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Tissue Cultured Marijuana ? You Bet !

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#182 is a fine specimen!   thanks for clarifying the chuting for me. I was getting jealous ! lol

 

Have you communicated with others who culture cannabis? in your state or others?

 

There are Tissue Culture Clubs, some cool people, some not so much. Most are not working with cannabis but can provide some rare plant genetics otherwise. I have a collection of hybrid tobacco's from around the world. I maintain it for some reason no sure of, but tobacco callus is crazy easy to work with.   My first was from seed I bought online and the collection grew from there.

 

have you cultured more than cannabis ?

 

peace

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Mitotic inhibitor

 

From Wikipedia, the free encyclopedia

 

The structure of paclitaxel, a widely used mitotic inhibitor.

A mitotic inhibitor is a drug that inhibits mitosis, or cell division. These drugs disrupt microtubules, which are structures that pull the cell apart when it divides. Mitotic inhibitors are used in cancer treatment, because cancer cells are able to grow and eventually spread through the body (metastasize) through continuous mitotic division and so are more sensitive to inhibition of mitosis than normal cells. Mitotic inhibitors are also used in cytogenetics (the study of chromosomes), where they stop cell division at a stage where chromosomes can be easily examined.[1]

 

Mitotic inhibitors are derived from natural substances such as plant alkaloids, and prevent cells from undergoing mitosis by disrupting microtubule polymerization, thus preventing cancerous growth. Microtubules are long, ropelike proteins that extend through the cell and move cellular components around. Microtubules are long polymers made of smaller units (monomers) of the protein tubulin. Microtubules are created during normal cell functions by assembling (polymerizing) tubulin components, and are disassembled when they are no longer needed. One of the important functions of microtubules is to move and separate chromosomes and other components of the cell for cell division (mitosis). Mitotic inhibitors interfere with the assembly and disassembly of tubulin into microtubule polymers. This interrupts cell division, usually during the mitosis (M) phase of the cell cycle when two sets of fully formed chromosomes are supposed to separate into daughter cells.[2][3]

 

Examples of mitotic inhibitors frequently used in the treatment of cancer include paclitaxel, docetaxel, vinblastine, vincristine, and vinorelbine.[1] Colchicine and griseofulvin are mitotic inhibitors used in the treatment of gout and toenail fungus, respectively.

Since chromosome segregation is driven by microtubules, colchicine is also used for inducing polyploidy in plant cells during cellular division by inhibiting chromosome segregation during meiosis; half the resulting gametes, therefore, contain no chromosomes, while the other half contains double the usual number of chromosomes (i.e., diploid instead of haploid, as gametes usually are), and lead to embryos with double the usual number of chromosomes (i.e., tetraploid instead of diploid). While this would be fatal in most higher animal cells, in plant cells it is not only usually well tolerated, but also frequently results in larger, hardier, faster-growing, and in general more desirable plants than the normally diploid parents; for this reason, this type of genetic manipulation is frequently used in breeding plants commercially.

 

When such a tetraploid plant is crossed with a diploid plant, the triploid offspring are usually sterile (unable to produce fertile seeds or spores), although many triploids can be propagated vegetatively. Growers of annual triploid plants not readily propagated must buy fresh seed from a supplier each year. Many sterile triploid plants, including some tree and shrubs, are becoming increasingly valued in horticulture and landscaping because they do not become invasive species. In certain species, colchicine-induced triploidy has been used to create "seedless" fruit, such as seedless watermelons (Citrullus lanatus). Since most triploids do not produce pollen themselves, such plants usually require cross-pollination with a diploid parent to induce fruit production.

 

Colchicine's ability to induce polyploidy can be also exploited to render infertile hybrids fertile, for example in breeding triticale (× Triticosecale) from wheat (Triticum spp.) and rye (Secale cereale). Wheat is typically tetraploid and rye diploid, with their triploid hybrid infertile; treatment of triploid triticale with colchicine gives fertile hexaploid triticale.

 

When used to induce polyploidy in plants, colchicine cream is usually applied to a growth point of the plant, such as an apical tip, shoot, or sucker. Also, seeds can be presoaked in a colchicine solution before planting. Another way to induce polyploidy is to chop off the tops of plants and carefully examine the regenerating lateral shoots and suckers to see if any look different.[40] If no visual difference is evident, flow cytometry can be used for analysis.

 

Doubling of plant chromosome numbers also occurs spontaneously in nature, with many familiar plants being fertile polyploids. Natural hybridization between fertile parental plants of different levels of polyploidy can produce new plants at an intermediate level, such as a triploid produced by crossing between a diploid and a tetraploid, or a hexaploid produced by crossing between a tetraploid and an octoploid http://bfy.tw/4ezC.

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Grassmatch, just to clarify, the callus I'm talking about that grew the leaves is in the photo I posted On Feb 16, not my profile photo. My profile photo was an explant put directly into shoot generation formula about three weeks prior and then transferred to rooting formula; still waiting for it to root.

mrD,

 

I wouldn't worry too much, roots are on their way. I really don't have any clue why here are none so far, but the sample looks healthy to me. I've also had callus that refused to do anything but divide for years, even with the same mix as dozens next to it. no idea, but its fun.

 

find that gene gun yet?

 

so far I'm killing cultures with colchicine. I've seen some successful work with seed in this manner but not cultures. Its new to me. No idea what ratio's to even start with and since I cannot yet tell a polyploidy from a norm in vitro the exp will take a long time to grow out each survivor, which there have been none so far.

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Grassmatch, interesting post on mitotoc inhibitors. Probably not something I'll do for a while, if ever, but fascinating, nonetheless. I remember hybrid corn we used to plant on the farm back before GMOs were the thing...the seeds weren't fertile. That article explains it (I think). Good luck with your pursuits. Yeah, I've read about the gene gun in a few different places...hell I've got to get a culture through to full plant status!

 

I've only messed with cannabis culture except for taking a couple cuttings off of my wife's bonsai tree. They are doing well so far. It would probably be beneficial to do some other things to put a bit more fun into it. I do have my eye on one of my neighbor's camellia bushes that blossoms prolifically...I mean over the top solid red every spring. Going to snatch some from that as soon as it starts shooting this spring.

 

There are no others in this area doing tissue culture that I'm aware of. I've spoken with enough to folks that if there were others pursuing this I would have heard through the grapevine. I'm talking once in a while with a guy in CA who has had good luck...he's been doing TC and hydro for decades. You probably know him, or know of him. Will probably take a drive down there at some point to see what he's up to. The only real connection I've had with others in this field is on these forums, and most, I've found out, are not very genuine.

 

If I can perfect this process, I'll have more business than I can handle. There are two growers I'm in with that will build a lab when that happens. Each of them will be growing several thousand plants a year of legal rec, via indoor grows. Medicinal grows are going to be going away because the state is trying to push it out...they cannot tax it. Medicinal varieties will still be grown and sold to dispensaries but not under a medicinal license. A grower can only have one or the other...medicinal or rec. I'm also loosely associated with another group of several growers. The one thing they all have in common is they want CLEAN starts with no PM or russet mites. TC can offer that without dousing them in chemicals. Personally, I have no interest in growing...only TC.

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carrots and tobacco are the fastest I've worked with, and where my street lessons came from. You can bring these to fruit in a fraction of the time it takes-me, start, to finish, with cannabis.  Toying with the ratios of those recipes used is where I found success with cannabis finally.

 

I loved the "lab" style growing I had for a few years in hydro. The tissue culture experience is really just the ultimate hydro imo. My issue came when I began working with organic styles in hydro and found difficulty with all the off the shelf ferts, and worse with my own. I learned a ton but growing cannabis as clean as possible was important to me so I changed at the first harvested plant in a dirt pot...I was sold. I brought the same organic issues into culture with some of the same difficulties. I'd like to say I'm 100% organic each time I culture, but isn't true. I did kick my antibiotic habit a long time ago and that is an accomplishment for me. Those basal salts are spot on and don't fail, but try some sort of organic sterilized compost and yuck. I'm working with fermented fertilizers but its a real pain in the donkey.

 

I've good luck with willow water but no idea what the concentrations are so I wing it sometimes. The most consistent results come from hormones, basal salts and agar. my patience is diminishing with any other technique, but I still have the organic bonr, enough to spend hundreds of hours in trials.

 

I shared some boxes of cultures with a man in cali for awhile. I've got little related business aspirations now but hope to meet with many other culturists in the future.

I hear you on the growing part. I cant afford to buy what I need so I grow it instead, but am seriously considering only growing for myself. The TC was always just for me. End users don't care/sense the differences in tc/traditional propagation. Growers on the other hand will drive the industry to standards found in EVERY other commercially propagated crop eventually I think if cannabis becomes a national crop available recreationally nationwide. Monsanto did it with ALL of their commercial crops and I so no reason why they wouldn't already be working the field.  Commercial cannabis breeders are already enjoying the rewards of clean starts, its only a matter time you'll be a wealthy TC guru!

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Colchicine is a toxic chemical that is often used to induce polyploidy in plants.(crocus leaves, etc) Basically, the colchicine prevents the microtubule formation during cell division, thus the chromosomes do not pull apart like they normally do. The end result is a cell that now has double the number of chromosomes that it would normally have. If this cell divides again in the future, then the doubled number of chromosomes are passed to the offspring cells. Plants that have more than the normal two sets of chromosomes are termed “polyploidy” in general, although specific names are given to the certain chromosome numbers (e.g. tetraploid or 4N plants have four sets of chromosomes). Polyploid plants are generated in an effort to create new plants that have new characteristics. Sometimes the polyploidy plants are sickly and not viable, but sometimes the polyploid plants have larger leaves and flowers. Orchid growers will often sell polyploidy plants that are larger or have larger flowers. Often the polyploid nature of the plant is included in the cultivar name (G. species ‘big flower 4N’). Colchicine is also used to try to make fertile hybrids between species with different numbers of chromosomes.

http://www.carnivorousplants.org/howto/Propagation/Colchicine.php

I found the source after rereading the sites massive data base instructions for working with carnivorous plants

the site offers this unrelated snip for (gmo) enthusiasts. be sure to peruse the entirety though, lots of great culturing info here.

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Professor Thomas Cahill wrote this.  One of the great educators of our time. RIP

 

Colchicine is a toxic chemical that is often used to induce polyploidy in plants.  Basically, the colchicine prevents the microtubule formation during cell division, thus the chromosomes do not pull apart like they normally do.  The end result is a cell that now has double the number of chromosomes that it would normally have.  If this cell divides again in the future, then the doubled number of chromosomes are passed to the offspring cells.  Plants that have more than the normal two sets of chromosomes are termed “polyploidy” in general, although specific names are given to the certain chromosome numbers (e.g. tetraploid or 4N plants have four sets of chromosomes).

            Polyploid plants are generated in an effort to create new plants that have new characteristics.  Sometimes the polyploidy plants are sickly and not viable, but sometimes the polyploid plants have larger leaves and flowers.  Orchid growers will often sell polyploidy plants that are larger or have larger flowers.  Often the polyploid nature of the plant is included in the cultivar name (G. species ‘big flower 4N’).  Colchicine is also used to try to make fertile hybrids between species with different numbers of chromosomes.  The use of colchicine to make hybrids is well documented in ICPS cultural section of this web site.

            Unfortunately, colchicine is very toxic and hazardous to handle.  It is acutely toxic and has been responsible for many accidental poisonings by people or pets consuming the “autumn crocus” (Colchicum sp.) that are sometimes used in gardens.  In addition to its acute toxicity, it also causes chromosomal defects.  Overall, this chemical in it pure form is best handled in a chemistry lab with a fume hood with gloves and other personal protective equipment.  Once it is dissolved and diluted into water, it can be handled outside the fume hoods but gloves are still a necessity since the chemical may be adsorbed through the skin.  A more comprehensive review of colchicine toxicity can be found in the cultural of this web site .

The objective of this project was to assess what concentrations of colchicine are necessary to have biological effects on different carnivorous plant species.  Seeds of different CP species were obtained from the seed bank and divided into different treatments.  For all species, there was a “control group” and a 500 ppm colchicine treatment group.  Some species with abundant seed availability had additional treatment concentrations.  The number of germinations were then counted after it was clear the control group no longer had any new germinations, which was two months for most species.  The goal is to create tetraploid plants with different characteristics, but the confirmation of polyploidy requires chromosome counting, which is beyond the scope of this project.

Methods:

            Seeds from 5 different genera were obtained from the ICPS seed bank.  The seed from each species was homogenized to prevent inter-packet variation.  The seed was counted into equal number of seeds for each treatment.  For each treatment, the seed was divided into three equal portions which were treated separately.  The one exception was the Dionaea seed where each treatment was dosed together.

            The dosing procedure involved creating a concentrated colchicine solution by adding 0.51 g of colchicine into 100 mL of purified (HPLC-grade) water.  This created a “stock” solution of 5100 ppm colchicine by weight.  For each treatment, the stock solution was diluted to achieve the desired concentration of colchicine.  For example, the 510 ppm solution involved diluting 1 mL of the stock solution by adding 9 mL of purified water in a test tube.  The control group just had 10 mL of pure water added to it.  The seeds were added to the test tubes and they were allowed to soak in the solution for 96 hour.  The test tubes were wrapped in aluminum foil to protect the solutions from light since colchicine breaks down in light.  They were stored at room temperature for the 96 hours.  The solutions were shaken each day to make sure that the seeds were in contact with the solution since many of the seeds initially floated on the solution.  TheSarracenia seed were stratified in a refrigerator for 4 weeks before being dosed with colchicine.

            The seeds were planted into 4” (~10 cm) square pots that were filled with a mixture of 50 sand and 50% peat (“CP mix”) except for the Darlingtonia that were planted on pure sphagnum.  The pots were individually placed in plastic bags and then the solution with seeds in it was poured into the pots as to avoid coming in contact with the toxic solution.  The test tubes were rinsed with clean water and poured into the pots to wash out any remaining seeds into the pots.  The plastic bags were sealed and the pots were placed in a greenhouse or sunny windowsill for germination. 

            The seed germination was monitored weekly.  Once the number of new germinations in the control group ceased, then a final count of the number of seedlings was conducted.  The germination counts did include plants that were probably not viable in the long run.  These stunted seedlings were noted.  For example, Figure 1 shows the normal and stunted seedlings for Byblis liniflora.  Any seedling that died before the final count was not counted as a “germination”.  The pots were counted twice and the two counts were averaged to get the final number of seedlings.  The Darlingtoniapots were the exception in that they were scored the following spring about 6 months after germination.  The Darlingtoniascore does not include plants that died within the 6 month period.

ColchicineByblis.jpg 
Figure 1.  Panel A shows the control Byblis liniflora plants 9 days after planting.  Panel B shows the 510 ppm colchicine treated group, which are clearly stunted by the treatment.  None of the plants picture here were viable.

Results:

            The results from the colchicine treatment indicates that there is a high degree of inter-genera susceptibility to colchicine as applied in this dosing experiment (Table 1).  None of the Drosera species showed any visible effect from the colchicine; the germination was comparable to the control group and the seedlings looked normal.  These plants, due to their short life cycle, were monitored until some of the plants started blooming.  Almost none of the plants looked different from “normal” plants of the same species except that a few of the D. capensis looked a little more washed-out in terms of color, but this is qualitative at best.  Two of the D. binata plants looked larger than the control, so they were isolated and they will be evaluated for differences in growth rate.  If these plants seem to be larger in the long run, then chromosome counts will be conducted on them to verify if polyploidy were generated.

            On the other extreme, Byblis liniflora showed a very high degree of susceptibility to colchicine.  The germination and survival rate among seedlings was very low in the treated group.  Furthermore, the seedlings were clearly stunted and most were not viable (Figure 1). Some of the colchicine seedlings developed a few stunted leaves but then died.  These plants were probably polyploid, but the lack of plant viability prevented testing for chromosome counts. Any future attempts at making polyploid Byblis liniflora will need to use lower concentrations of colchicine.

            The Sarracenia leucophylla showed no observable effect from the colchicine treatment.  The treated group had effectively the same germination rate as the controls and the seedlings appeared to be normal.  Future experiments to induce polyploidy will need to use higher concentrations of colchicine.

            The Darlingtonia seedlings germinated and where then scored in the following spring approximately 6 months after germination, thus these scores represent both germination as early survival.  The data suggests that colchicine may have reduced the germination/seedling viability, but the rather small sample set (57 control seeds and 57 treated seeds) makes any comparison rather tenuous.

            Lastly, the Dionaea showed an effect from the colchicine treatment, but not as dramatic as the Byblis.  The germination rate of both of the treated groups was lower than the control group by almost a factor of two.  The higher colchicine treatment also started to generate clearly stunted plantlets that were comparable to the Byblis plantlets.  It would appear that 5100 ppm colchicine would be a good dosing concentration for future experiments since germination was still reasonable (albeit ½ of normal) and some of the plants were clearly affected.  Whether any of these plants will develop into anything of cultural interest remains to be seen.  If any of the plants are visibly different, then they will be tested for polyploidy.

            The ultimate objective of this experiment is to generate polyploid plants with desirable characteristics.  Since chromosome counts require considerable effort, only plants that appear to be different will be tested in the future.  The long development times for some species (e.g. SarraceniaDionaea) means that some species may take considerable time to show effects.  Although some plants in this test were abnormal (Byblis, Dionaea, Drosera), none of them have been confirmed as polypoid and abnormal Byblis were not viable.  Until some of the plants are confirmed as polyploid, the results of this experiment are best used as a “range-finding” experiment that determines what concentrations of colchicine are needed to produce effects without causing excessive seedling mortality.  Byblis needed dosing concentrations below 500 ppm while Darlingtonia and Dionaea produce biological effects at 500 ppm dosing concentration.  Drosera andSarracinea require dosing concentrations of greater than 1000 ppm or a longer dosing exposure time.

            Additional experiments may be conducted in the future to add more species to the colchicine susceptibility table and to generate new plants with different characteristics (hopefully positive!).  If such experiments are conducted in the same fashion again, they will be added to the table presented here.

 

Table 1. Germination success (%) for several species of carnivorous plants exposed to different concentrations of colchicine (in ppm) for 96 hours.  The number of seeds per each treatment group is given after the species name.

Species (number of seeds in each treatment)       Germination success (%) by
colchicine concentration in ppm   0 (control) 102 510 2550 5100

Byblis liniflora (n = 150)

20.6

---

1.3 a

---

---

 

 

 

 

 

 

Darlingtonia californica (n = 57)

33.3 (b)

 

17.5 b

 

 

 

 

 

 

 

 

Dionaea muscipula (n = 96)

38.5

---

22.3

---

18.2 (a)

 

 

 

 

 

 

Drosera binata (n = 150)

35.3

---

34.7

---

---

Drosera capensis ‘red’ (n = 300)

67.5

74.0

69.7

65.0

---

Drosera filiformis “Florida red” (n = 150)

54.0

---

50.0

---

---

 

 

 

 

 

 

Sarracinea leucophylla (n = 150)

48.7

---

50

---

---

(a) some germinating seedlings were clearly stunted and were not viable.
(b) The seedling success was scored in the following spring approximately 6 months after germination.

 

Thomas Cahill
Arizona State University

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Professor Thomas Cahill wrote this.  One of the great educators of our time. RIP

 

Colchicine is a toxic chemical that is often used to induce polyploidy in plants.  Basically, the colchicine prevents the microtubule formation during cell division, thus the chromosomes do not pull apart like they normally do.  The end result is a cell that now has double the number of chromosomes that it would normally have.  If this cell divides again in the future, then the doubled number of chromosomes are passed to the offspring cells.  Plants that have more than the normal two sets of chromosomes are termed “polyploidy” in general, although specific names are given to the certain chromosome numbers (e.g. tetraploid or 4N plants have four sets of chromosomes).

            Polyploid plants are generated in an effort to create new plants that have new characteristics.  Sometimes the polyploidy plants are sickly and not viable, but sometimes the polyploid plants have larger leaves and flowers.  Orchid growers will often sell polyploidy plants that are larger or have larger flowers.  Often the polyploid nature of the plant is included in the cultivar name (G. species ‘big flower 4N’).  Colchicine is also used to try to make fertile hybrids between species with different numbers of chromosomes.  The use of colchicine to make hybrids is well documented in ICPS cultural section of this web site.

            Unfortunately, colchicine is very toxic and hazardous to handle.  It is acutely toxic and has been responsible for many accidental poisonings by people or pets consuming the “autumn crocus” (Colchicum sp.) that are sometimes used in gardens.  In addition to its acute toxicity, it also causes chromosomal defects.  Overall, this chemical in it pure form is best handled in a chemistry lab with a fume hood with gloves and other personal protective equipment.  Once it is dissolved and diluted into water, it can be handled outside the fume hoods but gloves are still a necessity since the chemical may be adsorbed through the skin.  A more comprehensive review of colchicine toxicity can be found in the cultural of this web site .

The objective of this project was to assess what concentrations of colchicine are necessary to have biological effects on different carnivorous plant species.  Seeds of different CP species were obtained from the seed bank and divided into different treatments.  For all species, there was a “control group” and a 500 ppm colchicine treatment group.  Some species with abundant seed availability had additional treatment concentrations.  The number of germinations were then counted after it was clear the control group no longer had any new germinations, which was two months for most species.  The goal is to create tetraploid plants with different characteristics, but the confirmation of polyploidy requires chromosome counting, which is beyond the scope of this project.

Methods:

            Seeds from 5 different genera were obtained from the ICPS seed bank.  The seed from each species was homogenized to prevent inter-packet variation.  The seed was counted into equal number of seeds for each treatment.  For each treatment, the seed was divided into three equal portions which were treated separately.  The one exception was the Dionaea seed where each treatment was dosed together.

            The dosing procedure involved creating a concentrated colchicine solution by adding 0.51 g of colchicine into 100 mL of purified (HPLC-grade) water.  This created a “stock” solution of 5100 ppm colchicine by weight.  For each treatment, the stock solution was diluted to achieve the desired concentration of colchicine.  For example, the 510 ppm solution involved diluting 1 mL of the stock solution by adding 9 mL of purified water in a test tube.  The control group just had 10 mL of pure water added to it.  The seeds were added to the test tubes and they were allowed to soak in the solution for 96 hour.  The test tubes were wrapped in aluminum foil to protect the solutions from light since colchicine breaks down in light.  They were stored at room temperature for the 96 hours.  The solutions were shaken each day to make sure that the seeds were in contact with the solution since many of the seeds initially floated on the solution.  TheSarracenia seed were stratified in a refrigerator for 4 weeks before being dosed with colchicine.

            The seeds were planted into 4” (~10 cm) square pots that were filled with a mixture of 50 sand and 50% peat (“CP mix”) except for the Darlingtonia that were planted on pure sphagnum.  The pots were individually placed in plastic bags and then the solution with seeds in it was poured into the pots as to avoid coming in contact with the toxic solution.  The test tubes were rinsed with clean water and poured into the pots to wash out any remaining seeds into the pots.  The plastic bags were sealed and the pots were placed in a greenhouse or sunny windowsill for germination. 

            The seed germination was monitored weekly.  Once the number of new germinations in the control group ceased, then a final count of the number of seedlings was conducted.  The germination counts did include plants that were probably not viable in the long run.  These stunted seedlings were noted.  For example, Figure 1 shows the normal and stunted seedlings for Byblis liniflora.  Any seedling that died before the final count was not counted as a “germination”.  The pots were counted twice and the two counts were averaged to get the final number of seedlings.  The Darlingtoniapots were the exception in that they were scored the following spring about 6 months after germination.  The Darlingtoniascore does not include plants that died within the 6 month period.

ColchicineByblis.jpg 

Figure 1.  Panel A shows the control Byblis liniflora plants 9 days after planting.  Panel B shows the 510 ppm colchicine treated group, which are clearly stunted by the treatment.  None of the plants picture here were viable.

Results:

            The results from the colchicine treatment indicates that there is a high degree of inter-genera susceptibility to colchicine as applied in this dosing experiment (Table 1).  None of the Drosera species showed any visible effect from the colchicine; the germination was comparable to the control group and the seedlings looked normal.  These plants, due to their short life cycle, were monitored until some of the plants started blooming.  Almost none of the plants looked different from “normal” plants of the same species except that a few of the D. capensis looked a little more washed-out in terms of color, but this is qualitative at best.  Two of the D. binata plants looked larger than the control, so they were isolated and they will be evaluated for differences in growth rate.  If these plants seem to be larger in the long run, then chromosome counts will be conducted on them to verify if polyploidy were generated.

            On the other extreme, Byblis liniflora showed a very high degree of susceptibility to colchicine.  The germination and survival rate among seedlings was very low in the treated group.  Furthermore, the seedlings were clearly stunted and most were not viable (Figure 1). Some of the colchicine seedlings developed a few stunted leaves but then died.  These plants were probably polyploid, but the lack of plant viability prevented testing for chromosome counts. Any future attempts at making polyploid Byblis liniflora will need to use lower concentrations of colchicine.

            The Sarracenia leucophylla showed no observable effect from the colchicine treatment.  The treated group had effectively the same germination rate as the controls and the seedlings appeared to be normal.  Future experiments to induce polyploidy will need to use higher concentrations of colchicine.

            The Darlingtonia seedlings germinated and where then scored in the following spring approximately 6 months after germination, thus these scores represent both germination as early survival.  The data suggests that colchicine may have reduced the germination/seedling viability, but the rather small sample set (57 control seeds and 57 treated seeds) makes any comparison rather tenuous.

            Lastly, the Dionaea showed an effect from the colchicine treatment, but not as dramatic as the Byblis.  The germination rate of both of the treated groups was lower than the control group by almost a factor of two.  The higher colchicine treatment also started to generate clearly stunted plantlets that were comparable to the Byblis plantlets.  It would appear that 5100 ppm colchicine would be a good dosing concentration for future experiments since germination was still reasonable (albeit ½ of normal) and some of the plants were clearly affected.  Whether any of these plants will develop into anything of cultural interest remains to be seen.  If any of the plants are visibly different, then they will be tested for polyploidy.

            The ultimate objective of this experiment is to generate polyploid plants with desirable characteristics.  Since chromosome counts require considerable effort, only plants that appear to be different will be tested in the future.  The long development times for some species (e.g. SarraceniaDionaea) means that some species may take considerable time to show effects.  Although some plants in this test were abnormal (Byblis, Dionaea, Drosera), none of them have been confirmed as polypoid and abnormal Byblis were not viable.  Until some of the plants are confirmed as polyploid, the results of this experiment are best used as a “range-finding” experiment that determines what concentrations of colchicine are needed to produce effects without causing excessive seedling mortality.  Byblis needed dosing concentrations below 500 ppm while Darlingtonia and Dionaea produce biological effects at 500 ppm dosing concentration.  Drosera andSarracinea require dosing concentrations of greater than 1000 ppm or a longer dosing exposure time.

            Additional experiments may be conducted in the future to add more species to the colchicine susceptibility table and to generate new plants with different characteristics (hopefully positive!).  If such experiments are conducted in the same fashion again, they will be added to the table presented here.

 

Table 1. Germination success (%) for several species of carnivorous plants exposed to different concentrations of colchicine (in ppm) for 96 hours.  The number of seeds per each treatment group is given after the species name.

Species (number of seeds in each treatment)       Germination success (%) by

colchicine concentration in ppm   0 (control) 102 510 2550 5100

Byblis liniflora (n = 150)

20.6

---

1.3 a

---

---

 

 

 

 

 

 

Darlingtonia californica (n = 57)

33.3 (b)

 

17.5 b

 

 

 

 

 

 

 

 

Dionaea muscipula (n = 96)

38.5

---

22.3

---

18.2 (a)

 

 

 

 

 

 

Drosera binata (n = 150)

35.3

---

34.7

---

---

Drosera capensis ‘red’ (n = 300)

67.5

74.0

69.7

65.0

---

Drosera filiformis “Florida red” (n = 150)

54.0

---

50.0

---

---

 

 

 

 

 

 

Sarracinea leucophylla (n = 150)

48.7

---

50

---

---

(a) some germinating seedlings were clearly stunted and were not viable.

(b) The seedling success was scored in the following spring approximately 6 months after germination.

 

Thomas Cahill

Arizona State University

Hey I didnt know you were such a fart smeller!  oops sorry,  Smart Feller!

 

How do you remember all of that?  :lolu:

 

Peace

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Genetically Modified Bacteria Producing THC

google translation(loose)

 

 

image-65282-panoV9free-qemd-65282.jpgic_lupe.png

REUTERS / Eric Heritage / USDA

E. coli bacteria: Manipulated version produced THC cannabis ingredient

Dortmund - Cannabis is an intoxicant which is used for centuries as a medicine. Even modern medicine expects a lot of the active ingredient called tetrahydrocannabinol, THC shortly. In particular, seriously ill, suffering from multiple sclerosis, have spasticity or get a chemotherapy for cancer could benefit from it. Soon there might be cannabis on prescription in Germany , should be implemented to become known on Monday plans of CDU, CSU and FDP will.

The question would then, however, how to handle the increasing demand. "In Germany, currently about 20 kg THC per year are manufactured for medical use," said Oliver Kayser, professor of Technical Biochemistry at the TU Dortmund. "But the demand is around a ton."

a method Kayser's team has now developed in the genetically engineered bacteria produce THC. "We have the biosynthesis quasi copied from the plant into a microorganism," Kayser explained in an interview with SPIEGEL ONLINE. The THC-production by another undertaking had succeeded for the first time worldwide.

THC is produced in Germany from fiber hemp, whose cultivation and imports are legal. "Since the fibers contain less than 0.2 percent THC, the production process is correspondingly expensive," said Kayser. From the cannabis plant, which can contain up to 25 percent THC, the active ingredient for legal reasons in Germany can not be recovered.

Drug 2016 at the earliest on the market

In the Dortmunder method developed in the past five years the intestinal bacterium Escherichia coli is used. Him, the isolated genes, which are responsible in the cannabis plant for the THC education implanted. Accordingly increases the bacteria can then produce by fermentation THC.

http://www.spiegel.de/wissenschaft/mensch/biosynthese-bakterien-produzieren-cannabis-wirkstoff-a-712513.html

 

 

 

image-65282-panoV9free-qemd-65282.jpgic_lupe.png

REUTERS / Eric Heritage / USDA

E. coli bacteria: Manipulated version produced THC cannabis ingredient

Dortmund - Cannabis is an intoxicant which is used for centuries as a medicine. Even modern medicine expects a lot of the active ingredient called tetrahydrocannabinol, THC shortly. In particular, seriously ill, suffering from multiple sclerosis, have spasticity or get a chemotherapy for cancer could benefit from it. Soon there might be cannabis on prescription in Germany , should be implemented to become known on Monday plans of CDU, CSU and FDP will.

The question would then, however, how to handle the increasing demand. "In Germany, currently about 20 kg THC per year are manufactured for medical use," said Oliver Kayser, professor of Technical Biochemistry at the TU Dortmund. "But the demand is around a ton."

a method Kayser's team has now developed in the genetically engineered bacteria produce THC. "We have the biosynthesis quasi copied from the plant into a microorganism," Kayser explained in an interview with SPIEGEL ONLINE. The THC-production by another undertaking had succeeded for the first time worldwide.

THC is produced in Germany from fiber hemp, whose cultivation and imports are legal. "Since the fibers contain less than 0.2 percent THC, the production process is correspondingly expensive," said Kayser. From the cannabis plant, which can contain up to 25 percent THC, the active ingredient for legal reasons in Germany can not be recovered.

Drug 2016 at the earliest on the market

In the Dortmunder method developed in the past five years the intestinal bacterium Escherichia coli is used. Him, the isolated genes, which are responsible in the cannabis plant for the THC education implanted. Accordingly increases the bacteria can then produce by fermentation THC. Together with a pharmaceutical company, the University of Dortmund is now planning to set up a company for THC production by the new process. Kayser expects that this THC may come as drug 2016 at the earliest on the market.

With the groundwork for industrial production of THC using the bacteria biotechnology working group to Kayser's colleague Andreas Schmid is busy. Already, the researchers believe that they can significantly reduce production costs. Koste kilo legally produced THC currently around 50,000 euros, Kayser expects will be around 2500 euros.

But whether it actually soon cannabis are in Germany on prescription, yet seems certain - because the plans of the black-yellow coalition have not only triggered enthusiasm in Berlin. The Drug Commissioner of the Federal Government, Mechthild Dyckmans (FDP) has designated the project as an important step for seriously ill people. The working group "Cannabis as Medicine" (ACM) contrast spoke of deception.

For the affected people the time being change nothing, criticized the ACM chairman Franjo Grotenhermen. He considers the new regulations are inadequate. The coalition had merely decided that the drugs may be authorized when a pharmaceutical company submits an application. However, at present there is only one such request for a preparation against multiple sclerosis. "Patients with other diseases have also not have access to appropriate medication."

Dyckmans welcomed the new rules, however. Cannabis containing medicines have a proven benefit for patients in pain. Also Eugen Brych of the German Hospice Foundation supported the project in principle: "Because it is disproportionately difficult to obtain cannabis as a medicine, currently many pain patients are forced into illegality." So far there are nationwide only 40 patients receiving such preparations from the pharmacy.

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Interesting science, though a bit difficult to decipher.

 

Things sure have changed in the Deutschland. Granted, Cannabis was in very short supply during my extended visit a few decades ago. Fortunately hashish was not and that's where we got our THC from. It was much more reasonable than the flowers also @ $1.50/gm or up to $900/kg. Most varieties were available with Afghan a personal favorite, also quite acceptable was Red or Blonde Lebanese. Occasionally connoisseur blends would appear though usually in much smaller quantities, like finger hash from India and Nepalese Hash Balls.   

 

THC from ecoli? hey, to each his own...

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everyday foods -from ecoli

when cannabis is ONLY available as GMO, I quit!

thanks to the overwhelming Monsanto and gmo foods supporters in our country, that day may be sooner than we think .

top-10-gmo-foods.jpg

I can personally attest to the difficulty of avoiding gmo foodstuff in my daily diet. Ecoli feed is the standard to feed agrobacterium- used to vector the transgenic foreign genes into a new host. Supposedly the ecoli is killed off by the broad spectrum of antibiotics used. Unfortunately its shown these strange bacterium, agrobacterium, lives on in our guts, in our livestock guts too, resulting in a plethora of disease, including massive sores on most feedlot cows, unable to heal even with the growth hormones and antibiotics used daily.

  These Charts Show Every Genetically Modified Food People Already Eat in the U.S.

See all the GMOs you may already be eating  

Chipotle announced Monday that the chain will no longer serve food containing genetically modified organisms (GMO), raising the bar for transparency in the United States, where there’s no requirement to indicate the presence of GMO ingredients on food labels or in restaurants. Likewise, biotechnology companies aren’t required to report which genetically modified seeds are used in production.

Yet the use of GMOs is undoubtedly widespread. Since GMOs were approved for commercial use, and then first planted into U.S. soil in 1996, their production has increased dramatically. More than 90% of all soybean cotton and corn acreage in the U.S. is used to grow genetically engineered crops. Other popular and approved food crops include sugar beets, alfalfa, canola, papaya and summer squash. More recently, apples that don’t brown and bruise-free potatoes were also approved by the FDA.

 

Adoption of GMO Crops in the US, 1996-2014

1996199820002002200420062008201020122014% Farmland per Crop0102030405060708090100Bt corn80Bt cotton84Ht cotton91Ht soybeans94Ht corn89
Ht: Herbicide-tolerant  Bt: Insect-resistant

It's also instructive to look at permits granted by the United States Department of Agriculture (USDA) and the Food and Drug Administration (FDA) — the American GMO gatekeepers, together with the Environmental Protection Agency – though it's important to note that not all the issued permits are for crops that are approved for commercial use.

To produce crops commercially, biotech companies apply for "deregulated status", the green light from the USDA to plant and distribute without restriction.

The bar chart below shows all deregulated crops, sized by the number of genetic varieties approved for each. The ten crops in green are currently produced in the United States, and described in detail in the list below.

USDA Approved Genetically Modified Crops

Produced in US Not currently produced
appleroseplumsugarbeettobaccoflaxcichorium intybusalfalfacanolaricepapayabeetsquashpotatorapeseedtomatocottonsoybeancorn05101520253011111112222226710162033
corn.png
1. Corn

Genetically modified corn turns up in many different products in the U.S. — and corn on the cob is the least of it. This crop is used to produce many different ingredients used in processed foods and drinks, including high-fructose corn syrup and corn starch. But the bulk of the GM corn grown around the world is used to feed livestock. Some is also converted into biofuels.

33 Genetically Modified Varieties
Applicant Phenotype Date Effective
AgrEvo Phosphinothricin Tolerant and Male Sterile 3/16/99 Pioneer Male Sterile and Phosphinothricin Tolerant 5/14/98 AgrEvo Phosphinothricin Tolerant and Moth and butterfly Resistant 5/8/98 Monsanto Herbicide Tolerant 11/18/97 Monsanto Herbicide Tolerant and European Corn Borer Resistant 5/27/97 DeKalb European Corn Borer Resistant 3/28/97 Northrup King European Corn Borer Resistant 7/18/96 Monsanto European Corn Borer Resistant 3/11/96 Plant Genetic Systems Male SterileMS3 2/22/96 DeKalb Glufosinate Tolerant 12/19/95 Monsanto Moth and butterfly Resistant 8/22/95 AgrEvo Glufosinate Tolerant 6/22/95 Ciba Seeds Moth and butterfly Resistant 5/17/95 Dow 2,4-D and ACCase-Inhibitor Tolerant 9/22/14 Bayer/Genective Herbicide Tolerant 9/25/13 Monsanto Male Sterile 9/25/13 Pioneer Insect Resistant and Glufosinate Tolerant 6/20/13 Stine Seed Herbicide Tolerant 5/3/13 Syngenta Rootworm Resistant 2/27/13 Monsanto Drought Tolerant 12/27/11 Pioneer Male Sterile, Fertility Restored, Visual Marker 6/28/11 Syngenta Thermostable Alpha-amylase 2/15/11 Syngenta Moth and butterfly Resistant 4/20/10 Pioneer Herbicide & Imidazolinone Tolerant 12/9/09 Monsanto European Corn Borer Resistant 7/24/08 Syngenta Corn Rootworm Protected 3/16/07 Monsanto High Lysine 1/23/06 Monsanto Corn Rootworm Resistant 12/14/05 Dow Corn Rootworm Resistant 9/23/05 Dow Moth and butterfly Resistant & Phosphinothricin Tolerant 10/20/04 Monsanto Corn Rootworm Resistant 10/8/02 Mycogen c/o Dow & Pioneer Moth and butterfly Resistant Phosphinothricin Tolerant 6/14/01 Monsanto Herbicide Tolerant 9/29/00
soybean.png
2. Soybeans

The second largest U.S. crop after corn, GM soy is used primarily in animal feed and in soybean oil—which is widely used for processed foods and in restaurant chains. In fact, soybean oil accounts for 61% of Americans' vegetable-oil consumption. It's also often used to make an emulsifier called soy lecithin, which is present in a lot of processed foods, including dark chocolate bars and candy.

20 Genetically Modified Varieties
Applicant Phenotype Date Effective
AgrEvo Phosphinothricin Tolerant 11/23/98 AgrEvo Phosphinothricin Tolerant 6/8/98 Du Pont High Oleic Acid Oil 5/7/97 AgrEvo Glufosinate Tolerant 7/31/96 Monsanto Herbicide Tolerant 5/18/94 Monsanto Dicamba Tolerant   Monsanto Lepidopteran-Resistant SoybeanMON 87751 10/17/14 Dow 2, 4-D, Herbicide and Glufosinate Tolerant 9/22/14 Dow 2,4-D and Glufosinate Tolerant 9/22/14 Bayer/Syngenta HPPD and Glufosinate Tolerant 7/18/14 Dow Insect Resistant 4/17/14 BASF Imidazolinone Tolerant 3/18/14 Monsanto Increased Yield 11/7/13 Bayer and M.S. Technologies Herbicide and Isoxaflutole TolerantFG72 8/21/13 Monsanto Stearidonic Acid Produced 7/13/12 Monsanto Improved Fatty Acid Profile 12/16/11 Monsanto Insect Resistant 10/12/11 Pioneer High Oleic Acid 6/8/10 Pioneer Herbicide & Acetolactate Synthase Tolerant 7/24/08 Monsanto Herbicide Tolerant 7/23/07
cotton.png
3. Cotton

Much of GM cotton is turned into cottonseed oil, which is used for frying in restaurants and in packaged foods like potato chips, oily spreads like margarine, even things like cans of smoked oysters. Some parts of the plant are also used in animal feed, and what's left over can be used to create food fillers such as cellulose.

16 Genetically Modified Varieties
Applicant Phenotype Date Effective
Calgene Bromoxynil Tolerant and Moth and butterfly Resistant 4/30/97 Du Pont Sulfonylurea Tolerant 1/25/96 Monsanto Herbicide Tolerant 7/11/95 Monsanto Moth and butterfly Resistant 6/22/95 Calgene Bromoxynil Tolerant 2/15/94 Monsanto Dicamba and Glufosinate Tolerant 1/20/15 Bayer Glufosinate Tolerant, Moth and butterfly Resistant 2/22/13 Bayer Glufosinate Tolerant, Moth and butterfly Resistant 10/12/11 Syngenta Moth and butterfly Resistant 9/29/11 Bayer CropScience Herbicide Tolerant 5/22/09 Syngenta Moth and butterfly Resistant 7/6/05 Monsanto Herbicide Tolerant 12/20/04 Mycogen/Dow Moth and butterfly Resistant 7/15/04 Mycogen/Dow Moth and butterfly Resistant 7/15/04 Aventis Phosphinothericin Tolerant 3/10/03 Monsanto Moth and butterfly Resistant 11/5/02
potato.png
4. Potatoes

A new kid on the block, the very recently approved GM crop is resistant to bruising and may produce less of a cancer-causing chemical, called acrylamide, when exposed to high heat. It has not yet seen adoption in the food supply, but is expected to be.

6 Genetically Modified Varieties
Applicant Phenotype Date Effective
Monsanto Colorado Potato Beetle and Potato Virus Y Resistant 2/25/99 Monsanto Potato Leafroll Virus & Colorado Potato Beetle Resistant 12/3/98 Monsanto Colorado Potato Beetle Resistant 5/3/96 Monsanto Coleopteran Resistant 3/2/95 J.R. Simplot Low-Acrylamide Potential, Reduced Black Spot BruiseE12, E24, F10, F37, J3, J55, J78, G11, H37, H50 11/10/14 Monsanto Potato Leafroll Virus & Colorado Potato Beetle Resistant 7/17/00
papaya.png
5. Papaya

Bred to withstand ringspot virus, which can destroy papaya plants, these genetically engineered 'Rainbow Papayas' were first commercially produced in the late 1990s. Much of the yield is grown in Hawaii.

2 Genetically Modified Varieties
Applicant Phenotype Date Effective
Cornell U Papaya Ringspot Virus Resistant 9/5/96 University of Florida Papaya Ringspot Virus Resistant 9/1/09
squash.png
6. Squash

Zucchini and yellow summer squash have been commercially available in the U.S. since the mid- to late-'90s, though GM squash accounts for just 25,000 acres of farmland, by some estimates.

2 Genetically Modified Varieties
Applicant Phenotype Date Effective
Asgrow Cucumber Mosaic Virus, Watermelon Mosaic Virus 2, and Zucchini Yellow Mosaic Virus Resistant 6/14/96 Upjohn Watermelon Mosaic Virus and Zucchini Yellow Mosaic Virus Resistant 12/7/94
canola.png
7. Canola

GM canola is used to make oil for cooking, as well as margarine. It's also used to produce emulsifiers that are used in packaged foods. By some estimates, 90% of canola grown in the U.S. and Canada is GM.

2 Genetically Modified Varieties
Applicant Phenotype Date Effective
Monsanto Herbicide Tolerant 9/25/13 Pioneer Herbicide Tolerant 7/18/13
alfalfa.png
8. Alfalfa

In a controversial decision in 2011, the FDA approved the commercial use of GM alfalfa that contains a gene making it resistant to herbicide. The crop is used mainly as hay for cattle.

2 Genetically Modified Varieties
Applicant Phenotype Date Effective
Monsanto/Forage Genetics Reduced Lignin 11/10/14 Monsanto & Forage Genetics Herbicide Tolerant 1/28/11, 6/14/05
apple.png
9. Apples

Another newly approved crop, this apple from a Canadian biotech company does not brown even after it's been sliced. It recently received FDA approval. The agency said it is safe to eat, which means they may appear on supermarket shelves.

1 Genetically Modified Variety
Applicant Phenotype Date Effective
Okanagan Non-Browning 2/18/15
sugarbeet.png

10. Sugar Beets

More than half the granulated sugar in the United States comes from GM sugar beets, which have been in production since 2008. Though their use was temporarily halted due to safety concerns, production resumed in 2011.

1 Genetically Modified Variety

Applicant Phenotype Date Effective

Monsanto and KWS SAAT AG Herbicide Tolerant July 20, 2012, February 8, 2011 (Partial), March 4, 2005

http://time.com/3840073/gmo-food-charts/

Edited by grassmatch

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GMO Vaccinations-from ecoli
 
Genetic engineering, also called genetic modification, is the direct manipulation of an organism's genome using biotechnology. New DNA may be inserted in the host genome by first isolating and copying the genetic material of interest using molecular cloning methods to generate a DNA sequence, or by synthesizing the DNA and then inserting this construct into the host organism. Genes may be removed, or "knocked out," using a nuclease. Gene targeting is a different technique that uses homologous recombination to change an endogenous gene, and can be used to delete a gene, remove exons, add a gene, or introduce point mutations.

Genetic engineering alters the genetic makeup of an organism using techniques that remove heritable material, or that introduce DNA prepared outside the organism either directly into the host or into a cell that is then fused or hybridized with the host. This involves using recombinant nucleic acid (DNA or RNA) techniques to form new combinations of heritable genetic material, followed by the incorporation of that material either indirectly through a vector system or directly through micro-injection, macro-injection and micro-encapsulation techniques.

In medicine, genetic engineering has been used to mass-produce insulin, human growth hormones, follistim (for treating infertility), human albumin, monoclonal antibodies, antihemophilic factors, vaccines,and many other drugs. Vaccination generally involves injecting weak live, killed, or inactivated forms of viruses or their toxins into the person being immunized. Genetically engineered viruses are being developed that can still confer immunity, but lack the infectious sequences. Mouse hybridomas, cells fused together to create monoclonal antibodies have been humanised through genetic engineering to create human monoclonal antibodies.
Genetically modified viruses

Scientist studying the H5N1 influenza virus to design a vaccine.

The process of genetic engineering involves splicing an area of a chromosome, a gene, that controls a certain characteristic of the body. The enzyme endonuclease is used to split a DNA sequence and to split the gene from the rest of the chromosome. For example, this gene may be programmed to produce an antiviral protein. This gene is removed and can be placed into another organism. For example, it can be placed into a bacteria, where it is sealed into the DNA chain using ligase. When the chromosome is once again sealed, the bacteria is now effectively re-programmed to replicate this new antiviral protein. The bacteria can continue to live a healthy life, though genetic engineering and human intervention has actively manipulated what the bacteria actually is.

Despite the early success demonstrated with the hepatitis B vaccine, no other recombinant engineered vaccine has been approved for use in humans. It is unlikely that a recombinant vaccine will be developed to replace an existing licensed human vaccine with a proven record of safety and efficacy. This is due to the economic reality of making vaccines for human use. Genetically engineered subunit vaccines are more costly to manufacture than conventional vaccines, since the antigen must be purified to a higher standard than was demanded of older, conventional vaccines. Each vaccine must also be subjected to extensive testing and review by the FDA, as it would be considered a new product. This is costly to a company in terms of both time and money and is unnecessary if a licensed product is already on the market. Although recombinant subunit vaccines hold great promise, they do present some potential limitations.

In addition to being less reactogenic, recombinant subunit vaccines have a tendency to be less immunogenic than their conventional counterparts. This can be attributed to these vaccines being held to a higher degree of purity than was traditionally done for an earlier generation of licensed subunit vaccines. Ironically, the contaminants often found in conventional subunit vaccines may have aided in the inflammatory process, which is essential for initiating a vigorous immune response. This potential problem may be overcome by employing one of the many new types of adjuvants that are becoming available for use in humans. Recombinant subunit vaccines may also suffer from being too well-defined, because they are composed of a single antigen. In contrast, conventional vaccines contain trace amounts of other antigens that may aid in conferring an immunity to infectious agents that is more solid than could be provided by a monovalent vaccine. This problem can be minimized, where necessary, by creating recombinant vaccines that are composed of multiple antigens from the same pathogen.


Source: Boundless. “Genetically Engineered Vaccines.” Boundless Microbiology. Boundless, 26 May. 2016. Retrieved 09 Jul. 2016 from https://www.boundless.com/microbiology/textbooks/boundless-microbiology-textbook/microbial-genetics-7/transgenic-organisms-94/genetically-engineered-vaccines-502-10778/

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  • Insulin-from ecoli  !!

 

 

  • vaccines
  • antivenoms
  • bacteria derived toxins
  • Immunoglobulins
  • monoclonal antibodies
  • allergens
  • blood products and clotting factors
  • hormones such as insulin, growth hormone,
  • enzymes such as pancreatins
  • heparins

 

 

GMOs in Food and Medicine: An Overview

By Richard Green (Independent Expert) on Thursday, August 28, 2014 - 22:44

Category: Future of GMO, Science and GMO Basics

 

 

 

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Originally posted on SkeptiForum.

When you search online for “GMOs” (Genetically Modified Organisms) the results that come up speak to GM crops, but GMOs are more than just plants. The technology of genetic modification or genetic engineering was first developed in the early 1970s, commercialized in pharmaceutical applications in the early 1980s, and then agricultural applications in the early 1990s.

The technology has been around for 40 years. It is hardly new. Perhaps if you compare it to the internal combustion engine it is new, but compared to something as recent and ubiquitous as flat screen HDTVs, DVRs, and Wi-Fi-friendly touch screen devices like iPhones and Tablets, it is a time tested technology.

In medicine, genetic engineering (GE) is used to make biopharmaceutical drugs. Various organisms are engineered for use as factories to produce the drug product. Bacteria are the preferred option, as they are the easiest to grow and scale-up for production, but depending on the complexity of the drug’s molecular structure, other organisms such as yeasts, mammalian cellsetc., can also be used to express the drug product. The first GE drug approved for use was insulin. By the year 2000, there were over 100 GE drugs on the market. Currently, people’s lives are changed every day by drugs like RemicadeEpoAvastin, and Neulasta.

With the notable exception of Golden Rice, the agricultural applications for GE have been predominantly directed at the practical concerns of farming, such as controlling destructive insects and weeds. Herbicide resistant and insect resistant crops have given farmers more choices.

geneticengineeringfoodmedicine.jpg

While the techniques used to modify the organisms are similar, the intent couldn’t be more different. Pharmaceutical companies are looking to manufacture drugs that are intended to have deliberate effects on the biochemistry of their targets. Agricultural companies are adding traits that will help farmers or benefit consumers (e.g. non-browning apple), without affecting the safety of the crop. In agriculture, the goal is to have no greater effect on our health and metabolism than the crop’s non-GE variety. Again Golden Rice is the exception, as it is designed to have a higher nutritional content than its non-GE counterpart.

GE drugs and crops do have some commonalities. Both are the result of very long and careful screening processes to find the right molecules/proteins and the genes that code for them. For a drug product, the candidates are molecules with the potential to treat or cure a disease or condition. The intent in agriculture is to find proteins/molecules that will provide a useful trait to a plant and does not cause harm to people or the environment. The next step is to take the candidate genes and insert them in the appropriate host organism/crop species, and after 40 years of refinement this is now the most routine part of the process. Then begins the painstaking operation of selecting the organism or plant that is expressing exactly what you want the way you want it. Which leads to the last bit GE crops and drugs have in common; both go through a multi-yearapproval process.

The approval process for drug products is to ensure that the drug does what it is designed to do with minimal side effects. Because these compounds are designed to have such profound biological impacts, they have a more in depth approval process. They must not only be shown to be safe, but also to be effective for the condition they are targeted to treat.

The approval process for agriculture is only concerned with safety and substantive equivalence. The process confirms that the crop is as safe to eat or use as its non-GE counterpart, and that it is not an unintended risk to the environment. Currently only a few genetic changes are made to a crop which ultimately results in the expression of well understood proteins. That makes checking for known allergens and digestibility a relatively straightforward process. These safeguards are doing their job, as the scientific consensus is quite clear, the currently approved GE crops pose no greater health or environmental concerns than their non-GE counterparts.

While genetic engineering is used in both agriculture and medicine, it is far more controversial in agriculture. Here is an explanation that helped shape my point of view: intellectually, I can grasp that adding or silencing a few well-characterized genes out of thousands is a drop in the genome bucket, but for me it makes it a bit more real to think of it in terms of people. Just look at the variety among us. Variations between our thousands of genes are why we are all different from each other, but even with those differences, we are all human. It is similar with plants. Changing one, or as we get better, a few genes, in the plant genome is barely a blip compared to the normal diversity between individuals.

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GM microorganisms are used to make vitamin B2 (riboflavin), vitamin C (ascorbic acid), xanthan (a thickener), citric acid, and enzymes used in cheeses, breads and baked goods, alcoholic beverages, and juice.

 

 

And when it comes to the safety of these genetically engineered crops, Phil Angell, Monsanto's director of corporate communications back in 1998 probably expressed it best when he told Michael Pollan that:

"Monsanto should not have to vouch for the safety of biotech food. Our interest is in selling as much of it as possible. Assuring its safety is FDA's job.
"

In a dramatic move that shows that the U.S. FDA is softening its stand on bio-pharmed treatments, the agency has approved the country's first genetically modified plant intended for the treatment of a human disease.

An Israeli firm grew human disease enzymes in carrot cells, and produced a treatment for Gaucher disease that they say shows improvement comparable to a treatment derived from hamster cells. The drug goes by the name Elelyso.

As reported by Popsci, the ability to manipulate the genes of plant cells isn't new, but until now concerns about human biologics have kept them from gaining traction with the FDA. I don't even want to think about the potential ramifications of this decision. Many may not know this, but Monsanto, well-known as the leader in biotechnology and genetically engineered foods, is also invested in the medical industry.

Will Biotech Drugs Become the Next Big Battle?

In 1995, The Upjohn Company—a pharmaceutical company founded in Michigan—merged with the Swedish pharmaceutical and biotech company Pharmacia AB, to form Pharmacia & Upjohn.

In 2000, Pharmacia & Upjohn merged with Monsanto Company, at which time the name was changed to Pharmacia. The drug divisions, including Monsanto's old Searle drug division, were retained in Pharmacia, while the agricultural divisions became a wholly owned subsidiary of Pharmacia.

A short while later, Pharmacia spun off this agricultural/biotech subsidiary into a "new Monsanto" company, with the agreement that the "new" Monsanto would indemnify Pharmacia against certain liabilities that could be incurred from judgments against Solutia—yet another Monsanto-owned company that creates a variety of plastic materials, which was sued by Alabama residents over long-term PCB contamination.

Pfizer then bought Pharmacia 2002, and today also owns the remainder of Upjohn. Bayer has also acquired certain assets.

As you can see, the past and present connections between all of these mega-corporations are dizzying in their complexity.

Monsanto, as a whole, has such a long history of questionable behavior; I shudder to think what might occur once it gets into the game of genetically engineered drug-crops, which is now right around the corner. We already know that coexistence between conventional or organic and genetically engineered crops is impossible, due to the spread of pollen and seed.

gmo products have snuck into our daily lives, like it or not, most  happily consume these products and support this technology regardless of purported dangers.

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How Pharmaceutical Companies Turn Genetically Modified Livestock Into Drug Factories

By Katharine Gammon Posted July 29, 2010
 
1 of 4
  • Pigs.jpg?itok=8vgfGqK9

    Pigs

    Drug
    Human factor (HF) VIII and IX,a¨the main treatments fora¨hemophilia A and B, which affect 1 in 4,000 and 1 in 20,000 men, respectively, worldwide

    Animal Advantage
    Compatibility. Pigs' biochemistry is nearly identical to humans', so the proteins they produce are less likely to be rejected. Pigs make up to 50 times as much HFIX per gallon of milk as is usually found in human blood, and perhaps 600 times as much HFVIII.

    Status
    HFIX is in preclinical animal trials led by the University of Nebraska. HFVIII preclinical trials could start next year.

  • Goats.jpg?itok=GAdAReRJ

    Inga Spence/Photo Researchers

    Goats

    Drug
    ATryn (recombinant antithrombin), an anticoagulant drug taken by some of the sufferers of clotting disorders, which affect up to 1 in 2,000 people

    Animal Advantage
    Efficiency. By the time a transgenic goat is 18 months old, it can produce 211 gallons of ATryn-laced milk a year. Cows can churn out more, but they take twice as long to reach milk-producing age.

    Status
    FDA-approved and available now from GTC Biotherapeutics

  • Rabbits.jpg?itok=kknP1MlW

    Scott Bauer/Agricultural Research Service

    Rabbits

    Drug
    Rhucin, a drug for the 1 in 30,000 people afflicted with hereditary angioedema, a condition in which the immune system causes painful swelling of the skin, intestines and mouth. Also in tests for reducing post-transplant organ inflammation

    Animal Advantage
    Speed. Because Rhucin is injected directly into the bloodstream, it can be administered in lower doses.Quick-breeding, inexpensive rabbits that provide 1,680 milligrams a day in their milk can easily meet demand.

    Status
    Dutch company Pharming recently finished clinical trials in Europe and expects approval there this month.

  • placeholder.gif

    Cows

    Drug
    Biohormon, a synthetic version of human growth hormone

    Animal Advantage
    Volume. Demand for HGH is high, and multiple daily injections are the norm. Cows produce the greatest volume of milk of any transgenic livestock: A Biohormon cow can make 26 grams of HGH per gallon of milk.

    Status
    Argentinean company a¨Bio Sidus is starting clinical trials as soon as this winter

  •  
  •  

Farms have always provided a steady supply of milk, cheese and meat. Now add medicine to the list. Last year, the Food and Drug Administration approved the first medication produced in genetically modified livestock—ATryn, an anticoagulant grown in goats—and now several drug companies have launched their own animal-made medications. The drugs work as well as the ones synthesized in labs; only the process of making them is different. Scientists insert a human gene for a medically useful protein into an animal embryo's DNA and place the embryo in a surrogate mother. Once the animal matures, a technician can extract the protein from its milk in greater quantities than can be produced in a lab.

Raising a goat is far easier than standard methods of creating protein-based medicines, such as growing batches of altered cells or pulling proteins from donated human blood. It's also cost-effective. William Heiden, the CEO of GTC Biotherapeutics, the makers of ATryn, says that running a farm instead of building a dedicated production line saves his company hundreds of millions of dollars. And scaling up production is as easy as letting the animals mate. Some scientists warn that transgenic animals could introduce modified DNA into other herds, but these livestock could also drastically cut the prices of lifesaving drugs. Here, a look into a "pharmer's" medicine cabinet. http://www.popsci.com/science/article/2010-07/down-pharm

Edited by grassmatch

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In the second century A.D., during the Han dynasty, a Chinese author and alchemist known as Wei Boyang is believed to have written: “Gold is the most valuable thing in all the world because it is immortal and never gets rotten. Alchemists eat it, and they enjoy longevity.” Nearly two millennia later, the precious metal may live up to the hype: It’s part of a cutting-edge approach to prolong the lives of cancer patients.

 

Though they didn’t quite capture the details, Wei Boyang and other ancients who associated gold’s long-lasting luster with good health were surprisingly prescient. Gold’s immortality—the fact that it doesn’t interact with most compounds and thus doesn’t corrode—makes it essentially nontoxic to the body. This characteristic gives it huge potential value in medicine, for mundane procedures like dental fillings and, in the era of nano­technology, for diagnosing and treating deadly diseases.

“There are an enormous number of people using gold nanoparticles,” says Chad Mirkin, a chemist at Northwestern University whose own studies focus on how the particles could help turn off genes that cause disease. “We’re talking hundreds and hundreds of researchers around the world.”

One surprising approach comes out of research conducted at Rice University in Texas, along with the MD Anderson Cancer Center and other institutes. Oncologists are now injecting cancer patients with ultra-tiny, gold-wrapped spheres. The nano­particles, each smaller than a red blood cell, accumulate in a tumor after slipping out of the bloodstream through little holes in the tumor’s rapidly growing vessels. Once there, the gold waits—until an oncologist blasts it with near-infrared light.

 
 

Despite gold’s shiny quality, the spheres are made to absorb rather than reflect certain wavelengths of light, a property used against the cancer cells. “We artificially contaminate the tumor,” says Sunil Krishnan of MD Anderson. The nanoparticles convert the light into heat, and as temperatures in the tumor climb above 104 degrees Fahrenheit, the cancer cells deform, shrivel and then disintegrate.

In experiments in mice, Krishnan is zapping the scraps of pancreatic cancer remaining after a tumor is removed surgically. But clinical trials in people, including for cancers of the head, neck and lungs, are targeting tumors without surgery.

Although gold can be expensive, some potential therapies use as little as 3 percent of the amount in a typical wedding band. Instead, the main obstacle will probably be rigorous safety tests. “One of the tenets of nano is that everything that is miniaturized is different,” says Mirkin. So researchers need to confirm that new gold-based treatments are friendly to the body.

If so, a sly little ditty written by a 17th-century herbalist who also recognized the curative powers of gold may prove true today:

 

“For gold is cordial, And that’s the reason,

Your raking misers live so long a season.”

 

http://www.smithsonianmag.com/innovation/how-doctors-harnessing-power-gold-fight-cancer-180949436/?no-ist

 

 

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Applications and Advantages of Tissue Culture in Cannabis Cultivation

Large scale agriculture is at its most efficient when it has a reliable, uniform supply of juvenile plantlets to grow from. Cannabis culture is no different, and starting vegetation phase of growth from already hardened plantlets (as opposed to seeds) saves time and resources otherwise spent on attempting to propagate non-viable or substandard / undesirable seeds within a group. An additional advantage in regulated environments is that the entirety of permitted canopy space can be devoted to growing plants instead of maintaining vegetative mothers and clones. Clonal (that is, genetically identical) plantlets – derived either from mother plant cuttings, or other methods as discussed below – add the even more important advantage of maintaining uniform strain genetics, leading to predictable growth behaviour and mature plant characteristics such as THC and CBD content, terpene profiles, and other strain specific traits which are associated with a strain name and any downstream product branding. Also, clonal plants all reach milestone stages within a much narrower window than plants derived from seeds. Seeds, in contrast, are by their very biological nature genetic reassortants of their parental stocks, making each seed unique; useful for long term selective breeding programs where variety to select from is of use, but not at all desirable once a lineage is selected and uniform product is desired.

Cannabis cultivators have of course known this for a long time. While seeds are used for some smaller scale growth applications, established growth operations almost all work via clonal mother plant cuttings. This ensures product uniformity, but is relatively labor intensive and requires significant physical space for the cutting and hardening stages. Mother plants have a finite lifespan and limited number of cuttings they may provide, making the method relatively slow to scale up. Finally, traditional mother and cut clone systems are susceptible to all of the same plant pathogens (such as Fusarium spp. or powdery mildew) that can infect vegetating and flowering cannabis crops. Infestation of mother plants and clones can in worst case scenarios lead to the complete loss of strain stock – a serious risk for the grower.

A technique widely used in other branches of industrial scale agriculture has the potential to address all of these shortcomings of classical mother and cut clones, and do so economically and with vast scalability. As you’ve probably guessed from the title, this approach is tissue culture (“TC”). Tissue culture from plants involves taking a small amount of plant tissue which is induced to return to- and maintained in- a primitive stage (in the form of ‘calluses’). These cells can be propagated indefinitely in defined synthetic media; when plantlets are desired, some of these cells are taken out and then induced to differentiate into all of the different tissue that makes up a complete plant. The first benefit from the method is that these cells can be maintained in large quantities in very small volumes and can be expanded to hundreds of thousands of cells very quickly, and again, in very small volumes. The second benefit is in scalability; hundreds of thousands of plantlets can be induced in far less time and effort than creating mere hundreds of cuttings. A crucial third benefit of TC over traditional cloning methods is that by stringent control of the maintenance conditions, appropriate growth media additives, and proper handling techniques combined with good facility sanitation practices, strains can be maintained and propagated free of detectable pathogens (TC methods require aseptic handling and the presence of pathogens are readily detected and eliminated).

So now we appreciate that TC can generate vast numbers of identical, healthy “calluses”; how do we go about turning these into plants that we recognize as plants, suitable to move forward to vegetation and flowering stages?

The process generally involves taking callus stock, expanding the number of cells in liquid media as single cells, then plating individual cells in their own culture chambers, then add the appropriate growth factors and hormones that induce them to differentiate into all of the normal tissue types that make up complete plants; as the cells divide they will differentiate into roots, stems, and leaves. Once this differentiation proceeds, they grow into complete plantlets. When these plantlets reach around the 6-8” tall stage, these are then ready to pass on for handling as normal juvenile plants, without any differences in process for subsequent growth.

An added benefit of TC processes is that calluses can be maintained long term without differentiation, at very low cost and low space requirements. Particularly for an operation wanting to maintain multiple strain types for long periods, but able to bring them into production quickly and in large numbers, TC is an economical and reliable solution.

In summary, TC is a well-established method for the retention, clonal expansion, and on-demand production of vast numbers of intrinsically healthy, uniform juvenile plants while offering cost and space savings over traditional methods of any appreciable scale. TC methods are the industry standard for a wide range of agricultural or ornamental plant species – from blueberries to strawberries to birch trees – and going forward as cannabis production becomes mainstream industry and requires the associated scale and uniformity, TC methods will become the obvious method of choice for forward-thinking producers. Whether that comes via development of in-house expertise and infrastructure, or through contracting services out to established TC nurseries, is a matter of choice; but one or the other will surely become the gold standard for cannabis stock maintenance in the very near future.

 

https://www.newcannabisventures.com/

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An Exclusive Look at Cannabis Tissue Culture, a Transformative Technology – Part 1 July 18, 2016 at 4:37 pm  

Meristematic-logo.jpg

Why Tissue Culture is Key to the Future of the Cannabis Industry

Part I: An Introduction to Cannabis T.C.

Guest post by Michael Stevens, Founding Partner at Meristematic Labs

Over the past decade, the scale and sophistication of the Cannabis Industry in the United States has developed dramatically.  This aggressive transition from isolated small level cultivators to full-scale industrial horticulture has bought about a new set of problems for both government regulators and the cannabis business community.  These problems have become new opportunities for innovative companies looking to push the limits of what is possible with cannabis cultivation.  One of the most significant, changes coming to the cannabis industry is the application and use of Plant Tissue Culture Technologies.

What exactly is Plant Tissue Culture?  The term, is usually defined as the controlled aseptic culturing of whole plants, plant cells, tissues, organs and protoplasts. Aseptic meaning, free from contaminates like insects and other external microorganisms. The procedure for introducing plant cells into culture is relatively simple.

Cannabis-Culture-in-Testtube.jpgA small section of the plant called the explant, is cleaned and placed in to a sterilized vessel containing a media substrate of agar, sugars and growth regulating chemicals.  In the case of Cannabis, micro nodes are taken from the original stock plant and placed in sterilized test tubes to grow.  After 10-14

days the node has developed into a juvenile cannabis plantlet, with no roots.  Given the proper conditions and fresh growth media, a plantlet can be indefinitely divided into other explants for new cultures.  These new cultures can then be moved into a cannabis depository for storage, placed into bioreactors, used for experimental trials or transferred into rooting media & acclimated for cultivation in the greenhouse environment.   The process of tissue culturing is generally described in four distinct stages.

  • Stage #1 Establishment of Aseptic Culture
  • Stage #2 Multiplication
  • Stage #3 In-vitro Rooting
  • Stage #4 Acclimation

Tissue culture was first used on a larger scale by the orchid industry in the early 1950’s.  A2-Ghost-orchid-6-pack-7-13-09-_2.jpgThe technology proved to be useful for propagating orchids with seed stock that is difficult to germinate.  Today, Plant Tissue Culture technologies are used to preserve endangered species like the Ghost Orchid (Dendrophylax lindenii).  Work done by Dr. Michael Kane & Dr. Hoang Nguyen at the University of Florida is using tissue culture to grow Ghost Orchids in the lab.  Once fully grown the orchids are reintroduced back to their native habitat in the Florida Panther National Wildlife Refuge.

The culturing of plant cells in-vitro is the first step in a number of specialized agri-industrial and biotechnological processes.  Living cultures can be use for micro-propagation, haploid production, somatic embryo-genesis, synthetic seed production callus cell production, protoplast isolation and much more.  The early stages of plant cryopreservation uses tissue cultured cells & special sugars to store the living cells at -190ºC.  Micro-propagation uses tissue-cultured cells grown in bioreactors to produce large numbers of identical plant shoots for starter plants.  Specialized plant cells like anther, callus and polyploidy cells can be produced and isolated using Plant Tissue Culture.   It is simply a matter of time until these types of high-level agri-technologies are applied to the cannabis industry.

LJ-Culture-meristematic.jpgOnly in the past few years have cultivators and VCs taken cannabis tissue culture seriously.  As real demand increases for cannabis and cannabis related products, cultivators of all sizes will need large quantities of clean, healthy, true to type starter plants.  Intellectual property based around cannabis phenotype development and developed plant traits will rely heavily on tissue culture and cryopreservation as methods of possession of phenotypes.  One of the safest ways for medically significant cannabis phenotypes to be transported and exchanged is as tissue culture in sealed vessels.   Both government regulators and cannabis industry professionals need to be educated about the benefits of this technology so it can continue to develop.

 

https://www.newcannabisventures.com/an-exclusive-look-at-cannabis-tissue-culture-a-transformative-technology-part-1/

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In the second century A.D., during the Han dynasty, a Chinese author and alchemist known as Wei Boyang is believed to have written: “Gold is the most valuable thing in all the world because it is immortal and never gets rotten. Alchemists eat it, and they enjoy longevity.” Nearly two millennia later, the precious metal may live up to the hype: It’s part of a cutting-edge approach to prolong the lives of cancer patients.

 

Guess I will just have to swap out my evening glass of wine for a shot of Goldschlager

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you guys saw that 4210 modified the MMMA twice on plants , right?

 

(g) “Marihuana plant” means any plant of the species Cannabis sativa L.

 

(j) “Plant” means any living organism that produces its own food through photosynthesis and has observable root formation or is in growth material.

 

is whatever gel you use considered growth material?

 

stay within your plant counts, everyone.

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you guys saw that 4210 modified the MMMA twice on plants , right?

 

(g) “Marihuana plant” means any plant of the species Cannabis sativa L.

 

(j) “Plant” means any living organism that produces its own food through photosynthesis and has observable root formation or is in growth material.

 

is whatever gel you use considered growth material?

 

stay within your plant counts, everyone.

yup.

 

agar/alginate  used in dental offices to make molds for dentures/implants. I've grown in a sponge, styrofoam, wood chips, water, air, fog, so sure, growth material is anything supporting growth I imagine.  jello worked for decades for me, until the net turned on.

The undifferentiated dna stored in culture are not considered plant material, but bio material. It does not produce its own food while having observable root formation and will not report roots or leaves until initiated to do so.  Some of my cultures are simply root formations, no leaves.

 

I can also store cuttings in animated suspension  in the same gel/different formulation for years in a test tube. No roots form, no growth, but it is plant material. I found it is legal to mail these cultures while following the bio rules. harmless and non human  and not on a list of terrorist bio materials makes that part easy.

 

edit for clarification=  sample cuttings stored in suspended animation are not allowed to be mailed, by us anyways.

the undifferentiated dna samples are allowed to be mailed with light restrictions.

cannabis "artificial seeds", similar to other cultures,  are also able to be mailed with the same light restrictions.

 

rules could change from day to day, consult legal beagles for up to date information and changes before mailing anything cannabis related.

Edited by trichcycler

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Why so many mix ups and losses at labs and sperm banks? Unregulated?

 

Gee, My sperm is to valuable and I worked to hard to let it slip through someone else's fingers.

Edited by beourbud

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