scholarly journals PROPERTIES OF NATIVE PROTEIN-CONTAINING ANTIGENS OF STREPTOCOCCUS PNEUMONIAE

2019 ◽  
Vol 1 (1) ◽  
pp. 22-28
Author(s):  
D. S. Vorobyev ◽  
I. B. Semenova ◽  
Yu. V. Volokh ◽  
E. E. Romanenko ◽  
A. P. Baturo ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Chemical Composition ◽  
Immune Serum ◽  
Rabbit Serum ◽  
Protective Activity ◽  
Native Protein ◽  
In Vivo Experiments ◽  
Sds Electrophoresis

Aim. The study of immunochemical and immunobiological properties of native protein-containing antigens of pneumococcus. Materials and methods. The study was carried out on the strains of the Collective Usage Center «Collection of Mechnikov Res. Inst. for Vaccine and Sera». In the work studied the chemical composition, the molecular weight of the obtained antigens in SDS-electrophoresis and antibody titers in ELISA. Protective activity of protein-containing antigens of pneumococcus was determined in experiments of active protection of mice. Results. Protein-containing antigens of pneumococcus were isolated from S. pneumoniae serotypes 3, 6B, 10A, 14, 19F, 23F and 36. The chemical composition of the preparations contained from 16 to 35% protein. In SDS-electrophoresis in polyacrylamide gel it was established that the molecular weight of protein-containing antigens of pneumococcus ranged from 14 to 116 kDa. Using ELISA shows the cross-activity of native antigens. Virtually all drugs reacted with antimicrobial rabbit serum obtained to serotype 19F (p≤0.05). Serotype serum 14 was less active and only protein-containing pneumococcal antigens obtained from 14 and 19F serotypes (p≤0.05) interacted with it. In the precipitation test according to Ouchterlony it was confirmed that preparations of serotypes 3, 6B, 14, 19F and 36 reacted with rabbit immune serum obtained for S. pneumoniae 19F serotype. In immunoblotting it was found that protein-containing antigens of pneumococcus isolated from serotypes 3, 6B, 10A, 14, 19F and 36 were associated with monoclonal antibodies to pneumococcal protein — pneumolysin. In vivo experiments it was shown that protein-containing antigens of pneumococcus protected animals from intraperitoneal infection of S. pneumoniae in homologous and heterologous systems (p≤0.05). Conclusion. The revealed immunochemical and cross-protective activity of protein-containing antigens of pneumococcus in vitro and in vivo experiments allows to select drugs derived from serotypes 6B, 10A, 19F and 36, as the most promising for further study of the intraspecific protective activity of individual native proteins of pneumococcus. 

Binding of 63Ni(II) to Ultrafiltrable Constituents of Rabbit Serum In Vivo and In Vitro

1975 ◽  
Vol 21 (4) ◽  
pp. 521-527 ◽  
Author(s):  
Noritake Asato ◽  
Maria van Soestbergen ◽  
F William Sunderman
Keyword(s):  
Rabbit Serum ◽  
Total Serum ◽  
In Vivo Experiments ◽  
The Mean ◽  
Layer Chromatography ◽  
In Vivo Administration ◽  
Mean Concentration

Abstract Binding of 63Ni(Il) to ultrafiltrable constituents of rabbit serum was studied (a) after in vitro incubation (2 h, 37 °C) of rabbit serum with 63NiCl2 (10-100 µmol/liter), and (b) at intervals (0.25-2 h) after in vivo administration of 63NiCl2 (40-160 µmol/kg body wt, i.v.). Serum ultrafiltrates were fractionated by thin-layer chromatography, and the separated compounds made visible by autoradiography and by ninhydrin staining. Several (≃5) ultrafiltrable 63Ni-complexes were demonstrable as distinct radiodense 63Ni-bands with chromatographic mobilities corresponding to those of ninhydrin-positive bands. Unbound 63Ni(II) was not detected in serum ultrafiltrates in either the in vitro or in vivo experiments. In sera (n = 10) incubated in vitro with 63Ni(II) (10 µmol/ liter), the mean percentage of ultrafiltrable 63Ni was 36% (range = 33-38) of total serum 63Ni. In contrast, in sera (n = 10) obtained 2 h after i.v. injection of 63Ni(II) (40 µmol/kg), the mean concentration of total serum 63Ni was 10.8 µmol/liter (range = 6-14), and the mean percentage of ultrafiltrable 63Ni was 15% (range = 9-21) of total serum 63Ni. The disparity between the percentages of ultrafiltrable 63Ni obtained in vitro and in vivo was obviated when the in vivo experiments were performed in rabbits bilaterally nephrectomized, with ligated common bile ducts. This investigation confirms the existence of several nickel receptors in serum ultrafiltrates and substantiates the role of ultrafiltrable complexes in the excretion of nickel.

Promising in vitro Anti-Toxoplasma gondii effects of commercial Chitosan

2020 ◽  
Vol 20 ◽  
Author(s):  
Bahman Rahimi Esboei ◽  
Masoud Keighobadi ◽  
Hajar Ziaei Hezarjaribi ◽  
Mahdi Fakhar ◽  
Ahmad Daryani ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Control Group ◽  
Low Molecular Weight ◽  
In Vitro Activity ◽  
Obligate Intracellular ◽  
In Vivo Experiments ◽  
Ic50 Values ◽  
Acute Toxoplasmosis

Background: Toxoplasmosis is a disease that results from infection with an obligate intracellular T. gondii parasite, one of the world's most common parasites. Considering the complications of chemical drugs and the need for an appropriate drug combination for treatment of toxoplasmosis and also considering the antimicrobial potential of chitosan, as a natural source, this study was aimed to evaluate in vitro activity of commercial chitosan (CC) on T. gondii. Methods: In this experimental study, the tachyzoites of T. gondii was collected from the peritoneal exudates from infected Balb/c mice. The tachyzoites were diluted in phosphate buffer saline (PBS). Chitosan with low molecular weight was commercially purchased. Then, at concentrations of 10, 50, 100 and 200 µg/mL and after 30, 60, 120 and 180 minutes the viability of tachyzoites were determined by using trypan blue 0.1%. Anti-T.gondii activity of CC in all concentration was significantly higher than pyrimethamine as control group (P=0.05). Results: The concentration of 200 µg/mL of CC had the highest effects and killed 30.5, 52, 59 and 81.5% of tachyzoites after 30, 60, 120 and 180 minutes. Moreover, IC50 values of CC were 515, 171, 12.5 and <10 μg/mL in comparison with pyrimethamine as 58.82 μg/mL for 30, 60, 120, and 180 min of exposure time. Conclusion: Our results indicate chitosan in low molecular weight had potent activity against T. gondii tachyzoites and could be an appropriate candidate for treatment of at least acute toxoplasmosis, certainly, after complementary in vivo experiments.

scholarly journals Monoclonal antibody with hemagglutination, immobilization, and neutralization activities defines an immunodominant, 47,000 mol wt, surface-exposed immunogen of Treponema pallidum (Nichols).

1984 ◽  
Vol 160 (5) ◽  
pp. 1404-1420 ◽  
Author(s):  
S A Jones ◽  
K S Marchitto ◽  
J N Miller ◽  
M V Norgard
Keyword(s):  
Treponema Pallidum ◽  
Immune Serum ◽  
Host Cells ◽  
Rabbit Serum ◽  
Functional Activities ◽  
Molecular Weights ◽  
Rabbit Igg ◽  
Major Surface

Radioimmunoprecipitation (RIP) analyses performed on 125I-surface-labeled Treponema pallidum cells using various immune sera revealed the presence of six major surface antigens (immunogens) with apparent molecular weights of 47 K, 36 K, 34 K, 32 K, 29 K, and 13 K. Among these, the 47 K surface antigen was most abundant. Radioimmunoprecipitation assays using 125I-labeled T. phagedenis biotype Reiter or immunoblot analyses using the same strain, failed to reveal the presence of the 47 K mol wt antigen in the representative nonpathogenic treponeme. Preabsorption of anti-T. pallidum immune rabbit serum (IRS) with the Reiter organism did not remove anti-T. pallidum antibodies from immune serum that reacted with the 47 K mol wt immunogen or other immunogens of T. pallidum present in the characteristic antigenic profile. Monoclonal antibodies (mAb) directed specifically against the 47 K mol wt immunogen of T. pallidum also failed to react with an analogous 47 K mol wt component in Treponema phagedenis biotype Reiter, further suggesting the unique presence of this antigen in pathogenic treponemes. The presence of the 47 K mol wt surface immunogen in pathogenic treponemes other than T. pallidum subspecies pallidum was also observed (43). Anti-47 K immunogen mAb was nonreactive against rabbit IgG or IgM. mAb directed specifically against the 47 K mol wt immunogen of T. pallidum was examined for strategic functional activities. It was found to be reactive in the microhemagglutination assay for T. pallidum antibodies, the T. pallidum immobilization test, and was found to be capable of significant blockage of attachment of virulent T. pallidum to host cells in tissue culture. Additional significant biological activity for the anti-47 K mol wt immunogen mAb was revealed through results of the in vitro-in vivo neutralization test of Bishop and Miller, in which a 99% or 100% neutralizing activity was demonstrated. The combined data of this study suggest that the 47 K mol wt immunogen of T. pallidum represents an abundant, immunodominant, surface-exposed immunogen possessing potential biological importance in the pathogenesis and immunology of T. pallidum infection. These studies serve to establish the first functionally defined immunogen for T. pallidum, which may represent the major immunogen of the organism.

scholarly journals Non-invasive monitoring of low molecular weight biomarkers relevant to skin inflammation and cancer

2021 ◽  
Author(s):  
◽  
Skaidre Jankovskaja
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Skin Inflammation ◽  
Basic Knowledge ◽  
Skin Disorders ◽  
Porcine Skin ◽  
Non Invasive ◽  
In Vivo Experiments

Development of skin inflammation and cancer in viable epidermis and dermis involve slow molecular weight (LMW) metabolites. We hypothesize that these LMW compounds can be collected on the surface of the skin and used for non-invasive diagnostics of skin disorders. Keeping in mind that substantial transdermal penetrationis achieved only for molecules of < 500 Da, we focused on topical monitoring of LMW biomarkers. In this thesis we investigated non-invasive, topical methods for monitoring LMW biomarkers by relevant in vitro and in vivo experiments. The LMW biomarkers were: - reactive oxygen species (ROS), specifically, hydrogen peroxide, H2O2 - amino acids and their derivatives, i.e., tryptophan (Trp), kynurenine (Kyn; a Trpderivative), phenylalanine (Phe), and tyrosine (Tyr; a Phe derivative). Initially, we have carried out in vitro experiments using dermatomed porcine skin and cell cultures. We characterized permeability of the biomarkers through skin and assessed methods of their monitoring. By using Prussian white particles, deposited on porcine skin, we demonstrated that hydrophilic biomarkers, such as H2O2, permeate the skin mainly through hair follicle pathways (Paper I). In paper II, we have showed that the enzymes transforming Trp to the inflammation and cancer biomarker Kyn, are expressed in the basal layer of epidermis. The magnitude of changes of the Trp/Kyn ratio in the cell culture model was assessed. In paper III, we have characterized Trp and Kyn permeability through skin in vitro, concluding that their permeabilities through stratum corneum are comparable. By in vivo experiments outlined in Paper IV, we have demonstrated the feasibility of topical, non-invasive sampling of Trp and Kyn, in relation to other amino acids. Kyn detection was compromised by its low abundance on the skin. In paper V, we performed a proof-of-concept study in vivo and confirmed that non-invasive sampling of Trp and amino acids of similar abundance, such as Phe and Tyr, is more robust. We concluded that Phe/Trp ratio might be equally good biomarker of skin disorders as a predicted Trp/Kyn ratio. Summarizing, the results of this thesis provide basic knowledge for deeper clinical studies of non-invasive, topical sampling of hydrophilic LMW biomarkers of skin inflammation and cancer.

Retard-insulins formed by complexing with polyamino acids

1988 ◽  
Vol 53 (11) ◽  
pp. 2944-2951 ◽  
Author(s):  
Günter Losse ◽  
Wolfgang Naumann ◽  
Heike Raddatz ◽  
Joachim Koszowicz
Keyword(s):  
Molecular Weight ◽  
Blood Glucose ◽  
Glutamic Acid ◽  
Physicochemical Parameters ◽  
Ionic Interactions ◽  
Glucose Lowering ◽  
In Vivo Experiments ◽  
Polyamino Acid

Polyglycine, polyalanine, polyleucine, poly-α-glutamic acid, poly-γ-glutamic acid and poly-α-lysine were complexed with insulin under non-denaturating conditions. The liberation behaviour of the hormone was investigated in vivo and in vitro in dependence on the insulin content, molecular weight, ionic interactions and hydrophobicity of the polyamino acid. The in vitro results were confirmed by the in vivo experiments with animals. They clearly pointed out the influence of physicochemical parameters on the bioavailability of insulin. Complexes of poly-α-lysine and polyglycine were shown to be the most suitable retard forms, producing significant blood glucose lowering effects over 12 hours.

Effects of Unfractionated and Low Molecular Weight Heparins on Platelet Thromboxane Biosynthesis “In Vivo”

1994 ◽  
Vol 72 (06) ◽  
pp. 942-946 ◽  
Author(s):  
Raffaele Landolfi ◽  
Erica De Candia ◽  
Bianca Rocca ◽  
Giovanni Ciabattoni ◽  
Armando Antinori ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Unfractionated Heparin ◽  
Low Molecular Weight Heparin ◽  
Primary Source ◽  
In Vivo Studies ◽  
Low Molecular Weight ◽  
Heparin Administration ◽  
To Receive

SummarySeveral “in vitro” and “in vivo” studies indicate that heparin administration may affect platelet function. In this study we investigated the effects of prophylactic heparin on thromboxane (Tx)A2 biosynthesis “in vivo”, as assessed by the urinary excretion of major enzymatic metabolites 11-dehydro-TxB2 and 2,3-dinor-TxB2. Twenty-four patients who were candidates for cholecystectomy because of uncomplicated lithiasis were randomly assigned to receive placebo, unfractionated heparin, low molecular weight heparin or unfractionaed heparin plus 100 mg aspirin. Measurements of daily excretion of Tx metabolites were performed before and during the treatment. In the groups assigned to placebo and to low molecular weight heparin there was no statistically significant modification of Tx metabolite excretion while patients receiving unfractionated heparin had a significant increase of both metabolites (11-dehydro-TxB2: 3844 ± 1388 vs 2092 ±777, p <0.05; 2,3-dinor-TxB2: 2737 ± 808 vs 1535 ± 771 pg/mg creatinine, p <0.05). In patients randomized to receive low-dose aspirin plus unfractionated heparin the excretion of the two metabolites was largely suppressed thus suggesting that platelets are the primary source of enhanced thromboxane biosynthesis associated with heparin administration. These data indicate that unfractionated heparin causes platelet activation “in vivo” and suggest that the use of low molecular weight heparin may avoid this complication.

Toxicity of Heparinoids, with Special Reference to the Precipitation of Fibrinogen

1964 ◽  
Vol 12 (01) ◽  
pp. 232-261 ◽  
Author(s):  
S Sasaki ◽  
T Takemoto ◽  
S Oka
Keyword(s):  
Molecular Weight ◽  
Dextran Sulfate ◽  
Anticoagulant Activity ◽  
Molecular Structures ◽  
Severe Reaction ◽  
Clinical Use ◽  
Molecular Weights ◽  
Close Relationship

SummaryTo demonstrate whether the intravascular precipitation of fibrinogen is responsible for the toxicity of heparinoid, the relation between the toxicity of heparinoid in vivo and the precipitation of fibrinogen in vitro was investigated, using dextran sulfate of various molecular weights and various heparinoids.1. There are close relationships between the molecular weight of dextran sulfate, its toxicity, and the quantity of fibrinogen precipitated.2. The close relationship between the toxicity and the precipitation of fibrinogen found for dextran sulfate holds good for other heparinoids regardless of their molecular structures.3. Histological findings suggest strongly that the pathological changes produced with dextran sulfate are caused primarily by the intravascular precipitates with occlusion of the capillaries.From these facts, it is concluded that the precipitates of fibrinogen with heparinoid may be the cause or at least the major cause of the toxicity of heparinoid.4. The most suitable molecular weight of dextran sulfate for clinical use was found to be 5,300 ~ 6,700, from the maximum value of the product (LD50 · Anticoagulant activity). This product (LD50 · Anticoagulant activity) can be employed generally to assess the comparative merits of various heparinoids.5. Clinical use of the dextran sulfate prepared on this basis gave satisfactory results. No severe reaction was observed. However, two delayed reactions, alopecia and thrombocytopenia, were observed. These two reactions seem to come from the cause other than intravascular precipitation.

Neutralization of a Low Molecular Weight Heparin (LHN-1) and Conventional Heparin by Protamine Sulfate in Rats

1986 ◽  
Vol 56 (03) ◽  
pp. 318-322 ◽  
Author(s):  
V Diness ◽  
P B Østergaard
Keyword(s):  
Molecular Weight ◽  
Low Molecular Weight Heparin ◽  
Thrombus Formation ◽  
Experimental Models ◽  
Protamine Sulfate ◽  
Low Molecular Weight ◽  
Antithrombotic Effect ◽  
Higher Doses

SummaryThe neutralization of a low molecular weight heparin (LHN-1) and conventional heparin (CH) by protamine sulfate has been studied in vitro and in vivo. In vitro, the APTT activity of CH was completely neutralized in parallel with the anti-Xa activity. The APTT activity of LHN-1 was almost completely neutralized in a way similar to the APTT activity of CH, whereas the anti-Xa activity of LHN-1 was only partially neutralized.In vivo, CH 3 mg/kg and LHN-1 7.2 mg/kg was given intravenously in rats. The APTT and anti-Xa activities, after neutralization by protamine sulfate in vivo, were similar to the results in vitro. In CH treated rats no haemorrhagic effect in the rat tail bleeding test and no antithrombotic effect in the rat stasis model was found at a protamine sulfate to heparin ratio of about 1, which neutralized APTT and anti-Xa activities. In LHN-1 treated rats the haemorrhagic effect was neutralized when APTT was close to normal whereas higher doses of protamine sulfate were required for neutralization of the antithrombotic effect. This probably reflects the fact that in most experimental models higher doses of heparin are needed to induce bleeding than to prevent thrombus formation. Our results demonstrate that even if complete neutralization of APTT and anti-Xa activities were not seen in LHN-1 treated rats, the in vivo effects of LHN-1 could be neutralized as efficiently as those of conventional heparin. The large fall in blood pressure caused by high doses of protamine sulfate alone was prevented by the prior injection of LHN-1.

Studies on Metabolism of Urokinase and Mechanism of Thrombolysis by Urokinase

1979 ◽  
Vol 42 (03) ◽  
pp. 885-894 ◽  
Author(s):  
Tatsuo Ueno ◽  
Norio Kobayashi ◽  
Tadashi Maekawa
Keyword(s):  
Molecular Weight ◽  
Plasma Clearance ◽  
Radioactive Material ◽  
Optimal Dosage ◽  
Plasma Clot ◽  
Optimal Dosage Regimen ◽  
Arterial Thrombus ◽  
Contact Experiment

SummaryPharmacokinetics of intravenously injected 125I-labeled urokinase (125I-UK) of a molecular weight of 33,000 daltons in normal rabbits and patients with various diseases were investigated. The plasma clearance of 125I-UK in rabbits was described by a biexponential curve within six hours with a half-life of 8 minutes, 2.3 hours, respectively. The radioactivity in the liver and kidneys 15 minutes after iv injection with 125I-UK was 9.6% and 14.0% of the radioactivity injected, respectively. Approximately 80% of the total radioactive material injected was excreted in the urine in 18 hours. No increase in activator activity in the urine was observed after a large amount of UK injection. Activity uptake of 125I-UK by experimentally induced arterial thrombus was little. Lysis of the stasis thrombus was produced by injecting 7.5 × 104 IU of UK in only one out of 8 rabbits. In vitro contact experiment revealed that transfer of 125I-UK to plasma clot is slow (24 hours for 10% of 125I-UK by plasma clot). In 4 patients plasma clearance of 125I-UK was essentially similar to that in rabbits. From the results obtained optimal dosage regimen of UK administration for complete thrombolysis in vivo was discussed.

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