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supracategorical fear information revealed by aversively conditioning multiple categories.fear-generalization is a critical function for survival, in which an organism extracts information from a specific instantiation of a threat (e.g., the western diamondback rattlesnake in my front yard on sunday) and learns to fear - and accordingly respond to - pertinent higher-order information (e.g., snakes live in my yard). previous work investigating fear-conditioning in humans has used functional magnetic resonance imaging (fmri) to demonstrate that activity patterns representing stimuli fr ...202033135598
toxicological profile of medically relevant crotalus species from mexico and their neutralization by a crotalus basiliscus/bothrops asper antivenom.specimens of the crotalus genus represent a potential snakebite problem in mexico, and despite the great number of species of crotalus present in this country, only a few of them are relevant from a medical point of view. crotalus envenomed patients can present a range of signs and symptoms, depending on the species involved, and their treatment is indistinctly with either of the anti-viperid antivenoms available in the mexican public health system. one of these antivenoms is produced by immuniz ...202032345455
toxin transcripts in crotalus atrox venom and in silico structures of toxins.the western diamondback rattlesnake (crotalus atrox) is a common and widespread north american pit viper species, and its venom possesses medical applications. in this research, we identified 14 of the most common transcripts encoding 11 major venom toxins including transcripts for a three-finger toxin (3ftx) from the crude venom of c. atrox. in silico three-dimensional (3d) structures of 9 venom toxins were predicted by using deduced toxin amino acid sequences and a computer programme-modeller. ...202032774833
repurposing cancer drugs batimastat and marimastat to inhibit the activity of a group i metalloprotease from the venom of the western diamondback rattlesnake, crotalus atrox.snakebite envenomation causes over 140,000 deaths every year, predominantly in developing countries. as a result, it is one of the most lethal neglected tropical diseases. it is associated with incredibly complex pathophysiology due to the vast number of unique toxins/proteins present in the venoms of diverse snake species found worldwide. here, we report the purification and functional characteristics of a group i (pi) metalloprotease (camp-2) from the venom of the western diamondback rattlesna ...202032397419
the origin and diversification of a novel protein family in venomous snakes.the genetic origins of novelty are a central interest of evolutionary biology. most new proteins evolve from preexisting proteins but the evolutionary path from ancestral gene to novel protein is challenging to trace, and therefore the requirements for and order of coding sequence changes, expression changes, or gene duplication are not clear. snake venoms are important novel traits that are comprised of toxins derived from several distinct protein families, but the genomic and evolutionary orig ...202032366667
comparison of thromboelastography versus conventional coagulation tests in simulated crotalus atrox envenomation using human blood.pit viper bites are a source of significant morbidity and mortality. pit viper bites can cause venom-induced consumptive coagulopathy (vicc), typically evaluated with laboratory-based conventional coagulation tests (ccts). however, ccts require a laboratory and average 1 h to conduct. thromboelastography (teg) provides real-time, point-of-care tests of coagulation that are fast and require no separate laboratory facilities, which could be advantageous in both hospital and austere settings. howev ...202031833475
crotalus atrox disintegrin reduces hemorrhagic transformation by attenuating matrix metalloproteinase-9 activity after middle cerebral artery occlusion in hyperglycemic male rats.hemorrhagic transformation after ischemic stroke is an independent predictor for poor outcome and is characterized by blood vessel rupture leading to brain edema. to date, no therapies for preventing hemorrhagic transformation exist. disintegrins from the venom of crotalus atrox have targets within the coagulation cascade, including receptors on platelets. we hypothesized that disintegrins from c. atrox venom can attenuate hemorrhagic transformation by preventing activation of matrix metalloprot ...202030242872
venom composition of adult western diamondback rattlesnakes (crotalus atrox) maintained under controlled diet and environmental conditions shows only minor changes.many species of snakes produce venom as a chemical means of procuring potentially fractious prey. studies have increasingly focused on venom compositional variation between and within individual snakes of the same species/subspecies, with significant differences often being observed. this variation in composition has been attributed to differences in age, season, diet, and environment, suggesting that these factors could help explain the inter- and intra-specific variation found in some snake ve ...201930954451
role of scale wettability on rain-harvesting behavior in a desert-dwelling rattlesnake.during storms in the southwestern united states, several rattlesnake species have been observed drinking rain droplets collected on their dorsal scales. this process often includes coiling and flattening of the snake's body, presumably to enhance water collection. here, we explored this rain-harvesting behavior of the western diamond-backed rattlesnake (crotalus atrox) from the perspective of surface science. specifically, we compared surface wettability and texture, as well as droplet impact an ...201931867507
rattlesnakes must drink: meal consumption does not improve hydration state.water is critical to survival, yet free-standing water is often rare in deserts and seasonally dry environments. thus, many dry-adapted species utilize either metabolic (that produced from metabolism) or dietary (that found in food) water to meet their hydric needs. it is suspected that desert reptiles can fulfill their hydric needs solely through dietary water intake. however, food consumption does not improve the hydration state of gila monsters (heloderma suspectum), a binge-feeding desert li ...201931135328
mechanisms underpinning the permanent muscle damage induced by snake venom metalloprotease.snakebite is a major neglected tropical health issue that affects over 5 million people worldwide resulting in around 1.8 million envenomations and 100,000 deaths each year. snakebite envenomation also causes innumerable morbidities, specifically loss of limbs as a result of excessive tissue/muscle damage. snake venom metalloproteases (svmps) are a predominant component of viper venoms, and are involved in the degradation of basement membrane proteins (particularly collagen) surrounding the tiss ...201930695027
the neutralization efficacy of expired polyvalent antivenoms: an alternative option.the expense of production and distribution of snakebite antivenom, as well as its relatively infrequent use, has caused antivenom to be increasingly difficult to obtain and ultimately producing an alarming global shortage. unused, expired antivenom may represent a significant, untapped resource to ameliorate this crisis. this study examines the efficacy of expired antivenom over time using in vitro, whole blood clotting, and platelet function statistics. representatives from three years for four ...201931229628
snake venom extracellular vesicles (svevs) reveal wide molecular and functional proteome diversity.proteins constitute almost 95% of snake venom's dry weight and are produced and released by venom glands in a solubilized form during a snake bite. these proteins are responsible for inducing several pharmacological effects aiming to immobilize and initiate the pre-digestion of the prey. this study shows that proteins can be secreted and confined in snake venom extracellular vesicles (svevs) presenting a size distribution between 50 nm and 500 nm. svevs isolated from lyophilized venoms collected ...201830104604
suspected anaphylaxis and lack of clinical protection associated with envenomation in two dogs previously vaccinated with crotalus atrox toxoid.to describe the clinical presentation of two canines present in anaphylactic shock secondary to rattlesnake envenomation. in both cases, there was no previous documented previous envenomation event and the initial sensitization required for anaphylactic response is believed to be secondary to crotalus atrox toxoid vaccine.201829248468
penetrating ocular injury by western diamondback rattlesnake. 201830119804
catroxmp-ii: a heme-modulated fibrinogenolytic metalloproteinase isolated from crotalus atrox venom.it has been recently demonstrated that the hemotoxic venom activity of several species of snakes can be inhibited by carbon monoxide (co) or a metheme forming agent. these and other data suggest that the biometal, heme, may be attached to venom enzymes and may be modulated by co. a novel fibrinogenolytic metalloproteinase, named catroxmp-ii, was isolated and purified from the venom of a crotalus atrox viper, and subjected to proteolysis and mass spectroscopy. an ion similar to the predicted sing ...201829761254
in vitro effects of crotalus atrox snake venom on chick and mouse neuromuscular preparations.the neuromuscular effect of venoms is not a major clinical manifestation shared between rattlesnakes native to the americas, which showed two different venom phenotypes. taking into account this dichotomy, nerve muscle preparations from mice and chicks were used to investigate the ability of crotalus atrox venom to induce in vitro neurotoxicity and myotoxicity. unlike crotalic venoms of south america, low concentrations of c. atrox venom did not result in significant effects on mouse neuromuscul ...201829604435
characterization of the rabbit as an in vitro and in vivo model to assess the effects of fibrinogenolytic activity of snake venom on coagulation.several in vitro investigations have demonstrated that anticoagulant effects of fibrinogenolytic snake venom metalloproteinases have been abrogated in human plasma by modifying fibrinogen with iron (fe) and carbon monoxide (co) to prevent catalysis or by directly inhibiting these enzymes with co. to translate these findings, we chose to assess the rabbit as a model of envenomation with crotalus atrox venom. it was determined with thrombelastography that 15 times the concentration of venom noted ...201828696521
crotalus atrox venom exposed to carbon monoxide has decreased fibrinogenolytic activity in vivo in rabbits.envenomation by haemotoxic enzymes remains a significant source of human morbidity and mortality worldwide, with administration of long-acting or multiple doses of antivenom first-line therapy. however, coagulopathy can still occur and recur. of interest, it has been recently demonstrated that direct, isolated exposure of snake venom enzymes with fibrinogenolytic activity to carbon monoxide (co) abrogates venom-mediated loss of coagulation in human plasma. these observations of co inhibition of ...201828691277
when less means more: dehydration improves innate immunity in rattlesnakes.immune function can vary based on availability of resources, and most studies of such influences have focused on the co-investment of energy into immune and other physiological functions. when energy resources are limited, trade-offs exist, which can compromise immunity for other functions. as with energy, water limitation can also alter various physiological processes, yet water has received little consideration for its possible role in modulating immune functions. we examined the relationship ...201728404727
crotalus atrox venom preconditioning increases plasma fibrinogen and reduces perioperative hemorrhage in a rat model of surgical brain injury.perioperative bleeding is a potentially devastating complication in neurosurgical patients, and plasma fibrinogen concentration has been identified as a potential modifiable risk factor for perioperative bleeding. the aim of this study was to evaluate preconditioning with crotalus atrox venom (cv-pc) as potential preventive therapy for reducing perioperative hemorrhage in the rodent model of surgical brain injury (sbi). c. atrox venom contains snake venom metalloproteinases that cleave fibrinoge ...201728102287
effects of purified human fibrinogen modified with carbon monoxide and iron on coagulation in rabbits injected with crotalus atrox venom.while snake venom derived enzymes, such as the thrombin-like activity possessing ancrod, have been used to treat thrombotic disease by defibrinogenating patients, the therapeutic potential of fibrinogenolytic snake venom enzymes, such as those derived from crotalus atrox, have not been fully explored. however, one of the potential risks of administering fibrinogenolytic enzymes to effect defibrinogenation is hemorrhage secondary to hypofibrinogenemia. the present investigation sought to determin ...201728889321
insight into the roles of selection in speciation from genomic patterns of divergence and introgression in secondary contact in venomous rattlesnakes.investigating secondary contact of historically isolated lineages can provide insight into how selection and drift influence genomic divergence and admixture. here, we studied the genomic landscape of divergence and introgression following secondary contact between lineages of the western diamondback rattlesnake (crotalus atrox) to determine whether genomic regions under selection in allopatry also contribute to reproductive isolation during introgression. we used thousands of nuclear loci to st ...201728616190
iron protects porcine plasma coagulation kinetics from degradation by crotalus atrox venom.while the administration of antivenom to treat hemotoxic snake bite injury remains the gold standard of therapy, we have demonstrated that modifying human fibrinogen with iron and carbon monoxide renders it resistant to fibrinogenolytic snake venom enzymes. in order to translate these findings into a possible biometal-based therapy complementary to antivenom administration, a preclinical model that possesses fibrinogen that closely mimics the human molecule in response to iron and carbon monoxid ...201728726133
effect of iron and carbon monoxide on fibrinogenase-like degradation of plasmatic coagulation by venoms of four crotalus species.annually, thousands suffer poisonous snake bite, often from defibrinogenating species. iron and carbon monoxide (co) improve coagulation kinetics by modulation of fibrinogen as demonstrated in various agkistrodon species and crotalus atrox. thus, we sought to determine whether pretreatment of plasma with iron and co could attenuate venom-mediated catalysis of fibrinogen obtained from four common crotalus species with known fibrinogenase activity. human plasma was pretreated with ferric chloride ...201726845427
opossum peptide that can neutralize rattlesnake venom is expressed in escherichia coli.an eleven amino acid ribosomal peptide was shown to completely neutralize western diamondback rattlesnake (crotalus atrox) venom in mice when a lethal dose of the venom was pre-incubated with the peptide prior to intravenous injection. we have expressed the peptide as a concatenated chain of peptides and cleaved them apart from an immobilized metal affinity column using a protease. after ultrafiltration steps, the mixture was shown to partially neutralize rattlesnake venom in mice. preliminary e ...201727718338
pathogenesis of hemorrhage induced by rattlesnake venom. an electron microscopic study.the pathogenesis of hemorrhage induced by rattlesnake (crotalus atrox) venom was investigated at the electron microscopic level. swiss-webster white mice were injected intramuscularly with one-fifth the ld(50) dose of whole venom. muscle samples were obtained by biopsy 2 minutes, 30 minutes and 3 hours after venom injection. capillaries in the endomysium were in various stages of degeneration. endothelial cells of damaged capillaries contained dilatated endoplasmic reticulum and perinuclear spac ...20164843388
evaluation of antimicrobial and antiviral activities of different venoms.animals' venomous secretions or peptides obtained from those secretions are used in the development of new therapeutic agents. the aims of this study were to investigate antimicrobial and antiviral activity of four different venoms obtained from the frog named bufo arenarum and the snakes crotalus atrox, causus rhombeatus and naja melanoleuca.201626743517
molecular cloning and characterization of cdnas encoding metalloproteinases from snake venom glands.snake venom metalloproteinases (svmps) are a superfamily of zinc-dependent proteases and participate in a number of important biological, physiological and pathophysiological processes. in this work, we simultaneously amplified nine cdnas encoding different classes of metalloproteinases from glands of four different snake species (agkistrodon contortrix laticinctus, crotalus atrox, crotalus viridis viridis and agkistrodon piscivorus leucostoma) by rt-pcr with a pair of primers. among the encoded ...201619799929
the deep origin and recent loss of venom toxin genes in rattlesnakes.the genetic origin of novel traits is a central but challenging puzzle in evolutionary biology. among snakes, phospholipase a2 (pla2)-related toxins have evolved in different lineages to function as potent neurotoxins, myotoxins, or hemotoxins. here, we traced the genomic origin and evolution of pla2 toxins by examining pla2 gene number, organization, and expression in both neurotoxic and non-neurotoxic rattlesnakes. we found that even though most north american rattlesnakes do not produce neuro ...201627641771
soluble p-selectin rescues viper venom-induced mortality through anti-inflammatory properties and psgl-1 pathway-mediated correction of hemostasis.venomous snakebites are lethal and occur frequently worldwide each year, and receiving the antivenom antibody is currently the most effective treatment. however, the specific antivenom might be unavailable in remote areas. snakebites by viperidae usually lead to hemorrhage and mortality if untreated. in the present study, challenges of rattlesnake (crotalus atrox) venom markedly increased the circulating soluble p-selectin (sp-sel) level, but not p-selectin (p-sel, selp(-/-)) mutants, in wild-ty ...201627779216
identification of lys49-pla2 from crude venom of crotalus atrox as a human neutrophil-calcium modulating protein.we fortuitously observed a human neutrophil intracellular free-calcium concentration ([ca(2+)]i) increasing activity in the commercially available phosphodiesterase i (pde i), which is actually dried crude venom of crotalus atrox. as this activity was not observed with another commercially available pure pde i, we tried to find out the causative molecule(s) present in 'crude' pde, and identified lys49-phospholipase a2 (lys49-pla2 or k49-pla2), a catalytically inactive protein which belongs to th ...201626937214
responses of infrared-sensitive tectal units of the pit viper crotalus atrox to moving objects.rattlesnakes perceive ir radiation with their pit organs. this enables them to detect and strike towards warm-blooded prey even in the dark. in addition, the ir sense allows rattlesnakes to find places for thermoregulation. animate objects (e.g., prey) tend to move and thus cause moving ir images across the pit membrane. even when an object is stationary, scanning head movements of rattlesnakes will result in moving ir images across the pit membrane. we recorded the neuronal activity of ir-sensi ...201626906281
iron and carbon monoxide attenuate degradation of plasmatic coagulation by crotalus atrox venom.hypofibrinogenemia is an important clinical consequence following envenomation by crotalus species, usually attenuated or prevented by administration of antivenom. it has been determined that iron and carbon monoxide (co) enhance fibrinogen as a thrombin substrate, likely secondary to conformational changes in molecular structure. we tested the hypothesis that pretreatment of plasma with iron and co could attenuate the effects of exposure to crotalus atrox venom. human plasma was exposed to 0 to ...201626575491
iron and carbon monoxide attenuate crotalus atrox venom-enhanced tissue-type plasminogen activator-initiated fibrinolysis.in addition to degrading fibrinogen as a source of consumptive coagulopathy, rattlesnake venom has also been demonstrated to enhance fibrinolysis and degrade alpha-2-antiplasmin. the goals of this investigation was to characterize the kinetic fibrinolytic profile of crotalus atrox venom in the absence and presence of tissue-type plasminogen activator (tpa), and to also ascertain if iron and carbon monoxide (co, a positive modulator of alpha-2-antiplasmin) could attenuate venom-enhanced fibrinoly ...201626575490
zebrin ii / aldolase c expression in the cerebellum of the western diamondback rattlesnake (crotalus atrox).aldolase c, also known as zebrin ii (zii), is a glycolytic enzyme that is expressed in cerebellar purkinje cells of the vertebrate cerebellum. in both mammals and birds, zii is expressed heterogeneously, such that there are sagittal stripes of purkinje cells with high zii expression (zii+), alternating with stripes of purkinje cells with little or no expression (zii-). the patterns of zii+ and zii- stripes in the cerebellum of birds and mammals are strikingly similar, suggesting that it may have ...201525692946
incipient speciation with biased gene flow between two lineages of the western diamondback rattlesnake (crotalus atrox).we used mitochondrial dna sequence data from 151 individuals to estimate population genetic structure across the range of the western diamondback rattlesnake (crotalus atrox), a widely distributed north american pit viper. we also tested hypotheses of population structure using double-digest restriction site associated dna (ddradseq) data, incorporating thousands of nuclear genome-wide snps from 42 individuals. we found strong mitochondrial support for a deep divergence between eastern and weste ...201525534232
problems with mitigation translocation of herpetofauna.mitigation translocation of nuisance animals is a commonly used management practice aimed at resolution of human-animal conflict by removal and release of an individual animal. long considered a reasonable undertaking, especially by the general public, it is now known that translocated subjects are negatively affected by the practice. mitigation translocation is typically undertaken with individual adult organisms and has a much lower success rate than the more widely practiced conservation tran ...201525040040
comparison of the protective effect of a commercially available western diamondback rattlesnake toxoid vaccine for dogs against envenomation of mice with western diamondback rattlesnake (crotalus atrox), northern pacific rattlesnake (crotalus oreganus oreganus), and southern pacific rattlesnake (crotalus oreganus helleri) venom.to evaluate effectiveness of a commercially available toxoid manufactured from western diamondback (wd) rattlesnake (crotalus atrox) venom against envenomation of mice with wd, northern pacific (np) rattlesnake (crotalus oreganus oreganus), and southern pacific (sp) rattlesnake (crotalus oreganus helleri) venom.201525710764
use of medicinal fauna in mexican traditional medicine.mexico has great biodiversity of fauna. the use of fauna with medicinal properties is a common practice since pre-hispanic times. in the last decade, there has been an interest in ethnozoological studies in mexico. therefore, more studies are needed in order to gather information regarding the use of fauna with medicinal properties in méxico. ethnozoological studies are necessary in order to discover new medications for human health. this review presents current information in terms of ethnozool ...201424440438
in silico molecular interaction analysis of ltnf peptide-lt10 with snake venom enzymes.lethal toxin neutralizing factor (ltnf) isolated from opossum (didephis virginiana) has been shown to exhibit anti-venom and anti-allergic property. the small synthetic peptide- lt10 derived from n-terminal of ltnf also showed this property in vivo. we applied molecular modeling, docking and molecular dynamic (md) simulation techniques to compute the interaction of lt10 peptide with few snake venom enzymes, namely pla2 from naja naja and atrolysin -c from crotalus atrox. our in silico interactio ...201424654849
thrombin-dependent incorporation of von willebrand factor into a fibrin network.attachment of platelets from the circulation onto a growing thrombus is a process involving multiple platelet receptors, endothelial matrix components, and coagulation factors. it has been indicated previously that during a transglutaminase reaction activated factor xiii (fxiiia) covalently cross-links von willebrand factor (vwf) to polymerizing fibrin. bound vwf further recruits and activates platelets via interactions with the platelet receptor complex glycoprotein ib (gpib). in the present st ...201425381443
organotopic organization of the primary infrared sensitive nucleus (lttd) in the western diamondback rattlesnake (crotalus atrox).pit vipers (crotalinae) have a specific sensory system that detects infrared radiation with bilateral pit organs in the upper jaw. each pit organ consists of a thin membrane, innervated by three trigeminal nerve branches that project to a specific nucleus in the dorsal hindbrain. the known topographic organization of infrared signals in the optic tectum prompted us to test the implementation of spatiotopically aligned sensory maps through hierarchical neuronal levels from the peripheral epitheli ...201424989331
mating systems, reproductive success, and sexual selection in secretive species: a case study of the western diamond-backed rattlesnake, crotalus atrox.long-term studies of individual animals in nature contribute disproportionately to our understanding of the principles of ecology and evolution. such field studies can benefit greatly from integrating the methods of molecular genetics with traditional approaches. even though molecular genetic tools are particularly valuable for species that are difficult to observe directly, they have not been widely adopted. here, we used molecular genetic techniques in a 10-year radio-telemetric investigation ...201424598810
snake venom glutaminyl cyclases: purification, cloning, kinetic study, recombinant expression, and comparison with the human enzyme.among various snake venom components, glutaminyl cyclase (vqc) is one of the least understood protein family and none of its members has been purified or characterized. here we confirmed the presence of vqc activity in a wide spectrum of venom species via enzymatic assay using a synthetic fluorogenic substrate. we have also cloned novel vqc cdnas from seven species including crotalus atrox. the results revealed more than 96% sequence similarities among vqcs and ∼75% sequence identities between v ...201424814012
skin lipid structure controls water permeability in snake molts.the role of lipids in controlling water exchange is fundamentally a matter of molecular organization. in the present study we have observed that in snake molt the water permeability drastically varies among species living in different climates and habitats. the analysis of molts from four snake species: tiger snake, notechis scutatus, gabon viper, bitis gabonica, rattle snake, crotalus atrox, and grass snake, natrix natrix, revealed correlations between the molecular composition and the structur ...201424157843
cdna cloning of a snake venom metalloproteinase from the eastern diamondback rattlesnake (crotalus adamanteus), and the expression of its disintegrin domain with anti-platelet effects.a 5' truncated snake venom metalloproteinase was identified from a cdna library constructed from venom glands of an eastern diamondback rattlesnake (crotalus adamanteus). the 5'-rapid amplification of cdna ends (race) was used to obtain the 1865 bp full-length cdna sequence of a snake venom metalloproteinase (camvmpii). camvmpii encodes an open reading frame of 488 amino acids, which includes a signal peptide, a pro-domain, a metalloproteinase domain, a spacer, and an rgd-disintegrin domain. the ...201323313448
antibody responses to natural rattlesnake envenomation and a rattlesnake toxoid vaccine in horses.antivenom antibody titers following administration of rattlesnake venom for antivenom production in horses are well documented; however, antivenom antibody titers following natural rattlesnake envenomation in horses are not. antibody titers produced in response to the commercially available rattlesnake venom vaccine are also not published. our study objectives were to measure antivenom antibody titers in rattlesnake-bitten horses and compare them to titers in horses vaccinated with the rattlesna ...201323515015
molecular basis for prey relocation in viperid snakes.vertebrate predators use a broad arsenal of behaviors and weaponry for overcoming fractious and potentially dangerous prey. a unique array of predatory strategies occur among snakes, ranging from mechanical modes of constriction and jaw-holding in non-venomous snakes, to a chemical means, venom, for quickly dispatching prey. however, even among venomous snakes, different prey handling strategies are utilized, varying from the strike-and-hold behaviors exhibited by highly toxic elapid snakes to t ...201323452837
gape size, its morphological basis, and the validity of gape indices in western diamond-backed rattlesnakes (crotalus atrox).maximum gape is important to the ecology and evolution of many vertebrates, particularly gape-limited predators, because it can restrict the sizes and shapes of prey that can be eaten. although many cranial elements probably contribute to gape, it is typically estimated from jaw length or jaw width, or occasionally from a combination of these two measures. we measured maximum gape directly for 18 individuals of the western diamond-backed rattlesnake, crotalus atrox. we measured each individual's ...201323108999
irreversible inactivation of snake venom l-amino acid oxidase by covalent modification during catalysis of l-propargylglycine.snake venom l-amino acid oxidase (sv-laao, a flavor-enzyme) has attracted considerable attention due to its multifunctional nature, which is manifest in diverse clinical and biological effects such as inhibition of platelet aggregation, induction of cell apoptosis and cytotoxicity against various cells. the majority of these effects are mediated by h2o2 generated during the catalytic conversion of l-amino acids. the substrate analog l-propargylglycine (lpg) irreversibly inhibited the enzyme from ...201323772385
directional sensitivity in the thermal response of the facial pit in western diamondback rattlesnakes (crotalus atrox).recent work published in the accompanying paper used a combination of 3d morphological reconstruction to define optical spread functions and heat transfer physics to study how external heat energy would reach the sensory membrane within the facial pit of pitvipers. the results from all of the species examined indicated asymmetric directional sensitivity, e.g. the pit would preferentially respond to stimuli located below and behind the snake. the present study was intended as a test of these find ...201222786639
resurrexit, sicut dixit, alleluia. snake venomics from a 26-year old polyacrylamide focusing gel.a 26-year-old dried polyacrylamide gel, cast in presence of an immobilized ph gradient and containing focused proteins from the venoms of a northern black-tailed rattlesnake (crotalus molossus molossus), and of a western diamondback rattlesnake (crotalus atrox) has been screened in order to see the feasibility of extracting the proteins, analyzing them by mass spectrometry (ms) and assessing their integrity. nine gel bands were excised along the ph 3-10 gradient and the gel segments reswollen in ...201221985938
relationship between the structure and the enzymatic activity of crotoxin complex and its phospholipase a2 subunit: an in silico approach.crotoxin, one of the major toxins of south american rattlesnake crotalus durissus subspecies, is an heterodimeric complex composed of two distinct subunits: a basic phospholipase a(2) (pla(2), cb) and an acidic nontoxic catalytically inactive protein, crotapotin (ca). it's well known that cb has a high enzymatic activity; however the molecular aspects that determine this fact remain unknown. in this study, an in silico approach was used to predict the ca structure by homology modeling, and the c ...201222481077
synthesis and some properties of constrained short-chain phosphatidylcholine analogues: (+)- and (-)-(1,3/2)-1-o-(phosphocholine)2,3-o- dihexanoylcyclopentane-1,2,3-triol.reported herein is the synthesis of (+)- and (-)-(1,3/2)-1-o-(phosphocholine)-2,3-o-dihexanoylcyclopentane-1,2, 3-triol. these are the enantiomers of a contrained analogue of dihexanoylphosphatidylcholine in which the glycerol backbone is replaced by all-trans cyclopentane-1,2,3-triol. evidence is presented to demonstrate that the (-)-enantiomer is a substrate for phospholipase a2 (pla2) (crotalus atrox) while the (+)-enantiomer is not. this strict enantiomeric (and positional) specificity was e ...20123370720
in vitro evaluation of total venom-antivenin immune complex formation and binding parameters relevant to antivenin protection against venom toxicity and lethality based on size-exclusion high-performance liquid chromatography.total venom-antivenin immune complex formation and binding parameters relevant to antivenin protection against venom toxicity and lethality can be evaluated using size-exclusion high-performance liquid chromatography (se-hplc). simple integration of regions within se-hplc elution profiles was used to compare binding characteristics of crotalidae polyvalent immune fab (ovine) antivenin (fabav) and crotalus atrox (western diamondback rattlesnake; c. atrox), c. varidis varidis (prairie rattlesnake; ...201121392522
crotalidae polyvalent immune fab: in patients with north american crotaline envenomation.crotalidae polyvalent immune fab is an antivenom comprising purified, sheep-derived, fab igg fragments and is indicated for use in patients with north american crotaline envenomation. crotalidae polyvalent immune fab is produced using four north american snake venoms: crotalus atrox, crotalus adamanteus, crotalus scutulatus, and agkistrodon piscivorus. intravenous crotalidae polyvalent immune fab was effective in patients aged =10 years who had minimal or moderate envenomation by a north america ...201121443271
inactivation of crotalus atrox venom hemorrhagic activity by direct current exposure using hens' egg assay.the hemotoxic venoms of viperidae and crotalidae are responsible for most of the evenomations in the united states, west africa, india, south-east asia, new guinea, and latin america. we previously reported that a short exposure of crotalus atrox venom to direct electric current (dc) from a low-voltage generator, in solution, causes consistent and irreversible inactivation of venom phospholipase a(2) and metalloproteases. here we report by in vivo assay on chicken embryos at stage 18 of developm ...201121800403
zootherapeutic practices in aquismón, san luis potosí, méxico.animal derived products have been a source of medicinal compounds since ancient times. this work documents the use of animal species in traditional medicine in the municipality of aquismón, san luis potosí, méxico.201121963568
folk medicinal use of fauna in mapimi, durango, méxico.to document the use of animal species in traditional medicine from the municipality of mapimi, durango, méxico.201120937375
isolation and identification of a snake venom metalloproteinase inhibitor from california ground squirrel (spermophilus beecheyi) blood sera.california ground squirrels (spermophilus beecheyi) show blood-based defenses to a variety of toxins in the venom of the northern pacific rattlesnake (crotalus oreganus oreganus). in this study we demonstrate the presence of an effective snake venom metalloproteinase inhibitor (svmpi) in s. beecheyi. the blood sera of california ground squirrels were effective at reducing the metalloproteinase activity of northern pacific (c. o. oreganus) and prairie rattlesnake (crotalus viridis viridis) venoms ...201121903126
neutralization of haemorrhagic activity of viper venoms by 1-(3-dimethylaminopropyl)-1-(4-fluorophenyl)-3-oxo-1,3-dihydroisobenzofuran-5-carbonitrile.viper envenomation undeniably induces brutal local manifestations such as haemorrhage, oedema and necrosis involving massive degradation of extracellular matrix at the bitten region and many a times results in dangerous systemic haemorrhage including pulmonary shock. snake venom metalloproteases (svmps) are being considered to be the primary culprits for the venom-induced haemorrhage. as a consequence, the venom researchers and medical practitioners are in deliberate quest of svmp inhibitors. in ...201121729242
specificity analysis of the c-type lectin from rattlesnake venom, and its selectivity towards gal- or galnac-terminated glycoproteins.the rattlesnake (crotalus atrox) venom lectin is a readily-prepared decameric c-type lectin, specific for gal and galnac. glycan microarray analysis showed it reacted with a wide range of glycans, chiefly recognizing sets of compounds with galβ1-4glcnac (lacnac), α-gal or α-galnac non-reducing termini. its array profile was therefore distinctly different from those of four previously studied mammalian c-type lectins with the same gal/galnac monosaccharide specificity, and it was more broadly rea ...201121805077
comparative study of anticoagulant and procoagulant properties of 28 snake venoms from families elapidae, viperidae, and purified russell's viper venom-factor x activator (rvv-x).snake venoms consist of numerous molecules with diverse biological functions used for capturing prey. each component of venom has a specific target, and alters the biological function of its target. once these molecules are identified, characterized, and cloned; they could have medical applications. the activated clotting time (act) and clot rate were used for screening procoagulant and anticoagulant properties of 28 snake venoms. crude venoms from daboia russellii siamensis, bothrops asper, bot ...201020677373
pilot studies of pressure-immobilization bandages for rattlesnake envenomations.pressure-immobilization bandages sequester venom in extremities and are recommended for snakebites without local toxicity. pilot studies were performed to determine the time of onset of toxicity and efficacy of pressure-immobilizations bandages in a porcine model of rattlesnake envenomation.201019888893
human cytokine response to texas crotaline envenomation before and after antivenom administration.the aim of this study was to characterize the human cytokine response to texas crotaline envenomation before and after antivenom administration.201020887908
rock squirrel (spermophilus variegatus) blood sera affects proteolytic and hemolytic activities of rattlesnake venoms.rock squirrels (spermophilus variegatus) from two sites in south central new mexico, where prairie (crotalus viridis viridis) and western diamondback (crotalus atrox) rattlesnakes are common predators, were assayed for inhibition of rattlesnake venom digestive and hemostatic activities. at statistically significant levels rock squirrel blood sera reduced the metalloprotease and hemolytic activity of venoms from c. v. viridis and c. atrox more than venom from an allopatric snake species, the nort ...201021184770
rosmarinic acid in argusia argentea inhibits snake venom-induced hemorrhage.a methanolic extract of argusia (or messerschmidia or tournefortia) argentea (boraginaceae) significantly inhibited hemorrhage induced by crude venom of trimeresurus flavoviridis. the extract was then separated according to antivenom activity by using silica gel column chromatography and hplc equipped with an octadecylsilanized silica gel (ods) column to afford rosmarinic acid (ra) (1) as an active principle. ra (1) significantly inhibited the hemorrhagic effect of crude venoms of t. flavoviridi ...201020512530
crotalid snake venom subproteomes unraveled by the antiophidic protein dm43.snake venoms are mixtures of proteins and peptides with different biological activities, many of which are very toxic. several animals, including the opossum didelphis aurita, are resistant to snake venoms due to the presence of neutralizing factors in their blood. an antihemorrhagic protein named dm43 was isolated from opossum serum. it inhibits snake venom metalloproteinases through noncovalent complex formation with these enzymes. in this study, we have used dm43 and proteomic techniques to e ...200919267469
subcutaneous crotaline fab antivenom for the treatment of rattlesnake envenomation in a porcine model.this study was designed to investigate whether the local, subcutaneous injection of crotaline fab antivenom (crofab) at the rattlesnake envenomation site would result in less extremity edema when compared to intravenous (i.v.) antivenom infusion alone.200919153852
cysteine-rich secretory proteins in snake venoms form high affinity complexes with human and porcine beta-microseminoproteins.beta-microseminoprotein (msp), a 10 kda protein in human seminal plasma, binds human cysteine-rich secretory protein-3 (crisp-3) with high affinity. crisp-3 is a member of the family of crisps, which are widespread among animals. in this work we show that human as well as porcine msp binds catrin, latisemin, pseudecin, and triflin, which are crisps present in the venoms of the snakes crotalus atrox, laticauda semifasciata, pseudechis porphyriacus, and trimeresurus flavoviridis, respectively. the ...200919341830
structural modification of proteins by direct electric current from low voltage.the interaction of direct electric current (dc) and proteins is a little explored topic. we had reported that exposure of crotalus atrox venom to dc caused irreversible inactivation of phospholipase a(2) and metalloprotease and that the eukaryote adenylate kinases (ak) precipitate in nondenaturing gel electrophoresis. ak1 displays an elevated percent difference of charged versus polar amino acid content (ch-po 14). commercial ak1 and other 17 enzymes with various ch-po values were exposed in sol ...200919827034
exploring the venom proteome of the western diamondback rattlesnake, crotalus atrox, via snake venomics and combinatorial peptide ligand library approaches.we report the proteomic characterization of the venom of the medically important north american western diamondback rattlesnake, crotalus atrox, using two complementary approaches: snake venomics (to gain an insight of the overall venom proteome), and two solid-phase combinatorial peptide ligand libraries (cpll), followed by 2d electrophoresis and mass spectrometric characterization of in-gel digested protein bands (to capture and "amplify" low-abundance proteins). the venomics approach revealed ...200919371136
nanoscale probing reveals that reduced stiffness of clots from fibrinogen lacking 42 n-terminal bbeta-chain residues is due to the formation of abnormal oligomers.removal of bbetal-42 from fibrinogen by crotalus atrox venom results in a molecule lacking fibrinopeptide b and part of a thrombin binding site. we investigated the mechanism of polymerization of desbbeta1-42 fibrin. fibrinogen trinodular structure was clearly observed using high resolution noncontact atomic force microscopy. e-regions were smaller in desbbeta1-42 than normal fibrinogen (1.2 nm +/- 0.3 vs. 1.5 nm +/- 0.2), whereas there were no differences between the d-regions (1.7 nm +/- 0.4 v ...200919289066
[rattlesnake bite poisoning, health damage and treatment in children].in mexico, we do not know the exact number of cases and the magnitude of health damage conditioned by poisonous rattlesnake bites, and little relative information exists in pediatric population.200914574752
recurrent hemorrhage after western diamondback rattlesnake envenomation treated with crotalidae polyvalent immune fab (ovine).recurrent coagulopathy has been observed in patients after rattlesnake envenomation treated with crotalidae polyvalent immune fab (ovine) [fabav]. while recurrent coagulopathy is well documented in the literature, clinically significant sequelae have not been reported. we present a case of recurrent thrombocytopenia after western diamondback envenomation treated with fabav, resulting in an extensive recurrent local hemorrhage.200818608290
tail morphology in the western diamond-backed rattlesnake, crotalus atrox.the shaker muscles in the tails of rattlesnakes are used to shake the rattle at very high frequencies. these muscles are physiologically specialized for sustaining high-frequency contractions. the tail skeleton is modified to support the enlarged shaker muscles, and the muscles have major anatomical modifications when compared with the trunk muscles and with the tail muscles of colubrid snakes. the shaker muscles have been known for many years to consist of three large groups of muscles on each ...200818553368
on the quaternary structure of a c-type lectin from bothrops jararacussu venom--bj-32 (bjcul).bj-32 (also known as bjcul) is a c-type lectin from the venom of bothrops jararacussu with specificity for beta-galactosides and a remarkable ability to agglutinate several species of trypanosomatids. our objective was to study the oligomerization state of native bj-32 by using different biophysical and computational methods. small-angle x-ray light scattering (saxs) experiments disclosed a compact, globular protein with a radius of gyration of 36.72+/-0.04a and molecular weight calculated as 14 ...200818948130
measuring agreement and discord among hemagglutination inhibition assays against different ophidian paramyxovirus strains in the eastern massasauga (sistrurus catenatus catenatus).at present, the hemagglutination inhibition (hi) assay is the sole commercially available serologic method available to detect exposure to ophidian paramyxovirus (opmv) in snakes. during 2006, 26 eastern massasaugas (sistrurus catenatus catenatus) were collected, and blood was sampled to determine their opmv status. samples from each snake were divided into 3 aliquots and tested by using commercially available hi assays against the 4 opmv isolates used in the 3 laboratories that offer the servic ...200818816997
inhibition of lung tumor colonization and cell migration with the disintegrin crotatroxin 2 isolated from the venom of crotalus atrox.disintegrins are low molecular weight proteins (4-15 kda) with an rgd binding region at their binding loop. disintegrin and disintegrin-like proteins are found in the venom of four families of snakes: atractaspididae, elapidae, viperidae, and colubridae. this report describes the biological activity of a disintegrin, crotatroxin 2, isolated by a three-step chromatography procedure from the venom of the western diamondback rattlesnake (crotalus atrox). the intact molecular mass for crotatroxin 2 ...200818387648
direct treatment of mouse or human blood with soluble 5'-nucleotidase inhibits platelet aggregation.adenosine signaling is known to inhibit platelet aggregation. extracellular adenosine mainly stems from enzymatic phosphohydrolysis of precursor nucleotides via ecto-5'-nucleotidase. previous studies suggest that soluble 5'-nucleotidase (5'-nt) derived from crotalus atrox venom may be clinically beneficial in vascular leakage, myocardial, renal, and intestinal ischemia, or acute lung injury. however, the effects of 5'-nt treatment on platelet aggregation remain unknown. we examined the direct ef ...200818511695
hemorrhagic activity of the vascular apoptosis-inducing proteins vap1 and vap2 from crotalus atrox.vascular apoptosis-inducing proteins (vaps) from hemorrhagic snake venom are apoptosis-inducing toxins that target vascular endothelial cells. we now show that vap1 and vap2 from crotalus atrox have hemorrhagic activity in mouse skin following intradermal injection. following intravenous injection, vap2 induced hemorrhage in the lung, intestine and kidney. although the hemorrhagic activity was relatively weak, these apoptosis toxins may play a role in the complex mechanism of snake venom-induced ...200818657564
fatty acid analyses may provide insight into the progression of starvation among squamate reptiles.fasting-induced changes in fatty acid composition have been reported to occur within the body lipids of several types of animals; however, little is known about the changes in fatty acid profiles exhibited by reptiles subjected to prolonged fasting. this study characterizes the fatty acid profiles of six reptile species subjected to sublethal periods of fasting lasting 0, 56, 112, and 168 days. analyses of fatty acid methyl esters (fames) conducted on the total body lipids of rattlesnakes (crota ...200818657629
stable isotopes may provide evidence for starvation in reptiles.previous studies have attempted to correlate stable isotope signatures of tissues with the nutritional condition of birds, mammals, fishes, and invertebrates. unfortunately, very little is known about the relationship between food limitation and the isotopic composition of reptiles. we examined the effects that starvation has on delta13c and delta15n signatures in the tissues (excreta, carcass, scales, and claws) of six, distantly related squamate reptiles (gaboon vipers, bitis gabonica; ball py ...200818613003
expression of mrnas coding for vap1/crotastatin-like metalloproteases in the venom glands of three south american pit vipers assessed by quantitative real-time pcr.snake venom metalloproteases encompass a large family of toxins, with approximately 200 members already catalogued, which exhibit a diversity of structures and biological functions. from this relatively large number, only a dozen examples of apoptosis-inducing metalloproteases, like vap1 and 2 from the venom of crotalus atrox, are known. since most vap1-like toxins ever characterized were purified from the venom of viperidae species inhabiting diverse places on earth, we investigate the expressi ...200818926840
characterization and cdna cloning of aminopeptidase a from the venom of gloydius blomhoffi brevicaudus.the aminopeptidase activities of snake venoms from gloydius blomhoffi brevicaudus, gloydius halys blomhoffii, trimeresurus flavoviridis, bothrops jararaca and crotalus atrox were investigated. aminopeptidase a (apa), aminopeptidase b and aminopeptidase n activities were present in all snake venoms. the strongest apa activity was found in venom from g. blomhoffi brevicaudus. the susceptibility to metallopeptidase inhibitors and the ph optimum of the partially purified enzyme from g. blomhoffi bre ...200717383704
phylogeographic structure and historical demography of the western diamondback rattlesnake (crotalus atrox): a perspective on north american desert biogeography.the western diamondback rattlesnake (crotalus atrox) is a prominent member of north american desert and semi-arid ecosystems, and its importance extends from its impact on the region's ecology and imagery, to its medical relevance as a large deadly venomous snake. we used mtdna sequences to identify population genetic structure and historical demographic patterns across the range of this species, and relate these to broader patterns of historical biogeography of desert and semi-arid regions of t ...200716934495
crofabtrade mark total anti-venom activity measured by se-hplc, and anti-pla(2) activity assayed in vitro at physiological ph.one problem in the development and refinement of anti-venoms is ascertaining both overall anti-venom reactivity and which key toxins are neutralized. here we show by se-hplc that the in vitro reaction of crofab anti-venin with crotalus atrox venom asymptotically nears completion (>95%) by 11 min at 4 degrees c by following the change in area under chromatographic peaks. the peaks for reactants decrease and the formation of high molecular weight complexes increases with time. to assay the large n ...200717261322
inactivation of phospholipase a2 and metalloproteinase from crotalus atrox venom by direct current.to achieve our aim of understanding the interactions between direct current and enzymes in solution, we exposed reconstituted crotalus atrox venom to direct electric current by immersing two platinum thread electrodes connected to a voltage generator (between 0 and 8 v) into a reaction mixture for a few seconds. then, we assayed the residual activity of phospholipases a(2) (pla(2)),metalloproteinases, and phosphodiesterases, abundant in crotaline snake venoms and relevant in the pathophysiology ...200717366544
use of silac for exploring sheddase and matrix degradation of fibroblasts in culture by the piii svmp atrolysin a: identification of two novel substrates with functional relevance.snake venom metalloproteinases (svmps) in viperid venoms primarily function to give rise to local and systemic hemorrhage following snake envenomation. years of research on these toxins, both in vitro and in vivo, indicate that they function by disrupting capillary basement membranes, stromal matrix and cell-cell and cell-matrix contacts to allow escape of capillary contents under pressure. however, most of these studies used either defined substrates in vitro or were limited by relevant antibod ...200717543881
soft epidermis of a scaleless snake lacks beta-keratin.beta-keratins are responsible for the mechanical resistance of scales in reptiles. in a scaleless crotalus snake (crotalus atrox), large areas of the skin are completely devoid of scales, and the skin appears delicate and wrinkled. the epidermis of this snake has been assessed for the presence of beta-keratin by immunocytochemistry and immunoblotting using an antibody against chicken scale beta-keratin. this antibody recognizes beta-keratins in normal snake scales with molecular weights of 15-18 ...200717664165
prey envenomation does not improve digestive performance in western diamondback rattlesnakes (crotalus atrox).although the toxic properties of snake venoms have been recognized throughout history, very little is known about the adaptive significance of these powerful mixtures. this study examined the popular hypothesis that prey envenomation enhances digestion by influencing the energetic costs of digestion and assimilation, gut passage time, and apparent assimilation efficiency (assim) in western diamondback rattlesnakes (crotalus atrox), a species whose venom is recognized for its comparatively high p ...200717671964
reduction of rattlesnake-venom-induced myonecrosis in mice by hyperbaric oxygen therapy.hyperbaric oxygen therapy (hbot) at 1, 2, and 2.75 atmospheres absolute (ata) was used to treat rattlesnake (crotalus atrox) venom-induced tissue damage and edema in thigh muscles of mice. tissue damage was evaluated by double-blind histopathologic examination: tissue edema was determined by measuring tissue water content. a total of 10 intermittent exposures to oxygen over a period of 4 days at 2 and 2.75 ata did not influence the resolution of venom-induced tissue edema, whereas tissue damage ...20072045641
snakes survive starvation by employing supply- and demand-side economic strategies.animals vary widely in their abilities to tolerate extended periods of food limitation. although some snakes are known for their unique ability to survive periods of inanition that last up to 2 years, very little is known about the biological mechanisms that allow them to do this. consequently, the present study examined physiological, compositional, and morphological responses to 168 days of starvation among three distantly related snake species (i.e., ball python, python regius; ratsnake, elap ...200717644357
proximate developmental mediators of sexual dimorphism in size: case studies from squamate reptiles.sexual dimorphism in size (sexual size dimorphism; ssd) is nearly ubiquitous, but the relative importance of genetic versus environmental control of ssd is not known for most species. we investigated proximate determinants of ssd in several species of squamate reptiles, including three species of sceloporus lizards and the diamond-backed rattlesnake (crotalus atrox). in natural populations of these species, ssd is caused by sexual differences in age-specific growth. males and females, however, m ...200721672836
cdna cloning and some additional peptide characterization of a single-chain vascular apoptosis-inducing protein, vap2.vascular apoptosis-inducing proteins (vaps) from hemorrhagic snake venom are apoptosis-inducing toxins targeting vascular endothelial cells. well-characterized vaps consist of disulfide-bridged double chains (ddvaps). the authors previously described a single-chain vap (scvap), vap2 from crotalus atrox, which also induces apoptosis in endothelial cells (masuda et al., 1998, european journal of biochemistry, 253, 36-41). the authors report here the whole cdna sequences and some additional peptide ...200717497365
crystal structures of catrocollastatin/vap2b reveal a dynamic, modular architecture of adam/adamalysin/reprolysin family proteins.catrocollastatin/vascular apoptosis-inducing protein (vap)2b is a metalloproteinase from crotalus atrox venom, possessing metalloproteinase/disintegrin/cysteine-rich (mdc) domains that bear the typical domain architecture of a disintegrin and metalloproteinase (adam)/adamalysin/reprolysin family proteins. here we describe crystal structures of catrocollastatin/vap2b in three different crystal forms, representing the first reported crystal structures of a member of the monomeric class of this fam ...200717485084
shotgun protein sequencing: assembly of peptide tandem mass spectra from mixtures of modified proteins.despite significant advances in the identification of known proteins, the analysis of unknown proteins by ms/ms still remains a challenging open problem. although klaus biemann recognized the potential of ms/ms for sequencing of unknown proteins in the 1980s, low throughput edman degradation followed by cloning still remains the main method to sequence unknown proteins. the automated interpretation of ms/ms spectra has been limited by a focus on individual spectra and has not capitalized on the ...200717446555
mechanisms controlling venom expulsion in the western diamondback rattlesnake, crotalus atrox.although many studies have documented variation in the amount of venom expended during bites of venomous snakes, the mechanistic source of this variation remains uncertain. this study used experimental techniques to examine how two different features of the venom delivery system, the muscle surrounding the venom gland (the compressor glandulae in the rattlesnake) and the fang sheath, could influence venom flow in the western diamondback rattlesnake, crotalus atrox. differential contraction of th ...200717094108
the ontogeny of contractile performance and metabolic capacity in a high-frequency muscle.high-performance muscles such as the shaker muscles in the tails of western diamond-backed rattlesnakes (crotalus atrox) are excellent systems for studying the relationship between contractile performance and metabolic capacity. we observed that shaker muscle contraction frequency increases dramatically with growth in small individuals but then declines gradually in large individuals. we tested whether metabolic capacity changed with performance, using shaker muscle contraction frequency as an i ...200716380925
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