Bothrops Lanceolatus Research Articles

Screening of antimicrobial activity in venom: Exploring key parameters

The escalating challenge of antibiotic resistance significantly threatens global health, underscoring the critical need for new antimicrobial agents. Venoms, increasingly recognized as reservoirs of bioactive compounds with diverse pharmacological effects, have been the focus of recent research. This work evaluates the use of various screening methodologies in assessing the antimicrobial activities of 185 venoms against some gram positive and gram negative bacteria, including E. coli ATCC 8739, B. subtilis ATCC 6633, P. aeruginosa ATCC 9027, and S. aureus ATCC 6538P species and explores the influence of settings on the findings. Furthermore, the research explored the possibility of purifying antimicrobial molecules from venoms through HPLC. Several fractions demonstrated antimicrobial activity against the tested strains.Our results reveal that the measured antimicrobial efficacy of venoms varies according to:i) venom concentration, ii) the detection method, including microdilution and radial diffusion assays, and iii) the choice of culture medium, specifically LB or MH.This strategy has allowed us, for the first time, to identify antimicrobial activity in: i) Bitis arietans venom against P. aeruginosa ATCC 9027, ii) Naja nubiae and Bothrops lanceolatus against B. subtilis ATCC 6633, P. aeruginosa ATCC 9027, and S. aureus ATCC 6538P, and iii) Hadogenes zuluanus, Mesobuthus caucasicus, Nebo hierichonticus, Opistophthalmus wahlbergii scorpions, and Mylabris quadripunctata beetles against S. aureus ATCC 6538P.These findings highlight venoms potential as effective antimicrobial resources and improve our understanding of key factors critical for an accurate detection of venoms antimicrobial properties.

A murine experimental model of the pulmonary thrombotic effect induced by the venom of the snake Bothrops lanceolatus.

The venom of Bothrops lanceolatus, a viperid species endemic to the Lesser Antillean Island of Martinique, induces thrombosis in a number of patients. Previous clinical observations indicate that thrombotic events are more common in patients bitten by juvenile specimens. There is a need to develop an experimental model of this effect in order to study the mechanisms involved. The venoms of juvenile and adult specimens of B. lanceolatus were compared by (a) describing their proteome, (b) assessing their ability to induce thrombosis in a mouse model, and (c) evaluating their in vitro procoagulant activity and in vivo hemostasis alterations. Venom proteomes of juvenile and adult specimens were highly similar, albeit showing some differences. When injected by the intraperitoneal (i.p.) route, the venom of juvenile specimens induced the formation of abundant thrombi in the pulmonary vasculature, whereas this effect was less frequent in the case of adult venom. Thrombosis was not abrogated by the metalloproteinase inhibitor Batimastat. Both venoms showed a weak in vitro procoagulant effect on citrated mouse plasma and bovine fibrinogen. When administered intravenously (i.v.) venoms did not affect classical clotting tests (prothrombin time and activated partial thromboplastin time) but caused a partial drop in fibrinogen concentration. The venom of juvenile specimens induced partial alterations in some rotational thromboelastometry parameters after i.v. injection. When venoms were administered i.p., only minor alterations in classical clotting tests were observed with juvenile venom, and no changes occurred for either venom in rotational thromboelastometry parameters. Both juvenile and adult venoms induced a marked thrombocytopenia after i.p. injection. An experimental model of the thrombotic effect induced by B. lanceolatus venom was developed. This effect is more pronounced in the case of venom of juvenile specimens, despite the observation that juvenile and adult venom proteomes are similar. Adult and juvenile venoms do not induce a consumption coagulopathy characteristic of other Bothrops sp venoms. Both venoms induce a conspicuous thrombocytopenia. This experimental model reproduces the main clinical findings described in these envenomings and should be useful to understand the mechanisms of the thrombotic effect.

Conservation of Coluber lanceolatus as the valid name of the Martinique fer-de-lance (no matter who its author is)

The fer-de-lance of Martinique is a pitviper of medical importance and conservation concern. It was described by La Cepède in 1789 as Coluber lanceolatus but as a result of a ruling of the Commission (Opinion 1463) this nomen is unavailable. Although a proposed new use of the Plenary Power by the Commission could restore the availability of Coluber lanceolatus La Cepède, 1789, this possibility remains hypothetical and under the Code-compliant interpretation, C. lanceolatus was made available by Bonnaterre in 1790 who proposed a homonymous poieonym. Coluber lanceolatus, which, as Bothrops lanceolatus, is currently universally accepted as the valid nomen of the fer-de-lance, is at risk because of the existence of two older nomina that apply to this taxon, Vipera cærulescens Laurenti, 1768 and its replacement name Coluber glaucus Gmelin, 1789. Consequently, we here provide the evidence necessary to demonstrate that Vipera cærulescens and Coluber glaucus are nomina oblita and to designate Coluber lanceolatus, under both the current proposed interpretations of its authorship, as a nomen protectum. Coluber lanceolatus Bonnaterre, 1790 has precedence over Coluber brasiliensis and C. tigrinus, two nomina with which it is considered synonymous and which were made available in the same work. However, precedence was established as a result of determination by the First Reviser only as recently as 2021. The case of Bothrops lanceolatus highlights that even well-established and widely accepted nomina may be potentially vulnerable to destabilisation by long-forgotten synonyms and illustrates the particular nomenclatural complications that can arise from valid Bonnaterre nomina that are homonymous with long-used but now unavailable La Cepède names.

Increased Binding of von Willebrand Factor to Sub-Endothelial Collagen May Facilitate Thrombotic Events Complicating Bothrops lanceolatus Envenomation in Humans.

Consumption coagulopathy and hemorrhagic syndrome exacerbated by blood anticoagulability remain the most important causes of lethality associated with Bothrops snake envenomation. Bothrops venom also engages platelet aggregation on the injured endothelium via von Willebrand factor (vWF) interactions. Besides platelet aggregation, some Bothrops venom toxins may induce qualitative thrombopathy, which has been in part related to the inhibition of vWF activation. We tested whether B. lanceolatus venom impaired vWF to collagen(s) binding (vWF:CB) activity. Experiments were performed with B. lanceolatus crude venom, in the presence or absence of Bothrofav, a monospecific B. lanceolatus antivenom. Venom of B. lanceolatus fully inhibited vWF to collagen type I and III binding, suggesting venom interactions with the vWF A3 domain. In contrast, B. lanceolatus venom increased vWF to collagen type VI binding, suggesting the enhancement of vWF binding to collagen at the vWF A1 domain. Hence, B. lanceolatus venom exhibited contrasting in vitro effects in terms of the adhesive properties of vWF to collagen. On the other hand, the antivenom Bothrofav reversed the inhibitory effects of B. lanceolatus venom on vWF collagen binding activity. In light of the respective distribution of collagen type III and collagen type VI in perivascular connective tissue and the sub-endothelium, a putative association between an increase in vWF:CB activity for collagen type VI and the onset of thrombotic events in human B. lanceolatus envenomation might be considered.

New Insights into Immunopathology Associated to Bothrops lanceolatus Snake Envenomation: Focus on PLA2 Toxin.

The systemic increase in inflammatory mediator levels can induce diverse pathological disorders, including potentially thrombus formation, which may be lethal. Among the clinical conditions in which the formation of thrombi dictates the patient's prognosis, envenomation by Bothrops lanceolatus should be emphasized, as it can evolve to stroke, myocardial infarction and pulmonary embolism. Despite their life-threatening potential, the immunopathological events and toxins involved in these reactions remain poorly explored. Therefore, in the present study, we examined the immunopathological events triggered by a PLA2 purified from B. lanceolatus venom, using an ex vivo human blood model of inflammation. Our results showed that the purified PLA2 from the venom of B. lanceolatus damages human erythrocytes in a dose dependent way. The cell injury was associated with a decrease in the levels of CD55 and CD59 complement regulators on the cell surface. Moreover, the generation of anaphylatoxins (C3a and C5a) and the soluble terminal complement complex (sTCC) indicates that human blood exposure to the toxin activates the complement system. Increased production of TNF-α, CXCL8, CCL2 and CCL5 followed complement activation. The venom PLA2 also triggered the generation of lipid mediators, as evidenced by the detected high levels of LTB4, PGE2 and TXB2. The scenario here observed of red blood cell damage, dysfunctions of the complement regulatory proteins, accompanied by an inflammatory mediator storm, suggests that B. lanceolatus venom PLA2 contributes to the thrombotic disorders present in the envenomed individuals.

Antibiotic susceptibility of cultivable microbiota from the oral cavity of captive Bothrops atrox and Bothrops lanceolatus: Implications for the treatment of snakebite-associated infections in the French departments of America

IntroductionSecondary infection is a frequent complication after bites by Bothrops atroxin French Guiana or B. lanceolatus in Martinique. Knowledge of the bacteria present in snake mouths is a valuable aid for determining probabilistic antibiotherapy after Bothrops bite. The objectives of this study were to describe the cultivable bacteria of the oral microbiota of the specimens of B. atrox and B. lanceolatus kept in captivity, and to study their susceptibility to antibiotics. MethodsFifteen B. atrox and 15 B. lanceolatus were sampled. Bacterial cultures were performed and each morphotype on plates was identified using MALDI-TOF mass spectrometry. Antibiotic susceptibility was studied using the agar disk diffusion method, with possible determination of the MICs. ResultsOne hundred and twenty-two isolates were identified: 52 isolates and 13 species in B. atrox, 70 isolates and 23 species in B. lanceolatus. The main species were Providencia rettgeri, Morganella morganii, Pseudomonas aeruginosa, Staphylococcus xylosus, and Paeniclostridium sordellii (only in B. lanceolatus mouths). For B. atrox, 96% of isolates were susceptible to piperacillin/tazobactam, cefepime, imipenem and meropenem, 94% to ciprofloxacin and 76% to cefotaxime and ceftriaxone. For B. lanceolatus, 97% of isolates were susceptible to meropenem, 96% to cefepime, 93% to imipenem and piperacillin/tazobactam, 80% to ciprofloxacin, and 75% to cefotaxime and ceftriaxone. Many isolates were resistant towards amoxicillin/clavulanate. ConclusionAmong currently recommended antibiotics, cefepime and piperacillin/tazobactam seem more suitable than cefotaxime or ceftriaxone in the event of a Bothrops bite. Ciprofloxacin may also be considered for B. atrox.

Bothrops lanceolatus snake venom impairs mitochondrial respiration and induces DNA release in human heart preparation.

Envenomations by Bothrops snakebites can induce overwhelming systemic inflammation ultimately leading to multiple organ system failure and death. Release of damage-associated molecular pattern molecules (DAMPs), in particular of mitochondrial origin, has been implicated in the pathophysiology of the deregulated innate immune response. To test whether whole Bothrops lanceolatus venom would induce mitochondrial dysfunction and DAMPs release in human heart preparations. Human atrial trabeculae were obtained during cannulation for cardiopulmonary bypass from patients who were undergoing routine coronary artery bypass surgery. Cardiac fibers were incubated with vehicle and whole Bothrops lanceolatus venom for 24hr before high-resolution respirometry, mitochondrial membrane permeability evaluation and quantification of mitochondrial DNA. Compared with vehicle, incubation of human cardiac muscle with whole Bothrops lanceolatus venom for 24hr impaired respiratory control ratio and mitochondrial membrane permeability. Levels of mitochondrial DNA increased in the medium of cardiac cell preparation incubated with venom of Bothrops lanceolatus. Our study suggests that whole venom of Bothrops lanceolatus impairs mitochondrial oxidative phosphorylation capacity and increases mitochondrial membrane permeability. Cardiac mitochondrial dysfunction associated with mitochondrial DAMPs release may alter myocardium function and engage the innate immune response, which may both participate to the cardiotoxicity occurring in patients with severe envenomation.

A complex biogeographic history of diversification in Neotropical lancehead pitvipers (Serpentes, Viperidae)

AbstractBased on the literature, we had predicted that the diversification within the Neotropical snake genusBothropsoccurred along a latitudinal gradient from north to south, with diversification into unoccupied niches through ecological opportunity, not correlated with geoclimatic events. Using a dated phylogeny and estimating likelihoods of ancestral states at cladogenesis events, we reconstructed ancestral areas and assessed major events of the diversification ofBothropsclades, and we also discuss systematic implications for this group. Based on the phylogeny we produced,B. lojanuswas not considered as part of the genusBothropssince the results recovered this species nested within theBothrocophiasclade. We infer that the diversification of the MioceneBothrops pictusandBothrops alternatusclades may be related to the uplift of the western slopes of the Andes and the Argentinian Patagonian Andes, respectively. The PlioceneBothrops taeniatusandBothrops osborneiclades may be related to the uplift of the eastern and northern Andes, respectively. The Plio‐PleistoceneBothrops neuwiediclade may be related to the habitat transitions from a warmer and forested environment to a cooler and open landscape; theBothrops jararaca(i.e. island endemic species) andBothrops lanceolatusclades to over‐water dispersal with island speciation; andBothrops atroxclade to the appearance of the Panamanian land bridge. We found that a multitemporal and multidirectional history of diversification may be correlated with geoclimatic and dispersalist events. We argue that the vacant niche hypothesis by itself does not explainBothropsdiversification.

Infectious Complications Following Snakebite by Bothrops lanceolatus in Martinique: A Case Series.

.Infections secondary to snakebite occur in a number of patients and are potentially life-threatening. Bothrops lanceolatus bites in Martinique average 30 cases per year and may result in severe thrombotic and infectious complications. We aimed to investigate the infectious complications related to B. lanceolatus bite. A retrospective single-center observational study over 7 years (2011–2018) was carried out, including all patients admitted to the hospital because of B. lanceolatus bite. One hundred seventy snake-bitten patients (121 males and 49 females) were included. Thirty-nine patients (23%) presented grade 3 or 4 envenoming. Twenty patients (12%) developed wound infections. The isolated bacteria were Aeromonas hydrophila (3 cases), Morganella morganii (two cases), group A Streptococcus, and group B Streptococcus (one case each). Patients were treated empirically with third-generation cephalosporin (or amoxicillin–clavulanate), aminoglycoside, and metronidazole combinations. Outcome was favorable in all patients. The main factor significantly associated with the occurrence of infection following snakebite was the severity of envenoming (P < 0.05). Our findings clearly point toward the frequent onset of infectious complications in B. lanceolatus–bitten patients presenting with grade 3 and 4 envenoming. Thus, based on the bacteria identified in the wounds, we suggest that empiric antibiotic therapy including third-generation cephalosporin should be administered to those patients on hospital admission.

Third-generation antivenomics analysis of the preclinical efficacy of Bothrofav® antivenom towards Bothrops lanceolatus venom.

Bothrops lanceolatus inflicts severe envenomings in the Lesser Caribbean island of Martinique. Bothrofav®, a monospecific antivenom against B. lanceolatus venom, has proven highly effective at the preclinical and clinical levels. Here, we report a detailed third-generation antivenomics quantitative analysis of Bothrofav®. With the exception of poorly-immunogenic peptides, Bothrofav® immunocaptured all the major protein components. These results, along with previous preclinical and clinical observations, underscore the high neutralizing efficacy of the antivenom against B. lanceolatus venom.

Oral Microbiota of the Snake Bothrops lanceolatus in Martinique.

In Martinique, Bothrops lanceolatus snakebite, although relatively uncommon (~30 cases/year), may result in serious complications such as systemic thrombosis and local infections. Infections have been hypothesized to be related to bacteria present in the snake’s oral cavity. In this investigation, we isolated, identified, and studied the susceptibility to beta-lactams of bacteria sampled from the oral cavity of twenty-six B. lanceolatus specimens collected from various areas in Martinique. Microbiota from B. lanceolatus oral cavity was polymicrobial. Isolated bacteria belonged to fifteen different taxa; the most frequent being Aeromonas hydrophyla (present in 50% of the samples), Morganella morganii, Klebsiella pneumoniae, Bacillus spp., and Enterococcus spp. Analysis of antibiotic susceptibility revealed that 66.7% of the isolated bacteria were resistant to amoxicillin/clavulanate. In contrast, the majority of isolated bacteria were susceptible to the third-generation cephalosporins (i.e., 73.3% with cefotaxime and 80.0% with ceftazidime). Microbiota from B. lanceolatus oral cavity is polymicrobial with bacteria mostly susceptible to third-generation cephalosporins but rarely to amoxicillin/clavulanate. In conclusion, our findings clearly support that first-line antibiotic therapy in the B. lanceolatus-bitten patients, when there is evidence of infection, should include a third-generation cephalosporin rather than amoxicillin/clavulanate.

The Ex vivo Eye Irritation Test (EVEIT) model as a mean of improving venom ophthalmia understanding

Snakes belonging to the genus Naja (Elapid family), also known as “spitting cobras”, can spit venom towards the eyes of the predator as a defensive strategy, causing painful and potentially blinding ocular envenoming. Venom ophthalmia is characterized by pain, hyperemia, blepharitis, blepharospasm and corneal erosions. Elapid venom ophthalmia is not well documented and no specific treatment exists. Furthermore, accidental ejection of venom by non-spitting vipers, as Bothrops, also occurs. The Ex vivo Eye Irritation Test model (EVEIT) has enabled important progress in the knowledge of chemical ocular burns. Considering the lack of experimental animal model, we adapted the EVEIT to study venom ophthalmia mechanisms. Ex vivo rabbit corneas were exposed to venoms from spitting (Naja mossambica, Naja nigricollis) and non-spitting (Naja naja, Bothrops jararaca and Bothrops lanceolatus) snakes, and rinsed or not with water. The corneal thickness and the depth of damage were assessed using high-resolution optical coherence tomography (HR-OCT) imaging and histological analysis. All Naja venoms induced significant corneal edema, collagen structure disorganization and epithelial necrosis. Corneas envenomed by African N. mossambica and N. nigricollis venoms were completely opaque. Opacification was not observed in corneas treated with venoms from non-spitting snakes, such as the Asian cobra, N. naja, and the vipers, B. jararaca and B. lanceolatus. Moreover, Bothrops venoms were able to damage the epithelium and cause collagen structure disorganization, but not edema. Immediate water rinsing improved corneal status, though damage and edema could still be observed. In conclusion, the present study shows that the EVEIT model was successfully adapted to set a new experimental ex vivo animal model of ophthalmia, caused by snake venoms, which will enable to explore new therapies for venom ophthalmia.

Preclinical evaluation of the neutralizing ability of a monospecific antivenom for the treatment of envenomings by Bothrops lanceolatus in Martinique

Bothrops lanceolatus is an endemic viperid species in the Lesser Caribbean island of Martinique. Envenomings by this species are characterized by local and systemic effects, among which the development of thrombosis in various organs is the most severe complication. An experimental toxicological characterization of this venom was performed using in vivo mouse tests and various in vitro assays. The venom induced lethal, local and systemic hemorrhagic, edema-forming, myotoxic, thrombocytopenic, proteinase and phospholipase A2 activities. The preclinical efficacy of a batch of monospecific Bothrofav® antivenom currently in use in Martinique was assessed. The antivenom was highly effective in the neutralization of all activities tested, in agreement with its described clinical efficacy. This batch of antivenom showed a higher preclinical efficacy as compared to a previous batch used in the past.

Snake Bites by Bothrops lanceolatus in Martinique.

Bothrops lanceolatus, a member of the Viperidae family and Crotalinae subfamily, is the only snake causing envenomation in Martinique, which is the only place it exists in the world. Any bite by this snake may result in severe thrombotic complications including cerebral, pulmonary, and myocardial infarction and affecting the patient's vital or functional prognosis. The exact composition of the venom as well as the mechanisms of toxicity involved are still debated today. The first specific antivenom immunotherapy (AVI), Bothrofav1, manufactured in 1991, reduced mortality and morbidity significantly. In 2004, however, an increase in the number of cases of Bothrops lanceolatus envenomation-induced ischemic strokes despite early AVI administration, raised questions about its effectiveness and encouraged the development of a second AVI, Bothrofav2, which became available in February 2011. Causes of the failures with the first AVI remain hypothetical, and the effectiveness of the second is under evaluation. This article reviews the published data available on this rare envenomation, present in one of our French overseas departments and discusses its fascinating scientific and medical issues.

Enzymatic and Pro-Inflammatory Activities of Bothrops lanceolatus Venom: Relevance for Envenomation.

Bothrops lanceolatus, commonly named ‘Fer-de-Lance’, is an endemic snake of the French Caribbean Island of Martinique. Envenomations by B. lanceolatus present clinical aspects characterized by systemic thrombotic syndrome and important local inflammation, involving edema and pain but limited hemorrhage. To investigate mechanisms of venom-induced inflammation, B. lanceolatus venom was characterized, its cross-reactivity with bothropic antivenom explored, its cytotoxicity on human keratinocytes and vascular cells, and the production of cytokines and chemokines were analyzed. We used electrophoretic separation, zymography, colorimetric or fluorimetric enzymatic assays, and immunochemical assays. Therapeutic South American bothropic antivenom cross-reacted with B. lanceolatus venom and completely or partially abolished its PLA2, hyaluronidase, and proteolytic activities, as well as its cytotoxicity for keratinocytes. The substrate specificity of B. lanceolatus venom proteases was emphasized. B. lanceolatus venom cytotoxicity was compared to the B. jararaca venom. Both venoms were highly cytotoxic for keratinocytes (HaCaT), whereas B. lanceolatus venom showed particularly low toxicity for endothelial cells (EAhy926). Patterns of cytokine and chemokine production by cells exposed to the venoms were highly pro-inflammatory. Thus, the results presented here show that B. lanceolatus venom toxins share important antigenic similarities with South American Bothrops species toxins, although their proteases have acquired particular substrate specificity. Moreover, the venom displays important cytotoxic and pro-inflammatory action on human cell types such as keratinocytes and endothelial cells, which are important players in the local and systemic compartments affected by the envenomation.

Living with or Eradicating Poisonous Snakes in Densely Populated Caribbean Islands—A Socio-Ecological Challenge for the French West Indies

The lancehead (Bothrops lanceolatus) is a poisonous snake endemic to Mar-tinique, an island in the Lesser Antilles arc. Today, this snake is on the verge of extinction. The recorded number of snakes killed yearly between 1970 and 2002 decreased by 97%. Despite the production of an antivenom in 1993, the local authorities set up a financial reward to encourage the eradication of lancehead snakes. Today the local population still perceives the snake as a threat, due to its fatal venom and its fierce behaviour. The case study of the lancehead in Martinique highlights the need to develop integrated strategies to conserve species of poisonous snakes on densely populated islands. This requires innovative, cross-sectoral strategies that involve decision-makers working along with multidisciplinary scientists. Approaching the complexity of ecosystems through the socio-ecological prism implies, conversely, linking up the domains of science and technology, life and environmental sciences, and human and social sciences, through interactions based on sharing common assumptions.

A case of bilateral occipital lobe infarcts following Indian tree viper bite.

A case of bilateral occipital lobe infarcts following Indian tree viper bite.

Endemic or exotic: the phylogenetic position of the Martinique Volcano Frog Allobates chalcopis (Anura: Dendrobatidae) sheds light on its origin and challenges current conservation strategies

Amphibian faunas of the Lesser Antilles are depauperate, with only a few species being endemic and generally threatened. Allobates chalcopis from the island of Martinique is a particularly enigmatic case being the only known dendrobatid endemic to an oceanic island. This species has previously been suggested as being introduced to Martinique. The question of its true origin remained unresolved because no individuals were found since its formal description in the 1990s. Twenty years after the last observation of the species, we succeeded in finding an isolated population of Allobates chalcopis in Martinique. The rediscovery allowed us to investigate the species’ phylogenetic position, confirming that it is nested within a clade of lowland Amazonian Allobates but nonetheless distantly related to any known species of the group. The arrival of the species in Martinique likely corresponds to an overseas dispersal from South America during the late Miocene, as previously hypothesized for Bothrops lanceolatus and Leptodactylus fallax; two other species endemic to Martinique and surrounding islands. However, the species was not found in its type locality 500 m a.s.l. but 300 m higher in altitude, in herbaceous areas of the summit of Montagne Pelée. The possible range reduction and population decline in combination with the evidence of endemicity of the species highlights the need for a reassignment of the current Red List status. Furthermore, a refined conservation strategy is needed to guarantee the long-term viability of Allobates chalcopis in its native range.

Osteopontin, a chemotactic protein with cytokine-like properties, is up-regulated in muscle injury caused by Bothrops lanceolatus (fer-de-lance) snake venom

Osteopontin (OPN) is a chemotactic, adhesive protein whose receptors include some integrins and matrix proteins known to have role in inflammatory and repair processes. We examined the time course of OPN expression at acute and chronic stages after intramuscular injection of Bothrops lanceolatus venom in rats. Additionally, we examined the expression of CD68 (a marker for phagocytic macrophages) and the myogenic factors, myoD and myogenin. There was a biphasic upregulation of OPN (6–48 h and 3–14 days post-venom), i.e., during acute inflammation and myogenic cell proliferation and differentiation phases. OPN was detected in CD68 + macrophages, fibroblasts, normal and damaged myofibers, myoblasts and myotubes. Myogenin was expressed in the cytoplasm (atypical pattern) and nucleus of myoblasts and myotubes from 18 h to 7 days, after which it was expressed only in nuclei. Macrophage numbers, OPN and myogenin expression were still elevated at 7, 14 and 7 days. At 3 days, when OPN achieved the peak, some clusters of myoblasts were within regions of intense collagen deposition. Fibrosis may represent limitation for repairing processes and may explain the small diameter of regenerated fibers at 21 days post-venom. The expression of OPN in the course of venom-induced damage and regeneration suggests stages-specific mediation role along the whole process.

Homoöpathische Therapie beim apoplektischen Insult: Arnica, Nux vomica, Lachesis, Crotalus horridus, Naja

The acute stroke needed - at once - a staging and proceeding. In addition to the medicamental management (Alteplase) in the first hour of the stroke incident the homoeopathic treatment in addition can show very successful results . Beside the so-called „antispastic remedy” nux vomica the homeoepathic snake remedies like lachesis, crotalus horridus, naja and bothrops lanceolatus are recommended.