A high level of Plant Vacuolar Protease, activated after moderate and high doses of radiation, possible playing role as radiation toxin and can induce development of Acute Radiation Syndromes in mammals after ingestion.

1. Radiation Effects.
Plants Immunity.
Toxicity of Plants after
irradiation.
Dmitri Popov MD (Canada), PhD (Russia-Canada)
Advanced Medical Technology and Systems Inc.
intervaccine@gmail.com

2. Radiation Effects. Toxicity of Plants after
irradiation.
• DOI: 10.13140/RG.2.1.3185.6400
• A plant vacuolar protease, VPE, mediates radiation-induced
hypersensitive cell death.
• A high level of Plant Vacuolar Protease, activated after moderate
and high doses of radiation, possible playing role as radiation toxin
and can induce development of Acute Radiation Syndromes in
mammals after ingestion.

3. Plant Immune System.
• “ Many plant-associated microbes are pathogens that impair plant growth and
reproduction. Plants respond to infection using a two-branched innate immune
system. The first branch recognizes and responds to molecules common to many
classes of microbes, including non-pathogens. The second responds to pathogen
virulence factors, either directly or through their effects on host targets. These
plant immune systems, and the pathogen molecules to which they respond,
provide extraordinary insights into molecular recognition, cell biology and
evolution across biological kingdoms. A detailed understanding of plant immune
function will underpin crop improvement for food, fibre and biofuels production”
• Nature 444, 323-329 (16 November 2006) | doi:10.1038/nature05286
• The plant immune system
• Jonathan D. G. Jones1 & Jeffery L. Dangl2

4. Plants immunity.
• Plant radiation resistance protects plants from radiation in two ways:
mechanisms and by radiation-induced responses of the immune
system.
• Relative to a susceptible plant, radiation resistance is the reduction of
radiation damage on or in the plant, while the term radiation
tolerance describes plants that exhibit little disease damage despite
substantial radiation levels.
• Plant Radiation Toxins (possible Vacuolar Processing Enzyme) can
induce acute radiation disease of mammals.
• Irradiated living plants with active mitosis can be toxic up to 30 days
after irradiation.

5. Vacuolar processing enzyme (VPE).
• “Apoptotic cell death in animals is regulated by cysteine proteinases called
caspases. Recently, vacuolar processing enzyme (VPE) was identified as a
plant caspase. VPE deficiency prevents cell death during hypersensitive
response and cell death of limited cell layers at the early stage of
embryogenesis. VPE plays an essential role in the regulation of the lytic
system of plants during the processes of defense and development. VPE is
localized in the vacuoles, unlike animal caspases, which are localized in the
cytosol. Thus, plants might have evolved a regulated cellular suicide
strategy that, unlike animal apoptosis, is mediated by VPE and the
vacuoles.”
• Curr Opin Plant Biol. 2005 Aug;8(4):404-8.
• Vacuolar processing enzyme: an executor of plant cell death.
• Hara-Nishimura I1, Hatsugai N, Nakaune S, Kuroyanagi M, Nishimura M.

6. Radiation Effects. Toxicity of Plants after
irradiation.
• Programmed cell death (PCD) occurs in animals and plants under
various stresses and during development. Recently, vacuolar
processing enzyme (VPE) was identified as an executioner of plant
PCD.
• “A cellular suicide strategy of plants: vacuole-mediated cell death”
• Apoptosis 2006; 11: 905–911 C 2006 Springer Science + Business
Media, LLC. Manufactured in The United States. DOI:
10.1007/s10495-006-6601-1. N. Hatsugai et al.

7. Radiation Effects. Toxicity of Plants after
irradiation.
• Recently, vacuolar processing enzyme (VPE) was identified as an
executioner of plant PCD. VPE is a cysteine protease that cleaves a
peptide bond at the C-terminal side of asparagine and aspartic acid.
• “A cellular suicide strategy of plants: vacuole-mediated cell death”
• Apoptosis 2006; 11: 905–911 C 2006 Springer Science + Business
Media, LLC. Manufactured in The United States. DOI:
10.1007/s10495-006-6601-1. N. Hatsugai et al.

8. Radiation Effects. Toxicity of Plants after
irradiation.
• Programmed cell death (PCD) is an active, genetically controlled
• process leading to selective elimination of unwanted or damaged cells
in eukaryotes. PCD is essential for growth and development of
multicellular organisms as well as for proper response to environment
(Gechev et al., 2006; Lam, 2004).

9. Radiation Effects. Toxicity of Plants after
irradiation.
• Plant PCD is associated with a number of developmental processes
including embryo formation, degeneration of the aleurone layer
during monocot seed germination, differentiation of tracheary
elements in water-conducting xylem tissues, formation of root
aerenchyma and epidermal trichomes, anther tapetum degeneration,
floral organ abscission, pollen self-incompatibility, remodeling of
some types of leaf shape, and leaf senescence. (Gechev et al., 2006;
Thomas and Franklin-Tong, 2004).
• Programmed Cell Death in Plants: New Insights into Redox Regulation
• and the Role of Hydrogen Peroxide
• Ilya Gadjev,1,* Julie M. Stone,† and Tsanko S. Gechev*

10. Radiation Effects. Toxicity of Plants after
irradiation.
• Hydrogen peroxide (H2O2) and other reactive oxygen species (ROS) have
become recognized to be key modulators of PCD of as well as many other
biological processes such as growth, development, and stress adaptation
(Gechev et al., 2006).
• Free Radic Biol Med. 2012 Jul 15;53(2):260-70. doi:
10.1016/j.freeradbiomed.2012.04.033. Epub 2012 May 8.
• Ionizing radiation induces mitochondrial reactive oxygen species
production accompanied by upregulation of mitochondrial electron
transport chain function and mitochondrial content under control of the
cell cycle checkpoint.
• Yamamori T1, Yasui H, Yamazumi M, Wada Y, Nakamura Y, Nakamura
H, Inanami O.

11. Radiation Effects. Toxicity of Plants after
irradiation.
• Although specific ROS receptors/sensors remain largely elusive,
downstream components of H2O2 and ROS signal transduction
networks controlling plant PCD have been identified, including
protein kinases, protein phosphatases, and transcription factors.
• The majority of these are restricted to plants, with only a few genes
having close homologues in animals. (Gechev et al., 2006).

12. Radiation Effects. Toxicity of Plants after
irradiation.
• Hydrogen peroxide is produced in all cellular compartments as a
result of reactions of energy transfer, electron leakage from saturated
electron transport chains, and the activities of various oxidases and
peroxidases (Apel and Hirt, 2004).

13. Radiation Effects. Toxicity of Plants after
irradiation.
• Programmed cell death (PCD) is a process by which cells in many
organisms die. The basic morphological and biochemical features of
PCD are conserved between the animal and plant kingdoms.
Cysteine proteases have emerged as key enzymes in the regulation of
animal PCD.

14. Radiation Effects. Toxicity of Plants after
irradiation.
• The discovery that cell death is a tightly regulated (programmed)
process has stirred a great deal of interest in its mechanisms.
Studies of animal systems have shown that the execution of
programmed cell death (PCD) or apoptosis is controlled by a multistep
signaling pathway (McConkey and Orrenius, 1994; Stewart, 1994).
• In plants, PCD has been implicated in xylogenesis (Fukuda,
1996; Groover et al., 1997), in some forms of senescence,
and in the hypersensitive response to pathogens and environmental
stresses (Greenberg, 1996; Mittler and Lam,
1996; Lamb and Dixon, 1997).

15. Radiation Effects. Toxicity of Plants after
irradiation.
• Although a detailed understanding of how plant cells die is still largely
unknown, recent studies have shown that the apoptotic pathways of
the animal and plant kingdoms are morphologically and biochemically
similar (Greenberg, 1996; Levine et al., 1996; Wang et al., 1996).

16. Radiation Effects. Toxicity of Plants after
irradiation.
• Specifically, the morphological hallmarks of apoptosis include
cytoplasmic shrinkage, nuclear condensation, and membrane
blebbing (Earnshaw, 1995; Martins and Earnshaw, 1997); the
biochemical events involve calcium influx, exposure of
phosphatidylserine and activation of specific proteases and DNA
fragmentation, first to large 50-kb fragments and then to nucleosomal
ladders (McConkey and Orrenius, 1994; Stewart, 1994; Wang et al.,
1996; O’Brien et al., 1998).

17. Radiation Effects. Toxicity of Plants after
irradiation.
• The Involvement of Cysteine Proteases and Protease Inhibitor Genes in the
Regulation of Programmed Cell Death in Plants.
• The Plant Cell, Vol. 11, 431–443, March 1999, www.plantcell.org © 1999
American Society of Plant Physiologists.

18. Radiation Effects. Toxicity of Plants after
irradiation.
• VPE processing system mediates a cellular suicide strategy in plants. In
animals, dying cells are packaged into apoptotic bodies
• and then engulfed by phagocytes. In contrast, because plants do not have
phagocytes and the cells are surrounded by rigid cell walls, plant
• cells must degrade their materials by themselves. VPE, which has caspase-
1-like activity, is accumulated after perception of death signals such
• as pathogen infection. VPE is involved in activation of the target proteins to
provoke disintegration of the vacuolar membranes. Consequently,
• the vacuolar hydrolytic enzymes leave the vacuole for the cytosol and
degrade cellular components. Plants have evolved a death strategy that
• is mediated by the VPE processing system, which is not seen in animals.
A cellular suicide strategy of plants: vacuole-mediated cell death.
N. Hatsugai et al. DOI: 10.1007/s10495-006-6601-1

19. Radiation Effects. Toxicity of Plants after
irradiation.
• The genome of an organism is under constant attack from endogenous and
exogenous DNA damaging factors, such as reactive radicals, radiation, and
genotoxins. Therefore, DNA damage response systems to sense DNA
damage, arrest cell cycle, repair DNA lesions, and/or induce programmed
cell death are crucial for maintenance of genomic integrity and survival of
the organism. Genome sequences revealed that, although plants possess
many of the DNA damage response factors that are present in the animal
systems, they are missing some of the important regulators, such as the
p53 tumor suppressor. These observations suggest differences in the DNA
damage response mechanisms between plants and animals. In this review
the DNA damage responses in plants and animals are compared and
contrasted. In addition, the function of SUPPRESSOR OF GAMMA
RESPONSE 1 (SOG1), a plant-specific transcription factor that governs the
robust response to DNA damage, is discussed.
Biology 2013, 2, 1338-1356; doi:10.3390/biology2041338

20. Radiation Effects. Toxicity of Plants after
irradiation.
• Can irradiated plant’s cells used for feeding induce radiation disease
of mammals?
• Yes.

21. Radiation Effects. Toxicity of Plants after
irradiation.
• A careful analysis by FDA of all Army data present (including 31 loose-
leaf notebooks of animal feeding test results) showed significant
adverse effects produced in animals fed irradiated food...
• http://www.mercola.com/article/irradiated/irradiated_research.htm
• In the course of legalizing the irradiation of beef, chicken, pork, fruit,
vegetables, eggs, juice, spices and sprouting seeds -- a process that
has spanned nearly 20 years -- the U.S. Food and Drug Administration
has dismissed or ignored a substantial body of evidence suggesting
that irradiated food may not be safe for human consumption.
• http://www.mercola.com/article/irradiated/irradiated_research.htm

22. Radiation Effects. Toxicity of Plants after
irradiation.
• What were these adverse effects?
• A decrease of 20.7 percent in surviving weaned rats.
• A 32.3 percent decrease in surviving progeny of dogs.
• Dogs weighing 11.3 percent less than animals on the control diets...
Carcinomas of the pituitary gland, a particularly disturbing finding
since this is an extremely rare type of malignant tumor."
• Food irradiation: An FDA report. FDA Papers, Oct. 1968.

23. Radiation Effects. Toxicity of Plants after
irradiation.
• Fatal Internal Bleeding in Rats (I)
• "A significant number of rats consuming irradiated beef died from
internal hemorrhage within 46 days, the first death of a male rat
coming on the 11th day of feeding. This rat became sluggish on the
8th day of the regimen and started refusing food. He continued to be
morbid during the next two days, did not eat any food, lost weight
and appeared anemic. He was found dead on the 11th day.
• Vitamin K deficiency in rats induced by feeding of irradiated beef.
• Journal of Nutrition, 69:18-21, 1959. (Cosponsored by the Surgeon
General of the US Army)

24. Radiation Effects. Toxicity of Plants after
irradiation.
• Fatal Internal Bleeding in Rats (II)
• "Hemorrhagic death had occurred in all males fed irradiated diets by
day 34... There is evidence to suggest that inefficient absorption of
vitamins, i.e. vitamin K, from the intestinal tract may contribute to a
deficiency state." [Note: Vitamin K plays a major role in blood
clotting.]
• Influence of age, sex, strain of rat and fat soluble vitamins on
hemorrhagic syndromes in rats fed irradiated beef.
• Federation Proceedings, 19:1045-1048, 1960. (Cosponsored by the
Surgeon General of the US Army)

25. Radiation Effects. Toxicity of Plants after
irradiation.
• Fetal Deaths in Mice
• "Freshly irradiated diets produced elevated levels of early deaths in
[mice fetuses]... The increase in early deaths would suggest that the
diet when irradiated has some mutagenic potential."
• Irradiated laboratory animal diets: Dominant lethal studies in the
mouse.
• Mutation Research, 80:333-345, 1981.
• http://www.mercola.com/article/irradiated/irradiated_research.htm

26. Radiation Effects. Toxicity of Plants after
irradiation.
• Toxic effects of irradiated foods. Nature, 211:302, 1966.
• A Thalidomide Warning (II)
• "Irradiating can bring about chemical transformations in food and food
components resulting in the formation of potential mutagens, particularly
hydrogen peroxide and various organic peroxides.
• It is now realized, especially since the thalidomide episode, that older
testing protocols do not detect the more subtle population hazards such as
mutagens and teratogens. In view of the serious consequences to the
human population which could arise from a high level of induced
mutations, it is desirable that protocols for irradiated food should include
in vivo tests on mammals for possible mutagenicity."
• Mutagenicity and cytotoxicity of irradiated foods and food components.
• http://www.mercola.com/article/irradiated/irradiated_research.htm

27. Radiation Effects. Toxicity of Plants after
irradiation.
• Bulletin of the World Health Organization, 41:873-904,
1969. (Cosponsored by the US Atomic Energy Commission and Food and
Drug Administration)
• A Host of Problems
• "Numerous studies have been carried out to ascertain whether cytotoxic
effects occur when un irradiated biological test systems are cultured or fed
with irradiated media or food. In such studies, adverse physiological
growth retardation and inhibition, cytological cell division inhibition and
chromosome aberrations and genetical effects have been observed in a
wide range of test systems, ranging from bacteriophages to human cells...
The available data suggest that a variety of free radicals may act as the
toxic and mutagenic agents.“
• http://www.mercola.com/article/irradiated/irradiated_research.htm

28. Radiation Effects. Toxicity of Plants after
irradiation.
• Cytotoxic and mutagenic effects of irradiated substrates and food
material. Radiation Botany, 11:253-281, 1971.
• A Cancer Warning
• "An increase in concentration of a mutagen in food by irradiation
will increase the incidence of cancer. It will take four to six decades to
demonstrate a statistically significant increase in cancer due to
mutagens introduced into food by irradiation. When food irradiation
is finally prohibited, several decades worth of people with increased
cancer incidence will be in the pipeline.“
• http://www.mercola.com/article/irradiated/irradiated_research.htm

29. Radiation Effects. Toxicity of Plants after
irradiation.
• Growth, reproduction, survival and histopathology of rats fed beef
irradiated with electrons. Food Research, 20:193-214, 1955.
• Chromosomal Damage to Human Cells (I)
• "Irradiated sucrose solutions were extremely toxic to human white
blood cells. Cell divisions were inhibited. Degenerated cell divisions
were observed and the chromosomes were grossly damaged. The
DNA was clumped or the chromosomes appeared shattered or
pulverized. In contrast, treatment with un irradiated sucrose at the
same concentration had no apparent effect on the mitotic rate and
the chromosomes were not visibly damaged.“
• http://www.mercola.com/article/irradiated/irradiated_research.htm

30. Radiation Effects. Toxicity of Plants after
irradiation.
• Cytotoxic and radiomimetic activity of irradiated culture medium on
human leukocytes. Current Science, 16:403-404, 1966.
• Toxic Chemical Formed in Food Containing Fat (I)
• "When food containing fat is treated by ionizing radiation, a group of
2-alkylcyclobutanones [toxic chemicals] is formed. To date, there is no
evidence that the cyclobutanones occur in unirradiated food. In vitro
experiments using rat and human colon cells indicate that 2-
dodecylcyclobutanone (2-DCB)... is clearly cytotoxic and genotoxic.“
• http://www.mercola.com/article/irradiated/irradiated_research.htm

31. Radiation Effects. Toxicity of Plants after
irradiation.
• Radiation Toxins – Effects of Radiation Toxicity, Molecular
Mechanisms of Action, Radiomimetic Properties and Possible
Countermeasures for Radiation Injury.
• http://www.intechopen.com/books/current-topics-in-ionizing-
radiation-research/radiation-toxins-molecular-mechanisms-of-
toxicity-and-radiomimetic-properties-

32. Radiation Effects. Toxicity of Plants after
irradiation.
• Some results under way.

33. Radiation Effects. Toxicity of Plants after
irradiation.