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PubMed1984

Characterization of hemorrhagic principles from Trimeresurus gramineus snake venom.

In addition to alpha-fibrinogenase (hemorrhagin I, HR1), a potent hemorrhagic principle (hemorrhagin II, HR2) was purified from Trimeresurus gramineus venom. It was homogeneous as judged by SDS-polyacrylamide gel electrophoresis. HR2 was a single peptide chain containing 10% carbohydrate with a molecular weight of 81,500. It possessed 669 amino acid residues per molecule, while HR1 contained only 203 amino acid residues per molecule with a molecular weight of 23,500. Both hemorrhagins possessed proteolytic activities toward fibrinogen, casein and azocoll. However, the proteolytic activities of HR1 were much more potent than those of HR2. They were devoid of TAME-esterase and phospholipase A2 activities which were found in crude venom. beta-Mercaptoethanol and antivenin completely inhibited the hemorrhagic activities of HR1 and HR2, while epsilon-aminocaproic acid, trasylol, p-bromophenacyl bromide, phenylmethanesulfonyl fluoride and soybean trypsin inhibitor did not. EDTA completely inhibited the hemorrhagic, fibrinogenolytic and caseinolytic activities of HR1. EDTA also completely inhibited the caseinolytic and fibrinogenolytic activities of HR2, but only partially inhibited its hemorrhagic activity. Subsequent addition of Zn2+ (5 mM) reversed the EDTA-induced inhibitory effect on the hemorrhagic activity of HR1. However, ZN2+ did not reverse the EDTA-induced inhibitory effect on the HR2-induced hemorrhagic activity. These hemorrhagins were found to be ZN2+-containing metalloproteinases. Therefore, the hemorrhagic activity of HR1 seems to be related to its proteolytic activity while that of HR2 seems to be unrelated to its proteolytic activity.

Animals↗
PubMed1989

On the significance of animal experiments in toxinology.

In increasing areas of the world, there is considerable controversy over the use of animals in scientific research. Ethical, scientific and economic reasons require a reduction in animal experiments as well as animal numbers, a refinement of existing methods and, wherever possible, a replacement of in vivo methods by in vitro test systems. By scoping on papers published in Toxicon between 1980 and 1986, the current situation of animal experimentation in toxinology is evaluated. Fundamental research and antivenin production are reviewed and special reference is given to the experiments involved in drug registration by the example of batroxobin, a purified serine proteinase from Bothrops moojeni venom used as a defibrinogenating agent. Although the ethical question is the most important motif, a reduction in the number of animal experiments can be achieved more pragmatically: the scientific community must learn to constantly ask whether the intention behind an experimental series justifies the use of animals or not. This learning process can be supported by special educational programmes at university level, by making financial support dependent on questionnaires concerning the planned animal experimentation, by including respective questions in the application files for grants, and by ethical guidelines that form an integral part in the instruction for authors of scientific journals.

Animal Experimentation↗
PubMed1993

Selection of Hymenoptera venoms for immunotherapy on the basis of patient's IgE antibody cross-reactivity.

BACKGROUND: Positive skin test results to multiple venoms in patients with Hymenoptera venom allergy may result from IgE antibody cross-reactivity among venom proteins. To avoid treatment with unnecessary and costly venoms, we have developed a venom RAST inhibition test that identifies individuals in whom a positive venom IgE RAST result is due to cross-reactive venom-specific IgE antibody. METHODS: Serum samples (n = 412) were collected over 5 years from patients with clinically characterized Hymenoptera venom allergy who had positive skin test results to more than one venom. Venom allergosorbent was added to serum containing IgE antivenin and buffer or 100 micrograms of homologous or heterologous venom. Bound IgE was detected with radiolabeled anti-human IgE. Intraassay variation less than 10% coefficient of variation and homologous venom inhibition greater than 80% were required for acceptance of data. A "cross-reactivity index" (CRI) was computed as a ratio of percent inhibition produced by heterologous versus homologous venom. RESULTS: Of the 412 sera-venom combinations analyzed, 41 (10%) were excluded because of incomplete homologous venom inhibition. Of the 371 remaining sera, 82% (n = 305) were studied for IgE anti-Polistes wasp venom (PWV) cross-reactivity with yellow jacket venom (YJV) and the other 66 for other venom specificity cross-inhibitions. Of the serum samples tested, 36.4% (111 of 305) contained IgE anti-PWV venom of which the binding to solid-phase PWV was inhibited with soluble YJV to a level that produced CRIs greater than 95%. We believe that this constitutes complete inhibition and demonstrates exclusively YJV cross-reactive antibody in these samples. The remaining 63.6% had CRIs from 0% to 95%, indicating IgE specific for a spectrum of unique and cross-reactive PWV allergens. Only 4.3% (13 of 305) had CRIs less than 5%, which is consistent with IgE restricted to PWV unique allergens. The degree of the IgE anti-PWV inhibition to solid-phase PWV by YJV was independent of the IgE anti-PWV level. CONCLUSIONS: This study shows that one third of patients with Hymenoptera venom allergy evaluated with positive YJV- and PWV-reactive IgE in the skin and/or serum were identified as candidates for exclusion of PWV from their immunotherapy regimen because their IgE anti-PWV was more than 95% cross-inhibitable with YJV. Cost analysis of the venom RAST inhibition test and a conventional 5-year Hymenoptera venom immunotherapy program indicates that this serologic evaluation is cost-effective.

Animals↗
PubMedDate not supplied

The incidence of wound infection following crotalid envenomation.

Many sources advocate the empiric use of antibiotics to prevent wound infection (WI) following crotalid (rattlesnake) envenomations. We undertook a prospective observational study to examine the incidence of infections following crotalid envenomation. We studied crotalid envenomations presenting to our institution with follow-up by direct examination or telephone consultation. All patients presenting to our institution from June 1990 to October 1991 with history and clinical evidence of crotalid envenomation were included in the analysis. Routine management of crotalid envenomation was undertaken in each case, including the use of antivenin, surgical debridement, and antibiotics only when indicated by signs and symptoms of infection. A total of 54 patients were observed during the study period. Twelve patients received prophylactic antibiotics begun either prior to transfer to our institution or following a surgical procedure, and were evaluated separately. Follow-up was obtained on 32 patients 7 or more days following envenomation. Only 1 patient from the study group developed clinical evidence of WI during the study period. We conclude that because of a low incidence of WI in this series, the routine use of prophylactic antibiotics in such patients may not be warranted.

Adult↗
PubMed2006

Local cutaneous arachnidism: a report of three cases and their management.

Bites from Loxosceles spiders can cause few symptoms to gangrenous skin necrosis or even death. To date, the treatment of the violin spider bite is largely unsatisfactory. Although no specific therapy exists, it has been suggested that heparin, steroids, dapsone, experimental antivenin and/or surgical excision may be beneficial. Three rare cases of suspected spider bite and their surgical treatment are reported. On the basis of geographical area, anamnesis and clinical symptoms, we suppose Loxoceles rufescens responsible for these bites.

Aged↗
PubMed2005

Three structurally related, highly potent, peptides from the venom of Parabuthus transvaalicus possess divergent biological activity.

The venom of South African scorpion Parabuthus transvaalicus contains a novel group of peptide toxins. These peptides resemble the long chain neurotoxins (LCN) of 60-70 residues with four disulfide bridges; however they are 58 residues long and have only three disulfide bridges constituting a new family of peptide toxins. Here we report the isolation and characterization of three new members of this mammal specific group of toxins. Dortoxin is a lethal peptide, bestoxin causes writhing in mice and altitoxin is a highly depressant peptide. Binding ability of these peptides to rat brain synaptosomes is tested. While the crude venom of P. transvaalicus enhances the binding of [(3)H] BTX to rat brain synaptosomes none of these individual toxins had a positive effect on binding. Although the primary structures of these toxins are very similar to birtoxin, their 3D models indicate significant differences. Dortoxin, bestoxin and altitoxin cumulatively constitute at least 20% of the peptide contained in the venom of P. transvaalicus and contribute very significantly to the toxicity of the venom of this medically important scorpion species. Therefore the amino acid sequences presented here can be used to make more specific and effective antivenins. Possible approaches to a systematic nomenclature of toxins are suggested.

Amino Acid Sequence↗
PubMed2005

The neutralizing effect of a polyclonal antibody raised against the N-terminal eighteen-aminoacid residues of birtoxin towards the whole venom of Parabuthus transvaalicus.

Scorpion venom is composed among other things of a large number of neurotoxic peptides affecting all major types of ion channels. The majority of the toxicity of the venom is attributed to the presence of these peptides. In our previous studies using a combination of HPLC and mass spectrometry, we showed that birtoxin like peptides are the major peptidic components of the venom of Parabuthus transvaalicus. These peptides are quite similar to each other differing by only few amino acid residues. In addition they all share a common N-terminus of eighteen amino acid residues. We hypothesize that neutralization of this domain will decrease the toxicity of the whole venom of P. transvaalicus. Polyclonal antibodies against the common N-terminal region of the peptides are generated. Here we show by bioassays that the polyclonal antibodies neutralize the venom of P. transvaalicus in a dose dependent manner and by mass spectrometry and western blotting that these peptides indeed react with the polyclonal antibodies. Previously antibodies generated against a single major toxic component of venom have proven to be an effective strategy for antivenin production. In the case of P. transvaalicus the generated antibody is against the majority of the peptidic fraction due to the presence of several highly similar and highly toxic components in this venom. We show that using the knowledge obtained through biochemical characterization studies it is possible to design very specific antibodies that will be useful for clinical applications against Parabuthus envenomation.

Animals↗
PubMed1997

Scorpion envenomation in Merida, Venezuela.

This study involves a review of the records of 64 children referred to the University of los Andes Hospital with the diagnosis of scorpion envenomation. The patients were divided into group 1 (local manifestations, 42.4%) and group 2 (systemic manifestations, 7.8%). Seven of these patients had severe cardiorespiratory complications, came from distinct geographical zones and received antivenin. Our results indicate that poisonous and extremely dangerous scorpions predominate in certain geographical zones within the state of Merida, Venezuela.

Adolescent↗
PubMed2000

Disseminated intravascular coagulation after cerastes vipera envenomation in a 3-year-old child: a case report.

This study presents a case of severe disseminated intravascular coagulation (DIC) in a 3-year-old child following envenomation by the snake, Cerastes vipera. A literature search revealed very few similar cases. We describe a child who was bitten in his left foot by a snake identified as a C. vipera. Initial symptoms were relatively benign. Local signs included a hemorrhagic vesicle at the site of the bite with marked swelling of the entire leg. Twenty-four hours later, the child developed severe bleeding due to DIC, which lasted 5 days and required repeated administration of blood and blood products and total exchange transfusion. The patient was discharged from the hospital after 7 days in good condition. To the best of our knowledge, severe DIC following envenomation by a C. vipera has not been previously described in the literature. Treatment was essentially supportive. The case report indicates that a specific antivenin against this snake's venom should be made available in our area.

Animals↗
PubMed1985

Noninvasive vascular studies in management of rattlesnake envenomations to extremities.

Twenty-five consecutive patients suffering rattlesnake envenomation to an extremity underwent noninvasive vascular arterial studies using contralateral extremities as controls. All but one received antivenin, and none underwent early surgical decompression. Pulse volume amplitudes in the envenomated extremities, reflecting pulsatile arterial blood flow, were increased in 24 of 25 patients (P less than .02) in spite of edema and discoloration. Ischemia requiring surgical procedures developed in three of seven patients with decreased skin temperatures, as compared to none of 18 with increased or mixed skin temperatures on the envenomated extremity (P less than .02). One of 24 survivors suffered a functional deficit. Most victims of rattlesnake bites have increased arterial pulsatile blood flow in an envenomated extremity and do well with medical therapy. Noninvasive arterial studies help select those who are more likely to require special surgical intervention.

Adolescent↗
PubMed1981

Bites and stings of uncommon arthropods. 2. Reduviids, fire ants, puss caterpillars, and scorpions.

Arthropod bites and stings may cause intense pain, sometimes out of proportion to the size of the lesion. Severe reactions may arise from prior exposure and buildup of antibodies. Diagnosis often can be made from the clinical presentation if the physician is familiar with the effects of the uncommon bites and stings. Some uncommon bites, such as those of reduviids, can be treated symptomatically, but others, such as those of poisonous scorpions, call for antivenin therapy.

Ants↗
PubMed2000

Inadequate stocking of antidotes in Taiwan: is it a serious problem?

OBJECTIVE: Insufficient hospital stock of a variety of poisoning antidotes is a worldwide problem. In an attempt to establish an antidote storage and distribution system for the response of the various poisoning accidents, we conducted a nationwide survey to characterize the current availability of selected antidotes and their anticipated need in Taiwan. MATERIALS AND METHODS: A questionnaire was mailed to 834 hospitals to gather information on the availability, anticipated need, and preferred purchase policy of 20 selected antidotes. A survey on the availability of cyanide antidote in 523 cyanide-handling facilities and their neighboring hospitals was also conducted. RESULTS: Hospitals of different size and service levels had a statistically significant difference in response rates. Except for pyridoxine, the availability and anticipated need for antidotes also varied significantly among different hospital groups. We found that physostigmine, cyanide antidote kit, BAL, EDTA, methylene blue, Vipera Russell formosensis antivenin, and botulism antitoxin were not available in most (>90%) hospitals. Interestingly, these antidotes are also among the most needed antidotes. Most hospitals preferred a government-ordered purchase of antidotes. In the survey of cyanide-processing facilities, a response rate of 24.1% was obtained and only 9.3% of these 107 facilities that both replied to the questionnaire and continued handling cyanide products had stocked cyanide antidote. It is noteworthy that cyanide antidote was also frequently lacking in the neighboring hospitals. CONCLUSIONS: The appropriate storage of antidotes in hospitals or workplaces in rural areas is instrumental in the timely treatment of certain poisonings, while nationwide unavailability is the critical problem. Raising awareness of the importance of antidotes by education, regular review of antidote storage, distribution plans, and appropriate legislation might provide solutions.

Antidotes↗
PubMed2003

Prospective study of morbidity associated with snakebite envenomation.

BACKGROUND: The morbidity associated with snakebite envenomation has not been well documented. METHOD: Using a standardized questionnaire all patients with snakebite reported to a regional poison center during the year 2001 were followed after hospital discharge by telephone until resolution of symptoms. RESULTS: One hundred and twenty-eight snakebite cases were reported, of which 16 (12.5%) were lost to follow-up and 31 (24.2%) reported no progression of symptoms beyond puncture and were deemed "dry bites." Eighty-one (63.3%) patients were followed for the duration of symptoms. Age ranged from 1 to 86 years with a mean of 32 years. There were 64 males (79%). The snakes were identified as copperhead (n = 57), unidentified venomous (n = 17), timber rattlesnake (n = 6), and cottonmouth (n = 1). All patients were initially evaluated in a hospital emergency department of which 51 were admitted. Nine patients received antivenin. Of the 37 patients who had a job, 33 lost a mean of 14 days of work (SD +/- 18.1). Mean duration of edema was reported as 11.4 days (S.D +/- 12). Recurrent edema frequently occurred with limb activity. Pain was scored on a scale of 1 to 10, with a mean score of 4.8 (SD +/- 2.7). Mean duration of pain was reported as 7.8 days (SD +/- 6.4). Thirty patients required accommodation for ambulation including crutches (n = 11), limp (n = 11), and no shoes or loose shoes (n = 14). Of the 26 patients bitten on the hand or finger, duration of reduced function persisted for a mean of 14.3 days (SD +/- 10.4) and reduction of hand strength persisted for a mean of 22 days (SD +/- 25.5). Five patients had poorly healing wounds at the bite site which persisted from 14 to 77 days with a mean of 45 days (SD +/- 22.8) CONCLUSION: In this study snakebite resulted in significant duration and extent of morbidity in a majority of patients.

Adolescent↗
PubMed1979

Reversibility and mode of action of Black Widow spider venom on the vertebrate neuromuscular junction.

Black widow spider venom (BWSV) stimulates transmitter release and depletes synaptic vesicles from muscles bathed in a sodium free medium containing 1 mM EGTA. However, frog neuromuscular junctions treated with BWSV in glucosamine Ringer's and post-treated with antivenin recover normal function. This suggests that probably the permanent block of neuromuscular transmission is due to changes in permeability of the nerve ending plasma membrane to cations such as Na+. When BWSV is applied in a medium lacking divalent cations and containing 1 mM EGTA, in most of the cases no effect is observed. We found that this inhibition can be overcome in three ways: (a) by adding divalent cations to the medium; (b) by increasing the tonicity of the medium with sucrose; (c) by raising the temperature of the medium. These results suggest that the lack of divalent cations influences the membrane fluidity. Moreover, in view of the report by Yahara and Kakimoto-Sameshima (1977. Proc. Natl. Acad. Sci. U.S.A. 74:4511--4515) that hypertonic media induce capping of surface receptors in lymphocytes and thymocytes, we think that these data further support the hypothesis that BWSV stimulates release by a dual mode of action; namely, it increases the nerve ending permeability to cations and also stimulates release directly via a process of redistribution of membrane components, a process which may also inhibit vesicle recycling.

Animals↗
PubMed1999

Toxicology cart for stocking sufficient supplies of poisoning antidotes.

Within a three-month period, the University of New Mexico Health Sciences Center (UNMHSC) encountered three toxicological emergencies in which antidotes were either unavailable or inadequately stocked. Patient A was a patient with ethylene glycol intoxication. The emergency department (ED) physician ordered a 10% ethanol infusion. The pharmacy staff was unable to locate the commercially available solution and had to compound the infusion, resulting in a delay in administration of the antidote. Patient B was a patient at an outside hospital with organophosphate exposure whose transfer to our ED was requested by the other hospital. The pharmacy staff was unable to locate the pralidoxime needed to treat this patient; therefore; the patient was not transferred. It was later discovered that the pralidoxime had been stored in a location inconsistent with the storage policy for this medication. Patient C was a patient with severe rattlesnake envenomation. The pharmacy staff could locate only half of the antivenin needed to treat this patient. In each of these three cases, it was necessary to compound the required medication or to obtain it from other local facilities. These cases underscore the need for pharmacies to stock adequate amounts of poisoning antidotes in one immediately accessible location. A similar problem with understocking of poisoning antidotes exists throughout the United States.

Antidotes↗
PubMed2005

Applications and optimization of immunization procedures.

Classical immunization protocols have produced an antibody-based humoral response that is very effective against susceptible infectious diseases. Immunization introduces an external substance to induce the host immune system to respond specifically. Typically an antigen is used, but DNA, or a primed, pre-existing leukocyte or antigen-presenting cell, can also be used. Immunization is currently being used or investigated for the prevention and treatment of infectious diseases, cancer, addictions, allergies, pregnancy, and autoimmune diseases. It is also being used to produce biologically active materials such as polyclonal and monoclonal antibodies, antivenins, and anti-toxins for treating a wide range of conditions. Animals have been integral to the development of immunization techniques, as producers of toxoids and antitoxins, as models (e.g., to validate materials and protocols used for immunization, to understand the impact of immunization itself on the immune system, and to help investigators devise methods for determining the efficacy of vaccines) and as beneficiaries themselves of vaccines and antitoxins. The choice of immunization protocols is complex, and results may be affected by many factors such as dose and concentration of antigen, choice of adjuvants, time between inoculation and response measurement, and method of detection. The immune system responses to an antigen are also complex and continue to develop with advancing age. Anatomical, physiological, and immune system differences between species influence responses to immunization, as do the purity and presentation of the antigens and adjuvants. When directly comparing results, animals should be sourced from the same supplier. This review highlights the many uses of immunization techniques and introduces important considerations for the choice of protocols and animal models.

Animals↗
PubMed1998

Inadvertent methamphetamine poisoning in pediatric patients.

With the increase in popularity of adult methamphetamine abuse in the state of Arizona, there has been an increase in the number of pediatric admissions from inadvertent methamphetamine poisoning. This report describes the signs and symptoms and the hospital course of pediatric patients inadvertently poisoned with methamphetamine. A nine-year retrospective chart review identified 18 pediatric patients poisoned only with methamphetamine. This review revealed that pediatric patients with methamphetamine poisoning commonly presented with the following signs and symptoms: tachycardia (18 patients), agitation (nine patients), inconsolable crying and irritability (six patients), and vomiting (six patients). The most common ancillary tests, excluding urine drug screens, used in formulating the diagnosis were computed tomography scans of the head (five patients) and lumbar punctures (three patients). Three patients received Centruroides sculpturatus antivenin intravenously. The most common complication of methamphetamine poisoning was rhabdomyolysis (two patients), and the average hospital stay for all patients was three days. This case series demonstrates that pediatric patients who ingest methamphetamine can present with signs and symptoms similar to those of an abdominal or neurologic pediatric emergency.

Adult↗
PubMed2006

[Antidotes availability in health services of Catalonia, Spain].

BACKGROUND AND OBJECTIVE: Acute poisoning can be treated in different hospital and extrahospital health services. Thus, availability of antidotes in hospital pharmacies, emergency departments and health centres play an important role in poisoning treatment. MATERIAL AND METHOD: This is a descriptive and transversal study of antidotes availability in different health services of Catalonia (Spain). From April to May 2005, a proper questionnaire was sent to 50 different health centres: 28 basic health centres (including medical ambulances) and 22 hospitals with a distinct medical complexity level, to assess the qualitative (QLA) and quantitative availability (QTA) of antidotes. RESULTS: QLA resulted to be 82% in extrahospital health centres and 50% in medical ambulances. QTA was 94% for ambulances, whereas, in health centres, differences were observed between rural (55%) and urban (33%) areas. Regarding basic general hospitals (level 1), reference hospitals (level II) and high technology hospitals (level III) the QLA showed values of 39, 54 and 55%, respectively, and the QTA came about 70, 54 y 54.5%, in that order. Furthermore, lack of adequate stocking was observed for compounded formulations, such as albumin, starch, ipecacuanha syrup or IV ethanol, and for essential antidotes as snake antivenin, digoxin immune Fab or hydroxocobalamine. CONCLUSIONS: There is understocking of poisoning antidotes throughout the health services of Catalonia. Heterogeneity was observed between services with a medical complexity level. The main deficiencies are found in antidotes that, to date, are available as compounded formulations or imported from other countries.

Acute Disease↗