Myriapod haemocyanin: the first three-dimensional reconstruction of Scolopendra subspinipes and preliminary structural analysis of S. viridicornis.

Haemocyanins (Hcs) are copper-containing, respiratory proteins that occur in the haemolymph of many arthropod species. Here, we report the presence of Hcs in the chilopode Myriapoda, demonstrating that these proteins are more widespread among the Arthropoda than previously thought. The analysis of transcriptome of S. subspinipes subpinipes reveals the presence of two distinct subunits of Hc, where the signal peptide is present, and six of prophenoloxidase (PPO), where the signal peptide is absent, in the 75 kDa range. Size exclusion chromatography profiles indicate different quaternary organization for Hc of both species, which was corroborated by TEM analysis: S. viridicornis Hc is a 6 × 6-mer and S. subspinipes Hc is a 3 × 6-mer, which resembles the half-structure of the 6 × 6-mer but also includes the presence of phenoloxidases, since the 1 × 6-mer quaternary organization is commonly associated with hexamers of PPO. Studies with Chelicerata showed that PPO activity are exclusively associated with the Hcs. This study indicates that Scolopendra may have different proteins playing oxygen transport (Hc) and PO function, both following the hexameric oligomerization observed in Hcs.

Epidemiologic Profile of Patients Transplanted With Hematopoietic Stem Cells in a Reference Service in the State of Rio Grande do Norte, Brazil.

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) consists of the intravenous infusion of healthy hematopoietic stem cells to restore the medullary and immunologic function of patients affected by a series of hematologic, oncologic, immunologic, malignant and nonmalignant inherited or acquired diseases, with the possibility of cure or increase of disease-free survival. OBJECTIVE: To characterize the epidemiologic profile and the cases of death of patients who underwent HSCT. METHODS: This is a cohort quantitative study, nested with a retrospective, descriptive, and analytical study of a hospital-based cohort that included the patients who underwent HSCT at a referral service in the state of Rio Grande do Norte, a region of northeastern Brazil. RESULTS: There was a slight male prevalence (52.94%), the age of the patients ranged from 2 to 73 years old, 18.38% were brown, 47.06% were married, 15.07% were students, 78.31% had a diagnosis of multiple myeloma, 93.38% developed gastrointestinal toxicities, all patients received chemotherapeutic treatment, 54.78% had allogeneic HSCT, and the cause of the most recorded deaths was septic shock (48.19%). CONCLUSIONS: This study showed relevant scientific evidence on the clinical and epidemiologic profile of patients who underwent HSCT. In general, sociodemographic data are similar to national and international research results.

Chronological Factors of the Process of Donation and Classification of Corneal Tissue.

INTRODUCTION: The quality of corneal tissue is influenced by several factors inherent to the recipient, donor, donation process, and transplantation that may primarily or secondarily interfere in the survival of a corneal graft. OBJECTIVE: The objective of this study was to identify the factors inherent to the donation process, specifically related to the harvesting and processing of tissue and to the donor, that may interfere with the quality of corneal tissue. MATERIALS AND METHODS: This was a cross-sectional, descriptive, and analytical study carried out from January to April 2015 at the Onofre Lopes University Hospital, Natal/RN, Brazil. A survey of the keratoplasties performed between the years 2010 and 2014 was carried out, totaling 258 donated corneas. RESULTS: The mean age of donors was 42.08 years, ranging from 2-80 years. As for quality, 64.45% of the corneas were classified as good, 23.05% as regular, 11.33% as poor, and 1.17% as excellent. The mean time between death and preservation was approximately 7 hours and 15 minutes, and the mean time elapsed until use was 10 days after immersion in preservation medium. DISCUSSION AND CONCLUSIONS: Factors inherent in the donation process that may interfere with the quality of the corneal graft correspond to factors of a chronological nature: donor age and time elapsed between death and preservation of the graft.

BCR-ABL1 transcript types showed distinct laboratory characteristics in patients with chronic myeloid leukemia.

In chronic myeloid leukemia (CML) two main types of messenger RNA (e14a2 and e13a2) can be produced by BCR-ABL1 gene rearrangement. Due to conflicting results, the clinical value of these transcripts remains controversial. The aim of this study was to identify associations of e14a2 and e13a2 transcripts with laboratory variables and also the response to treatment. This study included 203 adult patients with CML treated with Imatinib as first-line drug in a reference hematology center in Northeast Brazil. Clinical and laboratory data were obtained after informed consent. Samples were collected for RNA extraction and analyzed by reverse transcription-polymerase chain reaction (PCR), according to the international protocol BIOMED-1. The LeukemiaNet 2013 criteria were used to establish the molecular response. The frequency distribution of the BCR-ABL1 transcripts was e14a2 (64%), e13a2 (34%), and double positives (2%). The results showed a statistically significant association of the e14a2 transcript type with thrombocytosis (P = 0.0005) and the e13a2 with higher leukocyte count (P = 0.0491). In a subgroup of 44 patients, the molecular response to treatment with Imatinib was assessed by quantitative PCR at 3 months (BCR-ABL1 ≤ 10%), 6 months (BCR-ABL1 ≤ 1%), or 12 months (BCR-ABL1 ≤ 0.1%). Although patients with the transcript e14a2 showed higher frequency of good responses than patients with the transcript e13a2, this difference was not statistically significant. In agreement with published data, our results showed association of the BCR-ABL1 transcript e14a2 with thrombocytosis and the BCR-ABL1 transcript e13a2 with higher leukocytosis in patients with chronic myeloid leukemia.

Epidemiological Profile of Patients Undergoing Urgent Corneal Transplant in a Referral Center in Northeastern Brazil.

BACKGROUND: The purpose of the study was to characterize the epidemiological profile and average time on the waiting list of patients undergoing an urgent corneal transplantation (CT) in a referral center in the state of Rio Grande do Norte, northeastern Brazil. METHODS: This cross-sectional study included cases of urgent corneal transplants performed in a referral center from January 2010 to December 2014. Data were collected between January and March 2015. RESULTS: Seventy-four cases of emergency CT were analyzed. Of these, 69.4% were male, over the age range from 8 to 92 years. There were 38 transplants occurring in the right eye, 55.4% with tectonic purpose, 25.7% with optical purpose, and 18.9% therapy purpose. The main indications for patient inclusion in an emergency were the perforation (52.7%) and corneal ulcer unresponsive to medical treatment (33.8%). Interstitial keratitis was the most common diagnosis (74.3%), responsible for all prioritized cases in line for corneal ulcer unresponsive to treatment. The average length of the waiting line to perform the urgent CT was 9.03 days. CONCLUSIONS: In this study, there was a predominance of male patients and patients >50 years old, which corroborates the results of other studies with similar results. Perforation was the main indication to include the patient on the urgency list (52.7%). There is a need for further research to improve public health policies on the process of corneal transplantation as well as raising awareness of the importance of organ donation.

Interfaz entre formación en enfermería y actividades artístico-culturales en el espacio académico: visión de los estudiantes / The interface between the nursing formation process and artistic-cultural activities within the academic space: the students' perspective / A interface entre formação de enfermagem e atividades artístico-culturais no espaço acadêmico: visão dos estudantes

Objetivo Analizar la percepción de los estudiantes en relación con las contribuciones de su inserción en actividades artístico-culturales en el espacio universitario para la formación en enfermería. Método Investigación cualitativa, guiada por la epistemología de Fleck. Participaron 17 estudiantes de enfermería integrantes de movimientos artístico-culturales existentes en una institución de enseñanza superior, a saber: Grupo teatral, grupo musical y grupo de Maracatú. Los datos fueron recolectados por medio de entrevista semiestructurada y organizados por la técnica del discurso del sujeto colectivo. Resultados: Los estudiantes destacaron entre las ventajas de las actividades artístico-culturales el estímulo para estudiar, sentirse relajado y más preparado para las actividades académicas, así como propiciar el desarrollo de sensibilidad y competencias que puedan transformar los escenarios de la práctica clínica de enfermería. Fueron indicadas como dificultades, la credibilidad de estas actividades en la academia, frente a la lógica de la formación curativa. Conclusiones: Integrar actividades artísticas y culturales con la formación académica permite al alumno protagonizar modos renovados de pensar en el cuidado de enfermería y fortalecer conexiones con los diferentes conocimientos necesarios para la práctica del cuidado holístico.

Objective To analyze the students' perceptions of the impact which their participation in artistic-cultural activities within the university space has on their nursing-formation process. Method: This is a qualitative research guided by the epistemology of Fleck. Seventeen nursing students from a high level school, and also members of artistic-cultural movements such as theater, music and Maracatu groups, participated in this study. Data were collected through semi-structured interviews and then organized following the collective subject discourse technique. Results: Related to the positive influence of their artistic-cultural activities, the students highlighted their stimulus to study, feeling relaxed, and being more prepared for the academic activities including those which can help them develop competences and sensibilities to improve the clinical nursing practice scenarios. A remark on the academia who doubts the credibility of these activities and who follows a model in which health is only viewed as the absence of illness was also highlighted. Conclusions: The integration of artistic and cultural activities into the nursing formation process allows the students to develop renewed modes of thinking which can strengthen the links among their knowledge areas, and thus prepare them better for a holistic care practice

Objetivo Analisar a percepção dos estudantes em relação às contribuições da sua inserção em atividades artístico-culturais no espaço universitário para a formação em enfermagem. Método: Investigação qualitativa, guiada pela epistemologia de Fleck. Participaram 17 estudantes de enfermagem, integrantes de movimentos artístico-culturais existentes em uma instituição de ensino superior, a saber: Grupo teatral, grupo musical e grupo de Maracatú. Os dados foram coletados por médio da entrevista semiestruturada e organizados pela técnica de discurso do sujeito coletivo. Resultados Os estudantes destacaram de entre as vantagens das atividades artístico-culturais, o estímulo para estudar, sentir-se relaxado e mais preparado para as atividades acadêmicas, bem como propiciar o desenvolvimento de sensibilidade e competências que pudessem transformar os cenários da prática clínica de enfermagem. Foram indicadas como dificuldades, a credibilidade destas atividades na escola, perante à lógica da formação curativista. Conclusões Integrar atividades artísticas e culturais na formação acadêmica, permite ao aluno protagonizar modos renovados de pensar no cuidado de enfermagem e fortalecer conexões com os diferentes conhecimentos necessários para a prática do cuidado holístico.

Atividade antimicrobiana do óleo essencial de Lippia origanoides e Lippia rotundifolia frente a enterobactérias isoladas de aves / Antimicrobial activity of Lippia origanoides and Lippia rotundifolia oils against enterobacteria isolated from poultry

The aim of this study was to evaluate the antimicrobial activity of essential oils on enterobacteria Escherichia coli and Staphylococcus aureus isolated from poultry fecal samples in the cloaca from 49 laying hens. To analyze the antimicrobial sensibility an agar diffusion susceptibility test was performed and the Minimum Inhibitory Concentration (MIC) and the Minimum Bactericida Concentration (MBC) of Lippia origanoides and Lippia rotundifolia essential oils was determined. The concentrations used were 160, 80 and 40 L/mL. The L. origanoides essential oil showed antimicrobial effect from 40 L/mL dose against both microorganisms, with larger efficiency in E. coli. The L.rotundifolia essential oil was more efficient at the concentration of 160L/mL. Its effect was observed in all microorganisms. These results suggest that L. origanoides oil is more effective than L. rotundifolia oil in inhibiting the growth of microorganism isolated from poultry, although it also has a satisfactory antimicrobial effect. Results indicate the potential use of these plant's essential oils in poultry feed as an alternative to conventional antimicrobial products.

Whole-Genome Sequence of a Bordetella pertussis Brazilian Vaccine Strain.

Despite the reduction in incidence after vaccination, pertussis disease is still considered a public health problem worldwide, mainly due to recent and potential new outbreaks. We report here the complete genome of the Bordetella pertussis Butantan strain used in the Brazilian National Immunization Program as a whole-cell pertussis antigen to compose vaccines such as DTwP (diphtheria, tetanus, and whole-cell pertussis).

Atividade antimicrobiana do óleo essencial de Lippia origanoides e Lippia rotundifolia frente a enterobactérias isoladas de aves / Antimicrobial activity of Lippia origanoides and Lippia rotundifolia oils against enterobacteria isolated from poultry

The aim of this study was to evaluate the antimicrobial activity of essential oils on enterobacteria Escherichia coli and Staphylococcus aureus isolated from poultry fecal samples in the cloaca from 49 laying hens. To analyze the antimicrobial sensibility an agar diffusion susceptibility test was performed and the Minimum Inhibitory Concentration (MIC) and the Minimum Bactericida Concentration (MBC) of Lippia origanoides and Lippia rotundifolia essential oils was determined. The concentrations used were 160, 80 and 40 L/mL. The L. origanoides essential oil showed antimicrobial effect from 40 L/mL dose against both microorganisms, with larger efficiency in E. coli. The L.rotundifolia essential oil was more efficient at the concentration of 160L/mL. Its effect was observed in all microorganisms. These results suggest that L. origanoides oil is more effective than L. rotundifolia oil in inhibiting the growth of microorganism isolated from poultry, although it also has a satisfactory antimicrobial effect. Results indicate the potential use of these plant's essential oils in poultry feed as an alternative to conventional antimicrobial products(AU)

A new Factor Xa inhibitor from Amblyomma cajennense with a unique domain composition.

Bioactive compounds of great interest are found in the saliva of hematophagous organisms. While exploring a cDNA library derived from the salivary glands of the tick Amblyomma cajennense, a transcript that codes for a protein with unique structure (containing an N-terminal Kunitz-type domain and a C-terminus with no homology to any annotated sequences) was found. The recombinant mature form of this protein ( approximately 13.5kDa) was produced in Escherichia coli BL21 (DE3), and it was able to inhibit Factor Xa (FXa) and extend global blood clotting times in vitro and ex vivo. Static and dynamic predictions of its tertiary structure indicate regions that may be related to its FXa inhibitor function.

A new Factor Xa inhibitor from Amblyomma cajennense with a unique domain composition

Bioactive compounds of great interest are found in the saliva of hematophagous organisms. While exploring a cDNA library derived from the salivary glands of the tick Amblyomma cajennense, a transcript that codes for a protein with unique structure (containing an N-terminal Kunitz-type domain and a C-terminus with no homology to any annotated sequences) was found. The recombinant mature form of this protein (¡­13.5 kDa) was produced in Escherichia coli BL21 (DE3), and it was able to inhibit Factor Xa (FXa) and extend global blood clotting times in vitro and ex vivo. Static and dynamic predictions of its tertiary structure indicate regions that may be related to its FXa inhibitor function.

Bothrops insularis venomics: a proteomic analysis supported by transcriptomic-generated sequence data

A joint transcriptomic and proteomic approach employing two-dimensional electrophoresis, liquid chromatography and mass spectrometry was carried out to identify peptides and proteins expressed by the venom gland of the snake Bothrops insularis, an endemic species of Queimada Grande Island, Brazil. Four protein families were mainly represented in processed spots, namely metalloproteinase, serine proteinase, phospholipase A2 and lectin. Other represented families were growth factors, the developmental protein G10, a disintegrin and putative novel bradykinin-potentiating peptides. The enzymes were present in several isoforms. Most of the experimental data agreed with predicted values for isoelectric point and Mr of proteins found in the transcriptome of the venom gland. The results also support the existence of posttranslational modifications and of proteolytic processing of precursor molecules which could lead to diverse multifunctional proteins. This study provides a preliminary reference map for proteins and peptides present in Bothrops insularis whole venom establishing the basis for comparative studies of other venom proteomes which could help the search for new drugs and the improvement of venom therapeutics. Altogether, our data point to the influence of transcriptional and post-translational events on the final venom composition and stress the need for a multivariate approach to snake venomics studies.

A new anti-loxoscelic serum produced against recombinant sphingomyelinase D Results of preclinical trials

Envenomation by Loxosceles species (brown spider) can lead to local dermonecrosis and to serious systemic effects. The main toxic component in the venom of these spiders is sphingomyelinase D (SMase D) and various isoforms of this toxin are present in Loxosceles venoms. We have produced a new anti-loxoscelic serum by immunizing horses with recombinant SMase D. In the present study, we compared the neutralization efficacy of the new anti-loxoscelic serum and anti-arachnidic serum (the latter serum is used for therapy for loxoscelism in Brazil) against the toxic effects of venoms from spiders of the genus Loxosceles. Neutralization tests showed that anti-SMase D serum has a higher activity against toxic effects of L. intermedia and L. laeta venoms and similar or slightly weaker activity against toxic effects of L. gaucho than that of Arachnidic serum. These results demonstrate that recombinant SMase D can replace venom for anti-venom production and therapy.

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity.

Alkaline phosphatase (ALP) is important in calcification and its expression seems to be associated with the inflammatory process. We investigated the in vitro acute effects of compounds used for the prevention or treatment of cardiovascular diseases on total ALP activity from male Wistar rat heart homogenate. ALP activity was determined by quantifying, at 410 nm, the p-nitrophenol released from p-nitrophenylphosphate (substrate in Tris buffer, pH 10.4). Using specific inhibitors of ALP activity and the reverse transcription-polymerase chain reaction, we showed that the rat heart had high ALP activity (31.73 +/- 3.43 nmol p-nitrophenol.mg protein-1.min-1): mainly tissue-nonspecific ALP but also tissue-specific intestinal ALP type II. Both ALP isoenzymes presented myocardial localization (striated pattern) by immunofluorescence. ALP was inhibited a) strongly by 0.5 mM levamisole, 2 mM theophylline and 2 mM aspirin (91, 77 and 84%, respectively) and b) less strongly by 2 mM L-phenylalanine, 100 mL polyphenol-rich beverages and 0.5 mM progesterone (24, 21 to 29 and 11%, respectively). beta-estradiol and caffeine (0.5 and 2 mM) had no effect; 0.5 mM simvastatin and 2 mM atenolol activated ALP (32 and 36%, respectively). Propranolol (2 mM) tended to activate ALP activity and corticosterone activated (18%) and inhibited (13%) (0.5 and 2 mM, respectively). We report, for the first time, that the rat heart expresses intestinal ALP type II and has high total ALP activity. ALP activity was inhibited by compounds used in the prevention of cardiovascular pathology. ALP manipulation in vivo may constitute an additional target for intervention in cardiovascular diseases.

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity

Alkaline phosphatase (ALP) is important in calcification and its expression seems to be associated with the inflammatory process. We investigated the in vitro acute effects of compounds used for the prevention or treatment of cardiovascular diseases on total ALP activity from male Wistar rat heart homogenate. ALP activity was determined by quantifying, at 410 nm, the p-nitrophenol released from p-nitrophenylphosphate (substrate in Tris buffer, pH 10.4). Using specific inhibitors of ALP activity and the reverse transcription-polymerase chain reaction, we showed that the rat heart had high ALP activity (31.73 ± 3.43 nmol p-nitrophenol·mg protein-1·min-1): mainly tissue-nonspecific ALP but also tissue-specific intestinal ALP type II. Both ALP isoenzymes presented myocardial localization (striated pattern) by immunofluorescence. ALP was inhibited a) strongly by 0.5 mM levamisole, 2 mM theophylline and 2 mM aspirin (91, 77 and 84 percent, respectively) and b) less strongly by 2 mM L-phenylalanine, 100 mL polyphenol-rich beverages and 0.5 mM progesterone (24, 21 to 29 and 11 percent, respectively). â-estradiol and caffeine (0.5 and 2 mM) had no effect; 0.5 mM simvastatin and 2 mM atenolol activated ALP (32 and 36 percent, respectively). Propranolol (2 mM) tended to activate ALP activity and corticosterone activated (18 percent) and inhibited (13 percent) (0.5 and 2 mM, respectively). We report, for the first time, that the rat heart expresses intestinal ALP type II and has high total ALP activity. ALP activity was inhibited by compounds used in the prevention of cardiovascular pathology. ALP manipulation in vivo may constitute an additional target for intervention in cardiovascular diseases.

Transcriptome analysis of Loxosceles laeta (Araneae, Sicariidae) spider venomous gland using expressed sequence tags

The bite of spiders belonging to the genus Loxosceles can induce a variety of clinical symptoms, including dermonecrosis, thrombosis, vascular leakage, haemolysis, and persistent inflammation. In order to examine the transcripts expressed in venom gland of Loxosceles laeta spider and to unveil the potential of its products on cellular structure and functional aspects, we generated 3,008 expressed sequence tags (ESTs) from a cDNA library. Results: All ESTs were clustered into 1,357 clusters, of which 16.4% of the total ESTs belong to recognized toxin-coding sequences, being the Sphingomyelinases D the most abundant transcript; 14.5% include "possible toxins", whose transcripts correspond to metalloproteinases, serinoproteinases, hyaluronidases, lipases, C-lectins, cystein peptidases and inhibitors. Thirty three percent of the ESTs are similar to cellular transcripts, being the major part represented by molecules involved in gene and protein expression, reflecting the specialization of this tissue for protein synthesis. In addition, a considerable number of sequences, 25%, has no significant similarity to any known sequence. Conclusion: This study provides a first global view of the gene expression scenario of the venom gland of L. laeta described so far, indicating the molecular bases of its venom composition.

In vitro modulation of alkaline phosphatase activity of Saccharomyces cerevisiae grown in low or high phosphate medium

Our objective was to characterize the modulation of the activity of Saccharomyces cerevisiae alkaline phosphatases (ALPs) by classic inhibitors of ALP activity, cholesterol and steroid hormones, in order to identify catalytic similarities between yeast and mammalian ALPs. S. cerevisiae expresses two ALPs, coded for by the PHO8 and PHO13 genes. The product of the PHO8 gene is repressible by Pi in the medium. ALP activity from yeast (grown in low or high phosphate medium) homogenates was determined with p-nitrophenylphosphate as substrate, pH 10.4 (lPiALP or hPiALP, respectively). Activation of hPiALP was observed with 5 mM L-amino acids (L-homoarginine _ 186 percent, L-leucine _ 155 percent and L-phenylalanine - 168 percent) and with 1 mM levamisole (122 percent; percentage values, in comparison to control, of recovered activity). EDTA (5 mM) and vanadate (1 mM) distinctly inhibited hPiALP (2 and 20 percent, respectively). L-homoarginine (5 mM) had a lower activating effect on lPiALP (166 percent) and was the strongest hPiALP activator. Corticosterone (5 mM) inhibited hPiALP to 90 percent, but no effect was observed in low phosphate medium. Cholesterol, ß-estradiol and progesterone also had different effects on lPiALP and hPiALP. A concentration-dependent activation of lPiALP minus hPiALP was evident with all three compounds, most especially with ß-estradiol and cholesterol. These results do not allow us to identify similarities of the behavior of S. cerevisiae ALPs and any of the mammalian ALPs but allow us to raise the hypothesis of differential regulation of S. cerevisiae ALPs by L-homoarginine, ß-estradiol and cholesterol and of using these compounds to discriminate between S. cerevisiae lPiALP and hPiALP.

In vitro modulation of alkaline phosphatase activity of Saccharomyces cerevisiae grown in low or high phosphate medium.

Our objective was to characterize the modulation of the activity of Saccharomyces cerevisiae alkaline phosphatases (ALPs) by classic inhibitors of ALP activity, cholesterol and steroid hormones, in order to identify catalytic similarities between yeast and mammalian ALPs. S. cerevisiae expresses two ALPs, coded for by the PHO8 and PHO13 genes. The product of the PHO8 gene is repressible by Pi in the medium. ALP activity from yeast (grown in low or high phosphate medium) homogenates was determined with p-nitrophenylphosphate as substrate, pH 10.4 (lPiALP or hPiALP, respectively). Activation of hPiALP was observed with 5 mM L-amino acids (L-homoarginine - 186%, L-leucine - 155% and L-phenylalanine - 168%) and with 1 mM levamisole (122%; percentage values, in comparison to control, of recovered activity). EDTA (5 mM) and vanadate (1 mM) distinctly inhibited hPiALP (2 and 20%, respectively). L-homoarginine (5 mM) had a lower activating effect on lPiALP (166%) and was the strongest hPiALP activator. Corticosterone (5 mM) inhibited hPiALP to 90%, but no effect was observed in low phosphate medium. Cholesterol, ss-estradiol and progesterone also had different effects on lPiALP and hPiALP. A concentration-dependent activation of lPiALP minus hPiALP was evident with all three compounds, most especially with ss-estradiol and cholesterol. These results do not allow us to identify similarities of the behavior of S. cerevisiae ALPs and any of the mammalian ALPs but allow us to raise the hypothesis of differential regulation of S. cerevisiae ALPs by L-homoarginine, ss-estradiol and cholesterol and of using these compounds to discriminate between S. cerevisiae lPiALP and hPiALP.

Bothrops jararaca venom gland transcriptome: Analysis of the gene expression pattern

Bothrops jararaca is a pit viper responsible for the majority of snake envenoming accidents in Brazil. As an attempt to describe the transcriptional activity of the venom gland, ESTs of a cDNA library constructed from B. jararaca venom gland were generated and submitted to bioinformatics analysis. The results showed a clear predominance of transcripts coding for toxins instead of transcripts coding for proteins involved in cellular functions. Among toxins, the most frequent transcripts were from metalloproteinases (52.6%), followed by serine-proteinases (28.5%), C-type lectins (8.3%) and bradykinin-potentiating peptides (BPPs) (6.2%). Results were similar to that obtained from the transcriptome analysis of B. insularis, a phylogenetically close sister of B. jararaca, though some differences were observed and are pointed out, such as a higher amount of the hypotensive BPPs in B. insularis transcriptome (19.7%). Another striking difference observed is that PIII and PII-classes of metalloproteinases are similarly represented in B. jararaca in contrast to B. insularis, in which a predominance of PIII-class metalloproteinase, which present a more intense hemorrhagic action, is observed. These features may, in part, explain the higher potency of B. insularis venom. The results obtained can help in proteome studies, and the clones can be used to directly probe the genetic material from other snake species or to investigate differences in gene expression pattern in response to factors such as diet, aging and geographic localization.

Some aspects of the venom proteome of the Colubridae snake Philodryas olfersii revealed from a Duvernoy's (venom) gland transcriptome

We investigated the putative toxins of Philodryas olfersii (Colubridae), a representative of a family of snakes neglected in venom studies despite their growing medical importance. Transcriptomic data of the venom gland complemented by proteomic analysis of the gland secretion revealed the presence of major toxin classes from the Viperidae family, including serine proteases, metalloproteases, C-type lectins, Crisps, and a C-type natriuretic peptide (CNP). Interestingly, the phylogenetic analysis of the CNP precursor showed it as a linker between two related precursors found in Viperidae and Elapidae snakes. We suggest that these precursors constitute a monophyletic group derived from the vertebrate CNPs.