Questions related to Tobacco
In 2004, the Himalayan democracy of Bhutan embarked on a near-total ban on the manufacture, sale, and promotion of tobacco products. Modern-day neo-prohibitionists argue and assert that this approach to eliminate tobacco supply and demand will result in what they call a “tobacco-free society.” The empirical and scientific evidence from 2004 to 2021 in Bhutan indicates that the “tobacco-free society” approach has not worked. Instead, as determined in the peer-reviewed article ‘History of Bhutan’s Prohibition of Cigarettes: Implications for Neo-Prohibitionist and Their Critics.’ This has resulted in continued and significant tobacco use and robust black-market smuggling.
In 2021, as noted in the below article 'Bhutan Banned Smoking and It Didn't Go So Well,' Bhutan ended tobacco neo-prohibitionism due to significant ongoing tobacco use and black-market smuggling.
Instead, Bhutan embarked on peer-reviewed and scientifically certified anti-tobacco counter-marketing efforts and nicotine replacement therapies to counter tobacco's severe public health impact.
Is there anybody happy to be the reviewer of my paper (Frontiers in Psychiatry) on smoking frequency and life satisfaction and test whether self-rated health serves as a mediator between this association? I am happy to review back if your manuscript needs a reviewer. Please leave me your name, email address, and institution if you are interested.
Tolin S. A. wrote that "PVY has a wide host range among the Solanaceae, including 70 species of Nicotiana and is also found in annual and perennial weeds in 15 or more plant families. " (Tolin, S. A. (2008). Tobacco Viruses. Encyclopedia of Virology, 60–67. doi:10.1016/b978-012374410-4.00575-6).
But, I cannot find the list of these 15 plant families and their reaction to the infection. Is there any species of Prunis among host plants for PVY?
I want to demonstrate a transcriptional regulation by the coinfiltration with Agrobacterium in N.benthamiana the TF and a promotor sequence. But I don't know which strain of Agro is better for that GV3101 or 3010?
Hello guys, thank you for browsing my question.
I used ++NEBuilder HiFi DNA Assembly Master Mix (NEB-E2621S) to assemble up to 5 fragments products (2058bp insertions to 10953bp vector) and it failed.
I got three false positive colonies in (3 fragments assembling) O.N, and through miniprep got the plasmid and the digestion plus PCR shows bad results.
And after 48h I got a lot of colonies that cannot even grow in starters with antibiotics.
Please guide me the right way to success, thank you in advance!
Long and detailed version:
First of all, my cassette, the target gene is toxic to the dh5a, so I also ordered NEB® 5-alpha F' I q Competent E. coli (High Efficiency) to do the transformation.
I ordered primers which are 25bp long, and I used proofreading enzyme to gain the PCR product.
I digest and get the vector, it is pCAMBIA0390, for agro-transformation in plants.
So I got the PCR products 50ul, I ran 5ul on gel and found there are unspecific bands, so I cleaned both the digested vector and PCR product by the miniprep cleaning kit, then I got a very low concentration (around 2-10 ng/ml). All my PCR products are within 1200bp. And one of the PCR fragment has a lot of GC which is harder to get the PCR product, so I added a GC enhancer.
The biggest insertion (including vector are 5 fragments) is 2058bp, the whole plasmid with my insertions is supposed to be 10953 bp.
I added the whole reaction system as 20ul, and vector:insert = 1:1 according to their concentration (ng/ul). The PCR program was 60 minutes in 50c since I have 5 fragments.
Then I used 2ul and 18ul assembled products to do heat shock transformation to NEB® 5-alpha F' I q Competent E. coli (High Efficiency), and left the plates in several dilutions.
After 24h in 28c, there were 3 colonies appearing in 3 fragments assembling reactions. I grew the starters and did both digestion and PCR, no right products. The agar plates were kanamicin added, after 48h there were a lot of very small colonies that grew, and I tried to grow starter from them with kanamycin antibiotic, there were no colonies able to grow, the starters remained crystal clear.
So I was wondering if you know anything I did wrong or there is better condition for the assembling kit working (like digestion enzyme which acclaimed can work in 15min is always better to digest for 1h; and ligation which could be done within 0.5h but is it always better to do a 16h 16c.)
Thank you and looking forward to hearing from you.
- We expressed a maize CC-NBS-LRR protein in Nicotiana benthamiana by injecting the Agrobacterium strain GV3101 with a maize CNL protein into the leaves, cell death was observed after 3 days without the presence of the cognate pathogen effector, Does this happen often?
Can anyone suggest me how to avoid fungal infection on regeneration media while doing agrobacterium mediated transformation of arabidopsis leaves by co culture method?
For agro removal i prefer to use cefotaxime and that works well but i immediately get fungal contamination hence no further callus initiation. This has happened several times with me in case of tobacco and arabidopsis both.
The carcinogenic effect of smoking was finally proved in the 1960-/1970-ties. However, at that time DDT, Lindane and the like were sprayed onto the tobacco leaves, and the warming- and burning-products of these substances were inhaled by smokers.
Opposed to mice and rats used in trials, humans have been exposed to smoke from plant parts in thousands of generations. Humans are therefore likely to have evolved smoke resistance.
The eyelid could be a ”macro-example”. Has smoke resistance been shown in humans at a molecular level?
(I also asked this question as a discussion: https://www.researchgate.net/post/Is_the_carcinogenic_effect_of_smoking_solely_due_to_pesticides_Is_there_any_evidence_that_ecological_tobacco_causes_cancer_in_humans2
Please consider where your answer is most relevant. You can of course also answer both places.
This will be helpful to the readers of RG)
I have a very troublesome time when I trying to build the construct of my gene. Since the gene sequence of my target insert is toxic due to the leaky expression, I used the competent cell that ordered from NEB company: NEB® 5-alpha F' Iq Competent E. coli (High Efficiency) to reduce this problem, and I encountered trouble when I tried to extract the DNA from the starters.
I used traditional digestion to get my insert and the vector, the I did ligation. After the colonies grew on the ampicillin plats, I did colony PCR by the use of my insert primers, it shows the very good result that my insert is inside of the gene of the colonies' cell.
Then I used the colonies to grow the starters (O.N. in liquid LB+amp.). I also did the starter PCR, but in the comparison of the positive control (the plasmid where I get my insert from), which has strong right size band; the samples shows nothing.
After that, I redid everything, and miniprep (i.e., the DNA extraction). In addition, I digest the extracted plasmid. The intact plasmid looks 2kb smaller compared to what it should be, also the digested bands.
What could be the problem during the process? If the idea that using low-copy number e. coli is not really working, is there any other way that I can use to build the construct with this toxic gene? My aim is to transfer the gene into tobacco and produce protein.
Thank you so much and looking forward to hearing any constructive suggestions.
Good morning to everybody, I am working on article with tobacco and Arabidopsis genomic data and. Does anybody know database, server, or programm where I could find orthologs of Arabidopsis thaliana genes (e.g. At5g05630) in Nicotiana tabacum genome (e.g. Nitab4.5_0001047g0100)?
First picture is vector construction for express eGFP by using Agroinfiltration.
But no fluorescence was detected. So what was the problem.?
I used A. tumefaciens GV3101 strain and pGWB505 Binary gateway vector, and nicotiana benthamiana for inoculate plant.
And I observed inoculated leaf 7days post inoculation.
I guess the vector construction is wrong..
Please give me the advices.
My question is as the title.
I found that I cannot find an exact explanation on what are the bands that I see on this gel. I would suppose some of them are rRNAs, but which one is which?
From what I searched, there are four rRNAs for tobacco leaf, but why I have five bands?
I am looking for few common tobacco pathogens as well as other general plant fungal pathogens. If anyone will be willing to provide it then please contact me.
for my research I was wondering on the fate of glucose if I were to agroinfiltrate a high concentration of Glucose (eg 30mM) into the plant:
- will the glucose be translocated to sinks? I'm aware that normally it does not necessarily get shuttled as the "pure" monosaccharide form.
- If yes, does this only happen at night? Or are there other mechanisms that can operate to regulate the concentration?
Thanks in advance.
Hi I am trying to express my protein R in tobacco leaves and do a CoIP, but I can't detect the protein R in CoIP input.
I took the same tissue and added 8M Urea plus LDS protein buffer, protein R shows as a distinct band at the correct size. But when I grind the tissue and put in CoIP lysis buffer, after centrifugation, I put the input in LDS, I couldn't detect my protein, I only see a smear.
Does anyone know what went wrong?
We know very well that RNA silencing is a potential ways by which small plants such as tomato, tobacco etc protect themselves against viruses. I was just curious to know if RNA silencing is also equally effective against virus infection in big trees? Although, I am also not sure if viruses infect big trees.
Though tobacco is considered to have high levels of toxic compounds, nicotine being the most abundant, from my observations those who chew tobacco have up to 99% not suffering from dental problems. Comparatively with smoking tobacco those who chew are not at risk of lung cancer, or throat cancer.
I am planning a research project for next summer that investigates the effects of high temperature exposure for different lengths of time on the percent nicotine concentration in tobacco leaves. The setup is as follows:
Tobacco growing conditions
3 experimental groups of 10 or more will be grown in a chamber with a peak daytime temperature of 35oC and a minimum nighttime temperature of 26oC.
- group 1: 3 days exposure to heat
- group 2: 5 days exposure to heat
- group 3: 7 days exposure to heat.
3 control groups of 10 or more plants will be grown in a chamber with a peak daytime temperature of 30oC and a minimum nighttime temperature of 21oC.
- group 1: remove after 3 days
- group 2 remove after 5 days
- group 3 remove after 7 days
When the plants are removed from the temperature controlled growth chamber, I will dry the leaves in a 40 c oven until they are dry enough to powder. I plan to extract nicotine from leaf powder with methanol in an ultrasonic bath. Once I filter the resulting solution to remove particulate matter, I need to find percent nicotine. My school has a Hewlett-Packard GC-MS (NSF 9851032) with an autosampler. I am an undergrad with more background in bio than chem, and I am unfamiliar with gc-ms. Could I use this equipment to quantify nicotine in the leaves, or does this equipment only work for identification? If I can use this equipment for quantification, what general procedures should I follow? Any additional advice on methods would be much appreciated as well.
How can I extract nicotine in good yield from tobacco (and the good way for determination of it in extraction)?
I want to know if anyone knows some statistical simulation model that relates tobacco with childhood asthma. Recently we had a publication, but found very little information.
It is difficult to work on crops and chances of successful experiments are very less, that is why we prefer these model organisms to study the mechanism and then apply the same knowledge on other crops. So, according to you, for different traits and developmental studies which is better?
I am trying to produce His-Human-TGFB1 in tobacco leaf tissues using agrobacterium mediated transient expression system. The Purification is based on His-Tag Ni-NTA system. Protein is being purified but 60-80% of eluted protein consists of plant protein.
Conditions are as follows:
Note:His-TGFB1 is hydrophobic in nature; a Dimer; commercially available protein is recommended to reconstitute in Citrate buffer pH=3.0 or 4mM HCL
1. extraction buffer: 50mM Tris or 50mM PBS (pH=6.0), 300mM NaCl, 0.2% tritonX100, 1XPI, 5% glycerol, 10mM Imidazole, No DTT.
2. washing buffer is same as above except No PI, DTT,Triton. Imidazole con 25mM.
3. Elution is in same buffer with 300mM imidazole and no PI, DTT, Triton etc
I am looking forward for the suggestions. All are welcomed.
Thanks in advance
As health care advocates for children, pediatric staff and family medicine have to alert parents and older children about the risk of potential nicotine use among children and teenagers.
Several tobacco promotions may reach children in early age, either through social media or peers.
Many smoking habits and other forms of nicotine use begin in adolescence, hence the vital role of the health care system to prevent and proactively addresse this risk before such health risk problems arise in this vulnerable population.
As health care advocates:
What's the best way to address nicotine prevention among children and parents?
Tobacco is a deadly habit which is very common among all societies yet what we want to ask ourselves is: Does imposing price increments limit the habits among the smokers ?
It is a fair question to ask!
I'm trying to transfer my construct into plant (Nicotiana benthamiana) using Agrobacterium LBA 4044 it grow at the first few week good, but when transfer my explant into selection media with Kanamycin 100mg/L and Timentin320mg/L the edge of explants turn brown dose that mean the Antibiotics very high in concentration.? do know how can I solve this please?
By 2040, according to a new report by the Institute for Health Metrics and Evaluation, the Spanish are expected to have an average lifespan of 85.8 years, outliving even the Japanese, who have long headed the global longevity tables. And outliving those of us in the UK by almost 2.5 years.
Does anyone have a suggestion why this might be in a population noted for its consumption of alcohol and tobacco?
The idea of transferring C4 traits into C3 crops is not new and dates back to early years of this century when Japan Tobacco was granted a US patent for generation of PEP-CK type of C4 cycle in rice. The increasing levels of difficulty in meeting the food and fuel demand however, is again stemming up interest regarding C4 rice. With access to advanced technologies and monetarily self-sufficient international collaborations (like C4 consortium), much of the ground work like molecular tool development, unraveling the genetic infrastructure, collection and study of available genetic variability has been done. But still can we say that we will have first C4 rice cultivars by the next decade or so? or is this required in fact? I mean our C4 cultivars are not operating at full efficiency so why take our cereals C4, instead can't we just silence the photorespiration
Hello every one. I need help please!
I did transient expression in Nicotiana benthamiana using GV3101 Agrobactirum strain. The bacterial cultures which have GFP were co-inﬁltrated with equal amounts of an Agrobacterium suspension of a p19. After 5 days the plants were analysed by confocal microscopy. Unfortunately I couldn't find anything and there was no signal for GFP.
I did this two times now and it did not work at all.
Can anyone please give me any suggestion that might help.
My project is about the specific gene expression in tobacco hairy root tissue and I used NtREL1 promoter to produce transgenic tobacco plants with the ability of root specific expression. But my tobacco plants have problem in regeneration after transformation with the expression construct that contain this promoter. would you please help me in this respect? I would be grateful if you let me know your opinion about this issue.
I am overexpressing a plastial enzyme in tobacco, and recently I saw that the younger leaves of some plants are showing signs of chlorosis (see image).
Although I dont have a lot of experience growing tobacco plants (generally just to infiltrate with Agrobacterium), I am not sure this is a nutrient deficiency.
Any thoughs would be greatly appreciated.
I used agroinfiltration method to express YFP-tagged protein and FLAG-tagged protein in tobacco leaves, and after infiltration I observed the leaves by microscope, and the result showed the YFP protein expressed in the tobacco leaves. Then I tried to extract nucleus protein by various methods, but Western Bolt results showed that the antibody did not catch the protein, so I am wondering if there are some good protocols to extract nucleus protein from tobacco leaves.
I am trying to purify a recombinant protein (~15kD) from Tobacco. I am using Agroinfiltration based expression system. Expression is high, but when it comes to purification, I could not get Protein of Interest rather more non-specific bands. I extract protein in extraction buffer (50mM Tris-Cl, 150mM NaCl, 20mM imidazol, 0.1% tritonX100, 1mM DTT, protease inhibitor), and after getting soluble protein, it is passed through the Ni-NTA column and afterwards washed in (PBS, 30mM imidazole, Protease inhibitor). Final elution is done in PBS+250mM imidazole.
Please suggest me some strategy based on your experience. If someone is expert in this technique, their cooperation will be highly appreciated.
Thanks in Advance
Some time ago we started to have some problems with our BY-2 cell suspension cultures. The cells started to flocculate and form consistent cell masses that are very difficult to pipette when subculturing. We already renewed the medium and components but the phenotype remains. We also tried to start again from callus culture, but we also get the same aggregates. Does anyone ever experienced similar problems?
Thank you in advance.
Anybody has the plant cell line tobacco BY-2 cells in USA?
I need this cell line for experiments. I am Berkeley, and I would be grateful if you could send me the line!
Contact me through ResearchGate!
Does anyone know the molecular weight of component of N.glauca? which contain waxy and oil. I would appreciate it if someone could help to know the molecular weight of N.glauca. Thanks
Nicotine is ingredient of Tobacco/Cigarette-Tobacco. Is it possible to find nicotine and its compound in mixture of water and tobacco.
Recently, I am working on the electrolyte leakage assay in tobacco with leaf discs. When the discs were dissected from the leaf and kept in a tube, I put them in -7°C (salted water) directly for 3 hours. Strange thing is the discs from the same leaf show different electrolyte leakage (some is 10% and some 80%). I do not know why this could happen. Does someone have some experience of working on that and some explanation?
We started preparing our manuscript on "Prevailing social obstacles in keeping tobacco-free homes in urban areas: Realizing ways to overcome challenges", which was fully funded form Institute of Global Tobacco Control (IGTC), USA in 2016
Email Id: firstname.lastname@example.org
I want to identify factors associated with tobacco use among adolescents in a rural setting, and possible risk factors have been categorized into three: socio-demographic, behavioral, and environmental. Will it be right to create three models using hierarchical logistic regression so I can identify which model best predicts the outcome (tobacco initiation)?
I am trying to figure out how many Myzus persicae would infest a potato without top-down control (predator or parasitoid). In some papers, I have read that Myzus persicae is non-gregarious and therefore does not occur in a large number in a host plant. However, from my experience, a tobacco plant could hold quite a number of Myzus persicae and so could a chinese cabbage. Am I wrong to assume that there will be hundreds of aphids when there is no interference? Is Myzus persicae always gregarious or does it depend on the host plant it feeds on?
I am not getting shoot formation from callus. I have used 2-8mg/l BAP and ten times less concentration of IAA. I found transformation in 3-5% of calluses.
Only size of the callus increases in 16 hr light and eight hr dark periods. How could I solve this problem?
Do you know the relation between the pharmaceutical industry and WHO efforts against smokeless tobacco? Are there initiatives against nicotine dependence or only against smokeless tobacco? I am afraid that focusing only on smokeless tobacco opens pharmaceutical industry to dominate the nicotine dependence market (by selling nicotine substitution products) without addressing the key of the problem. Do you know some analysis/publication in this field?
We are trying to regenerate shoot from tobacco callus. for this purpose we have used different concentration of cytokinin. 1-8 mg/l BAp, kinetin, 1-8 mg/l BAP+kinetin and callus was put in 16 hour day and 8 hour night cycle but we didn't find any regeneration. callus become green and just increase in size.
then we use higher concentration of cytokinin that is BAP and very less concentration of auxin (1/4th conc. of cytokinin) that is 2,4-D. still we didn't get shoot regeneration. Conc. of BAP in this case is 2,6 and 8mg/l.
Is there any problem in the selection of phytohormones or there could be other reasons for plants were not regenerated.
kindly help me to solve this problem
I'm search a laboratory assay applicable in field (like immunochromatography) to determining tobacco passive exposure by urinary cotinine, with a cutoff concentration of 1-2 ng/mL.
Genotoxicity of tobacco and alcohol is a well known fact. These are carcinogenic in nature, too. Is there any impact of these on germ cells of human in general and determination of sex in particular ?
Stomatal aperture assays in abaxial epidermal peelings of tobacco leaves were done as described in Allen et al. (1999) and He et al. (2013). Peelings were induced to close all stomata in dark condition, and then a part of samples were kept in darkness, a part of samples were put in light condition to induce stomatal opening, and some samples were put in light with 50 µM of abscisic acid to induce stomatal closing (Schemes of the assay are in the following attached picture). Then I obtained microscopy images (10x, 20x and 40x) at three condition times (2 hours, 4h and 6h). I want to determinate the degree of stomatal aperture at those times in each treatment, but I don't know what kind of parameters I need to measure, and if there is any kind of equation that integrate those parameters. I imagine that important parameters can be: stomatal length and width, and stomatal pore length and with, but how can I end this story?
Thank you very much indeed.
I am a research student at haldia institute of technology (Haldia, east Medinipur, west Bengal, India). I would like to research on tobacco plant (Nicotiana tabacum). So I need tobacco seeds. Can anyone help me to get seeds of tobacco or else anyone can provide some tobacco seeds?
The current growth medium I use contains both NAA and 2'4'DA to prevent differentiation completely. What hormones are required to induce rooting? And what is the exact concentration?
I read some research article related to radioactive effects on using tobacco products in relation to.... I have some doubt kindly clear the following
1.why the 210Pb shows higher activity compare to 210Po in tobacco (cigarette, bidi brands)?
2. why cigarette ash content register more polonium 210 and lead 210 content ?
i am trying to isolate microorganisms from tobacco callus but i didn't find any microbial growth in nutrient agar,PDA with and without antibiotic.
are calluses free from microorganisms or there are some different media used for isolation of microorganisms from calluses.
can anybody have an idea about it.
I am in the middle of a research about analysis of cigarette smoke. And the instrument that i will use needs samples in liquid form.
I have run CAPM,3factor and 4factor regressions but i couldnt find significant results.Have you any idea of another measure?Someone told me to compare my portfolio with the benchmark characteristic that Wermers used to evaluate the mutual fund performance but it looks difficult to match my portfolio characteristics with these of the benchmark..
I need a Arabidopsis/Tobacco Y2H cDNA library to test against two bait protein(Virus proteins). I prepared cDNA library but with it in a Y2H assay there was no interaction seen against my bait proteins. Now I need the tobacco or arabidopsis cDNA library to check whether any interaction would happen with the bait protein.
I have just received fresh samples of tobacco leaves with white mold on them. So I am trying to ascertain the genetic diversity of these "fungi"
My work is on genetic characterization of S. gesnerioides in tobacco and I intend to use SSR markers for characterization studies. I need to check if there could be genetic differences between S. generioides collected from different locations within the same area and across regions since we are getting fresh reports of infestation in tobacco fields. I also need to collect S. gesnerioides from cowpea fields in areas where this strain was reportedly found and compare with the tobacco Striga
I want to transform tobacco by agro bacterium (GV strain), but my problem is that selection plates only contaminate. I used 500 mg/l cefotaxime in plate 250mg for washing but it can't remove it, and after some days the bacteria covers the explants and kills them. If anybody has any experience that can help me please let me know
It is clear that the experience of addiction to tobacco is different from the experience of addiction to alcohol, or something like cocaine. If we agree that addiction is socially constructed while also manifesting a physical or (dare I say) psychological dependency, then in what specific ways are commonly associated addictions (alcohol et. al.) really differentiated from tobacco. Intoxication of an altering of subjective experience may be the key to delineating one from the other. Thoughts?
Many smokers are using waterpipes worldwide, some of them to quit smoking. Smokers seem to perceive waterpipes as less harmful than cigarette smoking and think they will be able to quit waterpipes more easily than cigarettes. What's your experience/opinion about this?
The article reports the results of comprehensive analysis of dynamics in changes of chemical properties of Cavendish tobacco and organoleptic properties of its smoke during the cyclic process of additional thermal fermentation.
I'm looking for database on smokers, not smokers, former smokers by gender, age and type of disease, period 2012 - 2014 for the following countries: France, Switzerland, Austria and Slovenia. Preferably, the samples should have high number: more than 10,000 units.
Someone knows websites from which to download these databases?
I'm looking for database on smokers, not smokers, former smokers by gender, age and type of disease, period 2012 - 2014 for the following countries: France, Switzerland, Austria and Slovenia.
Preferably, the samples should have high number: more than 10,000 units.
Someone knows websites from which to download these databases?