Wednesday, 8 February 2017

(I) Homage to Dr. Clodomiro Picado. 45th years of the Clodomiro Picado Institute.

Here I would like to translate a video, the original in spanish on the:

45th years of the "Instituto Clodomiro Picado" 
Costa Rica

I do have some preferences, and this is one. To me this is one example of science, social involvement and dedication. 
Dr. Clodomiro Picado Twight was a humanist pioneer, not only toxinologist. He promote regulations to prevent snakebite deaths between the poor peasants of Costa Rica that 100 years later are still innovative and challenging. 
English is not my first language so please, take the good intention and I apologize before hand for all the mistakes. Thank you. 



This is an interview to Dr. José Mª Gutiérrez, in 2015  for a Costa Rica TV program called "ConCiencia". Very good play on words, that translated a part would be "With Science" and together becomes "Consciousness". 





1st PART

P: Hello, how are you? welcome to ConCiencia, today we have the pleasant visit of Dr. Jose Mª Gutiérrez Gutiérrez, who is an investigator at the Clodomiro Picado Institute, University of Costa Rica, and professor at the Faculty of Microbiology, at the University of Costa Rica.

Dr. Jose Mª Gutierrez at ConCiencia TV 
P: “Chema”, as affectionately we know you, welcome to our program.

DrJMG: Thank you so much. It is a pleasure to be here.

P: We are going to talk about the 45 years of the Clodomiro Picado Institute, founded in 1970. Please, tell us the historic sequence on how the Institute was founded in those times.

DrJMG: Yes, in fact, the Clodomiro Picado Institute is the corollary of a long term national project, that we could say starts with the pioneer work of Clodomiro Picado Twigt, who worked on the decade of 1910, 20 and 30, he died in 1944, and between the many topics that Cloromiro Picado investigated, studied, are the snakebites. He was worried for this problem that affects mainly the farmers of our country, the agriculture workers and…

P: Mainly the banana workers on those times…

DrJMG: Yes, and laborers in general, fincas, small farmers etc. As he saw that these people arrived to the hospital almost to die only in there as a result of a snake bite and that the country did not had an answer against it, then he devoted himself to scientifically study the problem and not only he study it but tried hard to found solutions for it. 
He realized that Brazil manufactured snake anti-venom serum since 1901, at the Butantan Institute, and then he reach an agreement with the Butantan Institute to import antivenom from Brazil to be used in Costa Rica. Furthermore, Clodomiro Picado brought snakebite issue at political levels because he promoted, together with the Heath Minister of the time, Dr. Solon Nuñez Frutos, a very pioneer law in the continent called "Law of defense against "Ofidismo"" that established several regulations with the aim that those peasants would have access to the anti-ophidic serum (snake anti-venom) 

*The word "ofidio" comes from the greec ὀφίδιον, diminutive of ὄφις, snake. 
Ofidismo would be the term used to define a snakebite. This term, is widely used is South America. In Spain the word "serpiente" is more common.

Serpiente comes from the latin "serpens", literally. It has an indoeuropean root as for example in hindi, the word used for snake is "sarpa". 

P: and this law is established…

DrJMG: In 1926…

P: and this law obliges the Government to…

Dr. Clodomiro Picado Twig

DrJMG:
 obliges the Government to have anti-venom, obliges for example the railroad drivers to transport the snakebitten people to the cities where there were hospitals and very interesting, obliges the farm owners to have anti-venom in the farm, and in the case a farm owner did not had the anti-venom and a laborer died from snakebite, the farm owner had to pay for two years the salary of this person to the family. So, really, Clodomiro moved in a very a wide range,  from basic science to the application of the knowledge with the snake anti-venoms up to the intervention into the political scope for the resolution on the problem. 

Now, Cloromiro Picado dies in 1944 leaving behind his tradition, with his followers like Alfonso Trejos Willis, Nanbadilla, Luis Bolaños and other people.  On the sixties, there is a resurgence of this topic in our country because the political sectors that prompted the reform of health sector wanted to wide the range of health services as part of the political movement. They where interested and thought that it was time for the country to have their own production of snake anti-venom, then, it happened a very particular conjuncture with the coalescence of the Costa Rica University, initially trough Dr. Jesús Mª Jiménez Porras, that was Biochemistry professor of the Medicine School, and later on Dr. Róger Bolaños that was the central figure in all this project, with the political support of the Health Ministry…

P: Róger Bolaños from the Microbiology Faculty… the Faculty was already existing as such…
Left: Dr. Clodomiro Picado Twight holding a snake.
Right: Dr. Róger Bolaños

DrJMG: Exactly, the Faculty was created in 1956 and Róger Bolaños was the Professor of Immunology of the Faculty of Microbiology and they come to him which, let it be added, was the son of Luis Bolaños, disciple of Clodomiro Picado so, there is a direct lineage that brings Róger Bolaños. It was also very important at that time the political will, isn’t it? And in this case this was reflected on the will of the Ministry of Health, Dr. Alvaro Aguilar Peralta, that clearly saw the situation and gave a decisive political support and a third component in this synergy was the presence of a North American military veterinary doctor that was working in the USA Embassy,
Dr. Herschell Flowers, who had experience with snakes and snake anti-venom in USA and was very motivated for this project, with the support of the USA Embassy.

So, Flowers together with Róger Bolaños and the political support of Dr. Aguilar Peralta… it was like a conjunction, the celestial bodies aligned, and this program was firmly supported. The program initially was called “Program of Snake Anti-venoms” and belonged to the Health Ministry. As a result of their work, at the beginning of 1967 the first batches of anti-venom: the poly-specific anti-venom against vipers and the anti-coral anti-venom were released. 
Polyvalent anti-venom still manufactured nowadays
From 1968 Róger Bolaños was the director of that Program. The Program was so successful, the anti-venoms started to be used on the country with a lot of success, clinically were much better than the imported anti-venoms, so the Health Ministry decided to transform the Program into an Institute and financed both by the Costa Rica government as well as by A.I.D. (Agency for International Development) of the USA, the first plants were built of what is now 
The anti-coral antivenom of the Institute
the Instituto Clodomiro Picado, on a land belonging to the Ministry of Health, in “Dulce Nombre de Coronado”. This was in 1970, in April 1970 the “Instituto” was inaugurated, then belonging to the Ministry of Health. In 1970 the Government changes, Dr. Aguilar Peralta participated on the foundation of the “Instituto” but then next minister is Dr. José Luis Orlich Bolmarsich on the Government of Mr. José Figueres Ferrer. On those years, Dr. Bolaños and the managers of the University take an important step, tremendously lucid, that was to promote thorough an agreement between the University and the Ministry of Health, that the Program would become part of the Costa Rica University. This took place in 1972. The most important is that becoming part of the University of Costa Rica, the “Instituto” assumes the academic and humanistic philosophy of the University of Costa Rica as the driving motor for its development, and this implies that the project is not only dedicated to manufacture the anti-venoms, as it was the original goal and as happen in many centers in South America for example, but that added to the production, there will be university activities of education, research and social involvement and it is precisely these “integrality” that when it is time to approach a complex health topic with many elements and components, what gives the strength and enthusiasm that allowed the “Instituto” to strongly develop on the decades that followed.

For further information you can refer to the article (Editorial):
Toxicon, Volume 54, Issue 7, 1 December 2009
Special Issue: Bothrops asper, 
From natural history to public health

Bothrops asper: Beauty and peril in the Neotropics, 
Editorial. pg 901 - 903
José Mª Gutiérrez. Instituto Clodomiro Picado. Facultad de Microbiología. Universidad de Costa Rica.
http://dx.doi.org/10.1016/j.toxicon.2009.06.012


Snakes have inspired fascination and fear to humans since ancient times, as reflected by the common presence of these reptiles in the mythology of many civilizations. Such sense of awe has been associated to the impact that snakebite envenomation has had worldwide, to the amazing life patterns and adaptations of these reptiles and, more recently, to the exciting discoveries that science has provided on their natural history and on the chemical composition and toxic effects of their venoms and isolated toxins. Moreover, progress in the knowledge of the structural and functional properties of venom components has paved the way for the use of these complex mixtures as sources of lead compounds for the development of diagnostic and therapeutic agents.

There are many species of venomous snakes, classified in the families Colubridae (sensu lato), Atractaspididae, Elapidae and Viperidae. A small group of them are responsible for most cases of human envenomations. Examples are the cobras, kraits and some viperids in Asia, the saw-scaled viper in the sub-Saharan Africa, rattlesnakes in North America, and some species of lance-head vipers in Central and South America. Among them, Bothrops asper inflicts the highest toll of envenomations in southern Mexico, Central America and northern regions of South America ( Gutiérrez, 1995 and Warrell, 2004). It is estimated that, in Central American countries, B. asper is responsible for more than 50% of the total number of cases, and for almost all fatalities due to snakebite envenomation ( Bolaños, 1982 and Gutiérrez, 1995). The impact of B. asper in public health and other aspects of life in this region has been well known since the times of precolumbian cultures, one of which named this species ‘Bec-Kara-Acá’, the ‘devil that brings death when it bites’ ( Picado, 1931).

A large amount of work has been performed on the biology and toxinology of this species. The pioneer in the study of B. asper and other Central American snakes was the renowned Costa Rican scientist Clodomiro Picado (1887–1944) who, after his advanced studies in France, developed a fruitful career in the clinical laboratory of San Juan de Dios Hospital, in Costa Rica ( Gutiérrez, 1986). Among his many areas of interest, the study of snakes, their venoms, the clinical features of envenomations, and the development of treatments occupied a leading place in his research agenda. His efforts resulted in many scientific publications and were summarized in the book Serpientes Venenosas de Costa Rica. Sus Venenos. Seroterapia Antiofídica ( Picado, 1931), which covers his studies of many years. Picado promoted the introduction of serotherapy, i.e. the use of antivenoms, in the treatment of snakebite envenomation in Costa Rica, through cooperation with Instituto Butantan (Brazil), which provided the antivenoms that were successfully used in Costa Rica for several decades (Picado, 1931).
Clodomiro Picado (1887–1944) holding an adult specimen 
of Bothrops asper close to his laboratory at Hospital 
San Juan de Dios, San José, Costa Rica (circa 1930).


In the 1960s, Jesús María Jiménez-Porras, a professor of Biochemistry at the School of Medicine, University of Costa Rica, performed innovative research in the biochemical characterization of snake venoms, including B. asper, then classified as B. atrox ( Jiménez-Porras, 1964). He unveiled a pattern of geographic and individual variation in the venom of this species. 
Then, an outstanding inter-institutional effort to confront the problem of snakebite envenomation in Costa Rica was promoted, with the participation of the Ministry of Health, the University of Costa Rica and the Embassy of the United States of America (Gutiérrez and Rojas, 1999). Thanks to this joint venture, known as Programa de Sueros Antiofídicos, Costa Rica succeeded in producing the first batches of antivenoms in Central America, and has since been self-sufficient in the supply of these immunobiologicals. In 1970, the Instituto Clodomiro Picado emerged from this program under the leadership of Róger Bolaños, a brilliant scientist and a dynamic technological entrepreneur who contributed to the study of snake venoms and consolidated the production of antivenoms in Costa Rica ( Bolaños, 1984). Many projects carried out at this institute, together with the work performed by researchers in other laboratories and countries, have generated a large body of knowledge on B. asper, its natural history, its venom and its impact in public health.Owing to the abundance of scientific literature on this species and to the fact that it is often dispersed in diverse sources, some of which are not easily accessible, it is difficult to have a comprehensive view of the available knowledge on this ecologically and medically relevant viperid. Moreover, the ever growing trend towards specialization in the study of snakes and venoms has brought a body of novel findings that creates the need of integration of such widespread knowledge. 

The aim of this special issue of Toxicon is to bring together, in a single publication, the available information on B. asper, on its natural history and intraspecies variation, on the biochemistry and proteomics of its venom, on the pathological and pathophysiological effects that B. asper venom induces at the experimental level, on the clinical features of human envenomations and their treatment, and on the search for novel therapeutic tools that could improve the management of these envenomations in the future. Mahmood Sasa, Dennis K. Wasko and William W. Lamar present an overview of the natural history of B. asper. Besides published data, they include unpublished observations by field herpetologists on the fascinating adaptations and life patterns of this species. Mónica Saldarriaga-Córdoba and colleagues investigate various factors influencing the variations in phenotypic parameters of B. asper, presenting valuable clues for interpreting intraspecies variability.

Yamileth Angulo and Bruno Lomonte review the proteins that have been isolated and characterized from the venom of B. asper, describing their structural features and their toxicological profile. Alberto Alape-Girón and colleagues provide a detailed review of the proteomics of B. asper venom, highlighting the complexity of its geographic and ontogenetic variation, and several implications of such variability. José María Gutiérrez and colleagues summarize the experimental studies on the local and systemic pathology and pathophysiology of envenomations by B. asper, reviewing the mechanisms of venom-induced tissue damage and the ensuing processes of tissue repair and regeneration. In addition, Catarina Teixeira et al. discuss the multifactorial inflammatory response elicited by B. asper venom.

Rafael Otero presents an overview of the complex and often severe clinical manifestations of envenomation by B. asper, as well as on the therapy of B. asper snakebite cases in South and Central America. Finally, Bruno Lomonte and colleagues review the studies on the search for inhibitors that could neutralize the toxic activities of B. asper venom, including antibodies, natural inhibitors and synthetic molecules. Their work highlights the potential usefulness of inhibitors for the improvement of the therapy of envenomations, especially concerning the halting of the progression of local tissue damage.

The collection of works included in this special issue offers a comprehensive view of the available knowledge on this venomous species. It also evidences gaps in our understanding of B. asper, and highlights the need to perform additional studies on subjects that range from natural history and venom composition and effects, to the public health and ecological impact of this species. A central goal of this issue is to promote a productive dialogue between toxinologists and specialists working on natural history and on the clinical aspects of envenomation. The detailed account of many aspects of the natural history of B. asper presents a number of intriguing questions on the adaptive role of venom. Likewise, the complex clinical alterations described in human envenomations by this species demand a deeper understanding on the mechanism of action of venom components, and on the possible synergistic effects of various toxins.

It is hoped that the integrative effort presented in this issue will foster the interest of scientists, technologists, public health workers and students in pursuing the study of unknown aspects and enigmas that remain open for understanding B. asper and related snakes. The gathering, in a single issue and for a single species, of scientists working in fields as diverse as herpetology, evolutionary biology, biochemistry, pharmacology, experimental pathology, immunology, clinical medicine and epidemiology, has been a highly stimulating and rewarding experience.

I deeply thank Alan Harvey and the editorial staff of Toxicon for supporting in many ways the publication of this issue. I am especially grateful as well to the authors who agreed to participate in this effort, and to the reviewers who, through their criticisms and suggestions, contributed to the improvement of the manuscripts and to the completion of this project. This special issue of Toxicon is dedicated to the memory of Clodomiro Picado and Róger Bolaños, pioneers of Toxinology and of the study of B. asper in Central America.



ॐ लोकाः समस्ताः सुखिनो भवन्तु ॥
Om Lokah Samasthah Sukhino Bhavantu
May all beings everywhere be happy and peaceful.


Tuesday, 7 February 2017

(V): VSofI: Hypnale hypnale. Sri Lanka polyspecific antivenom pre-clinical results. AVRI



As said in the last post, there is no antivenom available for Hump nosed pit viper (Hypnale Hypnale) envenomation. Good news is that in Sri Lanka, through a collaborative initiative, a polyspecific antivenom has been developed that includes Hypnale hypnale


Toxicon 122, October 2016, pg 152-159
Development of a new polyspecific antivenom for snakebite envenoming in Sri Lanka: Analysis of its preclinical efficacy as compared to a currently available antivenom

Mauren Villalta a, Andre s Sa nchez a, María Herrera a, Maria ngela Vargas a, A lvaro Segura a, Maykel Cerdas a, Ricardo Estrada a, Indika Gawarammana b, Dan E. Keyler c, d,Kimberly McWhorter c, Roy Malleappah c, Alberto Alape-Giro n a, Guillermo Leo n a,
Jose María Gutie rrez a, *a Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San Jos e, Costa Ricab Faculty of Medicine, University of Peradeniya, Sri Lanka
c Animal Venom Research International, Moreno Valley, CA, USA
d Department of Experimental and Clinical Pharmacology, University of Minnesota, Minneapolis, MN, USA 



(You can read full article or download the PDF from Researchgate)

ABSTRACT
A new whole IgG, freeze-dried, polyspecific antivenom was prepared from the plasma of horses immunized with the venoms of the snakes Daboia russelii, Echis carinatus, Hypnale hypnale, and Naja naja from Sri Lanka. The preclinical neutralizing ability of this antivenom against several toxic and enzymatic activities of these four venoms was analyzed, and compared with that of a batch of VINS antivenom (India) being currently used in Sri Lanka. The activities tested were: lethality, hemorrhagic, in vitro coagulant, proteinase and phospholipase A2. Both antivenoms neutralized, to a different extent, these activities of the venom of D. russelii, E. carinatus, and N. naja. In general, the polyspecific Sri Lankan antivenom was more effective than the Indian antivenom in the neutralization of the venoms of D. russelii and E. carinatus, whereas the Indian antivenom showed a higher efficacy against the venom of N. naja. Regarding H. hypnale, the new Sri Lankan antivenom was effective in the neutralization of all activities tested, whereas the Indian antivenom neutralized lethality but not hemorrhagic, coagulant, proteinase and PLA2 activities, in agreement with the fact that this venom is not included in the immunization mixture for this antivenom. Results suggest that the new polyspecific Sri Lankan antivenom has a satisfactory preclinical neutralizing profile and compares favorably with the Indian antivenom. This is ready to be tested in a clinical trial to evaluate its efficacy and safety in human victims of snakebite envenomings by D. russelii, E. carinatus and H. hypnale in Sri Lanka. 


To start with, the study describes the different profiles for the common venomous snakes in Sri Lanka like Daboia russelii, Echis carinatus, Hypnale Hypnale and Naja Naja. As there is no diagnostic tool to determine which is the snake responsible for the biting, a syndromic approach is the most common sense procedure to be followed. The authors describe the results  some wonderful studies done by Ariaratnam et al, 2009, 2008 on that subject. 
In Sri Lanka, the antivenoms used are manufactured in India, from indian snakes. Geographical variations are very important. The main reasons for developing their own antivenom are:
1) The need to use a more geographical congruent antivenom, prepared from the snakes of the territory. 
2) As Hypnale hypnale bites are very common and the possibility of death or chronic disability is now an accepted fact, this snake should be included in the antivenom.
3) A high incidence of adverse reactions with the indian antivenom.

The initiative consist on an international partnership:

  • Animal Venom Research International (AVRI), a nonprofit charity from USA

  • Instituto Clodomiro Picado, a public research and antivenom manufacturing center on the University of Costa Rica

  • University of Peradeniya and other public institutions in Sri Lanka. 


Superb view of Peradeniya University Campus





























The authors built a serpentarium after obtaining all the permits and sample venoms were sent to Instituto Clodomiro Picado for the development of the antivenom. 
After passing all the quality controls common practice in the Instituto Clodomiro Picado, the preclinical efficacy of the antivenom was measured following in-vitro and in-vivo assays based on the effect of the different venoms by themselves and after the incubation with the polyspecific antivenom developed. 
Results were compared to a lot of VINS antivenom (polyspecific antivenom from India against Naja Naja, D. Ruselii, E. Carinatus and B. Caeruleus).

All this is a process that has taken some years to develop and to put into practice. To me this is a very important initiative. It shows how sharing knowledge and resources is possible!



There is a preliminary article:
Toxicon Vol 69, July 2013, pg 90-97
Antivenom for snakebite envenoming in Sri Lanka: The need for geographically specific antivenom and improved efficacy
D.E. Keylera,b,*, I. Gawarammanaa,c, J.M. Gutiérrezd, K.H. Sellahewaa,e, K. McWhorter a, f, R. Malleappah a
a Animal Venom Research International, Moreno Valley, CA, USADepartment of Experimental and Clinical Pharmacology, University of Minnesota, Minneapolis, MN, USA c Faculty of Medicine, University of Peradeniya, Sri Lankad Instituto Clodomiro Picado, School of Microbiology, University of Costa Rica, San José, Costa Ricae Department of Medicine, Melaka Manipal Medical College, Malaysiaf School of Public Health, Loma Linda University, Loma Linda, CA, USA
ABSTRACT

Sri Lanka is a tropical developing island nation that endures significant economic and medical burden as a result of snakebite envenomation, having not only a high prevalence of envenomations, but also one of the highest incidence rates (200 snakebites/100,000 people/year) of venomous snakebite in the world (Kasturiratne et al., 2005). Ironically, the very snakes responsible for this human morbidity and mortality are a valuable biomedical and ecological national resource, despite the medical and economic consequences of envenomation. Currently, no snake antivenom is produced using venoms from native Sri Lankan snakes as immunogens, and there is a true need for an efficacious Sri Lanka, poly-specific snake antivenom. An approach to fulfilling this need via combining the scientific, technological and economical resources from Costa Rica and the United States with the knowledge and talent of Sri Lankan official governmental agencies, legal counsels, environmental, medical and veterinary academic institutions, and religious and cultural leaders has been initiated, coordinated and funded by Animal Venom Research International (AVRI), a nonprofit charity. This bridging of nations and the cooperative pooling of their resources represents a potential avenue for antivenom development in a developing country that suffers the consequences of few specific resources for the medical management of venomous snakebite. The desired final outcome of such an endeavor for Sri Lanka is, most importantly, improved medical outcomes for snakebite patients, with enhanced and expanded science and technology relating to snake venoms and antivenoms, and the collateral benefits of reduced economic cost for the country.


You can visit the web of AVRI, see the process, how this initiative started, have a virtual tour on the serpentarium they build, what do they do... very interesting.


http://www.usavri.org

The mission of AVRI is to aid in the development of more effective prevention and treatment protocols for animal envenomation (bites or stings from venomous animals) by mediating between involved parties, educating medical personnel and the public in order to reduce morbidity and mortality around the world.

The technology exists to improve pharmacotherapies, treatment protocols and quality of care, at an affordable cost, within the regions suffering most from animal envenomation. The underserved populations afflicted by animal envenomation are in dire need of external support to champion their cause and AVRI is the organization that can serve as their advocate. 


Furthermore, AVRI emphasizes animal conservation principles, with regard for all local laws, to minimize human/animal conflicts and prevent ecological imbalances. Venomous animals, while potentially harmful to humans, are also the victims of fear, myth and misinformation often leading to their being killed unnecessarily. These animals play an important role in the natural environment and need to be preserved. Through education and awareness, AVRI hopes to dispel fears, prevent harmful conflicts and preserve biodiversity.


Two TED talk on "creating an antivenom for SriLanka" by AVRI 


Published on Jan 30, 2014
TED Talk with Roy Malleappah, executive director of AVRI and Kimberly McWhorter,





Published on Mar 8, 2012
YOUTH TED talk by Kimberly McWhorter
Associate Executive Director, Kimberly McWhorter, MPH, CPH, DrPH(c) Board Certified in Public Health by the National Board of Public Health Examiners; Program Coordinator/Educator for Riverside County, California Community Health Agency; Program Planner; Grant writer; Recipient California State Association of Counties Merit Award for creation and implementation of an innovative local government program.



ॐ लोकाः समस्ताः सुखिनो भवन्तु ॥
Om Lokah Samasthah Sukhino Bhavantu
May all beings everywhere be happy and peaceful.