes due to significant changes in blood radioactivity during exercise and recovery.


1994 ◽  
Vol 72 (05) ◽  
pp. 745-749 ◽  
Author(s):  
Elza Chignier ◽  
Maud Parise ◽  
Lilian McGregor ◽  
Caroline Delabre ◽  
Sylvie Faucompret ◽  
...  

SummaryP-selectin, also known as CD62P, GMP140 or PADGEM, is present in platelet a-granules and endothelial cell Weibel-Palade bodies and is very rapidly expressed on the surface of these cells on activation. In this study, an anti P-selectin monoclonal antibody (LYP20) was used, in tandem with flow cytometry, to identify activated platelets at the site of induced vascular trauma or in peripheral blood. Moreover, electron microscopy was performed to characterize sites of vascular trauma and quantify the number of adhering platelets. The same induced vascular trauma was observed to result into animals responding in 2 different ways (Group I, Group II) following the degree of platelet activation. Five rats, out of 14 with induced vascular trauma, had more than half of their circulating platelets expressing P-selectin when drawn at the site of the trauma (67.4% ± 3.44) or in peripheral blood (78.5% ± 2.5) (Group I). In the remaining 9 animals a much smaller proportion of circulating platelets expressed P-selectin when assayed from trauma sites (18% ± 3.34) or in peripheral blood (18.0% ± 4.30) (Group II). Enhanced P-selectin expression by circulating platelets in Group I, compared to Group II, appears to be linked to the degree of activated platelets adhering at sites of trauma (171 ± 15 × 103 platelets versus 48 ± 31 × 103 platelets per mm2). In the 5 control animals, that were not operated on, platelets expressing P-selectin when drawn at the site of a mock trauma (7.0% ± 1.84) or in the peripheral blood (11.2% ± 3.30) showed little activation. In addition, no platelet adhesion was seen on the vascular bed of these animals. Results from this study show that analysis of P-selectin (CD62P) expression, in circulating platelets, is a valuable and rapid marker of platelet activation following severe vascular trauma induced in rats. However, activated platelets were not detected to the same extent in the peripheral blood of all animals having undergone vascular trauma. It is conceivable that platelets, depending on the degree of activation, may be actively sequestered in organs and prevented from circulating. Alternatively, P-selectin may be rapidly endocytosed, or not expressed, by activated circulating platelets depending on the type of agonists implicated in vivo activation.


2012 ◽  
Vol 5 (1) ◽  
pp. 37-43
Author(s):  
ABMM Alam ◽  
M Moniruzzaman ◽  
MB Alam ◽  
N Islam ◽  
F Khatoon ◽  
...  

Background: CIN has gained increased attention in the clinical setting, particularly during cardiac intervention but also in many other radiological procedures in which iodinated contrast media are used. There is at present good clinical evidence from well-controlled randomized studies that CIN is a common cause of acute renal dysfunction.Methodology: This was a prospective study conducted among the patients who underwent coronary angiography and percutaneous coronary intervention in the Department of Cardiology, Dhaka Medical College Hospital during January 2010 to December 2010. A total of 111 patients age range from 25 to 75 years were included in the study. Serum creatinine level at baseline and at the end of 48 hours was done in all these patients. Study population was divided into two groups according to development of acute kidney injury (AKI). Group-I = AKI, Group II = Not developed AKI. Results: AKI developed 11.7% of the study patient. DM and Preexisting renal insufficiency were significantly higher in group I patients. HTN was (61.5% Vs 44.9%) higher in group I but not significantly. History of ACE inhibitor/ARB, NSAID intake and LVEF <40% were significantly higher in group I patients. The mean±SD volume of CM (Contrast Media) were 156.9±44.8 ml and 115.4±30.0 ml in group I and group II respectively, which was significant. The mean±SD of serum creatinine after 48-72 hours of CAG/PCI was 1.4±0.37 mg/dl and 1.1±0.2 mg/dl in group I and group II respectively. The serum creatinine level increased significantly (p<0.05) after 48-72 hours of CAG/PCI in group I. In group II, S. creatinine level increased but not significant (p>0.05). Impaired renal function was found 76.9% and 2.0% in group I and group II respectively. DM, HTN, preexisting renal insufficiency, ACE inhibitor/ARB, NSAIDs, contrast volume (>150 ml), eGFR (<60 ml/min/ 1.73m2) and LVEF (<40%) are significantly (p0.05) associated for CIN development.Conclusion: CIN is an iatrogenic but preventable disorder results from the administration of contract media. Although rare in the general population, CIN occurs frequently in patients with underlying renal dysfunction and diabetes. In patients with pre angiographic normal renal function, the prevalence is low but in pre-existing renal impairment it may pose a serious threat. Thus risk factors are synergistic in their ability to predispose to the development of CIN. A careful risk-benefit analysis must always be performed prior to the administration of contrast media to patients at risk for CIN. DOI: http://dx.doi.org/10.3329/cardio.v5i1.12227 Cardiovasc. j. 2012; 5(1): 37-43


1980 ◽  
Vol 94 (3) ◽  
pp. 315-320 ◽  
Author(s):  
D. Schams ◽  
E. Stephan ◽  
R. D. Hooley

Abstract. Six Holstein bulls were housed in a climate-chamber under constant light regimen and after two weeks of preconditioning at 15°C, 60% relative humidity RH (day) and 12°C, 60% RH (night) were subjected to two weeks of heat exposure. This involved one week at 30°C and 60% RH (day) and 25°C and 60% RH (night) and a further week at 35°C, 60% RH (day) and 30°C, 60% RH (night). Three bulls were untreated (group I) and 3 bulls were treated (group II) just before and during heat exposure with a prolactin inhibitor to study the possible physiological role of prolactin on the regulation of water, potassium and sodium. Serum prolactin levels increased significantly (P < 0.01) in group I from the control value of 6 ng/ml to 33 and 44 ng/ml when the ambient temperature was increased (weeks 3 and 4) and then decreased to 21 and 12 ng/ml after reduction in temperature during weeks 5 and 6, respectively. For group II prolactin values decreased under the treatment with the prolactin inhibitor to 0.5 ng/ml and remained at this level throughout the experiment. GH levels were unaffected by heat treatment or by treatment with prolactin inhibitor. There were no differences between groups I and II in respiratory rate, pulse rate and rectal temperature. Water intake increased in both groups under heat exposure but decreased significantly afterwards only in group II. Differences in urinary excretion volume and blood serum osmolality were not significant. Urinary potassium and sodium excretion were unchanged in group II but increased with heat exposure in group I. During heat exposure 2 bulls of group II lost weight despite maintaining food intake.


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