Soldadura por uma das extremidades de dois cromossômios homólogos do tityus

In this paper the author describes a very interesting case of union of two homologous chromosomes of the scorpion Tityus bahiensis just by the opposite extremities. The two normal pairs of chromosomes behave as ordinarily, the members of each pair showing at times a slight disturbance in their regular parallelism. The complex chromosome, on the contrary, behaves itself as if it were devoid of kinetochores, that is, it does not orient like normal chromosomes nor reveal any kind of active movement. The fusion of the chromosomes has resulted from terminal breakage at the opposite ends, the correspondig fragments having been found unpaired in a cell in which two pairs of chromosomes were present. Consequently, the compound chromosome, like the normal ones, is provided with a kinetochore at each one of the free ends. Being thus a centric chromosome its behavior, or more exactly, its kinetic inactivity may be compared with that of the monovalents found elsewhere in meioses. It is due o the failure of a partner. The fusion of two homologous chromosomes has transformed them into a new chromosomal unit in whose corresponding parts the ability of pairing was entirely abolished. This result is in full contradiction with the theory of a point-to point attraction between homologous chromosomes attributed to particular power of the genes, since, if genes really exist, being placed in their original loci, they would promote the union side by side of the members of the compound chromosome. If an attraction loci-to-loci should prevail the compound chromosome would be bent as in Fig. 8, C or form a ring similar to the loops observed in the inverted segment of sailvary chromosomes of Drosophila, as represented in the Fig. 8, D and this, in accordance with the order of the loci resulting from an union of corresponding or opposite ends of the fused chromosomes, as indicated in the Fig, 8 A and B. The evidence in hand points to a fusion by non homologous extremities. The expected rings, however, have never been found in metaphase plates. From this fact the author concludes that there is no point-to-point attraction between chromosomes, a conclusion in full agreement with the behavior of Hemipteran chromosomes which, in spite of geing composed of two equivalent halves do not bend in order to adjust the corresponding loci. (Cf. the papers on Hemiptera published by the author in this volume).

Bothriurus pichicuy, nuevo escorpión chileno del grupo vittatus (Scorpiones, Bothriuridae)

Bothriurus pichicuy, a new scorpion species from the center coast of Chile, is described. It belongs to the vittatus species-group, and differs from the others species in the group by the pigmentation patterns of the prosoma, mesosoma and venter of metasoma, by the development and arrangement of the ventral keels of caudal segment V, and for its small size. The species is the only representative of the vittatus species-group that lives in sympatry with another Bothriurus species: B. coriaceus Pocock, 1893, from the coastal desert of Chile. Records of B. pichicuy came from the provinces of Petorca and Choapa.

Hiperglicemia na intoxicação escorpiônica experimental em cão

The production of hyperglycemia during the acute phase of scorpion poisoning produced by T. bahiensis in dogs is confirmed now. The highest degree on average, was reached 10 minutes after the injection of venom. In our hands, the previous bilateral adrenalectomy did not avoid the hyperglycemia. The average of the blood sugar level has been similar to that observed in dogs with adrenal glands, the highest blood sugar level was also registered after 10 minutes. The hyperglycemia obtained in adrenolectomized dogs is, probably, due to the liberation of Sympatin (Nor-adrenalin and adrenalin) as a consequence of the central excitation by the poison on the hepatic nerves and other ganglionar terminations of the Sympathetic Nervous System. Our present researches suggest that the venom has adrenergic action besides the central action.

Estudo do mecanismo da hiperglicemia e da hipertensão arterial, produzidas pelo veneno de escorpião, no cão

In the present paper we studied the mechanism of the hyperglycemia and hypertension evoked by the intravenous injection of scorpion venom (Tityus bahiensis) in the dog. We used 34 dogs, of both sex, weighing between 4.3 to 22 kg. These animals were divided in 3 groups and the following experiments were performed: in the first group (8 dogs) the animals were adrenalectomized after the intravenous injection of chlorpromazine; in the second group (16 dogs) the animals were injected with ganglionic blocking drugs (9.295 Ciba and hexamethonium); in the third group (10 dogs) the naimals were injected with dibenamine, and in 3 of them the adrenal glands were removed. The dogs of each group were injected intravenously with aqueous extract of 2 telsons of scorpion/kg; the average weight of each telson was 6,5 mg. The following results were obtained: 1) The hyperglycemia evoked by scorpion venom, in adrenalectomized dogs, was inhibited by chlorpromazine; 2) Ganglionic blocking drugs (9.295 Ciba and hexamthonium) were inefective as far as the hyperglycemic and pressor effects of venom are concerned; 3) In the animals treated with dibenamine, the venom produced a fall in blood pressure, both in the controle and in the adrenalectomized. The present experiments suggest that the scorpion venom has, besides the central action already described by other investigators, an adrenergic action, very similar to the adrenaline. On basis of our experiments we think that the adrenergic action is responsible, in part, by the productrion of hyperglycemia and hypertension.

Efeito de alguns curares naturais e da d-Tubocurarina retardando o tempo de coagulação e o tempo de protombina do sangue humano

In this paper the author points out to a old question of about 200 years ago in wich two kinds of opinions were discussed. BANCROFT and FONTANA in one hand atributes for the Indian arrow poison (curare) the propriety of uncoagulate the blood, and C. BEBNAHDJ, B. RODRIGUES and others made an contradictory opinion upon this subject. In our experiments, we utilized 4 curares samples from indians who lives near the Brazilian border at Colombia, the famous Ticunas poison, and the alkaloid d-Tubocurarine. These poisons were added in form of emulsion in saline to the blood and blood plasma in order to perform two kinds of experiments. In one serie of experiments we observed the effect of curare on human blood coagulation time according to LEE-WHITE technic puting 0.5 ml of the various poisons emulsions previously into the tube. By this method, we have found that the emulsion containing 0.1 g of the poison in 10 ml saline was the most effective (Table II), therefore we used this curare emulsion concentration in the other serie of experiments, in which we tested the action of these venoms on the human blood plasma prothrombins time, (Quick Technic) adding 0.1 ml of the saline poison emulsion to each 0.1 ml of human blood plasma. Results from these experiments can be seen on Table II. These experiments we have tried on one sample of human blood plasma plus the differents curares samples; and in another opportunity four samples of human blood plasma were tried with the curare from Ticunas indians (the most effective in this respect). Results from these experiments may be seen on Table III. All the poison tried in our experiments was previously tested on toads legs (B. crucifer) to verify his curares action. All times obtained with the experiments above, show highly significant results (P<001) when compared with the blood and blood plasma mixed with in the same volume of saline. Our results, point out that BANCROFT and FONTANA views upon the effect of curare on blood clothing time were correct. Curares enhance the blood clothing time "in vitro". But, in other hand, the work in that matter by NESI (6), and TISTHOUND (7) showing that d-Tubocurarine had no significant effect on blood clothing time of man and dogs "in vivo", made possible to conclude that the observations of C. BERNARD, B. RODRIGUES and others were also true. These discordance of opinions, we believe, may result as BANCROFT and FONTANA researches, were wade "in vitro" whereas C. Bernard, B. Rodrigues and others performed their experiments "in vivo".

Inflammatory effects of snake venom metalloproteinases

Metalloproteinases are abundant enzymes in crotaline and viperine snake venoms. They are relevant in the pathophysiology of envenomation, being responsible for local and systemic hemorrhage frequently observed in the victims. Snake venom metalloproteinases (SVMP) are zinc-dependent enzymes of varying molecular weights having multidomain organization. Some SVMP comprise only the proteinase domain, whereas others also contain a disintegrin-like domain, cysteine-rich, and lectin domains. They have strong structural similarities with both mammalian matrix metalloproteinases (MMP) and members of ADAMs (a disintegrin and metalloproteinase) group. Besides hemorrhage, snake venom metalloproteinase induce local myonecrosis, skin damage, and inflammatory reaction in experimental models. Local inflammation is an important characteristic of snakebite envenomations inflicted by viperine and crotaline snake species. Thus, in the recent years there is a growing effort to understand the mechanisms responsible for SVMP-induced inflammatory reaction and the structural determinants of this effect. This short review focuses the inflammatory effects evoked by SVMP.

Constituintes químicos e atividade antiedematogênica de Peltodon radicans (Lamiaceae)

Most of the snakebite incidents in the Amazon region involve Bothrops atrox, whose venom presents the most potent edematogenic and necrotic activities in the genus. This work describes the studies of isolation of the chemical constituents and antiedematogenic activity of the species Peltodon radicans (Lamiaceae), which is used in the treatment of snakebites and scorpion stings in the region. The extracts presented aliphatic hydrocarbons, 3beta-OH,beta-amirin (1), 3beta-OH,alpha-amirin (2), beta-sitosterol (3), stigmasterol (4), ursolic acid (5), 2alpha,3beta,19alpha-trihydroxy-urs-12-en-28-oic acid (tormentic acid, 6), methyl 3beta-hydroxy,28-methyl-ursolate (7), sitosterol-3-O-beta-D-glucopyranoside (8), and stigmasterol-3-O-beta-D-glucopyranoside (9). The flower extracts presented the higher antiedematogenic activity. This is the first report on the study of the flowers, stem, and roots of this plant.

Single-step purification of crotapotin and crotactine from Crotalus durissus terrificus venom using preparative isoelectric focusing

We describe the isolation of crotoxin, a presynaptic B-neurotoxin, as well as its subunits B (crotactine) and A (crotapotin) from lyophilized Crotalus durissus terrificus venom by a single-step preparative isoelectric focusing procedure. From 98 mg of dried venom protein 20.1 mg of crotactine and 13.1 mg of crotapotin were recovered in the first step of focalization and 4.2 mg in a second run. These values correspond to 35.7% of the total venom protein applied. Crotactine separated in the 9.3-7.0 pH range (tubes 1-6) and crotapotin in the 1.8-2.8 pH range (tubes 15-19) and both were homogeneous by SDS-PAGE and N-terminal amino acid analysis. Crotactine, a 12-kDa protein, presented hemolytic and phospholipase A2 activity. Thus, using isoelectric focusing we simultaneously purified both toxins in high yields. This method can be used as an alternative for the purification and characterization of proteins from other snake venoms under conditions in which biological activity is retained

Snake venom metalloproteinases and disintegrins: interactions with cells

Metalloproteinases and disintegrins are important components of most viperid and crotalid venoms. Large metalloproteinases referred to as MDC enzymes are composed of an N-terminal Metalloproteinase domain, a Disintegrin-like domain and a Cys-rich C-terminus. In contrast, disintegrins are small non-enzymatic RGD-containing cysteine-rich polypeptides. However, the disintegrin region of MDC enzymes bears a high degree of structural homology to that of the disintegrins, although it lacks the RGD motif. Despite these differences, both components share the property of being able to recognize integrin cell surface receptors and thereby to inhibit integrin-dependent cell reactions. Recently, several membrane-bound MDC enzymes, closely related to soluble venom MDC enzymes, have been described in mammalian cells. This group of membrane-anchored mammalian enzymes is also called the ADAM family of proteins due to the structure revealing A Disintegrin And Metalloproteinase domains. ADAMs are involved in the shedding of molecules from the cell surface, a property which is also shared by some venom MDC enzymes.

A single-step purification of bothropstoxin-1

Bothrops venoms are complex mixtures of components with a wide range of biological activities. Among these substances, myotoxins have been investigated by several groups. Bothropstoxin-1 (Bthtx-1) is a phospholipase A2-like basic myotoxin from Bothrops jararacussu. The purification of this component involves two chromatographic steps. Although providing a pure material, the association of these two steps is time consuming and a single-step method using high performance chromatography media would be useful. In the present study, we describe a single-step purification method for Bthtx-1. Bothrops jararacussu venom was dissolved in 1 ml buffer. After centrifugation, the supernatant was injected into a Resource-S cation exchange column connected to an FPLC system and eluted with a linear salt gradient. The complete procedure took 20 min, representing a considerable time gain when compared to a previously described method (Homsi-Brandenburgo MI et al. (1988) Toxicon, 26: 615-627). Bthtx-1 purity and identity, assessed by SDS-PAGE and N-terminal sequencing, resulted in a single band with a molecular mass of about 14 kDa and the expected sequence of the first 5 residues, S-L-F-E-L. Although the amount of protein purified after each run is lower than in the previously described method, we believe that this method may be useful for small-scale purifications.

Two related thrombin-like enzymes present in Bothrops atrox venom

This article describes the presence of two new forms of a thrombin-like enzyme, both with apparent molecular masses of 38 kDa, in Bothrops atrox venom. Both share the ability to cleave fibrinogen into fibrin and to digest casein. Both present identical Km on the substrate BApNA. Their N-terminal amino acid sequences are identical for 26 residues, sharing 80% homology with batroxobin and flavoxobin. Two groups of monoclonal antibodies (mAbs) raised against the purified enzyme forms recognized different epitopes of the putative corresponding enzymes present in B. atrox crude venom. On Western blotting analysis of B. atrox crude venom, mAbs 5DB2C8, 5AA10 and 5CF11, but not mAbs 6CC5 and 6AD2-G5, revealed two or more protein bands ranging from 25 to 38 kDa. By immunoprecipitation assays, the 6AD2-G5 mAb was able to precipitate protein bands of 36-38 kDa from B. atrox, B. leucurus, B. pradoi, B. moojeni, B. jararaca and B. neuwiedii crude venoms. Fibrinogen-clotting activity was inhibited when the same venom specimens were pre-incubated with mAb 6AD2-G5, except for B. jararaca and B. neuwiedii.

Extracellular matrix molecules as targets for brown spider venom toxins

Loxoscelism, the term used to describe lesions and clinical manifestations induced by brown spider's venom (Loxosceles genus), has attracted much attention over the last years. Brown spider bites have been reported to cause a local and acute inflammatory reaction that may evolve to dermonecrosis (a hallmark of envenomation) and hemorrhage at the bite site, besides systemic manifestations such as thrombocytopenia, disseminated intravascular coagulation, hemolysis, and renal failure. The molecular mechanisms by which Loxosceles venoms induce injury are currently under investigation. In this review, we focused on the latest reports describing the biological and physiopathological aspects of loxoscelism, with reference mainly to the proteases recently described as metalloproteases and serine proteases, as well as on the proteolytic effects triggered by L. intermedia venom upon extracellular matrix constituents such as fibronectin, fibrinogen, entactin and heparan sulfate proteoglycan, besides the disruptive activity of the venom on Engelbreth-Holm-Swarm basement membranes. Degradation of these extracellular matrix molecules and the observed disruption of basement membranes could be related to deleterious activities of the venom such as loss of vessel and glomerular integrity and spreading of the venom toxins to underlying tissues.

Actions of Crotalus durissus terrificus venom and crotoxin on the isolated rat kidney

Many studies have reported the occurrence of lethal acute renal failure after snakebites. The aim of the present investigation was to determine alterations in renal function produced by Crotalus durissus terrificus venom and crotoxin as well as the histological alterations induced by these venoms. Isolated kidneys from Wistar rats weighing 240 to 280 g were perfused with Krebs-Henseleit solution containing 6 g% of previously dialyzed bovine serum albumin. The effects of Crotalus durissus terrificus venom and crotoxin were studied on glomerular filtration rate (GFR), urinary flow (UF), perfusion pressure (PP) and percentage sodium tubular transport (%TNa+). The infusion of Crotalus durissus terrificus venom (10 µg/ml) and crotoxin (10 µg/ml) increased GFR (control80 = 0.78 ± 0.07, venom80 = 1.1 ± 0.07, crotoxin80 = 2.0 ± 0.05 ml g-1 min-1, P<0.05) and UF (control80 = 0.20 ± 0.02, venom80 = 0.32 ± 0.03, crotoxin80 = 0.70 ± 0.05 ml g-1 min-1, P<0.05), and decreased %TNa+ (control100 = 75.0 ± 2.3, venom100 = 62.9 ± 1.0, crotoxin80 = 69.0 ± 1.0 ml g-1 min-1, P<0.05). The infusion of crude venom tended to reduce PP, although the effect was not significant, whereas with crotoxin PP remained stable during the 100 min of perfusion. The kidneys perfused with crude venom and crotoxin showed abundant protein material in the urinary space and tubules. We conclude that Crotalus durissus terrificus venom and crotoxin, its major component, cause acute nephrotoxicity in the isolated rat kidney. The current experiments demonstrate a direct effect of venom and crotoxin on the perfused isolated kidney.

Effect of perimuscular injection of Bothrops jararacussu venom on plasma creatine kinase levels in mice: influence of dose and volume

The effect of dose and volume of a perimuscular injection of Bothrops jararacussu venom on myonecrosis of skeletal muscle was studied in mice. An increase of the venom dose (0.25 to 2.0 µg/g) at a given volume (50 µl) resulted in an increase in plasma creatine kinase (CK) levels 2 h after injection. Plasma CK activity increased from the basal level of 129.27 ± 11.83 (N = 20) to 2392.80 ± 709.43 IU/l (N = 4) for the 1.0 µg/g dose. Histological analysis of extensor digitorum longus muscle 4 h after injection showed lesion of peripheral muscle fibers, disorganization of the bundles or the complete degeneration of muscle fibers. These lesions were more extensive when higher doses were injected. Furthermore, an increase in volume (12.5 to 100 µl) by dilution of a given dose (0.5 µg/g) also increased plasma CK levels from 482.31 ± 122.79 to 919.07 ± 133.33 IU/l (N = 4), respectively. These results indicate that care should be taken to standardize volumes and sites of venom injections.

Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper

A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 µg) and fibrinogen (minimum coagulant dose = 4.2 µg) in vitro, and promotes defibrin(ogen)ation in vivo (minimum defibrin(ogen)ating dose = 1.0 µg). In addition, when injected intravenously in mice at doses of 5 and 10 µg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the `gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.