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PubMed1986

Differential effects of defined chemical modifications on antigenic and pharmacological activities of scorpion alpha and beta toxins.

Specific chemical modifications of scorpion alpha and beta toxins have been used to study the involvement of particular residues in both the pharmacological and the antigenic sites of these toxins. Modification by 1,2-cyclohexanedione of arginine-27 of a beta toxin, Centruroides suffusus suffusus toxin II, drastically decrease the antigenic activity without any influence on the pharmacological activity. Conversely, modification by the same reagent of arginine-2 of an alpha toxin, Androctonus australis Hector toxin III, led to a 100-times less pharmacologically potent derivative and did not induce a significant loss of antigenic activity. Excision of the N-terminal pentapeptide of another alpha toxin, Buthus occitanus mardochei toxin III, by pepsin digestion led to a non-toxic derivative retaining full antigenic activity. Thus, the N-terminal part of the conserved hydrophobic surface of the toxin is highly implicated in the pharmacological activity, whereas the region of arginine-27, located in the alpha helix situated on the back surface, opposite the conserved hydrophobic region, is fully implicated in the antigenic activity and is far from the pharmacological site. These results are good arguments in favor of the idea that in scorpion toxins the surfaces implicated in the pharmacological and the antigenic activities do not overlap. Since the antigenic sites are present in highly variable sequence the development of an efficient polyvalent serotherapy is questionable.

Amino Acids↗
PubMed1996

A conserved sequence region of scorpion toxins rendered immunogenic induces broadly cross-reactive, neutralizing antibodies.

Scorpion toxins constitute a family of proteins with a high degree of sequence diversity but a common mode of action. Neutralization of the toxic effects of scorpion stings by serotherapy is limited due to the various serotypes expressed by these proteins. We explored the possibility of raising antibodies to conserved parts of the toxins which could recognize several members of the family. We established the variability profile of a set of 25 scorpion toxin sequences, then evaluated systematically by peptide-scanning methods the antigenicity of one scorpion toxin. The most conserved regions were generally very poorly antigenic. One exception was the N-terminal region, which is both conserved and antigenic. Antibodies were raised in rabbits against an eight-residue synthetic peptide mimicking the N-terminal region. These peptide antibodies were cross-reactive with several scorpion toxins belonging to different serotypes and neutralized both the pharmacological effects (binding to rat brain synaptosomes) and the biological activity (toxicity in mice) of the parent toxin. The molecular model of the toxin indicates that antibody binding to residues 1-8 probably either masks some residue(s) of the N-terminus critical for the biological activity or overlaps with the epitope previously defined by neutralizing monoclonal antibody. These findings could open the way for new therapeutic strategies for the medical care of envenomations.

Amino Acid Sequence↗
PubMed2006

Gamma interferon, tumor necrosis factor alpha, and nitric oxide synthase 2, key elements of cellular immunity, perform critical protective functions during humoral defense against lethal pulmonary Yersinia pestis infection.

Pulmonary infection by Yersinia pestis causes pneumonic plague, a rapidly progressing and often fatal disease. To aid the development of safe and effective pneumonic plague vaccines, we are deciphering mechanisms used by the immune system to protect against lethal pulmonary Y. pestis infection. In murine pneumonic plague models, passive transfer of convalescent-phase sera confers protection, as does active vaccination with live Y. pestis. Here, we demonstrate that protection by either protocol relies upon both gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) cytokines classically associated with type 1 cellular immunity. In both protocols, abrogating IFN-gamma or TNF-alpha activity significantly decreases survival and increases the bacterial burden in pulmonary, splenic, and hepatic tissues. Neutralization of either cytokine also counteracts challenge-induced, vaccination-dependent upregulation of nitric oxide synthase 2 (NOS2). Moreover, genetic depletion of NOS2 suppresses protection conferred by serotherapy. We conclude that IFN-gamma, TNF-alpha, and NOS2, key elements of cellular immunity, perform critical protective functions during humoral defense against lethal pulmonary Y. pestis challenge. These observations strongly suggest that plague vaccines should strive to maximally prime both cellular and humoral immunity.

Animals↗
PubMed1985

Kinetics of growth and toxigenicity of Clostridium botulinum in experimental wound botulism.

An animal model of wound botulism was developed in mice using an inoculum of Clostridium botulinum type A spores. The number of C. botulinum in infected wounds was quantitated by culturing on egg yolk agar, and the level of C. botulinum toxin in infected wound tissue was measured by a bioassay in mice and by an enzyme-linked immunosorbent assay. All infected mice receiving no further treatment developed neuroparalytic symptoms consistent with botulism after an incubation period of ca. 48 h, and all of these animals died. Serotherapy with C. botulinum type A antitoxin initiated 24 h postchallenge reduced the mortality rate to 5%. Treatment with metronidazole 2 to 24 h postchallenge resulted in recovery rates of 40 to 91%.

Animals↗
PubMed1987

Protection of mice against experimental cryptococcosis by anti-Cryptococcus neoformans monoclonal antibody.

Humoral immunity does not play a prominent role during experimental cryptococcosis. However, previous studies have shown that immunoglobulin G (IgG) anti-Cryptococcus neoformans antibodies can mediate cell-dependent yeast killing in vitro. Therefore, the protective effect of a previously described monoclonal IgG1 anti-C. neoformans antibody (E1) administered intraperitoneally 24 h before intravenous infection with a C. neoformans serotype A strain was evaluated in mice. Heavily infected (3 X 10(6) cells) untreated mice died in 2.9 +/- 0.5 (standard deviation) days. Survival time was 17.9 +/- 1.6 days for mice treated with 100 micrograms of E1 and 3.0 +/- 0.7 days for mice treated with 100 micrograms of a monoclonal IgG1 anti-thyroglobulin antibody used as a control. Protection was dose dependent and required at least 10 micrograms of E1 (mean antibody concentration in serum +/- standard deviation, 6.6 +/- 2.3 micrograms/ml). Insufficient concentrations of IgG anti-C. neoformans antibody could explain previous negative results obtained with polyclonal immune serum. After infection with a smaller inoculum (5 X 10(3) to 5 X 10(4)), the protective effect of E1 was confirmed by the presence of fewer CFUs in the spleens and brains of treated mice than in those of controls. CFU were still detected in the brains of protected mice 5 days after infection, although soluble antigen was negative in sera. These results suggest that passive serotherapy with monoclonal IgG antibodies could participate in the prevention or treatment of experimental cryptococcosis.

Animals↗
PubMed2003

[Should the therapeutic effort in artificial nutrition be limited?].

BASIS: There exists general agreement among the experts in Bioethics in order not to maintain a treatment if it is considered that it is useless. However, there is no unanimity about if serotherapy, enteral nutrition (EN), and parenteral nutrition (PN) are really therapeutic measures or else they are measures that they are part of the concept of palliative care. OBJECTIVE: Know the opinion of users and health workers of National Health System (NHS) in these issues. DESIGN: Cross-sectional descriptive study. AREA: Primary and specialized care. METHODS: A survey was distributed to 219 individuals (84 users of the NHS, 135 health workers), that it included sociodemographic parameters, the opinion on different aspects of ethics and nutritional support, and the measures regarded as palliative care. RESULTS: The majority of the respondents think that the EN (70.8%; 95% IC: 64.2-76.8) and PN (54.8%; 95% IC: 47.9-61.6) are part of the basic care, with differences between the health personnel and the users of the NHS. 50.2% considers that there are no differences between applying or removing nutritional support. 71.3% of the health professionals answer that this decision depends on the medical, 60.5% of the users of the NHS thinks that it depends on the patient himself and/or its family members. All the evaluated measures are regarded as basic care, regardless of the characteristics of the respondent. CONCLUSIONS: Artificial nutrition is understood as a basic care, regardless of the situation and quality of life of the patient. While the health professionals think that the decision to initiate/to retire artificial nutrition depends on the physician, the users of the NHS consider that it depends on the patient/family in case of disability of this.

Adult↗
PubMed1985

Use of SM-1 monoclonal antibody and human complement in selective killing of small cell carcinoma of the lung.

SM-1 is a murine monoclonal antibody strongly reactive with a cell membrane antigen of small cell carcinoma (SCC) of the lung but unreactive with the membrane of most other carcinomas and normal tissues including normal bone marrow. We have found that in the presence of human complement, SM-1 antibody is highly cytotoxic to SCC cells. Using three treatments with antibody and complement, more than 99% of SCC cells in culture were lysed, as determined by the chromium release and clonogenic assays. Similar efficiency of SCC cell lysis was observed when one SM-1 antibody treatment was followed by three treatments with human complement. In contrast, there was little antibody-dependent lysis of non-small cell lung cancer cells, other carcinomas, and leukemia cell lines. The amount of chromium released from normal bone marrow cells treated with SM-1 antibody and complement was minimal and was mainly due to the effect of complement alone. Clonogenic assays, including colony-forming unit-granulocytic/monocytic, erythroid burst-forming unit, and colony-forming unit-granulocytic/erythroid/monocytic/megakaryocytic, also showed no significant SM-1 antibody-dependent cytotoxicity on normal bone marrow precursors. Since SM-1 antibody is selectively cytotoxic to SCC cells in the presence of human complement, it is a potentially useful agent for the selective eradication of tumor cell contamination in marrows of patients with metastatic small cell lung cancer and possibly for in vivo serotherapy.

Antibodies, Monoclonal↗
PubMed2003

Monoclonal LYM-1 antibody-dependent cytolysis by human neutrophils exposed to GM-CSF: auto-regulation of target cell attack by cathepsin G.

Murine monoclonal antibody (mAb) Lym-1 is an immunoglobulin G2a specific for certain human leukocyte antigen-DR variants expressed on the surface of malignant B cells. It has been proposed for serotherapy in patients with B lymphomas. We have previously shown that mAb Lym-1 synergizes with granulocyte macrophage-colony stimulating factor to promote Raji B-lymphoid cell lysis by human neutrophils via the intervention of neutrophil Fc receptors type II and D-mannose-inhibitable interactions between CD11b-CD18 integrins and CD66b glycoproteins. Here, we provide evidence that the process is oxygen-independent by inference related to the release of primary granules and is regulated by cathepsin G activity. The lysis was indeed reproduced by replacing normal neutrophils with cells from three patients suffering from chronic granulomatous disease, i.e., neutrophils genetically incapable of generating oxidants. Moreover, the lysis was inhibited by the serine protease inhibitor 3,4-dichloroisocoumarin and by Z-glycyl-leucyl-phenyl-chloromethyl ketone (Z-Gly-Leu-Phe-CMK), which blocks cathepsin G. Conversely, the lysis was unaffected by N-methoxysuccinyl-alanyl-alanyl-prolyl-alanyl-CMK (MeOSuc-Ala-Ala-Pro-Ala-CMK; elastase inhibitor) and MeOSuc-Ala-Ala-Pro-valine (Val)-CMK, which inhibits elastase and proteinase 3. The ability of neutrophils, engaged in cytolysis, to release cathepsin G was proved by detecting this enzymatic activity spectrophotometrically and immunocytochemically. Moreover, inhibition of cathepsin G activity by concentrations of Z-Gly-Leu-Phe-CMK, incapable of affecting elastase activity, was found to reduce the release of elastase and myeloperoxidase from neutrophils under conditions similar to those used for cytolytic assays. These findings suggest that neutrophils auto-regulate their lytic efficiency by controlling the exocytosis of primary granules via their cathepsin G activity.

Antibodies, Monoclonal↗
PubMedDate not supplied

Neostigmine in the treatment of snake accidents caused by Micrurus frontalis: report of two cases (1).

Antivenom in order to be effective in the treatment of coral snake accidents must be injected very soon after the bite owing to the rapid rate of absorption of the venom neurotoxins. As this is not always possible, other forms of treatment besides serotherapy must be employed to avoid asphyxia and death. Neostigmine and artificial respiration are used for this purpose. Neostigmine restores neuromuscular transmission if the venom-induced blockade results from a reversible interaction of its neurotoxins with the end-plate receptors. This is the mechanism of the neuromuscular blockade produced by the venom of M. frontalis snakes from centereastern and southern Brazil, and Argentine. Neostigmine is able, therefore, to antagonize the blockade, and has been shown to be very effective in the treatment of the experimental envenomation of dogs and monkeys. In the present communication, two cases of M. frontalis accidents treated with antivenom and neostigmine are reported. In both, neostigmine was successful in producing regression of the paralysis, confirming the effectiveness shown in the treatment of the poisoning induced in animals by M. frontalis venom.

Adult↗
PubMedDate not supplied

[Retrospective study on Latrodectus stings in Bahia, Brazil].

This work is a retrospective study of latrodectism in the State of Bahia, Brazil, from August 1980 to July 1990. The data concerning the accidents were obtained from file cards at the Antivenom Information Center of Bahia (AVICB). Latrodectus curacavienis was the ethiologic agent identified in 28% of the arachnid accidents. The major incidence was registered in urban area (57%) affecting men (70%) more than women, with 10 to 29 year-old age group (58%). Local pain (56%), erythematous papula (29%) and light oedema (17%) were the principal local symptoms. Pain in the limbs (29%), tremor and rigidities (29%), sweating (28%), limbs and arms paresthesia (21%) and abdominal pain (17%) were systemic ones. The treatment was mainly symptomatic (67%) and antivenin serum was used in 21% of the cases. After serotherapy, 64% of the patients left the hospital within less than 24 hours.

Adolescent↗
PubMed2006

Allogeneic hematopoietic stem cell transplantation from an alternative stem cell source in Fanconi anemia patients: analysis of 47 patients from a single institution.

We transplanted 47 patients with Fanconi anemia using an alternative source of hematopoietic cells. The patients were assigned to the following groups: group 1, unrelated bone marrow (N = 15); group 2, unrelated cord blood (N = 17), and group 3, related non-sibling bone marrow (N = 15). Twenty-four patients (51%) had complete engraftment, which was not influenced by gender (P = 0.87), age (P = 0.45), dose of cyclophosphamide (P = 0.80), nucleated cell dose infused (P = 0.60), or use of anti-T serotherapy (P = 0.20). Favorable factors for superior engraftment were full HLA compatibility (independent of the source of cells; P = 0.007) and use of a fludarabine-based conditioning regimen (P = 0.046). Unfavorable factors were > or = 25 transfusions pre-transplant (P = 0.011) and degree of HLA disparity (P = 0.007). Intensity of mucositis (P = 0.50) and use of androgen prior to transplant had no influence on survival (P = 0.80). Acute graft-versus-host disease (GVHD) grade II-IV and chronic GVHD were diagnosed in 47 and 23% of available patients, respectively, and infections prevailed as the main cause of death, associated or not with GVHD. Eighteen patients are alive, the Kaplan-Meyer overall survival is 38% at approximately 8 years, and the best results were obtained with related non-sibling bone marrow patients. Three recommendations emerged from the present study: fludarabine as part of conditioning, transplant in patients with < 25 transfusions and avoidance of HLA disparity. In addition, an extended family search (even when consanguinity is not present) seeking for a related non-sibling donor is highly recommended.

Acute Disease↗
PubMed1994

Traditional methods used for controlling animal diseases in Iran.

In ancient times in Iran, infectious diseases of animals and human beings were referred to as choleraic diseases. Rhazes (9th century), followed by Avicenna (10th century), Jorjani (11th century) and others, had specific opinions on the cause and effect relationship in these diseases, which recall the fermentation theory of Louis Pasteur. In ancient Iran, the methods adopted for veterinary procedures were those of general theoretical and practical medicine, including the humoral theory, accurate diagnosis, signs and symptoms, and the prescription of herbal and mineral medicines or substances of animal origin. If herbal treatment failed, cauterisation and surgery were used. When refractory and contagious infectious diseases occurred, animals were evacuated from the infected region, in order to preserve their health, with resort to the mercy of Allah (God) as a final remedy. Iranian scientists of ancient times had interesting views on rabies. A kind of serotherapy was used for treating persons bitten by rabid dogs. Vaccination was performed many centuries ago by using dried smallpox lesions. In Baluchistan (Iran), infants were encouraged to play with and touch the teats of cows affected with cowpox, in order to immunise the children against smallpox, and this was centuries before the discovery of smallpox vaccine by Edward Jenner. Camelpox was also used for human immunisation. In the case of caprine pleuropneumonia, an extract or juice was obtained from the lungs of affected animals and was inactivated by treatment with certain herbal medicines which had a disinfectant effect. A thread coated with this extract was passed through the ear of healthy goats to render them immune. The author lists various diseases and their treatment. This work forms part of detailed research by the author with reference to some 2,200 books and many ancient manuscripts on the history of veterinary science in Iran.

Animal Diseases↗
PubMed1995

Prevention of thromboses in human patients with Bothrops lanceolatus envenoming in Martinique: failure of anticoagulants and efficacy of a monospecific antivenom. Research Group on Snake Bites in Martinique.

Envenomation by the Bothrops lanceolatus, a snake found only in Martinique, leads to swelling and pain, and occasionally to systemic signs and/or coagulopathy. Severe thromboses at some distance from the site of the bite may appear within 48 hr. Uncertainties as to the actual development of thrombotic complications in patients appearing to be suffering from moderate poisoning and as to the availability and the toxicity of a monospecific antivenom (AVS) initially led us to reserve antivenom for the most severe cases, and to use anticoagulants to prevent thromboses in all patients. This approach was modified after we observed serious thromboses in patients with moderate poisoning. Of 50 adult snake bite cases hospitalized between June 1991 and August 1994, 11 developed serious thrombotic complications at 36 /+- 27 hr (mean +/- SD) (range 12-96) following envenomation, despite early preventive anticoagulant therapy. Those included pulmonary embolism (two cases), cerebral infarction (six cases), myocardial infarction (one case), and cerebral and myocardial infarctions (two cases). Sixteen patients were not treated with AVS: 10 of these recovered without complications and six developed systemic thrombosis causing permanent disability in three cases. Thirty were treated with an intravenous infusion of 2-6 vials of AVS given 2-48 hr after the bite. Of these, three died of cerebral infarction that developed before the initiation of serotherapy. All others recovered. Among patients treated with AVS, three presented with mild anaphylactic reactions, while one developed serum sickness that responded to steroids. These data indicate that preventive anticoagulant therapy is of limited efficacy in Martinique.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1984

Recombinant leukocyte A interferon: an active agent in advanced cutaneous T-cell lymphomas.

High-dose recombinant leukocyte A interferon (50 X 10(6) U/m2 body surface area, intramuscularly, three times weekly) was tested in a clinical trial involving patients with advanced cutaneous T-cell lymphomas to determine its effectiveness and toxicity. All 20 patients had advanced stages of disease refractory to two or more standard therapies. Objective partial remissions lasting 3 months to more than 25 months (median, 5 months) were documented in 9 patients. The major dose-limiting toxicity was a severe influenza-like syndrome with malaise, anorexia, depression, weight loss, and decreased performance status; this effect was reversible after dose reductions in all patients and did not recur with re-escalation of doses in 10 patients. This interferon preparation is highly effective in the treatment of advanced refractory cutaneous T-cell lymphomas, and new schedules to reduce toxicity and achieve complete responses, combined treatment with chemotherapy or serotherapy, and its use in earlier stages of disease should be investigated.

Adult↗
PubMedDate not supplied

[Epidemiological and clinical aspects of snake bites in the municipalities of the state of Amazonas, Brazil].

In the State of Amazonas, accidents with snakes are a public health problem. For this reasons, the objective of this work was to carry out a descriptive study of the snake accidents attended in the health units of 34 municipalities, one district and two border platoons in the State of Amazonas. The characteristics most commonly observed among those involved in snake accidents were: farmers (50. 4%), male (81.3%), belonging to the working age-group (72.1%), bitten on an upper limb (88.5%) by a "jararaca" (48.6%) or a "surucucu" (46.8%) in the rural part of the municipality (70.2%). The local signs and symptoms most frequently observed in those who received medical care more than 6 hours after the accident (57.3%) were edema (76.9%), pain (68.7%), erithema (10.2%) and hemorrhage (9. 3%). The systemic manifestation most frequently observed was hemorrhage (18.8%). Serotherapy was administered in only 65.9% of patients, the intravenous route being the route most commonly used to administer the antivenin (52.3%), while other non- recommended routes were widely used. In the majority of patients the antivenin given was antibotropic. The most frequent complications were: abscess 13.7%, necrosis 12.3%, secondary infection 8.3%, renal insufficiency 2.5% and gangrene 2.5%. The medical procedures most used in the treatment of these complications were drainage 52.6%, debridement 28.9%, amputation 10.5%, surgical cleaning 5.3% and peritoneal dialysis 2.6%. The fatality rate was 1%.

Adolescent↗
PubMed2000

[Apropos of a case of rabies occurring despite vaccination after exposure].

Rabies is endemic in Sub-Saharan Africa. Control is based on vaccination of subjects at high-risk for exposure and prompt post-exposure treatment. However the severity of some animal bites and early mismanagement can lead to development of rabies despite post-exposure treatment. This danger is illustrated by the case described herein involving a 35-year-old man who was bitten by a rabid dog and treated with 5 doses of antirabies vaccine. The wound, which consisted of two deep punctures on the forearm, was promptly washed with water and disinfected. Despite this treatment, characteristic clinical manifestations of rabies appeared one month after exposure and the patient died 48 hours later. This case raises the question of the causes for failure of the preventive recommendations proposed by the WHO for category III animal bites. In our case, two possible causes of failure and subsequent lethal outcome are that serotherapy was not associated and the incubation period was short. Other possible causes of rabies after post-exposure therapy are discussed with reference to the literature.

Adult↗
PubMed2000

[Pneumococcal antibiotic resistance].

In 1875, 7 years prior to the description of the Koch bacillus, Klebs visualized the first Streptococcus pneumoniae in a pleural fluid. Since then, this organism has played a determinant role in biomedical science. From a biological point of view, it was largely implicated in the development of passive and active immunization by serotherapy and vaccination, respectively. Genetic transformation was also first observed in S. pneumoniae, leading to the discovery of DNA. From a clinical point of view, S. pneumoniae is still today a prime cause of otitis media in children and of pneumonia in all age groups, as well as a predominant cause of meningitis and bacteremia. In adults, bacteremia is still entailed with a mortality of over 25%. Although S. pneumoniae remained very sensitive to penicillin for many years, penicillin-resistance has emerged and increased dramatically over the last 15 years. During this period of time, the frequency of penicillin-resistant isolates has increased from < or = 1% to frequencies varying from 20 to 60% in geographic areas as diverse as South Africa, Spain, France, Hungary, Iceland, Alaska, and numerous regions of the United States and South America. In Switzerland, the current frequency of penicillin-resistant pneumococci ranges between 5 and > or = 10%. The increase in penicillin-resistant pneumococci correlates with the intensive use of beta-lactam antibiotics. The mechanism of resistance is not due to bacterial production of penicillinase, but to an alteration of the bacterial target of penicillin, the so-called penicillin-binding proteins. Resistance is subdivided into (i) inter mediate level resistance (minimal inhibitory concentration [MIC] of penicillin of 0.1-1 mg/L) and (ii) high level resistance (MCI > or = 2 mg/L). The clinical significance of intermediate resistance remains poorly defined. On the other hand, highly resistant strains were responsible for numerous therapeutical failures, especially in cases of meningitis. Antibiotics recommended against penicillin-resistant pneumococci include cefotaxime, ceftriaxone, imipenem and in some instances vancomycin. However, penicillin-resistant pneumococci tend to present cross-resistances to all the antibotics of the beta-lactam family and could even become resistant to the last resort drugs mentioned above. Thus, in conclusion, the explosion of resistance to penicillin in pneumococci is a ubiquitous phenomenon which must be fought against by (i) a strict utilization of antibiotics, (ii) the practice of microbiological sampling of infected foci before treatment, (iii) the systematic surveillance of resistance profiles of pneumococci against antibiotics and (iv) the adequate vaccination of populations at risk.

Adult↗
PubMed1998

[Use of toxic fraction isolated from Algerian Androctonus australis hector scorpion venom for the assessment of anti-venom serum].

Today, serotherapy is the only specific treatment used against the scorpionique envenomation. This treatment was associated with other drugs as a symptomatic and supportive care. The anti-venom against scorpion stings is still prepared using the conventional manufacturing production. To evaluate the efficacy of the anti-venom, different protocols are used: The immunization of animals (rabbit and horse) with a whole toxic fraction prepared by molecular filtration was the first step to improve the anti-venom. Some observations also show the need for the quality control of this preparation of anti-venom. In this study, we describe the use the toxic fraction isolated from Androctonus australis Hector in the new protocol of anti-venom production. The quality control of the product is evaluated by using a combination of experimental immunological assay systems (ELISA, Hemaglutination, in vivo assessment of the neutralization of the lethal effect of the venom).

Algeria↗