scholarly journals Qualifying the T-2 Toxin-Degrading Properties of Seven Microbes with Zebrafish Embryo Microinjection Method

Toxins ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 460
Author(s):  
Edina Garai ◽  
Anita Risa ◽  
Emese Varga ◽  
Mátyás Cserháti ◽  
Balázs Kriszt ◽  
...  
Keyword(s):  
Sublethal Effects ◽  
Degradation Products ◽  
Rhodococcus Erythropolis ◽  
Bacterial Strains ◽  
Cell Stage ◽  
Bacterial Metabolites ◽  
Mycotoxin Degradation

T-2 mycotoxin degradation and detoxification efficiency of seven bacterial strains were investigated with zebrafish microinjection method in three steps ((1) determination of mycotoxin toxicity baseline, (2) examination of bacterial metabolites toxicity, (3) identification of degradation products toxicity). Toxicity of T-2 was used as a baseline of toxic effects, bacterial metabolites of strains as control of bacterial toxicity and degradation products of toxin as control of biodegradation were injected into one-cell stage embryos in the same experiment. The results of in vivo tests were checked and supplemented with UHPLC-MS/MS measurement of T-2 concentration of samples. Results showed that the Rhodococcus erythropolis NI1 strain was the only one of the seven tested (R. gordoniae AK38, R. ruber N361, R. coprophilus N774, R. rhodochrous NI2, R. globerulus N58, Gordonia paraffinivorans NZS14), which was appropriated to criteria all aspects (bacterial and degradation metabolites of strains caused lower toxicity effects than T-2, and strains were able to degrade T-2 mycotoxin). Bacterial and degradation metabolites of the NI1 strain caused slight lethal and sublethal effects on zebrafish embryos at 72- and 120-h postinjection. Results demonstrated that the three-step zebrafish microinjection method is well-suited to the determination and classification of different bacterial strains by their mycotoxin degradation and detoxification efficiency.

Stability-Indicating Determination of Tedizolid Phosphate in the Presence of its Active Form and Possible Degradants

2021 ◽  
Author(s):  
Eman M Moaaz ◽  
Ezzat M Abdel-Moety ◽  
Mamdouh R Rezk ◽  
Ahmed S Fayed
Keyword(s):  
Flow Rate ◽  
Phosphate Buffer ◽  
Mobile Phase ◽  
Degradation Products ◽  
Active Form ◽  
Pharmaceutical Formulations ◽  
Bacterial Strains ◽  
Stress Degradation ◽  
Tlc Plates

Abstract Tedizolid phosphate is an antibiotic prodrug that is metabolized into tedizolid which is used against various resistant bacterial strains. In this study, tedizolid phosphate was subjected to stress degradation conditions, namely, hydrolysis (neutral, acidic and alkaline), thermal, oxidative and photolytic ones. The prodrug was stable toward thermal and photolytic stress conditions, while it showed significant degradation upon applying oxidative and hydrolytic conditions. Two suggested chromatographic methods are described for separation and determination of tedizolid phosphate from the resulted degradation products. The first method is HPLC using Waters Xselect HSS C18 (250 × 4.6 mm, 5 μm) analytical column and mobile phase composed of phosphate buffer (50 mM, pH 6.5):acetonitrile (70:30, %v/v) pumped at flow rate of 1.0 mL/min with UV-detection at 300 nm. The second method is a TLC coupled with densitometric quantitation, precoated silica TLC-plates as a stationary phase and a mobile phase of methanol:butanol:ethyl acetate:ammonia (33%, w/v) (60:20:20:10,%v/v) were used. The chromatographed plates were scanned at 300 nm. The linearity was confirmed over concentration range of 1–100 μg/mL and 1–12 μg/band for HPLC and TLC-densitometric methods, respectively. Both methods were found to be suitable for determination of tedizolid phosphate in pure form and in its pharmaceutical formulations.

scholarly journals Cross-Feeding among Probiotic Bacterial Strains on Prebiotic Inulin Involves the Extracellularexo-Inulinase ofLactobacillus paracaseiStrain W20

2018 ◽  
Vol 84 (21) ◽  
Author(s):  
Markus C. L. Boger ◽  
Alicia Lammerts van Bueren ◽  
Lubbert Dijkhuizen
Keyword(s):  
Degradation Products ◽  
Lactobacillus Paracasei ◽  
Transport Systems ◽  
Glycoside Hydrolase Family ◽  
Bacterial Strains ◽  
Human Gut ◽  
Content Type ◽  
Beneficial Effects ◽  
Probiotic Lactobacilli

ABSTRACTProbiotic gut bacteria employ specific metabolic pathways to degrade dietary carbohydrates beyond the capabilities of their human host. Here, we report how individual commercial probiotic strains degrade prebiotic (inulin type) fructans. First, a structural analysis of commercial fructose oligosaccharide-inulin samples was performed. These β-(2-1)-fructans differ in termination by either glucose (GF) or fructose (FF) residues, with a broad variation in the degrees of polymerization (DPs). The growth of individual probiotic bacteria on short-chain inulin (sc-inulin) (Frutafit CLR), a β-(2-1)-fructan (DP 2 to DP 40), was studied.Lactobacillus salivariusW57 and other bacteria grew relatively poorly on sc-inulin, with only fractions of DP 3 and DP 5 utilized, reflecting uptake via specific transport systems followed by intracellular metabolism.Lactobacillus paracaseisubsp.paracaseiW20 completely used all sc-inulin components, employing an extracellularexo-inulinase enzyme (glycoside hydrolase family GH32 [LpGH32], also found in other strains of this species); the purified enzyme converted high-DP compounds into fructose, sucrose, 1-kestose, and F2 (inulobiose). The cocultivation ofL. salivariusW57 andL. paracaseiW20 on sc-inulin resulted in cross-feeding of the former by the latter, supported by this extracellularexo-inulinase. The extent of cross-feeding depended on the type of fructan, i.e., the GF type (clearly stimulating) versus the FF type (relatively low stimulus), and on fructan chain length, since relatively low-DP β-(2-1)-fructans contain a relatively high content of GF-type molecules, thus resulting in higher concentrations of GF-type DP 2 to DP 3 degradation products. The results provide an example of howin vivocross-feeding on prebiotic β-(2-1)-fructans may occur among probiotic lactobacilli.IMPORTANCEThe human gut microbial community is associated strongly with host physiology and human diseases. This observation has prompted research on pre- and probiotics, two concepts enabling specific changes in the composition of the human gut microbiome that result in beneficial effects for the host. Here, we show how fructooligosaccharide-inulin prebiotics are fermented by commercial probiotic bacterial strains involving specific sets of enzymes and transporters. Cross-feeding strains such asLactobacillus paracaseiW20 may thus act as keystone strains in the degradation of prebiotic inulin in the human gut, and this strain–exo-inulinase combination may be used in commercialLactobacillus-inulin synbiotics.

scholarly journals Evaluation of the Multimycotoxin-Degrading Efficiency of Rhodococcus erythropolis NI1 Strain with the Three-Step Zebrafish Microinjection Method

2021 ◽  
Vol 22 (2) ◽  
pp. 724
Author(s):  
Edina Garai ◽  
Anita Risa ◽  
Emese Varga ◽  
Mátyás Cserháti ◽  
Balázs Kriszt ◽  
...  
Keyword(s):  
Degradation Products ◽  
Synergistic Effects ◽  
Rhodococcus Erythropolis ◽  
Toxic Effects ◽  
Zebrafish Embryos ◽  
Combined Exposure ◽  
Step Method ◽  
Cell Stage ◽  
Antagonistic Effects ◽  
Very High

The multimycotoxin-degrading efficiency of the Rhodococcus erythropolis NI1 strain was investigated with a previously developed three-step method. NI1 bacterial metabolites, single and combined mycotoxins and their NI1 degradation products, were injected into one cell stage zebrafish embryos in the same doses. Toxic and interaction effects were supplemented with UHPLC-MS/MS measurement of toxin concentrations. Results showed that the NI1 strain was able to degrade mycotoxins and their mixtures in different proportions, where a higher ratio of mycotoxins were reduced in combination than single ones. The NI1 strain reduced the toxic effects of mycotoxins and mixtures, except for the AFB1+T-2 mixture. Degradation products of the AFB1+T-2 mixture by the NI1 strain were more toxic than the initial AFB1+T-2 mixture, while the analytical results showed very high degradation, which means that the NI1 strain degraded this mixture to toxic degradation products. The NI1 strain was able to detoxify the AFB1, ZEN, T-2 toxins and mixtures (except for AFB1+T-2 mixture) during the degradation experiments, which means that the NI1 strain degraded these to non-toxic degradation products. The results demonstrate that single exposures of mycotoxins were very toxic. The combined exposure of mycotoxins had synergistic effects, except for ZEN+T-2 and AFB1+ZEN +T-2, whose mixtures had very strong antagonistic effects.

scholarly journals Determination of Grading for Maxillary and Mandibular Tori- an in Vivo study

2018 ◽  
Vol 11 (2) ◽  
pp. 679-688
Author(s):  
Parithimar kalaignan ◽  
Jaya Shree Mohan ◽  
Arun Jayakumar
Keyword(s):  
Sample Size ◽  
Total Sample ◽  
Grading System ◽  
Total Sample Size ◽  
Torus Palatinus ◽  
Length Width ◽  
Mandibular Tori

To establish a definitive grading system & derive classification of maxillary &mandibular tori based on shape . A cluster sample of 72 patients who sought treatment at the dental college with maxillary and/or Mandibular tori were selected, alginate impression were made for each patient and the stone casts poured were evaluated based on the following criteria; shape, prominence and dimensions (length, width, height) to surface a new grading system for the tori. From a total sample size of 72 patients with tori, 38 (52.8%) patients were dentulous (either fully or partially dentulous), and remaining 34 (47.2%) patients were edentulous. A total of 5 patients only had both maxillary and mandibular tori simultaneously present at the time of inspection, while all other 67 patients had only maxillary tori. From the 72 patients who presented with torus palatinus, in relation to the tori shape, 10 (13.9%) were flat, 43 (59.7%) were lobular, 16(22.2%) were spindle shaped and only 3 (4.2%) were of nodular shape. From the 5 patients who presented with torus mandibularis, 3 presented with tori of mild prominence, and 2 others presented with a moderately prominent mandibular tori.In relation to torus palatinus dimension, the width of tori varied widely from 4.0mm to 25.0mm, while the length too had a wide variation from 10.0mm to 37.0mm. The height however hadn’t as much variation, ranging from 0.5mm to 5.0mm. A successful grading system was accomplished in relation to maxillary tori and its effect on the prognosis of maxillary denture stability and retention could be determined from this grading system; however this was not successful for mandibular tori due to insufficient sample size of patients in this category. The grading system devised in our study was reasonable for use by dental graduates, though the study had short comings in few aspects that were not looked upon on initiation, it would be a stepping stone towards more research in this field in relation to maxillary and mandibular tori.. From this study, it is anticipated that Prosthodontist can have a precise idea of the tori to prosthesis relationship when faced with such patients in their day to day practice.

Criteria for determination of lipid peroxidation in tissues: estimation in liver of mice intoxicated with carbon tetrachloride

1987 ◽  
Vol 65 (7) ◽  
pp. 1503-1506 ◽  
Author(s):  
Ryungsoon Song Kim ◽  
Frank S. LaBella
Keyword(s):  
Lipid Peroxidation ◽  
Degradation Products ◽  
Lipid Peroxides ◽  
Thiobarbituric Acid ◽  
Time Intervals ◽  
Microsomal Membranes ◽  
Liver Homogenates ◽  
Second Peak

The profiles of lipid peroxidation products in liver homogenates or microsomal membranes prepared from CCl4-intoxicated mice were determined by several commonly employed methods. The level of conjugated dienes peaked within 30 min and then decreased, suggesting the transitory nature of lipid peroxides in vivo. Values for thiobarbituric acid positive material peaked 30 min after CCl4 treatment, diminished thereafter for a time, and gradually rose to a new maximum at 24 h; the first peak appears to represent lipid peroxides and the second represents further degradation products including malondialdehyde. Fluorescence intensity (excitation, 360 nm; emission, 430 nm) was closely correlated with the second peak. Our findings support the involvement of lipid peroxidation in CCl4-induced hepatotoxicity in mice and emphasize the necessity for several analytical indices of lipid peroxidation performed at different time intervals.

A Molecular Approach to Immunoscintigraphy: A Study of the T-Antigen Conformation on the Surface of Tumors

1987 ◽  
Vol 26 (01) ◽  
pp. 1-6 ◽  
Author(s):  
S. Selvaraj ◽  
M. R. Suresh ◽  
G. McLean ◽  
D. Willans ◽  
C. Turner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Tumor Cell ◽  
T Antigen ◽  
Human Carcinomas ◽  
Animal Tumor ◽  
Synthetic Antigens

The role of glycoconjugates in tumor cell differentiation has been well documented. We have examined the expression of the two anomers of the Thomsen-Friedenreich antigen on the surface of human, canine and murine tumor cell membranes both in vitro and in vivo. This has been accomplished through the synthesis of the disaccharide terminal residues in both a and ß configuration. Both entities were used to generate murine monoclonal antibodies which recognized the carbohydrate determinants. The determination of fine specificities of these antibodies was effected by means of cellular uptake, immunohistopathology and immunoscintigraphy. Examination of pathological specimens of human and canine tumor tissue indicated that the expressed antigen was in the β configuration. More than 89% of all human carcinomas tested expressed the antigen in the above anomeric form. The combination of synthetic antigens and monoclonal antibodies raised specifically against them provide us with invaluable tools for the study of tumor marker expression in humans and their respective animal tumor models.

The Inhibition of Platelet Aggregation by an Aorta Intima Extract

1974 ◽  
Vol 32 (02/03) ◽  
pp. 417-431 ◽  
Author(s):  
A. du P Heyns ◽  
D. J van den Berg ◽  
G. M Potgieter ◽  
F. P Retief
Keyword(s):  
Platelet Aggregation ◽  
Degradation Products ◽  
Adenosine Diphosphate ◽  
Inhibitory Effect ◽  
Protective Role ◽  
Vascular Trauma ◽  
Wear And Tear ◽  
Sequential Addition ◽  
Aggregation Activity

SummaryThe platelet aggregating activity of extracts of different layers of the arterial wall was compared to that of Achilles tendon. Arterial media and tendon extracts, adjusted to equivalent protein content as an index of concentration, aggregated platelets to the same extent but an arterial intima extract did not aggregate platelets. Platelet aggregation induced by collagen could be inhibited by mixing with intima extract, but only to a maximum of about 80%. Pre-mixing adenosine diphosphate (ADP) with intima extracts diminished the platelet aggregation activity of the ADP. Depending on the relationship between ADP and intima extract concentrations aggregating activity could either be completely inhibited or inhibition abolished. Incubation of ADP with intima extract and subsequent separation of degradation products by paper chromatography, demonstrated a time-dependent breakdown of ADP with AMP, adenosine, inosine and hypoxanthine as metabolic products; ADP removal was complete. Collagen, thrombin and adrenaline aggregate platelets mainly by endogenous ADP of the release reaction. Results of experiments comparing inhibition of aggregation caused by premixing aggregating agent with intima extract, before exposure to platelets, and the sequential addition of first the intima extract and then aggregating agent to platelets, suggest that the inhibitory effect of intima extract results from ADP breakdown. It is suggested that this ADP degradation by intima extract may play a protective role in vivo by limiting the size of platelet aggregates forming at the site of minimal “wear and tear” vascular trauma.

Storage of Human Blood Platelets

1974 ◽  
Vol 32 (02/03) ◽  
pp. 405-416 ◽  
Author(s):  
M. R Hardeman ◽  
Carina J L. Heynens
Keyword(s):  
Storage Temperature ◽  
Platelet Rich Plasma ◽  
Blood Platelets ◽  
Storage Period ◽  
Serotonin Uptake ◽  
Hypotonic Shock ◽  
Glycolytic Intermediates

SummaryStorage experiments were performed at 4°, 25° and 37° C with platelet-rich plasma under sterile conditions. In some experiments also the effect of storing platelets at 4° C in whole blood was investigated.Before, during and after three days of storage, the platelets were tested at 37° C for their serotonin uptake and response to hypotonic shock. In addition some glycolytic intermediates were determined.A fair correlation was noticed between the serotonin uptake and hypotonic shock experiments. Both parameters were best maintained at 25° C. Also platelet counting, performed after the storage period, indicated 25° C as the best storage temperature. Determination of glycolytic intermediates did not justify any conclusion regarding the optimal storage temperature. Of the various anticoagulants studied, ACD and heparin gave the best results as to the serotonin uptake and hypotonic shock response, either with fresh or stored platelets. The use of EDTA resulted in the lowest activity, especially after storage.The results of these storage experiments in vitro, correspond well with those in vivo reported in the literature.

Is Quantitative Determination of Fibrin(ogen) Degradation Products and Thrombin-Antithrombin III Complexes Useful to Diagnose Deep Venous Thrombosis in Outpatients?

1989 ◽  
Vol 62 (04) ◽  
pp. 1043-1045 ◽  
Author(s):  
Paul F M M van Bergen ◽  
Eduard A R Knot ◽  
Jan J C Jonker ◽  
Auke C de Boer ◽  
Moniek P M de Maat
Keyword(s):  
Quantitative Determination ◽  
Venous Thrombosis ◽  
Deep Venous Thrombosis ◽  
Antithrombin Iii ◽  
Degradation Products ◽  
Family Doctor ◽  
Diagnostic Value ◽  
Impedance Plethysmography ◽  
Fibrin Degradation Products

SummaryWe studied the diagnostic value of recently introduced ELISA’s for the determination of thrombin-antithrombin III (TAT) complexes, fibrin degradation products (FbDP), fibrinogen degradation products (FgDP) and total degradation products (TDP) for deep venous thrombosis (DVT) in plasma of 239 consecutive outpatients, suspected for DVT by their family doctor. DVT was confirmed by impedance plethysmography in 60 patients. Using the 95th percentile range of 42 healthy volunteers the sensitivity for the detection of DVT was: 37% for TAT, 95% for TDP, 92% for FbDP and 90% for FgDP. Specificity was: 88% for TAT, 16% for TDP, 20% for FbDP and 25% for FgDP.We conclude that these assays are of little value in the diagnosis of DVT in outpatients.

Comparison of High Purity Factor IX Concentrates and a Prothrombin Complex Concentrate in a Canine Model of Thrombogenicity

1991 ◽  
Vol 66 (05) ◽  
pp. 609-613 ◽  
Author(s):  
I R MacGregor ◽  
J M Ferguson ◽  
L F McLaughlin ◽  
T Burnouf ◽  
C V Prowse
Keyword(s):  
High Purity ◽  
Time Course ◽  
Prothrombin Complex Concentrate ◽  
Degradation Products ◽  
Canine Model ◽  
Factor Ix ◽  
In Vitro Tests ◽  
Albumin Infusion

SummaryA non-stasis canine model of thrombogenicity has been used to evaluate batches of high purity factor IX concentrates from 4 manufacturers and a conventional prothrombin complex concentrate (PCC). Platelets, activated partial thromboplastin time (APTT), fibrinogen, fibrin(ogen) degradation products and fibrinopeptide A (FPA) were monitored before and after infusion of concentrate. Changes in FPA were found to be the most sensitive and reproducible indicator of thrombogenicity after infusion of batches of the PCC at doses of between 60 and 180 IU/kg, with a dose related delayed increase in FPA occurring. Total FPA generated after 100-120 IU/kg of 3 batches of PCC over the 3 h time course was 9-12 times that generated after albumin infusion. In contrast the amounts of FPA generated after 200 IU/kg of the 4 high purity factor IX products were in all cases similar to albumin infusion. It was noted that some batches of high purity concentrates had short NAPTTs indicating that current in vitro tests for potential thrombogenicity may be misleading in predicting the effects of these concentrates in vivo.

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