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Accepted posters (unedited)
(Presenters in Bold)
If your abstract has been accepted for presentation but it does not appear in the list below, please let us know as soon as possible by email at VenomsOxford@gmail.com.
Generation of an oligoclonal VHH mixture for in vivo neutralization of Elapid snake venoms
Nick J. Burlet1*, Shirin Ahmadi1*, Melisa Benard-Valle1, Camilla H. Dahl1, Tasja W. Ebersole1, Emily U. Gregg-Sørensen1, Alid Guadarrama-Martínez2, Kim Boddum3, Timothy P. Jenkins1, Alejandro Alagón2, Anne Ljungars1, and Andreas H. Laustsen1
1Department of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, Denmark
2Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México. México
3Sophion Bioscience A/S. Denmark
*Equal contributions
Every year, around 140,000 people die as a result of snakebite envenoming with many more suffering from long-term complications. While current plasma-derived antivenoms have saved countless lives, they also come with several drawbacks including poor safety profiles, a low content of therapeutically relevant antibodies, and batch-to-batch variation. To address some of these challenges, we explore single-domain antibodies (VHHs) that target medically relevant toxins as an alternative treatment approach. Our aim is to design an oligoclonal mixture of VHHs that can neutralize the lethality of 18 medically relevant Elapid snakes from sub-Saharan Africa. In this poster, I will describe how we discover neutralizing VHHs against specific toxins from medically relevant Elapidae snake species in sub-Saharan Africa. Specifically, I will discuss how we use proteomic data to identify key toxins, our design of phage display discovery campaigns for the selection of cross-reactive VHHs, and data from in vitro and in vivo assays utilized to characterize selected VHHs.
A comprehensive in vitro study to map the neurotoxic profile of black mamba (D. polylepis) venom
Iara A Cardoso1, Mark C Wilkinson1, Rohit N Patel1, Damien Redureau2, Loic Quinton2, Michael Morton3, Nicholas R Casewell1, Stefanie K Menzies1,4
1Centre for Snakebite Research and Interventions, Liverpool School of Tropical Medicine, Liverpool, UK
2Laboratory of Mass Spectrometry, MolSys Research Unit, University of Liège, Liège, Belgium
3ApconiX Ltd, Alderley Park, Cheshire, UK 4Biomedical and Life Sciences, Lancaster University, Lancaster, UK
Black mamba (Dendroaspis polylepis) is a highly venomous snake and ranks among the most medically significant species in sub-Saharan Africa. Its venom is known to be highly neurotoxic, mainly composed of Kunitz-type proteinase inhibitors (dendrotoxins), and toxins belonging to the three-finger toxin (3FTx) family (α-neurotoxins, muscarinic toxins, and fasciculins). The work presented here aimed to map the neurotoxic effects of D. polylepis venom toxins on their human targets. The venom was first fractionated through cation exchange chromatography (CX) and subsequent fractions were assessed with different in vitro assays. A cell-based assay of muscle-type nAChR (nicotinic acetylcholine receptor) function was used to evaluate the presence of α-neurotoxins. The fractions showing nAChR inhibition were then subjected to reversed phase chromatography and the individual toxins were identified by mass spectrometry. CX fractions were analysed by surface plasmon resonance, which provided their profile of binding to a recombinant AChBP (acetylcholine binding protein) and complemented the cell-based assay. Additionally, CX fractions were evaluated regarding their influence on human acetylcholinesterase activity, which can indicate the presence of fasciculins, and automated patch clamp demonstrated the ability of some CX fractions to inhibit Kv 1.1 (a voltage-gated potassium channel), pointing to the presence of dendrotoxins. Altogether, these results provide useful information about black mamba neurotoxins and their potent pre- and post-synaptic effects, highlighting their synergistic and diverse potential.
Metalloprotease Varience in Bothrops jararaca Venom: Proteomics and Activity
Enzo Paulesini de Sousa¹, Nathália da Costa Galizio¹, Jose Aparecido Ferreira De Lima³, Jarbas Prado Vidueiros², Sávio Stefanini Sant’Anna², Daniel Carvalho Pimenta³, Luís Roberto de Camargo Gonçalves¹, Karen de Morais-Zani¹
¹Laboratory of Pathophysiology, Butantan Institute, Brazil
²Laboratory of Herpetology, Butantan Institute, Brazil
³Laboratory of Biochemistry and Biophysics, Brazil
Snake venoms are the most studied animal toxins. The variability in venom composition has been described not only within and interspecies, but also according to sex, age and geographic distribution. The Bothrops genus is responsible for 90% of snakebites in Brazil, with their venom having three main activities: proteolytic or necrotizing, coagulant and hemorrhagic. The resulting clinical aspects from the envenoming from the bites of this genus can be characterized by important local manifestations, owing to the action of different families of toxins, including metalloproteases (SVMP). These enzymes are involved in several pathophysiological processes, such as the development of edema, degradation of extracellular matrix components, hemorrhage, degradation of factors of the coagulation cascade and platelet aggregation inhibition. In general, SVMPs are the most abundant component among Bothrops venoms, as is the case of the Bothrops jararaca species, in which it corresponds to about 25% of the venom. However, in a recent study carried out by our group, it was verified by high-performance liquid chromatography (HPLC) that some individuals of this species have poor amounts of SVMP, which reflects in a lower proteolytic activity on collagen. In addition, this variability regarding SVMP content may influence the pathophysiology of the envenomation caused by this species. Based on this finding, the objective of this work was to analyze 54 individuals of the species B. jararaca of state of São Paulo in order to establish possible venom phenotypes regarding SVMP abundance, and, therefore, contribute to the understanding of how the SVMP variability affects envenomation regarding to lethality and hemorrhagic activity. By HPLC analyses, it was possible to identify four possible venom phenotypes in relation to the content of SVMPs, and while 76% of the individuals displayed the two expected standard peaks related to this toxin family, 11% expressed only the first one, 4% only the second, and 9% showed no relevant HPLC peaks. These evidences were subsequently corroborated by SDS-PAGE, where the same four phenotypes identified according to electrophoretic profile and by mass spectrometry, where we could confirm the the different classes of SVMP. Subsequently, we performed different analyses with the venoms individually (or in pools) measuring the degradation of basement membrane components, their action on fibrinogenolysis, and their coagulant potential, identifying how the different phenotypes act on key aspects of the pathophysiology caused by Bothropic venom. We hope the current work will help our understanding regarding the SVMP role in the hemorrhagic effects, a severe factor in snakebite accidents. This work was supported by FAPESP (2020/07268-2; 2021/07627-5), CNPq (309995/2022-1), CAPES (88887.973680/2024-00), Secretaria de Estado da Saúde de São Paulo and Fundação Butantan.
Action of Marimastat, a broad-spectrum metalloprotease inhibitor, on the neuromuscular disorder by Bothrops alternatus (Viperidae: Crotalinae) venom in rodents
Samuel R. Dias1, Gabrielly C. Souza-Gomes1, Vitória S. Proença1, Isabele N. Oliveira1, Hugo A. Ghirotti1, Sofia N. S. Azevedo1, Poliana J. Demico1, Sofia N.S. Azevedo1, Kristian A. Torres-Bonilla2, Stephen Hyslop2, Sávio S. Sant’Anna3, Karen Morais-Zani3, Rogério Giuffrida1, Rafael S. Floriano1
1Laboratory of Toxinology and Cardiovascular Research, University of Western São Paulo (UNOESTE), Presidente Prudente, SP, Brazil
2Department of Translational Medicine, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, SP, Brazil
3Laboratory of Herpetology, Butantan Institute (IB), São Paulo, SP, Brazil
Envenomation by Bothrops snakes cause intense local damage and systemic disfunctions mostly mediated by proteases. Marimastat (MMT), an anticancer drug, has shown considerable ability in preventing snake venom-induced disturbances. Here, we assessed the action of MMT alone or associated with an anti-Bothrops antivenom (AV) on the neuromuscular disorder induced by Bothrops alternatus (BAv) in vitro. Proteolytic (caseinolytic) and esterasic activities were determined by spectrophotometry using artificial substrates (azocasein and DL-BAPNA, respectively). Mouse phrenic nerve-diaphragm preparation was mounted in an Ugo Basile SRL myographic system under 1 g tension, with twitches being indirectly evoked by supramaximal stimuli (0.2 Hz, 0.1 ms) and recorded using a DY1 force displacement transducer coupled to a DataCapsule-Evo and LabScribe4 software. MMT (0.03–3 mM) exhibited concentration-dependent inhibitory action on the caseinolytic activity of BAv, with minimum interference on the esterasic activity at 3 mM; MMT (0.3 mM) was chosen as minimum inhibitory concentration for neuromuscular analysis [DA440nm/min: 4.2 ± 0.1 (BAv 1 mg/ml) vs. 1.1 ± 0.1 (BAv + MMT 0.3 mM), p<0.05 vs. BAv alone, n=3]. BAv (10–100 mg/ml) produced concentration-dependent neuromuscular blockade, with the highest concentration (100 mg/ml) reaching 50% and 90% blockade in 48±7 and 89±13 min, respectively (p<0.05 vs. control preparation, n=4). MMT (0.3 mM) significatively delayed the BAv (100 mg/ml)-induced neuromuscular blockade, with 50% blockade occurring in 86±11 min (p<0.05 vs. BAv alone, n=4). MMT (0.3 mM) combined with AV (antivenom:venom ratio of 1:5 v/w) avoided the occurrence of 50% blockade in 120 min incubation. In conclusion, MMT exhibits potential application as an useful pharmacological strategy to prevent neuromuscular disorder caused by BAv venom.
Generation of chimeric antibodies from horses immunised with sub-Saharan African snake venoms
Rhys W. Dunphy1, Sandra Ergueta-Carballo1, Sheikh Arif1, Jonathan Rockman1, Andrés Sánchez2, Julien Slagboom3, Jeroen Kool3, José M. Gutiérrez2, John McCafferty1
1Department of Medicine, University of Cambridge, Cambridge, CB2 0QQ, UK
2Instituto Clodomiro Picado, Universidad de Costa Rica, Costa Rica
3AIMMS, Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, Amsterdam, the Netherlands
Snake envenomation kills over 100,000 people annually and cripples three times as many. Current treatment relies on animal-derived serum, which, although life-saving presents many drawbacks. Therefore, it is crucial to modernise snakebite treatment. This poster outlines our approach where we isolated lymphocytes from horses immunised with sub-Saharan African snake venoms. The variable heavy (VH) and variable light (VL) antibody sequences were extracted and combined into single-chain fragment variables (scFv). Afterwards, these scFv were cloned into a phagemid plasmid allowing the production of phage antibody libraries. The resulting libraries were selected on purified venom fractions and/or whole venom from sub-Saharan African snakes. Preliminary venom binding studies were then performed as a way of indicating the selections success. Promising selections had their scFv converted into a chimeric IgG format, comprised of horse VH & VL domains coupled to human constant heavy (CH) and constant light (CL) domains. These chimeric IgG were then expressed in Expi293 mammalian cells and assayed for binding. Next steps involve imminent examination of their neutralisation capacity in functional assays, which if effective will be further studied incorporating in vivo experiments.
Structural and electrophysiological characterization of presynaptically active peptides from Bothrops bilineatus smaragdinus (Viperidae: Crotalinae) venom
Rafael S Floriano1, Fernanda YGM Couceiro1, Francis L Pacagnelli1, Kristian A Torres-Bonilla2, Stephen Hyslop2, Bruno Lomonte3, Robert M Drummond4, Daniel C Pimenta5, Rafael J Borges6
1Laboratory of Toxinology and Cardiovascular Research, University of Western São Paulo, Presidente Prudente, SP, Brazil
2Department of Translational Medicine, Faculty of Medical Sciences, State University of Campinas, Campinas, SP, Brazil
3Clodomiro Picado Institute, University of Costa Rica, San José, Costa Rica
4Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow UK
5Laboratory of Biochemistry and Biophysics, Butantan Institute, São Paulo, SP, Brazil
6Center of Medicinal Chemistry, Center for Molecular Biology and Genetic Engineering, State University of Campinas, Campinas, SP, Brazil
The Amazonian viper Bothrops bilineatus smaragdinus venom (BBS-v) induces intense neuromuscular facilitation followed by irreversible blockade in mouse nerve phrenic-diaphragm (PND) preparation. In this study, we characterized two presynaptically active peptides from BBS-v based on mass spectrometry and neuromuscular electrophysiological approaches. BBS-v was fractionated by gel filtration using a TSKgel G2000SWXL column coupled to a HPLC Shimadzu SCL-10AVP system to obtain the fraction P8 responsible by inducing neuromuscular excitatory action in PND preparation. Fraction P8 was subsequently purified by reverse phase (RP)-HPLC using a Jupiter C18 column and resulting peptides being subjected to determination of intact mass and sequence by mass spectrometry in an ESI-IT-TOF Shimadzu, including residual neurotoxicity in vitro. Miniature (MEPPs) end-plate potentials were determined by conventional electrophysiology techniques in PND preparation. RP-HPLC analysis of fraction P8 revealed the presence of two peptides with 444.19 Da (P8-1) and 430.17 Da (P8-2) of intact mass. Peptides had their sequences determined in <EKW and <ENW (with <E = pyroglutamic acid) for P8-1 and P8-2, respectively. Fraction P8 (15 mg/ml) presented intense increase in the frequency of MEPPs [MEPPs/min: from 8.7±2.9 (basal) to 75.7±7.1 (t30 min), p<0.05, n=4]. Considering the proportions in which these peptides appear in 15 mg of P8/ml (~55% and ~45% for P8-1 and P8-2, respectively), only peptide P8-1 partially reproduced the increase in spontaneous acetylcholine release by the whole fraction [MEPPs/min: from 6.3±2.3 (basal) to 40.2±13 (t30 min), *p<0.05, n=4]; in addition, there was as intense excitatory action combining these proportions of P8-1 and P8-2 [MEPPs/min: from 10.2±3.4 (basal) to 58±13.7 (t30 min), p<0.05, n=4]. Peptides P8-1 and P8-2 synergically modulate the increase in cholinergic synaptic release, being involved in the B. b. smaragdinus venom-induced neuromuscular facilitation.
Action of a broad-spectrum metalloprotease inhibitor (Marimastat) on the neuromuscular activity of Lachesis muta muta (South American Bushmaster) venom in mouse phrenic nerve-diaphragm preparation
Vitória S Proença-Hirata1, Isabele N Oliveira1, Gabrielly C Souza-Gomes1, Samuel R Dias1, Hugo A Ghirotti1, Sofia NS Azevedo1, Poliana J Demico1, Nelson J Silva Jr.2, Kristian A Torres-Bonilla3, Stephen Hyslop3, Rogério Giuffrida1, Rafael S Floriano1
1Laboratory of Toxinology and Cardiovascular Research, University of Western São Paulo (UNOESTE), Presidente Prudente, SP, Brazil
2Graduate Program in Environmental Sciences and Health, Pontifical Catholic University of Goiás (PUC Goiás), Goiânia, GO, Brazil
3Department of Translational Medicine, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, SP, Brazil
Envenomation by Lachesis exhibits intense local myonecrosis, including haemostatic and cardiovascular disorders. Marimastat (MMT), developed as an anticancer drug, has shown ability in preventing snake venom-induced disturbances. Here, we assessed the action of MMT alone or associated with an anti-Bothrops/Lachesis antivenom (AV) on the neuromuscular disorder induced by Lachesis muta muta (LMMv) in vitro. Caseinolytic and esterasic activities were determined by spectrophotometry using artificial substrates (azocasein and DL-BAPNA, respectively). Mouse phrenic nerve-diaphragm preparation was mounted in an myographic system (1 g tension), with twitches being indirectly evoked by supramaximal stimuli (0.2 Hz, 0.1 ms) and recorded by a DY1 transducer coupled to a DataCapsule-Evo and LabScribe4 software. MMT (0.03–3 mM) exhibited potent inhibitory action on the LMMv caseinolytic activity, without altering the esterasic activity; MMT (0.03 mM) was chosen as minimum inhibitory concentration for neuromuscular assays [DA440nm/min: 5.4±0.38 (LMMv 1 mg/ml) vs. 1.2±0.37* (LMMv + MMT 0.03 mM), *p<0.05 vs. LMMv alone, n=3]. LMMv (10–100 mg/ml) produced slight neuromuscular facilitation within 10-20 min incubation, progressing to concentration-dependent blockade; the highest concentration (100 mg/ml) reached 50% and 90% blockade in 54±5 and 80±6 min, respectively (p<0.05 vs. control preparation, n=4). MMT (0.03 mM) efficiently delayed the LMMv (100 mg/ml)-induced neuromuscular blockade from 50 min incubation, avoiding the occurrence of 50% blockade up to 120 min recording (p<0.05 vs. LMMv alone, n=4). AV alone (antivenom:venom ratio of 1:3 v/w) did not prevent LMMv (100 mg/ml)-induced 50% blockade reached in 81±8 min (p<0.05 vs. LMMv alone, n=4) and did not offer additional protection combined with MMT (0.03 mm). In conclusion, MMT exhibits potential application as a replacement pharmacological alternative to antivenom therapy to prevent neuromuscular disorder by LMMv.
Into the untrodden world of centipede venoms: a case study of scolopendrid venoms
Aditi1,2, Pragyadeep Roy1, Jahnavi Joshi1,2
1CSIR-Centre for Cellular and Molecular Biology, Hyderabad, India
2Academy of Scientific and Innovative Research, CSIR-CCMB, Hyderabad, India
Venoms have evolved many times in the animal kingdom and are considered a key trait contributing to the evolutionary success of many groups. Understanding the evolutionary and ecological drivers of venom diversity is crucial, though it remains a less explored aspect of venom research. Among terrestrial predatory arthropods, centipedes represent some of the oldest living lineages of venomous animals, with ~ 3100 described extant species. The centipede family Scolopendridae comprises ~400 species worldwide, with high diversity in tropical regions. We examined venom diversity and its drivers within the family Scolopendridae. Venom diversity was documented using an integrated proteo-transcriptomic approach where individual venom profiles were generated for ~40 species of seven scolopendrid genera (n=166), representing ~80% of the genera level diversity. This helped identify and classify venom components with putative toxin names and functions and minimize toxin annotation error rates. The venom profiles of the members of Scolopendridae were predominantly neurotoxic in nature. Scolopendra (n=29) had the greatest values of hill numbers q0 (μ=47.11), q1 (μ=26.74) and q2 (μ=17.18), and the least was in Asanada. We also measured body size, latitude, longitude and elevational range for each species to examine if they explain venom diversity estimates. Our preliminary analyses using generalized linear regression (GLM) found a significant relationship between venom diversity and body size. We also performed PGLS at genera level with body size as the predictor variable, which showed a strong phylogenetic signal with ????=1.696 and R2 = 0.99. We are gathering data on other ecological predictors, such as diet, to understand the drivers of venom diversity and complexity.
Yeast surface display technology for the discovery of monoclonal antibodies against snake venom toxins
Camille Abada1, Mark C. Wilkinson1, Rohit N. Patel1, Nicholas R. Casewell1, Stuart Ainsworth2, Stefanie K. Menzies1,3
1Centre for Snakebite Research and Interventions, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool, L3 5QA, UK
2Department of Infection Biology and Microbiomes, Institute of Infection, Veterinary and Ecological 16 Sciences, University of Liverpool, Liverpool, L3 5RF
3Biomedical and Life Sciences, Lancaster University, Bailrigg, Lancaster LA4 4YW, UK
Snakebite envenoming is a neglected tropical disease, causing high rates of mortality and morbidity worldwide. Currently, antivenom is the standard treatment, however there is a pressing need for next generation antivenoms that offer improved therapeutic efficacy and safety. Here, we present early-stage data demonstrating the use of yeast surface display (YSD) technology to generate new monoclonal antibodies (mAbs) against snake venom toxins. This approach utilizes a nonimmune human library in which each yeast cell typically displays up to 1 x 105 copies of a unique single-chain antibody fragment (scFv) fused to a surface protein. ScFv are isolated through rounds of magnetic-activated cell sorting (MACS) followed by rounds of fluorescence-activated cell sorting (FACS), a high throughput screening technique. YSD is an advantageous platform utilizing eukaryotic protein-directed evolution, it is highly versatile, cheap and screening strategies are based on quantitative methods. Moreover, scFv candidates can be improved through iterative maturation processes, enriching biochemical properties such as affinity and stability. We initially targeted alpha-bungarotoxin (a-Bgtx) to validate our approach and successfully isolated diverse antibody clones. Neutralizing assays and cross-reactivity tests for these candidates are ongoing. Building on this initial selection success, we expanded our pipeline to develop antibodies targeting Ecarin – a key pathogenic toxin, belonging to the snake venom metalloproteinase family, that degrades prothrombin. Functional assays are underway to identify antibodies that can neutralise the effects of Ecarin, to potentially offer therapeutic benefits to address the global health challenge presented by snakebite.
Bitis arietans bite poisoning in Angola regarding a clinical case with compartment syndrome
Paula R S Oliveira1, Nelson Baldaia2, Cerezo H Gaspar3, Jéssica Campos4, Jose B Silva5
1Department of Biomedicine of the National Center for Scientific Research, Luanda, Angola
2,3,4Intensive Care Unit of the Main Military Hospital/Instituto Superior, Luanda, Angola
5Public Health Department of the Faculty of Medicine of the Agostinho Neto University, Luanda
Snake bite poisoning is a serious medical condition and a frequent medical-surgical emergency, which primarily affects rural communities in Africa, Latin America, Asia and New Guinea. They cause a high number of victims that can lead to death or permanent physical weakness. Throughout Southern and Eastern Africa, Bitis arietans is responsible for the majority of cases of serious poisoning and death. This article aims to present the first published clinical case that occurred in Angola regarding compartment syndrome caused by a Bitis arietans bite. He presented compartment syndrome of the right upper limb and profuse bleeding from the bite site with flexenular lesions at the level of the forearm, limited movement and painful palpation. Thumb with bleeding punctuates lesion and blackened skin, asymmetrical chest due to increased volume of the right pectoral region, painful on palpation and without subcutaneous emphysema. After a period of 92 days in hospital and undergoing 9 surgeries, the patient evolved satisfactorily, despite a prolonged stay, with surgical approximation of the edges of the surgical wound and a decision to be discharged with follow-up in a Surgery consultation. The multivariate spectrum of clinical complications developed and presented in this clinical case demonstrates that snakebites have unique characteristics that make their prevention and control challenging. Immediate inaccessibility to differentiated healthcare tends to increase complications and the probability of death, when combined with the fact that anti-venom is not produced or made available in our country. In Angola, snake bites are still not a notifiable disease, which makes it difficult to know the statistics of the problem and consequently better supplies of anti-venom, which are also not available in hospitals. Keywords: Compartment syndrome, Bitis arietans, Angola
Neurological complications in Russell’s Viper bite
Pallavi S Raut and Sadanand D Raut
Vighnahar Nursing Home, Narayangaon Pune, Maharashtra, India
We are providing emergency medical services to rural downtrodden community of Junnar Ambegaon region (Pune)since 1990. We Initiated “Mission- Zero Snake Bite Death” since 2005. Due to early & effective management of shock, cardio-respiratory failure, AKI, we have saved more than 12,000 snakebite victims with 0% mortality, minimum morbidity, incidence of AKI is minimum (15%), 4% required hemodialysis. Bite to needle time is minimized, due to massive public awareness. Total 336 patients admitted at Vighnahar Nursing Home during 2021-24 were studied, Cobra- 46 (13.40 %), Common krait 45 (13.39%), RV-241(71.7%), SSV -4 (1.19%). M-157 (46.7%), F-133(39.6%), Children-46 (13.71), RV bite patients admitted in shock, intracerebral bleed & cerebral infarcts. ventilator support 12 (6 %), AKI- 35 (15%) AKI, HD-10 (4.15%). Associated Co-Morbidity- 33 (13.41%), Neurotoxicity-42 (17%), Cardiotoxicity-77(31.26%). Case 1- 44 yrs M, H/O RV Bite while Working in sugarcane farm, admitted within 20 mins in shock, Gasping, unconscious, Convulsions, Hematuria, Bp- 187/127 mmHg, Spo2- 85, WBCT- 20 min Incomplete. Treated with NIV, shock management, Inj. ASVs 45 vials, D3- MRI- Rt Hemiparesis, CT- Subacute infarct/Encephalitis. D4- MRI- Hemorrhagic infracts, IC BLEED. Slow neurological improvement, memory, speech, power improved gradually after 1 month. well oriented & walking without support. Case 2- 65 yrs M H/O RV bite at Rt hand, admitted in shock, gasping, profuse sweating, restlessness, convulsions, gum & oral bleeding, unconscious, P-132 /min, BP 144/83 mm of Hg, Spo2 40 % WBCT –more than 35 min. Treated with shock management, Ventilatory support, Inj ASVs 60 vials, FFP 10, Fresh Blood transfusion, oral packing, other emergency medical management. MRI- multiple bilateral cerebral infarcts, Tracheostomy on D-13, D-23 started sluggishly respond to verbal commands with minimal movements of eye lids & all 4 limbs. We have achieved 100% survival with minimum complications.
Social representations of the fish Talassophryne nattereri in the community of Barra de Mamanguape, Brazil
Mell Shirley L. Torres, Grazzyane M. Dautro, André Luiz M. Pessanha, Karla P. de Oliveira Luna
Department of Biology, State University of Paraíba, Barúnas Street, Campina Grande – PB, Brazil
Envenomings by venomous animals in the sea, estuaries and rivers is an important public health occurrence in Brazil. In this context, the fish Thalassophryne nattereri constitutes an important health problem in fishing and seafood communities on the country’s coast. In this article, the social representations (SR) of fishermen and shellfish gatherers from Barra de Mamanguape (PB) were evaluated with the aim of understanding how relationships with this fish impact the community. To this end, instruments such as focus group and TALP were applied. Content analysis and the IRAMUTEQ® software were used to analyze the data. From the analysis of the results, it was possible to observe that fear is the main word evoked by fishermen, and pain mentioned by shellfish gatherers. Furthermore, it was possible to observe that community reports regarding poisoning, treatment, external anatomy and seasonality of the fish corroborate scientific data. From the research it is possible to indicate that this problem is important for the community, as it is closely linked to their work activity, but without specific treatment available in the country. Furthermore, the importance of the information evoked by fishermen and shellfish gatherers about the animal is highlighted, indicating a great local knowledge about the animal, its ways of life and envenomings.
Atroxobin, a snake venom thrombin-like enzyme from Peruvian Bothrops atrox, inhibits proliferation and migration by reducing mitochondrial metabolism in triple-negative breast cancer cells.
Félix A. Urra1,2,3, Yarela Herrera1,2,3, Yarlequé Armando2,4, Vivas-Ruiz Dan2,4
1Metabolic Plasticity and Bioenergetics Laboratory, Molecular and Clinical Pharmacology Program, Institute of Biomedical Sciences, Faculty of Medicine, University of Chile, Av. Independencia 1027,Santiago 7810000, Chile.
2Network for Snake Venom Research and Drug Discovery, Av. Independencia 1027, Santiago 7810000, Chile.
3MIBI: Interdisciplinary Group on Mitochondrial Targeting and Bioenergetics, Faculty of Medicine, University of Chile, Av. Independencia 1027, Santiago 7810000, Chile.
4Laboratorio de Biología Molecular, Facultad de Ciencias Biológicas, Universidad Nacional Mayor de San Marcos, Av. Venezuela Cdra 34 S/N, Ciudad Universitaria, Lima 01, Perú.
Snake venom thrombin-like enzymes (SVTLE) are functionally analogous to human alpha-thrombin, converting fibrinogen to fibrin. However, studies suggest these enzymes may act on the same targets with different results. Previously, our group reported the first SVTLE (pictobin from Bothrops pictus venom) that reduces the mitochondrial membrane potential and ATP synthesis, triggering mitochondrial fragmentation. Despite the above, the signalling pathways involved in this action of mitochondrial bioenergetics remain unclear. This work proposes that atroxobin, an SVTLE purified from Peruvian Bothrops atrox venom, acts on cell metabolism, inhibiting cell proliferation and migration in triple-negative breast cancer cell line MDA-MB-231. These cells were treated with atroxobin (25 µg/mL) for 24 h. Oxygen consumption rate (OCR) through the XFe96 Seahorse extracellular flux analyzer, mitochondrial membrane potential and cell cycle by flow cytometry, cell viability by MTT reduction, total live vs. dead cells by trypan blue staining, and finally, migration by transwell chambers were evaluated. Our results show that atroxobin decreases mitochondrial respiration, cell proliferation, and migration but does not affect MDA-MB-231 cell viability. Interestingly, the conditioned medium from atroxobin-treated MDA-MB-231 cells promotes the migration of THP-1 monocytes, possibly by inducing the release of inflammatory factors. Interestingly, the pre-treatment with dimethyl alpha-ketoglutarate (dm-αKg) reversed the effects of atroxobin on the proliferation and migration of MDA-MB-231 cells. This study contributes to understanding the anti-cancer mechanisms of an SVTLE. Acknowledgments: This work was funded by ANID-CHILE PhD fellowship-21222256, Anillo Grant-ACT210097, FONDEQUIP-EQM220164, FONDECYT-1241547, and VRIP-UNMSM/PERU Grant-PCONFIGI B23100451.
