scholarly journals Non-invasive ploidy determination in live fish by measuring erythrocyte size in capillaries

2021 ◽  
Vol 51 (3) ◽  
pp. 275-280
Author(s):  
Karel Halačka ◽  
Karel Janko ◽  
Lukáš Vetešník
Keyword(s):  
Flow Cytometry ◽  
Rapid Determination ◽  
Minimum Size ◽  
Gibel Carp ◽  
Small Fish ◽  
Reliable Determination ◽  
Live Fish ◽  
Blood Smears ◽  
The Individual

Information about ploidy is important in both commercial and conservation aquaculture and fish research. Unfortunately, methods for its determination, such as karyology, determination of the amount of DNA in a cell using microdensitometry or flow cytometry and/or measuring erythrocytes in a blood smear can be stressful or even destructive. Some of these methods are also limited by the relatively large minimum size of the individual being measured. The aim of this study was to test a new low-stress method of determining ploidy by measuring the size of erythrocytes in the capillaries of a fish, including small individuals. First, we examined diploid and triploid loach (Cobitis sp.) and gibel carp, Carassius gibelio (Bloch, 1782), using flow cytometry and blood smears, with these results being used as a control. Subsequently, we measured the size of erythrocytes in the caudal fin capillaries of anesthetized fishes of known ploidy under a light microscope. For both the loaches and gibel carp, direct observation of the mean erythrocyte size in epithelial fin capillaries provided a consistent and reliable determination of ploidy when compared with the controls based on flow cytometry and blood smears. This new method allows for rapid determination of ploidy in living small fish, where collection of tissue using other methods may cause excessive stress or damage. The method outlined here simply requires the measurement of erythrocytes directly in the bloodstream of a live fish, thereby making it possible to determine ploidy without the need for blood sampling. The method described is sufficiently efficient, less demanding on equipment than many other procedures, can be used by relatively inexperienced personnel and has benefits as regards animal welfare, which is especially important for fish production facilities or when dealing with rare or endangered species.

scholarly journals Non-invasive ploidy determination in live fish by measuring erythrocyte size in capillaries

2021 ◽  
Vol 51 (3) ◽  
pp. 275-280
Author(s):  
Karel Halačka ◽  
Karel Janko ◽  
Lukáš Vetešník
Keyword(s):  
Flow Cytometry ◽  
Rapid Determination ◽  
Minimum Size ◽  
Gibel Carp ◽  
Small Fish ◽  
Reliable Determination ◽  
Live Fish ◽  
Blood Smears ◽  
The Individual

Information about ploidy is important in both commercial and conservation aquaculture and fish research. Unfortunately, methods for its determination, such as karyology, determination of the amount of DNA in a cell using microdensitometry or flow cytometry and/or measuring erythrocytes in a blood smear can be stressful or even destructive. Some of these methods are also limited by the relatively large minimum size of the individual being measured. The aim of this study was to test a new low-stress method of determining ploidy by measuring the size of erythrocytes in the capillaries of a fish, including small individuals. First, we examined diploid and triploid loach (Cobitis sp.) and gibel carp, Carassius gibelio (Bloch, 1782), using flow cytometry and blood smears, with these results being used as a control. Subsequently, we measured the size of erythrocytes in the caudal fin capillaries of anesthetized fishes of known ploidy under a light microscope. For both the loaches and gibel carp, direct observation of the mean erythrocyte size in epithelial fin capillaries provided a consistent and reliable determination of ploidy when compared with the controls based on flow cytometry and blood smears. This new method allows for rapid determination of ploidy in living small fish, where collection of tissue using other methods may cause excessive stress or damage. The method outlined here simply requires the measurement of erythrocytes directly in the bloodstream of a live fish, thereby making it possible to determine ploidy without the need for blood sampling. The method described is sufficiently efficient, less demanding on equipment than many other procedures, can be used by relatively inexperienced personnel and has benefits as regards animal welfare, which is especially important for fish production facilities or when dealing with rare or endangered species.

Intraoperative flow cytometry analysis of glioma tissue for rapid determination of tumor presence and its histopathological grade

2013 ◽  
Vol 118 (6) ◽  
pp. 1232-1238 ◽  
Author(s):  
Takahiro Shioyama ◽  
Yoshihiro Muragaki ◽  
Takashi Maruyama ◽  
Takashi Komori ◽  
Hiroshi Iseki
Keyword(s):  
Flow Cytometry ◽  
Predictive Value ◽  
Rapid Determination ◽  
Characteristic Curve ◽  
Biopsy Sample ◽  
Surgical Biopsy ◽  
Diagnostic Efficacy ◽  
Histopathological Grade ◽  
Number Of Cells

Object Intraoperative histopathological investigation plays an important role during surgery for gliomas. To facilitate the rapid characterization of resected tissue, an original technique of intraoperative flow cytometry (iFC) was established. The objective in this study was evaluation of this technique's efficacy for rapidly determining tumor presence in the surgical biopsy sample and WHO histopathological grade of the neoplasm. Methods In total, 328 separate biopsy specimens obtained during the resection of 81 intracranial gliomas were analyzed with iFC. The evaluated malignancy index (MI) was defined as the ratio of the number of cells with greater than normal DNA content to the total number of cells. The duration of iFC in all cases was approximately 10 minutes. Each sample was additionally investigated histopathologically on frozen and permanent formalin-fixed paraffin-embedded tissue sections. The latter process was used as a “gold standard” control for evaluation of the diagnostic efficacy of iFC analysis. Results The MI differed significantly between neoplastic and perilesional brain tissue (25.3% ± 22.0% vs 4.6% ± 2.6%, p < 0.01). Receiver operating characteristic curve analysis revealed a corresponding area under the curve value of 0.941. The optimal cutoff level of the MI for identification of tumor in the biopsy specimen was 6.8%, which provided 0.88 sensitivity, 0.88 specificity, 0.97 positive predictive value, 0.60 negative predictive value, and 0.88 diagnostic accuracy. Additionally, the MI showed a significant association with WHO histopathological grades of glioma (p < 0.01), but its values in Grade II, III, and IV tumors overlapped prominently and were on average 13.3% ± 11.0%, 35.0% ± 21.8%, and 46.6% ± 23.1%, respectively. Conclusions Results of this study demonstrate that iFC with the determination of the MI may be feasible for rapidly determining glioma presence in a surgical biopsy sample.

scholarly journals Rapid determination of general cell status, cell viability, and optimal harvest time in eukaryotic cell cultures by impedance flow cytometry

2019 ◽  
Vol 103 (20) ◽  
pp. 8619-8629 ◽  
Author(s):  
Christian Opitz ◽  
Grit Schade ◽  
Silvan Kaufmann ◽  
Marco Di Berardino ◽  
Marcel Ottiger ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Cell Cultures ◽  
Rapid Determination ◽  
Eukaryotic Cell ◽  
Harvest Time ◽  
Optimal Harvest

Sweep Co-Distillation Cleanup of Fortified Edible Oils for Determination of Organophosphate and Chlorinated Hydrocarbon Pesticides

1967 ◽  
Vol 50 (3) ◽  
pp. 605-615
Author(s):  
R W Storherr ◽  
Edward J Murray ◽  
I Klein ◽  
Lynn A Rosenberg
Keyword(s):  
Fruits And Vegetables ◽  
Edible Oils ◽  
Rapid Determination ◽  
Organophosphate Pesticides ◽  
Nitrogen Flow ◽  
Distillation Method ◽  
Chlorinated Pesticides ◽  
Chlorinated Hydrocarbon ◽  
The Individual

Abstract The technique of sweep co-distillation developed for organophosphate pesticides in fruits and vegetables was expanded for application to organophosphate and chlorinated hydrocarbon pesticides in edible oils. A modified longer and wider Storherr tube and heating assembly was used in this work and 3 mm diameter glass beads replaced the glass wool packing used in crop work. The work on oils was performed at 248 ± 3°C with a nitrogen flow of 600 ml/min and injection of a sweeping solvent every 3 minutes for 30 minutes. Phosphate pesticides were analyzed by GLC with a potassium thermionic detector and chlorinated pesticides were determined by ECGLC, after a micro Florisil column was used to eliminate extraneous peaks. Recoveries of ten phosphate pesticides from nine edible oils ranged from 74 to 100% at fortification levels of 0.04 to 6.25 ppm; average recoveries for the individual pesticides were 90—99%. Average recoveries for the chlorinated pesticides ranged from a low of 75% for Tedion to a high of 98% for endrin and dieldrin. Sensitivity of 0.01 is readily obtainable for compounds such as parathion and heptachlor cpoxide. The sweep co-distillation method is suitable for the rapid determination of the 14 chlorinated and 10 organophosphate pesticides tested in edible oils

scholarly journals An automated system for selective and continuous measurements of vertical Thoron profiles for the determination of transport times near the ground

2013 ◽  
Vol 6 (1) ◽  
pp. 867-910
Author(s):  
D. Plake ◽  
I. Trebs
Keyword(s):  
Measurement System ◽  
Automated System ◽  
Transport Time ◽  
Reliable Determination ◽  
Plant Canopies ◽  
Highly Sensitive ◽  
Resistance Approach ◽  
Reactive Trace Gases ◽  
The Individual

Abstract. The quantification of in-canopy transport times is of a~major importance for the investigation of sources, sinks and net fluxes of reactive trace gases within plant canopies. The Damköhler number, which compares timescales of chemical reactions with transport times, is a widely applied measure to evaluate flux divergences. In this study, we present and evaluate a novel automated measurement system for selective vertical Thoron (Tn) profiles near the earth's surface and demonstrate its suitability for the direct and reliable determination of transport times within a natural grassland canopy. For the first time, we perform a rigorous determination of systematic and random uncertainties of Tn (and Rn) concentrations under field conditions for this type of measurement system. The obtained median precisions for three concentration classes (> 100 Bq m−3, 100–15 Bq m−3 < 15 Bq m−3) were 8.8%, 23.2% and 132.1% for Tn (and 16.6%, 25.0%, 99.2% for Rn). We calculate in-canopy transport times (τ) and propagate it's uncertainty from the individual errors of the Tn concentration measurements. A quality assessment of τ for the field experiment revealed a good data quality with 44% of the relative uncertainties below 50%. The occurrence of transport time uncertainties higher than 100% was caused by absolute Tn gradients lower than 70 Bq m−3, which was found for 22% of all determined transport times. In addition, the method was found to be highly sensitive to the Tn concentrations at the upper of two inlet heights (zu). Low values of CTnzu result in high absolute uncertainties of the transport time. A comparison of the measured τ with empirical parameterizations revealed the lowest scatter for the measured τ values. We found an excellent agreement of τ with transport times obtained by the in-canopy resistance approach used, e.g. in the SURFATM model during daytime, while the SURFATM model significantly overestimated transport times during nighttime.

scholarly journals Assessment of Enzyme Inhibition: A Review with Examples from the Development of Monoamine Oxidase and Cholinesterase Inhibitory Drugs

2017 ◽  
Author(s):  
Rona R. Ramsay ◽  
Keith F. Tipton
Keyword(s):  
Monoamine Oxidase ◽  
Enzyme Inhibition ◽  
Reliable Determination ◽  
Misleading Information ◽  
Inappropriate Use ◽  
Ic50 Value ◽  
The Individual ◽  
Careful Assessment ◽  
Target Effects

The actions of many drugs involve enzyme inhibition. This is exemplified by the inhibitors of monoamine oxidases (MAO) and the cholinsterases (ChE) that have been used for several pharmacological purposes. This review describes key principles and approaches for the reliable determination of enzyme activities and inhibition as well as some of the methods that are in current use for such studies with these two enzymes. Their applicability and potential pitfalls arising from their inappropriate use are discussed. Since inhibitor potency is frequently assessed in terms of the quantity necessary to give 50% inhibition (the IC50 value), the relationships between this and the mode of inhibition is also considered, in terms of the misleading information that it may provide. Incorporation of more than one functionality into the same molecule to give a multi-target-directed ligands (MTDLs) requires careful assessment to ensure that the specific target effects are not significantly altered and that the kinetic behaviour remains as favourable with the MTDL as it does with the individual components. Such factors will be considered in terms of recently developed MTDLs that combine MAO and ChE inhibitory functions.

scholarly journals Rapid determination of antifungal activity by flow cytometry.

1994 ◽  
Vol 32 (4) ◽  
pp. 1088-1091 ◽  
Author(s):  
L Green ◽  
B Petersen ◽  
L Steimel ◽  
P Haeber ◽  
W Current
Keyword(s):  
Flow Cytometry ◽  
Antifungal Activity ◽  
Rapid Determination

Conformational dependence of solvation energy of phosphates

1981 ◽  
Vol 46 (8) ◽  
pp. 1722-1733 ◽  
Author(s):  
Tomáš Bleha ◽  
Ján Mlýnek ◽  
Igor Tvaroška
Keyword(s):  
Continuous Model ◽  
Solvation Energy ◽  
Molecular Volume ◽  
Water Effect ◽  
Reliable Determination ◽  
Solvent Water ◽  
Dimethyl Phosphate ◽  
The Individual ◽  
Electrostatic Dispersion

Conformational dependence of solvation energy has been calculated for dimethyl phosphate, its monoanion and ion pair with the proton considering rotation about P-O bonds in the molecules. The solvent (water) effect has been expressed using the continuous model according to Sinanoglu. Contribution of individual terms (electrostatic, dispersion and cavity) of the solvation energy in the stabilization of individual conformers has been analyzed. Some of methodical problems of the calculation of solvation energy have been discussed and particularly the inevitability of reliable determination of torsional dependence of molecular volume has been stressed. A comparison of the three molecules has shown that different character of the solvation energy dependences is due to the electrostatic term and is connected with orientation of the group dipole moment of the central PO2 group in the individual molecules.

scholarly journals An automated system for selective and continuous measurements of vertical thoron profiles for the determination of transport times near the ground

2013 ◽  
Vol 6 (4) ◽  
pp. 1017-1030 ◽  
Author(s):  
D. Plake ◽  
I. Trebs
Keyword(s):  
Measurement System ◽  
Automated System ◽  
Transport Time ◽  
Reliable Determination ◽  
Plant Canopies ◽  
Highly Sensitive ◽  
Resistance Approach ◽  
Reactive Trace Gases ◽  
The Individual

Abstract. The quantification of in-canopy transport times is of major importance for the investigation of sources, sinks and net fluxes of reactive trace gases within plant canopies. The Damköhler number, which compares timescales of chemical reactions with transport times, is a widely applied measure to evaluate flux divergences. In this study we present and evaluate a novel automated measurement system for selective vertical thoron (Tn) profiles near the Earth's surface and demonstrate its suitability for the direct and reliable determination of transport times within a natural grassland canopy. For the first time, we perform a rigorous determination of systematic and random uncertainties of Tn (and Rn) concentrations under field conditions for this type of measurement system. The obtained median precisions for three concentration classes (> 100 Bq m−3, 100–15 Bq m−3, < 15 Bq m−3) were 8.8%, 23.2% and 132.1% for Tn (and 16.6%, 25.0%, 99.2% for Rn). We calculate in-canopy transport times (τ) and propagate their uncertainty from the individual errors of the Tn concentration measurements. A quality assessment of τ for the field experiment during a period of 51 days revealed good data quality with 44% of the relative uncertainties below 50%. The occurrence of transport time uncertainties higher than 100% was caused by absolute Tn gradients lower than 70 Bq m−3 m−1, which was found for 22% of all determined transport times. In addition, the method was found to be highly sensitive to the Tn concentrations at the upper of the two inlet heights (zu). Low values of CTnzu result in high absolute uncertainties of the transport time. A comparison with empirical parameterizations revealed a much lower scatter for the τ values determined from our measurements. We found an excellent agreement with τ values obtained by the in-canopy resistance approach used, e.g., in the SURFATM model during daytime, while the SURFATM model significantly overestimated transport times during nighttime.

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