Journal of Venomous Animals and Toxins Including Tropical Diseases最新文献

Background: PnPa11 and PnPa13 are synthetic peptides derived from Phoneutria nigriventer spider venom, which display antinociceptive and neuroprotective properties. In this work, we evaluated the safety of intravitreal use and the neuroprotective effect of these peptides.

Methods: The cytotoxicity and the antiangiogenic activity of these peptides were evaluated by the sulforhodamine-B method and chicken chorioallantoic membrane (CAM) assay, respectively. The in vivo safety was analyzed in Wistar rats that were intravitreally injected with different doses (0.50; 1.25; 2.50; 3.75 and 5.00 µg/mL) of these peptides (right eye, n = 6). The retinal function was assessed by electroretinography exams (ERG), intraocular pressure (IOP), and histological analyzes. In order to investigate the neuroprotective effect, Wistar rats received intravitreal injections (right eye, n = 6) of peptides at 1.25 µg/mL and then were exposed to blue LED light. In addition, the visual function and the retinal microstructure were verified.

Results: Cytotoxicity analyses demonstrated that the peptides did not present any toxicity over ARPE-19 (adult retinal pigmented epithelial) cell line and the antiangiogenic study highlighted that the peptides promoted the reduction of blood vessels. The intravitreal injection did not cause major changes, neither induced any irreversible damage. In the retinal degeneration assay, the ERG records demonstrated that the prior treatment with PnPa11 and PnPa13 protected the retina from damage. Morphological analyses confirmed the ERG findings. Immunoblotting analyses revealed that PnPa11 increased Erk1/2, NR2A, and NR2B retinal expression after the light stress model, but did not cause Akt1 activation, while PnPa13 prevented Erk1/2 and Akt1 dephosphorylation.

Conclusions: The intraocular administration of these peptides was well tolerated and presented protective activity against retinal degeneration, suggesting the potential use of these peptides as neuroprotectors in the ophthalmological field.

Background: Trimeresurus stejnegeri stejnegeri bite induces tissue swelling, pain, thrombocytopenia, rhabdomyolysis, and acute renal failure. However, the incidence of coagulopathy, factors associated with wound necrosis, and the appropriate management of this condition have not been well characterized yet.

Materials: This study included patients bitten by T. s. stejnegeri that were admitted to the study hospitals from 2001 to 2016. Patient characteristics, laboratory data, and management approaches were compared in victims with and without wound necrosis.

Results: A total of 185 patients were evaluated: three patients (1.6%) were asymptomatic; whereas tissue swelling and pain, local ecchymosis, wound necrosis, coagulopathy, thrombocytopenia, rhabdomyolysis, and renal impairment were present in 182, 53, 13, 15, 10, 1, and 3 patients, respectively. One patient died from coagulopathy and hemorrhagic shock. Antivenom was administered to all envenomed patients at a median time of 1.8 h after the bite. The median total dose of antivenom was five vials. Chi-square analysis showed that bitten fingers, using cold packs during first aid, presence of bullae or blisters, lymphangitis or lymphadenitis, local numbness and suspected infection to be significantly associated with wound necrosis. After adjustment using a multivariate logistic regression model, only cold packs as first aid, bulla or blister formation, and wound infection remained significant.

Conclusions: The main effects of T. s. stejnegeri envenomation are tissue swelling, pain, and local ecchymosis. We do not recommend the use of cold packs during first aid to reduce wound pain, as this may be a risk factor for wound necrosis. In addition, patients with bulla or blister formation should be carefully examined for subsequent wound necrosis. Antiplatelet use may worsen systemic bleeding. No severe rhabdomyolysis or renal failure was observed in this large case series, we therefore considered that they were not prominent effects of T. s. stejnegeri bite.

Background: The Amazon basin is one of the seven major geographical areas where scorpionism is recorded. In French Guiana, 90 stings per 100,000 inhabitants are registered per year. As the severity of cases is higher in children, descriptive studies are needed to have a better understanding of this pathology. The aim of the present study is to describe pediatric scorpionism in French Guiana.

Methods: We conducted a monocentric descriptive retrospective study on scorpion stings in all pediatric patients admitted to Cayenne General Hospital from January 1, 2002 to December 31, 2018.

Results: In this survey, 132 patients were included. Of them, 63% were male. Patients with general signs of envenomation were younger and lighter (p = 0.04). The picture was "one sting" (95.3%) by a "big" (47.6%), "black" (60%) and "small pincer" (58%) scorpion on the extremity of the body (84%). Stings occurred mainly during the day, while patients changed clothes. There was no envenomation during night. The monthly evaluation highlights that the number of stings and percentage of general signs of envenomation were closely connected to a composite variable including the variation of the level of rivers (p = 0.005). Cardiac symptoms were recorded in 82% of cases with general signs of envenomation. The presence of pulmonary; ear, nose, and throat (ENT); or gastrointestinal symptoms are related to major envenomation (p = 0.001, p = 0.01, and p = 0.02 respectively). Leukocytosis and glycemia increased according to the envenomation grade whereas serum potassium and alkaline reserve decreased. Forty-six patients needed hospitalization and seven of them required intensive care. No patient died nor presented sequelae at discharge from the hospital.

Conclusion: Pediatric scorpionism in French Guiana is closely associated with child activities and climatic conditions. Severe envenomation presented most of the time with cardiac, pulmonary, and gastrointestinal symptoms.

Background: Neuroimaging strategies are essential to locate, to elucidate the etiology, and to the follow up of brain disease patients. Magnetic resonance imaging (MRI) provides good cerebral soft-tissue contrast detection and diagnostic sensitivity. Inflammatory lesions and tumors are common brain diseases that may present a similar pattern of a cerebral ring enhancing lesion on MRI, and non-enhancing core (which may reflect cystic components or necrosis) leading to misdiagnosis. Texture analysis (TA) and machine learning approaches are computer-aided diagnostic tools that can be used to assist radiologists in such decisions.

Methods: In this study, we combined texture features with machine learning (ML) methods aiming to differentiate brain tumors from inflammatory lesions in magnetic resonance imaging. Retrospective examination of 67 patients, with a pattern of a cerebral ring enhancing lesion, 30 with inflammatory, and 37 with tumoral lesions were selected. Three different MRI sequences and textural features were extracted using gray level co-occurrence matrix and gray level run length. All diagnoses were confirmed by histopathology, laboratorial analysis or MRI.

Results: The features extracted were processed for the application of ML methods that performed the classification. T1-weighted images proved to be the best sequence for classification, in which the differentiation between inflammatory and tumoral lesions presented high accuracy (0.827), area under ROC curve (0.906), precision (0.837), and recall (0.912).

Conclusion: The algorithm obtained textures capable of differentiating brain tumors from inflammatory lesions, on T1-weghted images without contrast medium using the Random Forest machine learning classifier.

Background: Journal Impact Factor (JIF) has several intrinsic flaws, which highlight its inability to adequately measure citation distributions or indicate journal quality. Despite these flaws, JIF is still widely used within the academic community, resulting in the propagation of potentially misleading information. A critical review of the usefulness of JIF is needed including an overview of the literature to identify viable alternative metrics. The objectives of this study are: (1) to assess the usefulness of JIF by compiling and comparing its advantages and disadvantages; (2) to record the differential uses of JIF within research environments; and (3) to summarize and compare viable alternative measures to JIF.

Methods: Three separate literature search strategies using MEDLINE and Web of Science were completed to address the three study objectives. Each search was completed in accordance with PRISMA guidelines. Results were compiled in tabular format and analyzed based on reporting frequency.

Results: For objective (1), 84 studies were included in qualitative analysis. It was found that the recorded advantages of JIF were outweighed by disadvantages (18 disadvantages vs. 9 advantages). For objective (2), 653 records were included in a qualitative analysis. JIF was found to be most commonly used in journal ranking (n = 653, 100%) and calculation of scientific research productivity (n = 367, 56.2%). For objective (3), 65 works were included in qualitative analysis. These articles revealed 45 alternatives, which includes 18 alternatives that improve on highly reported disadvantages of JIF.

Conclusion: JIF has many disadvantages and is applied beyond its original intent, leading to inaccurate information. Several metrics have been identified to improve on certain disadvantages of JIF. Integrated Impact Indicator (I3) shows great promise as an alternative to JIF. However, further scientometric analysis is needed to assess its properties.

Background: South American rattlesnakes are represented in Brazil by a single species, Crotalus durissus, which has public health importance due to the severity of its envenomation and to its wide geographical distribution. The species is subdivided into several subspecies, but the current classification is controversial. In Brazil, the venoms of C. d. terrificus and C. d. collilineatus are used for hyperimmunization of horses for antivenom production, even though the distinction of these two subspecies are mostly by their geographical distribution. In this context, we described a comparative compositional and functional characterization of individual C. d. collilineatus and C. d. terrificus venoms from three Brazilian states.

Methods: We compared the compositional patterns of C. d. terrificus and C. d. collilineatus individual venoms by 1-DE and RP-HPLC. For functional analyzes, the enzymatic activities of PLA₂, LAAO, and coagulant activity were evaluated. Finally, the immunorecognition of venom toxins by the crotalic antivenom produced at Butantan Institute was evaluated using Western blotting.

Results: The protein profile of individual venoms from C. d. collilineatus and C. d. terrificus showed a comparable overall composition, despite some intraspecific variation, especially regarding crotamine and LAAO. Interestingly, HPLC analysis showed a geographic pattern concerning PLA₂. In addition, a remarkable intraspecific variation was also observed in PLA₂, LAAO and coagulant activities. The immunorecognition pattern of individual venoms from C. d. collilineatus and C. d. terrificus by crotalic antivenom produced at Butantan Institute was similar.

Conclusions: The results highlighted the individual variability among the venoms of C. durissus ssp. specimens. Importantly, our data point to a geographical variation of C. durissus ssp. venom profile, regardless of the subspecies, as evidenced by PLA₂ isoforms complexity, which may explain the increase in venom neurotoxicity from Northeastern through Southern Brazil reported for the species.

Background: The present study was carried out aiming to evaluate the impact of laryngeal sequelae on the quality of life of treated paracoccidioidomycosis (PCM) patients.

Methods: This cross-sectional study was conducted at the Otorhinolaryngology Outpatient Clinic of the University Hospital, Federal University of Mato Grosso do Sul, Brazil. Thirty-two PCM patients considered clinically and immunologically cured were included: 16 with laryngeal involvement during the active phase of the disease (laryngeal PCM group) and 16 without laryngeal involvement (control group). They were submitted to structured interview, otorhinolaryngology examination, videolaryngoscopy, videoendoscopic swallowing study, completed two questionnaires for voice self-assessment - Voice-related Quality of Life (V-RQOL) and Voice Handicap Index (VHI) - and were asked to score their voices on a scale from zero to 10 (self-assessment of vocal quality).

Results: Dysphonia was present in 50% of the cases. Patients with laryngeal PCM presented worse voice-related quality of life scores on the V-RQOL and poorer vocal quality self-assessment than the control group. No significant differences in the VHI were found between the groups. None of the participants developed dysphagic sequelae, although some minor changes were observed on videoendoscopic examination.

Conclusion: There were no dysphagia complaints and only a few mild changes were found on the fiberoptic endoscopic evaluation of swallowing, suggesting that this evaluation should be performed only in specific cases. Patients with laryngeal involvement presented worse V-RQOL and self-assessment voice quality. This study contributes to the current knowledge of the functional assessment of the larynx affected by PCM and the impact of dysphonia on quality of life.

Development of antibiotic resistance that leads to resurgence of bacterial infections poses a threat to disease-free existence for humankind and is a challenge for the welfare of the society at large. Despite research efforts directed towards treatment of pathogens, antibiotics within new improved classes have not emerged for years, a fact largely attributable to the pharmacological necessities compelling drug development. Recent reversion to the use of natural products alone or in combination with standard drugs has opened up new vistas for alternative therapeutics. The success of this strategy is evident in the sudden interest in plant extracts as additives/synergists for treatment of maladies caused by drug-resistant bacterial strains. Animal venoms have long fascinated scientists as sources of pharmacologically active components that can be exploited for the treatment of specific ailments and should be promoted further to clinical trials. In the present review, we outline the scope and possible methods for the applications of animal venoms in combination with commercial antibiotics to offer a better treatment approach against antibiotic-resistant infections.

Background: Spider venom is a potential source of pharmacologically important compounds. Previous studies on spider venoms reported the presence of bioactive molecules that possess cell-modulating activities. Despite these claims, sparse scientific evidence is available on the cytotoxic mechanisms in relation to the components of the spider venom. In this study, we aimed to determine the cytotoxic fractions of the spider venom extracted from Phlogiellus bundokalbo and to ascertain the possible mechanism of toxicity towards human lung adenocarcinoma (A549) cells.

Methods: Spider venom was extracted by electrostimulation. Components of the extracted venom were separated by reversed-phase high performance liquid chromatography (RP-HPLC) using a linear gradient of 0.1% trifluoroacetic acid (TFA) in water and 0.1% TFA in 95% acetonitrile (ACN). Cytotoxic activity was evaluated by the MTT assay. Apoptotic or necrotic cell death was assessed by microscopic evaluation in the presence of Hoechst 33342 and Annexin V, Alexa Fluor^TM 488 conjugate fluorescent stains, and caspase activation assay. Phospholipase A₂ (PLA₂) activity of the cytotoxic fractions were also measured.

Results: We observed and isolated six fractions from the venom of P. bundokalbo collected from Aurora, Zamboanga del Sur. Four of these fractions displayed cytotoxic activities. Fractions AT5-1, AT5-3, and AT5-4 were found to be apoptotic while AT5-6, the least polar among the cytotoxic components, was observed to induce necrosis. PLA₂ activity also showed cytotoxicity in all fractions but presented no relationship between specific activity of PLA₂ and cytotoxicity.

Conclusion: The venom of P. bundokalbo spider, an endemic tarantula species in the Philippines, contains components that were able to induce either apoptosis or necrosis in A549 cells.

Background: Liposomes are highly useful carriers for delivering drugs or antigens. The association of glycosylphosphatidylinositol (GPI)-anchored proteins to liposomes potentially enhances the immunogenic effect of vaccine antigens by increasing their surface concentration. Furthermore, the introduction of a universal immunoglobulin-binding domain can make liposomes targetable to virtually any desired receptor for which antibodies exist.

Methods: We developed a system for the production of recombinant proteins with GPI anchors and histidine tags and Strep-tags for simplified purification from cells. This system was applied to i) the green fluorescent protein (GFP) as a reporter, ii) the promising Plasmodium falciparum vaccine antigen PfRH5 and iii) a doubled immunoglobulin Fc-binding domain termed ZZ from protein A of Staphylococcus aureus. As the GPI-attachment domain, the C-terminus of murine CD14 was used. After the recovery of these three recombinant proteins from Chinese hamster ovary (CHO) cells and association with liposomes, their vaccine potential and ability to target the CD4 receptor on lymphocytes in ex vivo conditions were tested.

Results: Upon immunization in mice, the PfRH5-GPI-loaded liposomes generated antibody titers of 10³ to 10⁴, and showed a 45% inhibitory effect on in vitro growth at an IgG concentration of 600 µg/mL in P. falciparum cultures. Using GPI-anchored ZZ to couple anti-CD4 antibodies to liposomes, we created immunoliposomes with a binding efficiency of 75% to CD4⁺ cells in splenocytes and minimal off-target binding.

Conclusions: Proteins are very effectively associated with liposomes via a GPI-anchor to form proteoliposome particles and these are useful for a variety of applications including vaccines and antibody-mediated targeting of liposomes. Importantly, the CHO-cell and GPI-tagged produced PfRH5 elicited invasion-blocking antibodies qualitatively comparable to other approaches.