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PubMed1992

Strain-dependent restricted VH and VL usage by anti-bacterial levan monoclonal antibodies.

The immune response to polysaccharides is highly regulated and has several distinguishing features, including restricted clonotype and isotype expression. The basis for this highly restricted response is not fully understood. To address these questions in a systematic manner, we have generated a panel of 102 mAb from CBA/CaHN (CBA/Ca) and BALB/cAnN (BALB/c) mice after one and two injections of bacterial levan (BL), a beta(2----6)-linked polyfructosan with beta(2----1)-linked fructose branch points (inulin determinant, In). This panel of mAb was examined for isotype, fine specificity, VH and VL region gene family usage and relationships between these parameters. After one or two injections of BL in both strains, mAb were IgM and IgG3. Fine specificity and VH/VL gene family usage differed markedly, however, between the two strains. Only 4% (2/51) of CBA/Ca mAb recognized the In determinant, whereas 77% (40/51) of BALB/c mAb recognized this epitope. In both strains, VH usage was restricted and certain families were overrepresented. In CBA/Ca mice, the overall response to BL was dominated by VHJ558 (45%, 23/51), the largest VH family, but VH36-60 (27%, 14/51) and VHJ606 (25%, 13/51) were also highly utilized and overrepresented. In BALB/c mice, the overall response to BL was dominated by VHJ606 (79%, 39/49 designated), a relatively small VH family. More importantly, after a single immunization with BL one particular VH/VL pair (VHJ606/ kappa 11) was used by 88% (36/41) of BALB/c mAb and was associated with In reactivity. In summary, BALB/c and CBA/Ca responses to BL differ in fine specificity and VH/VL usage.

Animals↗
PubMed1992

Cyclosporine A treatment and evaluation of glomerular filtration rate in patients with a transplanted kidney.

According to some findings [Ross et al. 1987], the plasma concentration of creatinine (Pcr) is an inaccurate reflection of the glomerular filtration rate (GFR) in renal graft recipients with stabilized renal function, treated with cyclosporine A (CyA). In this study, we sought to determine whether the inaccuracy of GFR assessment on the basis of Pcr or creatinine clearance (Ccr) in these individuals is greater than in patients suffering from various chronic renal diseases untreated by any drugs during the examination. The study was performed in 30 renal graft recipients, treated with CyA in combination with azathioprine and prednisone. Further, 51 patients suffering from a chronic renal disease, mostly chronic glomerulonephritis or tubulointerstitial nephritis, were investigated. GFR was evaluated on the basis of polyfructosan clearance (CPF). A significant linear relation between Ccr and CPF (r = 0.829, p less than 0.001) was demonstrated in individuals with a transplanted kidney graft treated with CyA. A relationship of the same character was observed in the group of patients suffering from chronic renal diseases (r = 0.935, p less than 0.001). There is no statistically significant difference between the regression lines characterizing these relationships in both groups. A significant correlation of hyperbolic character between Pcr and CPF was found in both groups investigated (r = 0.693, p less than 0.001, and r = 0.741, p less than 0.001, respectively). The hyperbolic relationship noted in the studied groups did not differ significantly. These findings confirm a normal, or a mildly raised Pcr can be associated with a marked decrease in GFR, in some cases to a value as low as a third of the normal one.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed2005

[Chemical structure of fructosan from Condonopsis pilosula].

OBJECTIVE: To study the polysaccharide of Condonopsis pilosula. METHOD: The polysaccharide, CPP-1, was purified by DE-52 cellulose and Sephadex G-200 gel column chromatography. Purity and molecular weight of the polysaccharide were determined by gel permeation chromatography. Methylation analysis, periodate oxidation and degradation, IR, 1H-NMR and 13C-NMR methods were adopted to elucidate the chemical structure. RESULT: The molecular weight of CPP-1 was estimated to be 7.5 x 10(4), and the structure of CPP-1 was a beta-(2 --> 1) linked beta-D-fructosan. CONCLUSION: CPP-1 was a neutral homosaccharide.

Codonopsis↗
PubMedDate not supplied

Cosmeceutical properties of levan produced by Zymomonas mobilis.

Levan, a polysaccharide that can be produced by both plants and microorganisms, is a sugar polymer composed of fructose, with beta-2,6 linkages. Here, we have attempted to assess the possible use of levan produced by Zymomonas mobilis as a cosmeceutical ingredient. In service of this goal, we assessed a host of levan's properties, including its moisturizing effects, cell cytotoxicity, cell proliferation effects, and anti-inflammation effects. Levan exhibited a moisturizing effect that was almost exactly the same as that evidenced by hyaluronic acid, as well as a similar cell proliferation effect in human fibroblast and keratinocyte cell lines. Moreover, in our cell proliferation test, which was conducted using bio-artificial skin constructed via 3-dimensional (3-D) culture after the induction of primary skin inflammation with 0.05% sodium lauryl sulfate (SLS), cell viability in the presence of levan (0.01 mg/ml, 0.05 mg/ml) was determined to be higher than cell viability in the absence of levan. In our anti-inflammation test, which was also conducted using 3-D artificial skin, and which involved the measurement of a quantity of secreted interleukin-1alpha (IL-1alpha), a pre-inflammatory mediator induced by SLS, we determined that the quantity of IL-1alpha in the 3-D artificial skin treated with 0.01 mg/ml and 0.05 mg/ml of levan was less than that registered in a skin sample that had been treated only with SLS. In this study, we determined that levan exerted an anti-inflammatory effect against inflammatory reactions to skin irritants, and also that levan exerted a cell-proliferative effect in bio-artificial skin, thereby indicating its potential applicability as a cosmeceutical agent.

Adult↗
PubMed1991

Cross-reactions between idiotypes, Plasmodium falciparum derived peptides, dinitrophenyl and beta(2-->6) polyfructosan.

In this paper we seek evidence for the participation of the idiotype-anti-idiotype network in the polyclonal B-cell activation (PBA) associated to malaria. For this purpose we tested by an immunoradiometric assay a panel of nine monoclonal antibodies (including seven anti-idiotype antibodies) against three different (plasmodial or non plasmodial) heteroantigens: the 307 synthetic peptide (an epitope of a P. falciparum hepatic stage specific antigen) the (NANP)4 synthetic peptide (a repetitive epitope of the circumsporozoite protein of the P. falciparum sporozoite surface), and dinitrophenyl (DNP) molecule coupled to bovine serum albumin (BSA). Besides the anti-TNP-DNP antibody, the ABPC48 idiotype (directed against beta polyfructosan a fragment of levan molecule) and one anti-idiotype antibody reacted with DNP-BSA. Two other anti-idiotype antibodies (directed against idiotypes of antibodies specific of beta poly-fructosan and phosphorylcholine) were positive against the (NANP)4 antigen. Three antibodies reacted with the 307 antigen which was also recognized by the ABPC48. One of these antibodies was positive to both P. falciparum peptides tested. These preliminary results suggest the existence of crossreactions between plasmodial antigens and idiotypes of antibodies directed against other heterologous antigens. Thus, malaria induced cross-reactive antibodies could act as hetero and/or auto-antibodies explaining, at least partially, the malaria associated PBA phenomenon and modulating the specific immune response during the course of the infection.

Animals↗
PubMed1975

[Intracellular processes leading to the induction of insulin membrane receptors. Study on Acetabularia mediterranea].

Algal cells have been incubated for 7 to 10 days in darkness in sea water containing either actinomycin D or cycloheximide (10 mug/ml) or fructose glucose (5.5 mM). Measurements of insulin fixation to whole cells, plasma membranes or cytosol fractions after that treatment indicate that dark consumption of chloroplastic fructosans stored in photosynthetic conditions would lead to a "signal" inducing translation and transcription of nuclear DNA genetic message resulting in an increase of the number of insulin cellular receptor sites.

Acetabularia↗
PubMed1991

Partial characterization of the glucosyltransferases of an oral Streptococcus salivarius strain.

The oral Streptococcus salivarius strain S3 possessed extracellular glucosyltransferase (GTF) and cell-associated fructosyltransferase activities. Chromatofocusing column chromatography of crude GTF yielded multiple peak fractions, which were categorized into two types which catalysed (1) water-insoluble glucan synthesis and (2) water-soluble glucan formation. The GTF isozyme eluted in fraction 29 synthesized mutanase- and dextranase-sensitive insoluble glucan from sucrose, while fraction 81 GTF synthesized soluble glucan which was insensitive to both glucanases. The ability to agglutinate Streptococcus sobrinus cells was high for insoluble glucan but negligible for soluble glucan. The insoluble glucan-synthesizing activity of fraction 29 GTF was inhibited by the presence of dextran T10. Isoelectric focusing yielded a single activity band but SDS-polyacrylamide gel electrophoresis gave multiple bands for both GTF fractions 29 and 81. The extracellular crude GTF activity was inactivated by 0.1% SDS or 1% Tween 80, but the activity was completely restored by the addition of 1% Triton X-100.

Detergents↗
PubMed1990

Changes of lithium elimination in acute polyuric renal failure caused by cisplatin and mercuric chloride.

Changes in lithium elimination were studied under the condition of experimentally induced polyuric acute renal failure by cisplatin (6 mg/kg body wt) or HgCl2 (3 mg/kg body wt). The histologically proven lesions of tubuli were associated with the decrease of plasma clearance of lithium (C(Li)) and polyfructosan-S (CP(FS). The decrease of these clearance values was not proportional and the ratio C(Li)/CP(FS) (indicating renal fractional excretion of Li+) increased significantly (p less than 0.01). The increase of C(Li)/CP(FS) was related to the increase of renal fractional sodium excretion (FENa) (p less than 0.01). The results suggest that the impairment of tubular cells by cisplatin or HgCl2 caused the decrease of tubular reabsorption of Li+ and Na+. From the pharmacokinetic point of view, these experiments suggest that changes in tubular transport of drugs should be taken into account in their dosing adjustment in patients with acute polyuric tubular lesion.

Acute Kidney Injury↗
PubMed1990

[Validity of routinely used methods of testing glomerular filtration in pediatric practice].

The authors compare different methods used commonly in paediatric practice to assess glomerular filtration (GFR)--creatinine clearance and assessment of GFR by means of Schwartz formula with plasma clearance of polyfructosan S (corresponds to inulin plasma clearance). A group of 108 children aged 4-16 years (x = 10.95) was examined. The patients were divided into three groups by the magnitude of polyfructosan S clearance: greater than 100 ml/min/1.73 m2; 50-100 ml/min/1.73 m2 and less than 50 ml/min/1.73 m2. The authors correlated the clearance of polyfructosan S with creatinine clearance and assessment of GFR according to Schwartz. The method which proved to be most sensitive for detection of reduced GFR in the area of 50-100 ml/min/1.73 m2 was creatinine clearance with urine collection one hour after a previous water load (r = 0.748). In the stage of chronic renal failure with GFR less than 50 ml/min/1.73 m2 the correlation was close with the three-hour creatinine clearance (r = 0.957) and equally close was the correlation with GFR according to Schwartz (r = 0.885). The authors discuss the probability of detection of impaired GFR by commonly used methods and draw attention to the advantages of examination of plasma clearance by polyfructosan S.

Adolescent↗
PubMed1990

A molecular and structural analysis of the VH and VK regions of monoclonal antibodies bearing the A48 regulatory idiotype.

The results presented in this paper explore the molecular basis for expression of the A48 regulatory Id (RI). A48 RI+ mAb derived from idiotypically manipulated mice molecularly resembled the A48 and UPC 10 prototypes of this system by utilizing a VHX24-Vk10 combination. Id expression by these antibodies was not restricted by a particular D region sequence, JH, or JK segment, but quantitative differences in Id expression were associated with utilization of different members of the VK10 germ-line gene families. The VL sequences of these A48 RI+ mAb has identified amino acid residues lying in four different idiotope-determining regions which may contribute to the structural correlate of this Id. A comparative sequence analysis of the VH regions of these VHX24 utilizing A48 RI+ mAb with several A48 RI+ mAb utilizing VHJ558 or VH7183 VH genes as well as a hybrid transfectoma antibody derived from two A48 RI-, VHJ558 utilizing hybridomas, all suggested that four nonconsecutive positions which lie outside the idiotope-determining regions may contribute structural elements toward expression of this Id. The VH and VL regions of the A48RI+, VHX24-Vk 10+ mAb showed low to moderate levels of somatic mutation which showed different patterns of distribution between the complementary determining region (CDR) and framework regions in the H and L chains. Although the VK sequences contained 50% of the replacement mutations in the CDR, with a replacement/silent mutation ratio of 10, the CDR of the VH sequences contained only 31% of the replacement mutations with a replacement/silent mutation ratio of 0.69.

Amino Acid Sequence↗
PubMed1989

Expression of a cross-reactive idiotope on naturally occurring murine antibodies against 3-fucosyllactosamine, levan, and dextran.

We recently identified a cross-reactive Id (6C4) that is expressed on the H chain of many BALB/c mAb against the 3-fucosyllactosamine (3-FL) determinant, Gal(beta 1-4) (Fuc(alpha 1-3] GlcNAc-R. The VH segments of seven mAb that we recently sequenced are encoded by VH441, which also encodes VH segments of antibodies against galactan, levan, and dextran. To analyze the expression of the 6C4 Id on naturally occurring anti-carbohydrate antibodies, we isolated 6C4+ antibodies by affinity chromatography from pools of normal BALB/c serum. Approximately 20 to 30% of antibodies against 3-FL and levan, and all antibodies against dextran, were removed from the sera by passage over a column containing mAb 6C4. Absorption of the eluate with 3-FL beads removed anti-3-FL antibodies but not anti-dextran or anti-levan. The expression of a cross-reactive Id on naturally occurring antibodies against several carbohydrate Ag suggests that these antibodies may participate in an Id network. We also reported previously that BALB/c mice have naturally occurring anti-3-FL antibodies and respond well to immunization against this determinant, whereas C57BL/6 mice do neither. To examine the role of the Igh-C allotype in the regulation of the anti-3-FL response, we studied congenic strains of BALB/c and C57BL/6 mice. Both congenic strains produced anti-3-FL antibodies in response to immunization, but only C.B-20 mice exhibited naturally occurring antibodies. These data suggest that the naturally occurring and elicited antibody responses against 3-FL are differentially regulated.

Animals↗
PubMed1989

[Polyfructosan and para-aminohippuric acid clearance, comparison of 2 methods for determining glomerular filtration and plasma renal flow].

Glomerular Filtration Rate (GFR) and Renal Plasma Flow (RPF) were measured respectively, Polyfructosan and Para-aminohippuric acid clearances, in 13 patients with different degrees of renal insufficiency. Two different methods were used, the standard method and one without urine collection. The mean GFR (66.4 +/- 27.8 vs. 72.2 +/- 19.0 ml/m, r = 0.74) and RPF (431 +/- 204 vs 490 +/- 188 ml/m, r = 0.76) as well as Filtration Fraction (17.7 +/- 8.3 vs. 16.6 +/- 6.6%) were well correlated. These data show that within the normal range of GRF and RPF the method determining only plasma levels is as reliable as the standard method, giving the advantage to be less troublesome for patients and staff, requiring no bladder catheterization.

Adult↗
PubMed1977

Effects of prolonged administration of high molecular levan on arterial blood gases, oxygen transport and acid base balance in rabbits.

The effect of daily intraperitoneal injections of 100 mg of high molecular levan for 4-6 weeks on a number of physiological parameters was studied in rabbits. The parameters studied included whole blood pH, blood gases and acid-base balance. Control rabbits were similarly treated with physiological saline. In five rabbits T-50 was determined after levan treatment. None of the studied parameters was significantly affected by the levan treatment.

Acid-Base Equilibrium↗