scholarly journals Application of an electrometric method for measurement of in vitroinhibition of blood cholinesterases from sheep, goats and cattle by dichlorvos and carbaryl

2012 ◽  
Vol 51 (No. 2) ◽  
pp. 45-50
Author(s):  
Mohammad FK ◽  
Al-Baggou BK ◽  
Alias AS ◽  
Faris GA-M
Keyword(s):  
Cholinesterase Inhibition ◽  
Experimental Protocol ◽  
Electrometric Method ◽  
Before And After ◽  
Acetylthiocholine Iodide ◽  
Initial Ph ◽  
Cholinesterase Activities ◽  
In Vitro Inhibition ◽  
Substrate Addition

A modified electrometric method was described in sheep, goats and cattle and used to demonstrate in vitro inhibition of plasma and erythrocyte cholinesterase activities by the organophosphate and carbamate insecticides dichlorvos and carbaryl, respectively. A typical reaction mixture for the measurement of cholinesterase activity contained 3 ml distilled water, 3 ml barbital-phosphate buffer (pH 8.1), 0.2 ml plasma or erythrocytes and 0.1 ml acetylthiocholine iodide (7.5%) as a substrate. The mixture was incubated at 37<sup>&omicron;</sup>C for 30 min in sheep, 40&nbsp;min in goats and 20 min in cattle. The pH of the reaction mixture was determined by a pH meter before and after the incubation. The initial pH was measured before the substrate addition. The enzyme activity was expressed as ∆pH/incubation time = (pH1 &ndash; pH2) &ndash; ∆pH of blank. The method of inhibitor-cholinesterase incubation was used to measure the in vitro inhibition of plasma and erythrocyte cholinesterase activities. Dichlorvos in concentrations of 0.5 and 1 &mu;m inhibited plasma and erythrocyte cholinesterase activities by 24&ndash;85%, whereas carbaryl in concentrations of 5 and 10 &mu;m inhibited them by 50&ndash;89%. The results suggest that the described electrometric method could be efficiently used for detecting cholinesterase inhibition in ruminants, and further point to the value of the present experimental protocol of in vitro cholinesterase inhibition in preliminary toxicological examinations of anticholinesterase compounds

In Vitro Inhibition of Blood Cholinesterase Activities From Horse, Cow, and Rat by Tetrachlorvinphos

2003 ◽  
Vol 22 (6) ◽  
pp. 429-433 ◽  
Author(s):  
Subramanya Karanth ◽  
Carey Pope
Keyword(s):  
Rat Plasma ◽  
Organophosphorus Insecticide ◽  
Lower Sensitivity ◽  
Livestock Species ◽  
In Vitro Sensitivity ◽  
Horse Plasma ◽  
Cholinesterase Activities ◽  
In Vitro Inhibition ◽  
Blood Fraction

The organophosphorus insecticide tetrachlorvinphos (TCVP) is commonly used as a feed-through larvicide in many livestock species, including cattle and horses. Cholinesterase (Ch E) activity in blood (generally plasma or whole blood) is often employed to assess organophosphorus insecticide intoxication in animals as well as humans. In many species, including horse and man, plasma contains predominantly butyrylcholinesterase whereas red blood cells in all species express exclusively acetylcholinesterase. To evalulate the comparative interaction of TCVP with blood Ch Es in different species, we compared the in vitro sensitivity of Ch E activity in plasma and erythrocytes from horse, cow, and rat. Horse plasma Ch E was most sensitive (IC50, 30 minutes, 30°C = 97 n M), whereas horse erythrocyte Ch E activity was least sensitive (IC50 > 1 m M). In contrast, cow plasma Ch E showed lower sensitivity (IC50 = 784 μM) to inhibition by TCVP than erythrocyte Ch E (IC50 = 216 μM). Rat plasma and erythrocyte Ch E activities had relatively similar sensitivity to TCVP (IC50 = 54 μM and 78 μM, respectively). The results suggest that plasma and erythrocyte Ch E from horse, cow, and rat show marked species- and blood fraction-dependent differences in sensitivity to TCVP. Knowledge of such differences in sensitivity of blood Ch E activities to TCVP may be important in the clinical interpretation of intoxication with this pesticide across species.

scholarly journals Plasma and whole brain cholinesterase activities in three wild bird species in Mosul, IRAQ: In vitro inhibition by insecticides

2011 ◽  
Vol 4 (3) ◽  
pp. 144-148 ◽  
Author(s):  
Ashraf Alias ◽  
Muna Al-Zubaidy ◽  
Yaareb Mousa ◽  
Fouad Mohammad
Keyword(s):  
Wild Bird ◽  
Bird Species ◽  
Wild Birds ◽  
Whole Brain ◽  
Electrometric Method ◽  
Northern Iraq ◽  
Carbamate Insecticides ◽  
Brain Cholinesterase ◽  
Cholinesterase Activities

Plasma and whole brain cholinesterase activities in three wild bird species in Mosul, IRAQ:In vitroinhibition by insecticidesPlasma and brain cholinesterase activities were determined in three wild bird species to assess their exposure to organophosphate and carbamate insecticides which are used in agriculture and public health. In the present study, we used an electrometric method for measurement of cholinesterase activities in the plasma and whole brain of three indigenous wild birds commonly found in northern Iraq. The birds used were apparently healthy adults of both sexes (8 birds/species, comprising 3-5 from each sex) of quail (Coturnix coturnix), collard dove (Streptopelia decaocto) and rock dove (Columba livia gaddi), which were captured in Mosul, Iraq. The mean respective cholinesterase activities (Δ pH/30 minutes) in the plasma and whole brain of the birds were as follows: quail (0.96 and 0.29), collard dove (0.97and 0.82) and rock dove (1.44 and 1.42). We examined the potential susceptibility of the plasma or whole brain cholinesterases to inhibition by selected insecticides. The technique ofin vitrocholinesterase inhibition for 10 minutes by the organophosphate insecticides dichlorvos, malathion and monocrotophos (0.5 and 1.0 μM) and the carbamate insecticide carbaryl (5 and10 μM) in the enzyme reaction mixtures showed significant inhibition of plasma and whole brain cholinesterase activities to various extents. The data further support and add to the reported cholinesterase activities determined electrometrically in wild birds in northern Iraq. The plasma and whole brain cholinesterases of the birds are highly susceptible to inhibition by organophosphate and carbamate insecticides as determined by the described electrometric method, and the results further suggest the usefulness of the method in biomonitoring wild bird cholinesterases.

scholarly journals Acute Toxicity of Veterinary and Agricultural Formulations of Organophosphates Dichlorvos and Diazinon in Chicks

2011 ◽  
Vol 62 (4) ◽  
pp. 317-323 ◽  
Author(s):  
Muna Al-Zubaidy ◽  
Yaareb Mousa ◽  
Mohammad Hasan ◽  
Fouad Mohammad
Keyword(s):  
Acute Toxicity ◽  
Toxic Action ◽  
Cholinesterase Inhibition ◽  
Plasma Samples ◽  
Brain Cholinesterase ◽  
Cholinesterase Activities ◽  
The Brain ◽  
Lethal Doses

Acute Toxicity of Veterinary and Agricultural Formulations of Organophosphates Dichlorvos and Diazinon in ChicksFormulation components of organophosphate insecticidal preparations might affect their toxic action in animals. The objective of this study was to examine and compare the acute toxicity and cholinesterase inhibition in seven to 14-day-old chicks dosed orally with dichlorvos and diazinon in standard veterinary and agricultural formulations. The acute (24 h) oral median lethal doses (LD50) of the formulations were determined using the up-and-down method. Respective LD50 of dichlorvos of the veterinary and agricultural formulations in chicks were 11.1 mg kg-1 and 6.51 mg kg-1 and those of diazinon 6.4 mg kg-1 and 6.7 mg kg-1. Plasma and brain cholinesterase activities were measured by electrometry after in vivo and in vitro exposure to organophosphates. The chicks showed signs of cholinergic toxicosis within one hour of dosing. Dichlorvos (8 mg kg-1) and diazinon (4 mg kg-1) in the veterinary and agricultural formulation significantly reduced both plasma and brain cholinesterase activities in the chicks. The veterinary formulation of dichlorvos reduced plasma ChE by 60 % and agricultural by 40 % and brain ChE by 93 % and 87 %, respectively. In contrast, ChE inhibition by diazinon in the agricultural formulation of diazinon was stronger than by the veterinary formulation; 72 % vs. 64 % in plasma and 97 % vs. 80 % in the brain, respectively. The highest in vitro inhibitions were observed with dichlorvos in the agricultural formulation (50 %) in the brain samples and with diazinon in the agricultural formulation (52 %) in the plasma samples. While they exist, differences between formulations cannot be taken as a rule and further investigations should inventory the toxicity of standard veterinary and agricultural organophosphate formulations in addition to the known data for pure forms.

Electron microscopic study of the membrane glycoproteins in the rat kidney after cisplatin treatment

1989 ◽  
Vol 47 ◽  
pp. 912-913
Author(s):  
S.K. Aggarwal
Keyword(s):  
Alkaline Phosphatase ◽  
Rat Kidney ◽  
Light And Electron Microscopy ◽  
Kidney Tissue ◽  
Membrane Glycoproteins ◽  
Electron Microscopic ◽  
Before And After ◽  
Interstrand Cross Links ◽  
Cross Links

The proposed primary mechanism of action of the anticancer drug cisplatin (Cis-DDP) is through its interaction with DNA, mostly through DNA intrastrand cross-links or DNA interstrand cross-links. DNA repair mechanisms can circumvent this arrest thus permitting replication and transcription to proceed. Various membrane transport enzymes have also been demonstrated to be effected by cisplatin. Glycoprotein alkaline phosphatase was looked at in the proximal tubule cells before and after cisplatin both in vivo and in vitro for its inactivation or its removal from the membrane using light and electron microscopy.Outbred male Swiss Webster (Crl: (WI) BR) rats weighing 150-250g were given ip injections of cisplatin (7mg/kg). Animals were killed on day 3 and day 5. Thick slices (20-50.um) of kidney tissue from treated and untreated animals were fixed in 1% buffered glutaraldehyde and 1% formaldehyde (0.05 M cacodylate buffer, pH 7.3) for 30 min at 4°C. Alkaline phosphatase activity and carbohydrates were demonstrated according to methods described earlier.

Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay

1994 ◽  
Vol 52 ◽  
pp. 270-271
Author(s):  
Henry H. Eichelberger ◽  
John G. Baust ◽  
Robert G. Van Buskirk
Keyword(s):  
Cold Storage ◽  
Structural Integrity ◽  
Culture Media ◽  
Cell Structure ◽  
Natural State ◽  
Sodium Cacodylate ◽  
Ruthenium Tetroxide ◽  
Cacodylate Buffer ◽  
Before And After

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.

Inhibitory effects of bioaccessible anthocyanins and procyanidins from apple, red grape, cinnamon on α-amylase, α-glucosidase and lipase

2020 ◽  
pp. 1-9
Author(s):  
Pınar Ercan ◽  
Sedef Nehir El
Keyword(s):  
Inhibitory Activity ◽  
Digestive Enzymes ◽  
Positive Control ◽  
Anthocyanin Content ◽  
Inhibitory Effects ◽  
Total Anthocyanin ◽  
Total Anthocyanin Content ◽  
Before And After ◽  
Red Grape

Abstract. The goals of this study were to determine and evaluate the bioaccessibility of total anthocyanin and procyanidin in apple (Amasya, Malus communis), red grape (Papazkarası, Vitis vinifera) and cinnamon (Cassia, Cinnamomum) using an in vitro static digestion system based on human gastrointestinal physiologically relevant conditions. Also, in vitro inhibitory effects of these foods on lipid (lipase) and carbohydrate digestive enzymes (α-amylase and α-glucosidase) were performed with before and after digested samples using acarbose and methylumbelliferyl oleate (4MUO) as the positive control. While the highest total anthocyanin content was found in red grape (164 ± 2.51 mg/100 g), the highest procyanidin content was found in cinnamon (6432 ± 177.31 mg/100 g) (p < 0.05). The anthocyanin bioaccessibilities were found as 10.2 ± 1%, 8.23 ± 0.64%, and 8.73 ± 0.70% in apple, red grape, and cinnamon, respectively. The procyanidin bioaccessibilities of apple, red grape, and cinnamon were found as 17.57 ± 0.71%, 14.08 ± 0.74% and 18.75 ± 1.49%, respectively. The analyzed apple, red grape and cinnamon showed the inhibitory activity against α-glucosidase (IC50 544 ± 21.94, 445 ± 15.67, 1592 ± 17.58 μg/mL, respectively), α-amylase (IC50 38.4 ± 7.26, 56.1 ± 3.60, 3.54 ± 0.86 μg/mL, respectively), and lipase (IC50 52.7 ± 2.05, 581 ± 54.14, 49.6 ± 2.72 μg/mL), respectively. According to our results apple, red grape and cinnamon have potential to inhibit of lipase, α-amylase and α-glucosidase digestive enzymes.

Radiolabeled r-Hirudin as a Measure of Thrombin Activity at, or within, the Rabbit Aorta Wall In Vitro and In Vivo

1994 ◽  
Vol 71 (04) ◽  
pp. 499-506 ◽  
Author(s):  
Mark W C Hatton ◽  
Bonnie Ross-Ouellet
Keyword(s):  
Rabbit Aorta ◽  
Balloon Injury ◽  
Recombinant Hirudin ◽  
Aorta Wall ◽  
Thrombin Activity ◽  
Before And After ◽  
The Matrix

SummaryThe behavior of 125I-labeled recombinant hirudin towards the uninjured and de-endothelialized rabbit aorta wall has been studied in vitro and in vivo to determine its usefulness as an indicator of thrombin activity associated with the aorta wall. Thrombin adsorbed to either sulfopropyl-Sephadex or heparin-Sepharose bound >95% of 125I-r-hirudin and the complex remained bound to the matrix. Binding of 125I-r-hirudin to the exposed aorta subendothelium (intima-media) in vitro was increased substantially if the tissue was pre-treated with thrombin; the quantity of l25I-r-hirudin bound to the de-endothelialized intima-media (i.e. balloon-injured in vitro) correlated positively with the quantity of bound 131I-thrombin (p <0.01). Aortas balloon-injured in vivo were measured for thrombin release from, and binding of 125I-r-hirudin to, the de-endothelialized intimal surface in vitro; 125I-r-hirudin binding correlated with the amount of active thrombin released (p <0.001). Uptake of 125I-r-hirudin by the aorta wall in vivo was proportional to the uptake of 131I-fibrinogen (as an indicator of thrombin activity) before and after balloon injury. After 30 min in the circulation, specific 125I-r-hirudin binding to the uninjured and de-endo- thelialized (at 1.5 h after injury) aorta wall was equivalent to 3.4 (± 2.5) and 25.6 (±18.1) fmol of thrombin/cm2 of intima-media, respectively. Possibly, only hirudin-accessible, glycosaminoglycan-bound thrombin is measured in this way.

Effekte von Substanz P und Neurotensin auf die Kontraktilität des Ileums der Maus in vitro: Inhibition durch STW 5

2007 ◽  
Vol 45 (08) ◽  
Author(s):  
D Hagelauer ◽  
O Kelber ◽  
D Weiser ◽  
S Laufer ◽  
H Heinle
Keyword(s):  
Substanz P ◽  
In Vitro Inhibition
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