Adenosine deaminase (ADA), histamine, and IgE are endogenously present in animals. Research from this laboratory reported decreased levels of these substances in organs of mice as a consequence of sub-lethal injection of Naja kaouthia venom. This research reports that decreased ADA, histamine, and IgE levels were prevented by specific treatment and prolonged recovery periods. Adult Balb/c mice injected IM with sub-lethal venom dose were divided into five groups. Group 1 were injected with PBS; Group 2 with anti-cobra venom; and Group 3 with lethal toxin neutralizing factor (LTNF). Groups 4 and 5 were treated with IM or oral synthetic LT-10. After 24 hours, mice were sacrificed and organ homogenates were assayed for ADA, histamine, and IgE. Group 1 showed substantial reduction in levels of these substances. It was revealed that decreased levels were prevented by treatment with anti-cobra venom, LTNF, and LT-10. In a second series of experiments, venom-injected mice were sacrificed after 3, 7, and 10 days and organs assayed for ADA, histamine, and IgE levels. The recovery period to homeostasis for ADA, histamine, and IgE was 7 to 10 days.

The pharmacological substances adenosine deaminase (ADA), histamine, and IgE are endogenously present in animals. They are implicated in allergy and asthma and detectable in blood serum. This research reports the presence of ADA, histamine, and IgE at varying levels in almost all major organs of mice. This research further reports that intramuscular injection of sub-lethal dose of Naja kaouthia venom disrupted homeostasis and lowered the levels of ADA, histamine, and IgE in the organs of mice. Adult Balb/C male mice were injected with a half lethal dose of the venom. The mice were sacrificed at 2, 8, and 24 hours post-injection and different organs were collected. Organs were homogenized, centrifuged, and the supernatants were assayed for ADA, histamine, and IgE using respective antisera by immunological test enzyme-linked immunosorbent assay (ELISA). Organs from mice injected with PBS served as controls. No major decrease in the levels of ADA, histamine, and IgE was observed after 2 hours of venom injection. However, tremendous decreases in the levels of ADA, histamine, and IgE was observed in organs 24 h post-injection. The highest decrease for ADA was observed in the brain, liver, lung, muscle, and testis; for histamine, in the heart, muscle, lung, and testis; and for IgE, in the bone, heart, lung, muscle, and testis. This is a first-hand investigation showing the effect of envenomation on the pharmacokinetics of ADA, histamine, and IgE in organs.

Lethal Toxin Neutralizing Factor (N-LTNF), MW 63.0 kDa, was isolated from opossum serum. After trypsin digestion, the active domain of N-LTNF was isolated and sequenced. The synthetic peptide consisting of ten amino acids was designated as LT-10. N-LTNF and LT-10 inhibited the lethality of animal, plant and bacteria toxins when tested on mice non-immunologically. The antibodies against N-LTNF and LT-10 only reacted immunologically with toxins and not with non-toxic substances. Anti-LTNF and anti-LT-10 reacted immunologically by ELISA test with toxins that were not detected by mouse test, such as cholera toxin and digoxin. Anti-LTNF and anti-LT-10 failed to react immunologically with non-toxic substances, such as nerve growth factor and collagen. Currently, mouse bioassay is in use for toxin detection and assay. Binding affinity of IgG from anti-LT-10 showed a linear relationship with mouse bioassay by ELISA detection limit to some toxins only. This may be due to the fact that anti-LTNF and anti-LT-10 detected the toxins that were not lethal to mouse. Thus, anti-LTNF and anti-LT-10 can be useful in assaying toxins as an alternative to mouse bioassay.

Pharmacological substances such as adenosine deaminase (ADA), collagen, histamine, IgE, myoglobin, and nerve growth factor (NGF) are endogenously present in animals. Research from this laboratory reported decreased levels of ADA, histamine, IgE, and NGF in organs of mice injected with sub-lethal doses of cobra venom. The goal of this research is to observe the levels of ADA, collagen, histamine, IgE, myoglobin, and NGF in certain organs of mice injected with venom from the bee Apis mellifera. Adult Balb/c female mice IM injected with half lethal dose of bee venom were sacrificed after 2 and 8 hours for removal of organs. The homogenates of the organs were assayed by ELISA for ADA, collagen, histamine, IgE, myoglobin, and NGF using respective antisera. Organs from mice injected with PBS were used as controls. It was observed that there were decreased levels of ADA, collagen, histamine, IgE, myoglobin, and NGF in certain organs after 2 h and tremendous decrease after 8 h. This is the first report showing the pharmacokinetics of ADA, collagen, histamine, IgE, myoglobin, and NGF as consequence of honeybee envenomation.

It is well documented that several pharmacological substances are released within the victim's body after snakebite. These substances are also believed to be endogenously present in animals, specifically levels of myoglobin and histamine that are reported to rise after envenomation. However, there is no published data regarding the presence of these substances in venoms per se. This research reports the detection of myoglobin and histamine in snake, scorpion, honeybee, and toad venoms by immunological test. It is unlikely that the rise in levels of myoglobin and histamine is due to that added from the bite, since a single toxin devoid of such components is capable of elevating levels of these substances. Nonetheless, it is likely that the rise in levels of myoglobin and histamine after envenomation is due to the venom or toxin reacting with cells of various organs of the victim. Therefore, this phenomenon can be compared to cancer markers, which are endogeneously present in humans at low levels and elevated in cancerous states.

Lethal Toxin Neutralizing Factor (LTNF), isolated from opossum with having molecular weight 63 kDa, is a potent antidote for animal, plant, and bacterial toxins. This communication deals with the identification of a small fragment of LTNF eliciting the anti-lethal activity of animal, plant, and bacterial toxins when tested in mice. Purified LTNF was treated with trypsin to cause fragmentation at the arginine and lysine sites. The fragments were separated by high-performance liquid chromatography (HPLC) and were tested against anti-LTNF for binding affinity by enzyme-linked immunosorbent test (ELISA). The fragment showing the most binding to anti-LTNF was sequenced. Synthetic peptides consisting of 15 and 10 amino acids from the N-terminal were constructed and designated as LT-15, with amino acid sequence Leu-Lys-Ala-Met-Asp-Pro-Thr-Pro-Pro-Leu-Trp-Ile-Lys-Thr-Glu, and LT-10, with sequence Leu-Lys-Ala-Met-Asp-Pro-Thr-Pro-Pro-Leu. Death due to intramuscular (IM) injection of predetermined lethal doses of toxins derived from animal, plant, and bacteria was prevented treating the mice with synthetic peptides LT-15 and LT-10. The lethality was inhibited when the treatment was given before or after the toxin injection. Synthetic LTNF can be made in abundance and should become a universal therapy against intoxication caused by animal, plant, and bacteria.

Currently, the use of antivenoms is the only available treatment for envenomation caused by venomous animals namely, snake, scorpion, spider, tick and jelly fish. Antivenoms are generally produced in large animals, mostly in horses. A large percentage of the population is allergic to horse proteins. Several animals are known to be resistant to snakebites and the antihemorrhagic and anti-lethal components have been isolated from sera of opossum, mongoose, meerkat and hedgehog, as well as from venomous and non-venomous snakes. Anti-lethal factor named Lethal Toxin Neutralizing Factor (LTNF) has been isolated in purity from opossum (Didelphis virginiana) serum by high pressure liquid chromatography (HPLC). The molecular weight of LTNF is 63 kDa, and it does not form precipitation with venoms or toxins by immunodiffusion. Death due to intraperitoneal (IP) injection of a predetermined lethal dose of venom from major families of snakes, for instance Crotalidae, Elapidae, Viperidae and Hydrophiidae, is prevented in mice by subsequent IP inoculation of LTNF. Furthermore, LTNF neutralizes the lethality of scorpion and bee venoms and toxins from various animals, plants and bacteria. Thus, natural LTNF from opossum serum has potential as a universal therapy for envenomation caused by animals, plants and bacteria.

Cancer cell inhibitors, named Atroporin and Kaotree, having molecular weights of 35 kDa and 6 kDa have been isolated from the venoms of Crotalus atrox and Naja naja kaouthia, respectively, by fractionation on high pressure liquid chromatography. The purified Atroporin and Kaotree showed killing effects on various types of human (breast, colon, liver, ovary, etc.) and animal cancer cells in concentrations as low as 0.5µg/ml, and having no effect on normal mouse kidney, liver, spleen, and erythrocytes up to 5.0µg/ml. Both Atroporin and Kaotree prevent the formation of ascitic tumors caused by myeloma cells in Balb/C mice. In addition, both Atroporin and Kaotree showed regression of ascitic tumors formed by myeloma cells. Atroporin and Kaotree complement each other, as in combination they showed elevated anti-cancer activity in vitro and in vivo systems. However, Atroporin and Kaotree are immunologically distinct proteins showing no cross reactivity. Atroporin and Kaotree, individually or in combination, have the potential for cancer biotherapy.