Tetrachlorodibenzodioxin
This polychlorinated dibenzo-p-dioxin compound is a byproduct of various industrial processes and can accumulate in the food chain, posing a significant risk to human and animal health.
Researching the latest methods and protocols for studying Tetrachlorodibenzodioxin is crucial to understanding its mechanisms of action, exposure pathways, and potential mitigation strategies.
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Most cited protocols related to «Tetrachlorodibenzodioxin»
At proestrus as determined by daily vaginal smears, the female rats, (90 days) were pair-mated with male rats (120 days). On the next day, the females were separated and their vaginal smears were examined microscopically and if they were sperm-positive (day 0) the rats were tentatively considered pregnant and then weighed with a digital animal weighing balance to monitor increases in body weight. Vaginal smears were continued for monitoring diestrus status in these rats until day 7. On embryonic day 7 (E-7) these females were weighed to determine if there was a significant increase in (greater than about 10 g) body weight, to confirm pregnancy in sperm-positive females. These pregnant rats were then given daily intraperitoneal injections of any one of the following single chemicals or mixtures with an equal volume of sesame oil (Sigma) on days E-8 through E-14 of gestation [43] (link). Treatment groups were Control, Pesticide (Permethrin+DEET), Plastics (Bisphenol-A, DBP and DEHP), Dioxin (TCDD), and Jet Fuel (JP8 hydrocarbon). The pregnant female rats treated with various mixtures were designated as the F0 generation. When there was a drop in the litter size and the sex ratio of pups in F1 generation of Plastics group, another treatment group was included with only half the dose of Bisphenol-A, DBP and DEHP and this group was designated ‘Low Dose Plastics’ group. Doses, percent of oral LD50, and sources of chemicals for the compounds are given in
Each of the 30 boys, with his mother, was asked to complete a nurse-administered detailed questionnaire on medical history, diet, and lifestyle. The diet questions were used to measure the current and lifetime consumption of locally grown or raised foods. The question was worded, "Does your child eat any of the following foods from local sources (i.e. your own garden or farms or lakes in the Chapaevsk area)? Yes/No". There were separate questions for current intake and lifetime intake of each food item. The distances the boys lived from the Khimprom factory at the time of the study and during pregnancy were assessed by questionnaire based on maternal self-report as <2, 2–6, or >6 kilometers, and the distance at the time of the study was also estimated using ArcView GIS 3.0 mapping of addresses.
Based on the latitude/longitude provided by EPA, 382 facilities were within 10 km of residences in our analysis. We checked the accuracy of these facility locations by comparing the coordinates to locations determined through web-based aerial photographs and ancillary information (Google Inc. Mountain View, CA; Environmental Systems Research Institute, Redlands, CA, USA); locations were corrected if necessary. We verified locations for 340 (89%) facilities and excluded 42 facilities that we could not verify. The median distance between the original and corrected location ranged from 132 meters (coal-fired electric generating facilities) to 23 km (hazardous waste incinerators).
Most recents protocols related to «Tetrachlorodibenzodioxin»
Example 1
Each of the peptides having amino acid sequences of SEQ ID NOS: 1, 2, and 3 mixed with a coating buffer (20 mM sodium phosphate, pH 9.6) at a concentration of 1.8 mM was seeded on a plate for an enzyme-linked immunosorbent assay (ELISA) and cultured at 4° C. overnight. Subsequently, the peptide was washed with phosphate buffered saline with Tween-20 (PBST) and blocked with 3% of bovine serum albumin (BSA) for 2 hours at room temperature. After washing with PBST, 2 μM of 2,3,7,8-tetrachlorodibenzo-p-dioxin (hereinafter, referred to as TCDD) was added to each well and cultured at room temperature for 2 hours. Subsequently, after washing with PBST, treatment with anti-TCDD antibody conjugated with fluorescein isothiocyanate (FITC) was conducted at a ratio of antibody:PBST=1:100 and the resultant was cultured for 2 hours at room temperature. Then, after washing with PBST, an excitation 488 nm/emission 520 nm value was measured using a fluorescence meter, and the results are shown in
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Example 2
HaCaT cells, human keratinocyte cells, were seeded on a 6-well plate at a density of 3×105 cells/well and cultured overnight. Subsequently, 10 nM of TCDD and 50 μM of the peptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3 were added to the culture medium. After 30 minutes of reaction, the cells were treated for 1 hour and collected to obtain nuclei and cytoplasmic proteins separated from each other. Westin blotting was performed using an aryl hydrocarbon receptor (AhR) antibody (Santa Cruz Biotechnology, U.S.A.) to identify activated nuclear translocation of AhR, and the results are shown in
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Example 174
tert-Butyl 6-(8-methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine]-1′-carboxylate. tert-Butyl 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[4H-1,3-benzo-dioxine-2,4′-piperidine]-1′-carboxylate (0.80 g, 1.9 mmol), 7-bromo-8-methoxy-quinoline (0.45 g, 1.9 mmol), palladium(II) acetate (0.024 g, 0.11 mmol) and triphenylphosphine (0.10 g, 0.38 mmol) in 1,4-dioxane (30 mL), DMF (50 mL) was added aq. Na2CO3 (0.5 M) (6.0 mL, 3.0 mmol). The mixture was vacuum degassed then heated at 85° C. overnight. The mixture was diluted with EtOAc (200 mL) and water (100 mL) and extracted. The aqueous extract was washed with EtOAc (50 mL) and the combined organics were dried over Na2SO4, filtered and concentrated. The residue was dissolved in DCM, applied to a silica gel loading cartridge (5 g) and purified on silica gel (80 g, 0-40% EtOAc:hexanes) to afford tert-butyl 6-(8-methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine]-1′-carboxylate (0.38 g, 0.82 mmol, 43% Yield). LCMS m/z=463.
Step 2.
6-(8-Methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine]. A mixture of tert-butyl 6-(8-methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine]-1′-carboxylate (0.38 g, 0.82 mmol) and TFA (0.5 mL, 7 mmol) in DCM (10 mL) was stirred at RT for 24 h, then was diluted with DCM (20 mL) and NaOH (1M, 24 mL). The layers were separated and the aqueous phase was further extracted with DCM (2×20 mL). The combined organics were filtered through a phase separator, then dried over Na2SO4, filtered, and concentrated in vacuo to give a white foam. A small amount (50 mg) was purified by preparative HPLC to afford 6-(8-methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine] TFA salt (20 mg). Analysis: LCMS m/z=363; 1H NMR (400 MHz, DMSO-d6) δ: 9.02 (dd, J=4.4, 1.6 Hz, 1H), 8.71 (br s, 2H), 8.57 (d, J=7.5 Hz, 1H), 7.87 (d, J=8.5 Hz, 1H), 7.71-7.64 (m, 2H), 7.54 (dd, J=8.5, 2.3 Hz, 1H), 7.43 (d, J=2.0 Hz, 1H), 7.03 (d, J=8.5 Hz, 1H), 5.00 (s, 2H), 3.88 (s, 3H), 3.29-3.16 (m, 4H), 2.19-2.06 (m, 4H). The remainder was used in the next step without further purification.
Step 3.
6-(8-Methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine]-1′-carboxamide. A mixture of 6-(8-methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine] (0.198 g, 0.546 mmol), trimethylsilyl isocyanate (0.30 mL, 1.9 mmol), DIPEA (0.50 mL, 2.9 mmol), and DCM (10.0 mL) was stirred overnight. The solution was concentrated and the resulting material was diluted with DCM an put on a 5 g preload silica gel. The material was purified on silica gel chromatography (24 g, 0-10% EtOAc:hexanes) to afford 6-(8-methoxy-7-quinolyl)spiro[4H-1,3-benzodioxine-2,4′-piperidine]-1′-carboxamide (0.183 g, 0.451 mmol, 83%) as an off-white solid. Analysis: LCMS m/z=406 (M+1); 1H NMR (400 MHz, DMSO-d6) δ 8.95 (dd, J=4.0, 1.8 Hz, 1H), 8.38 (dd, J=8.3, 1.8 Hz, 1H), 7.77 (d, J=8.5 Hz, 1H), 7.61-7.47 (m, 3H), 7.36 (d, J=2.3 Hz, 1H), 6.97 (d, J=8.5 Hz, 1H), 6.04 (s, 2H), 4.95 (s, 2H), 3.94 (s, 3H), 3.52-3.37 (m, 4H), 1.91-1.77 (m, 3H), 1.88-1.77 (m, 1H).
Example 4
HaCaT cells, human keratinocyte cells, were seeded on a 6-well plate at a density of 3×105 cells/well and cultured overnight. Subsequently, 10 nM of TCDD and 50 μM of the peptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3 were added to the culture medium. After 30 minutes of reaction, the cells were treated for 24 hours and further treated with DCFH-DA for 30 minutes. Then, the cells were collected and subjected to FACS analysis to observe changes of average FL1 values, and the results are shown in
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Example 3
HaCaT cells, human keratinocyte cells, were seeded on a 6-well plate at a density of 3×105 cells/well and cultured overnight. Subsequently, 50 nM of TCDD and 50 μM of the peptide consisting of an amino acid sequence of SEQ ID NO: 1, 2, or 3 were added to the culture medium. After 30 minutes of reaction, the cells were treated for 5 minutes and immobilized with 4% paraformaldehyde for 30 minutes. Then, after washing three times, the cells were reacted with 0.5% Triton X-100 for 15 minutes and washed three times. Subsequently, the cells were blocked with 3% BSA for 1 hour and reacted with a primary antibody against TCDD conjugated with fluorescein isothiocyanate (FITC) (1:100) at 4° C. overnight. The cells were stained and mounted with 4,6-diamidino-2-phenylindole (DAPI) and observed with a fluorescence microscope. The results are shown in
As shown in
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More about "Tetrachlorodibenzodioxin"
This polychlorinated dibenzo-p-dioxin compound is a byproduct of various industrial processes and can accumulate in the food chain, posing a significant risk to human and animal health.
TCDD is also known as 2,3,7,8-Tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and dioxin.
Researching the latest methods and protocols for studying TCDD is crucial to understanding its mechanisms of action, exposure pathways, and potential mitigation strategies.
The use of solvents like DMSO, cell culture media containing FBS, and transfection reagents like Lipofectamine 2000 are common in TCDD research.
Molecular biology techniques like RNA extraction using TRIzol and RNeasy kits, as well as the addition of L-glutamine, are often employed.
Studying the effects of TCDD in comparison to other toxic compounds like Benzo[a]pyrene can provide valuable insights.
PubCompare.ai's AI-driven platform can help locate top protocols from literature, preprints, and patents, while providing intelligent comparisons to enhance reproducibility and accuracy.
Discover the optimal research methods and products for your TCDD studies and experienece the future of scientific discovery today.