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Abstract Background Neutrophil migration to an inflamed site constitutes the first line of the innate immune response against invading microorganisms. Given the crucial role of endogenous lectins in neutrophil mobilization and activation, lectins from exogenous sources have often been considered as putative modulators of leukocyte function. Lectins purified from snake venom have been described as galactoside ligands that induce erythrocyte agglutination and platelet aggregation. This study evaluated human neutrophil migration and activation by C-type lectin BJcuL purified from Bothrops jararacussu venom. Results Utilizing fluorescence microscopy, we observed that biotinylated-BJcuL was evenly distributed on the neutrophil surface, selectively inhibited by D-galactose. Lectin was able to induce modification in the neutrophil morphology in a spherical shape for a polarized observed by optical microscopy and exposure to BJcuL in a Boyden chamber assay resulted in cell migration. After 30 minutes of incubation with BJcuL we found enhanced neutrophil functions, such as respiratory burst, zymozan phagocytosis and an increase in lissosomal volume. In addition, BJcuL delays late apoptosis neutrophils. Conclusion These results demonstrate that BJcuL can be implicated in a wide variety of immunological functions including first-line defense against pathogens, cell trafficking and induction of the innate immune response since lectin was capable of inducing potent neutrophil activation.

Abstract Background Autogenous Iliac Crest Bone Graft (ICBG) has been the "gold standard" for spinal fusion. However, bone graft harvest may lead to complications, such as chronic pain, numbness, and poor cosmesis. The long-term impact of these complications on patient function and well-being has not been established but is critical in determining the value of expensive bone graft substitutes such as recombinant bone morphogenic protein. We thus aimed to investigate the long-term complications of ICBG. Our second aim was to evaluate the psychometric properties of a new measure of ICBG morbidity that would be useful for appropriately gauging spinal surgery outcomes. Methods Prospective study of patients undergoing spinal fusion surgery with autologous ICBG. The SF-36v2, Oswestry Disability Index, and a new 14-item follow-up questionnaire addressing persistent pain, functional limitation, and cosmesis were administered with an 83% response rate. Multiple regression analyses examined the independent effect of ICBG complications on physical and mental health and disability. Results The study population included 170 patients with a mean age of 51.1 years (SD = 12.2) and balanced gender (48% male). Lumbar fusion patients predominated (lumbar = 148; cervical n = 22). At 3.5 years mean follow-up, 5% of patients reported being bothered by harvest site scar appearance, 24% reported harvest site numbness, and 13% reported the numbness as bothersome. Harvest site pain resulted in difficulty with household chores (19%), recreational activity (18%), walking (16%), sexual activity (16%), work activity (10%), and irritation from clothing (9%). Multivariate regression analyses revealed that persistent ICBG complications 3.5 years post-surgery were associated with significantly worse disability and showed a trend association with worse physical health, after adjusting for age, workers' compensation status, surgical site pain, and arm or leg pain. There was no association between ICBG complications and mental health in the multivariate model. Conclusion Chronic ICBG harvest site pain and discomfort is reported by a significant percentage of patients undergoing this procedure more than three years following surgery, and these complications are associated with worse patient-reported disability. Future studies should consider employing a control group that does not include autologous bone graft harvest, e.g., a group utilizing rhBMP, to determine whether eliminating harvest-site morbidity does indeed lead to observable improvement in clinical outcome sufficient to justify the increased cost of bone graft substitutes.

Abstract The field of health-related quality of life (HRQOL) could benefit from a broadening of perspectives to include recent advancements in research on adaptation, positive psychology, and genetics. These advances shed new light on the extent to which HRQOL is changeable or fixed. The objective of this paper is to integrate these insights and to discuss their implications for HRQOL research. We describe the Hedonic Treadmill theory, which asserts that positive events only temporarily affect happiness since people quickly return to hedonic neutrality. New empirical evidence suggests important revisions of this theory, providing a more optimistic picture of the possibility for change. Advances in positive psychology show that relatively simple interventions have the power to induce a sustainable increase in levels of happiness. Finally, a small but growing number of studies have found independent genetic influences in well-being, life satisfaction, perceived health, and even HRQOL. Given the increasing empirical evidence that HRQOL can be sustainably enhanced and is in part genetically determined, it may be useful to consider HRQOL as a concept that has state (environmental) and trait (genetic) components. This distinction will allow us to explore new pathways of improving theory, methods, and clinical practice. The overarching novel questions concern the extent to which HRQOL components are environmentally or genetically determined, and which factors lead to lasting improvement. This distinction begs for new research approaches, such as time-sampling techniques and interdisciplinary research investigating the genetic variants of HRQOL. Distinguishing between those aspects that are amenable to change from those that are relatively fixed and stable will help better target specific support interventions.

Abstract The increasing evidence for response shift phenomena in quality of life (QOL) assessment points to the necessity to reconsider both the measurement model and the application of psychometric analyses. The proposed psychometric model posits that the QOL true score is always contingent upon parameters of the appraisal process. This new model calls into question existing methods for establishing the reliability and validity of QOL assessment tools and suggests several new approaches for describing the psychometric properties of these scales. Recommendations for integrating the assessment of appraisal into QOL research and clinical practice are discussed.

Abstract Mounting evidence for response shifts in quality of life (QOL) appraisal indicates the need to include direct measurement of the appraisal process itself as a necessary part of QOL assessment. We propose that directly assessing QOL appraisal processes will not only improve our ability to interpret QOL scores in the traditional sense, but will also yield a deeper understanding of the appraisal process in the attribution of and divergence in meaning. The published evidence for response shift is reviewed, and an assessment paradigm is proposed that includes the explicit measurement of QOL appraisal process parameters: 1) induction of a frame of reference; 2) recall and sampling of salient experiences; 3) standards of comparison used to appraise experiences; and 4) subjective algorithm used to prioritize and combine appraisals to arrive at a QOL rating. A QOL Appraisal Profile, which measures key appraisal processes, is introduced as an adjunct to existing QOL scales. The proposed theoretical model, building on the Sprangers and Schwartz (1999) model and highlighting appraisal processes, provides a fully testable theoretical treatment of QOL and change in QOL, suggesting hypothesized causal relationships and explanatory pathways for both cross-sectional and longitudinal QOL research.

Background: Almost all Tityus characterized toxins are from subgenera Atreus and Tityus , there are only a few data about toxins produced by Archaeotityus , an ancient group in Tityus genus., Methods: Tityus (Archaeotityus) mattogrossensis crude venom was fractionated by high performance liquid chromatography, the major fractions were tested in a frog sciatic nerve single sucrose-gap technique. Two fractions (Tm1 and Tm2) were isolated, partially sequenced by MALDI-TOF/MS and electrophysiological assayed on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells., Results: The sucrose-gap technique showed neurotoxicity in four fractions. One fraction caused a delay of action potential repolarization and other three caused a reduction in amplitude. An electrophysiological assay showed that Tm1 is active on HEK293 Nav 1.3, HEK293 Nav 1.6, DUM and DRG cells, and Tm2 on HEK293 Nav 1.3 and DRG cells, but not in HEK293 Nav 1.6. In addition, Tm1 and Tm2 did promote a shift to more negative potentials strongly suggesting that both are α-NaScTx., Conclusion: Although Tityus (Archaeotityus) mattogrossensis is considered an ancient group in Tityus genus, the primary structure of Tm1 and Tm2 is more related to Tityus subgenus. The patch clamp electrophysiological tests suggest that Tm1 and Tm2 are NaScTx, and also promoted no shift to more negative potentials, strongly suggesting that both are α-NaScTx. This paper aimed to explore and characterize for the first time toxins from the ancient scorpion Tityus (Archaeotityus) mattogrossensis., Competing Interests: Competing interests: The authors declare that they have no competing interests.

Genome-wide mutational screens are central to understanding the genetic underpinnings of evolved and engineered phenotypes. The widespread adoption of CRISPR-Cas9 genome editing has enabled such screens in many organisms, but identifying functional sgRNAs still remains a challenge. Here, we developed a methodology to quantify the cutting efficiency of each sgRNA in a genome-scale library, and in doing so improve screens in the biotechnologically important yeast Yarrowia lipolytica. Screening in the presence and absence of native DNA repair enabled high-throughput quantification of sgRNA function leading to the identification of high efficiency sgRNAs that cover 94% of genes. Library validation enhanced the classification of essential genes by identifying inactive guides that create false negatives and mask the effects of successful disruptions. Quantification of guide effectiveness also creates a dataset from which determinants of CRISPR-Cas9 can be identified. Finally, application of the library identified novel mutations for metabolic engineering of high lipid accumulation., (Copyright © 2019 International Metabolic Engineering Society. All rights reserved.)

Ductal pancreatic adenocarcinoma is by far the most common solid tumor of the pancreas. It has a very poor prognosis, especially in the more advanced stages which are no longer locally confined. Due to mostly unspecific symptoms, imaging is key in the diagnostic process. Because of the widespread use of imaging techniques, incidental findings are to a greater extent discovered in the pancreas, which subsequently entail further work-up. Ductal pancreatic adenocarcinoma can be mimicked by a large number of different lesions, such as anatomical variants, peripancreatic structures and tumors, rarer primary solid pancreatic tumors, cystic tumors, metastases or different variants of pancreatitis. Additionally, a number of precursor lesions can be differentiated. The correct classification is thus important as an early diagnosis of ductal pancreatic adenocarcinoma is relevant for the prognosis and because the possibly avoidable treatment is very invasive. All major imaging techniques are principally suitable for pancreatic imaging. In addition to sonography of the abdomen, usually the baseline diagnostic tool, computed tomography (CT) with its superior spatial resolution, magnetic resonance imaging (MRI) with its good soft tissue differentiation capabilities, possibly in combination with MR cholangiopancreatography (MRCP), endosonography with its extraordinary spatial resolution, conceivably with additional endoscopic retrograde CP or the option of direct biopsy and finally positron emission tomography CT (PET-CT) as a molecular imaging tool are all particularly useful modalities. The various techniques all have its advantages and disadvantages; depending on the individual situation they may need to be combined.

Envenomation by wasp stings is a public health preoccupation, and signals after stings have variable effects depending on the number of attacks and individual sensitivities. Even with the high rate of wasp sting cases, the study of phatophysiological effects of the envenomation is still very incipient. In this context, early and accurate assessment of this prognostic can aid in the reduction of the symptomatology and complete remission of the later symptoms. Then, the present study evaluated the toxicological effects caused by envenomation produced by Synoeca surinama, a wasp easily found in Neotropical regions. In vivo tests comprised the evaluation of LD50 (OECD 423), nociception, edema, myotoxic lesion and hemorrhage induction, in vitro tests were realized to evaluate hemolysis, contractile and coagulation alteration. The envenomation effects observed were dose- and time-dependent; the LD50 observed for S. surinama was 178 μg/kg, approximately 17 times more lethal than that of the honeybee. Moreover, a potent algesic and oedema effect, and weak hemorrhagic signal were observed after injection of the venom wasp. Assays in vitro showed that this venom is able to prolong the clotting time of plasma and to increase creatine kinase levels. Our results demonstrated that this venom induced serious local and systemic effects in mammals and, so, to avoid permanent damage to the patient, health professionals should carefully investigate each accident. Moreover, due to its high occurrence in Neotropical regions, ecological management, particularly in areas with free access of children and elderly, should be performed., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The unborn child requires special protection. In this context, the indication for an X-ray examination is to be checked critically. If thereupon radiation of the lower abdomen including the uterus cannot be avoided, the examination should be postponed until the end of pregnancy or alternative examination techniques should be considered. Under certain circumstances, either accidental or in unavoidable cases after a thorough risk assessment, radiation exposure of the unborn may take place. In some of these cases an expert radiation hygiene consultation may be required. This consultation should comprise the expected risks for the unborn while not perturbing the mother or the involved medical staff. For the risk assessment in case of an in-utero x-ray exposition deterministic damages with a defined threshold dose are distinguished from stochastic damages without a definable threshold dose. The occurrence of deterministic damages depends on the dose and the developmental stage of the unborn at the time of radiation. To calculate the risks of an in-utero radiation exposure a three-stage concept is commonly applied. Depending on the amount of radiation, the radiation dose is either estimated, roughly calculated using standard tables or, in critical cases, accurately calculated based on the individual event. The complexity of the calculation thereby increases from stage to stage. An estimation based on stage one is easily feasible whereas calculations based on stages two and especially three are more complex and often necessitate execution by specialists. This article demonstrates in detail the risks for the unborn child pertaining to its developmental phase and explains the three-stage concept as an evaluation scheme. It should be noted, that all risk estimations are subject to considerable uncertainties. • Radiation exposure of the unborn child can result in both deterministic as well as stochastic damage und hitherto should be avoided or reduced to a minimum.• In case of radiation a suitable hygiene consultation may be necessary.• For risk assessment a three-stage concept is applied, which, depending on the radiation exposure, estimates or calculates the dose for the unborn child.• The radiologist plays a crucial role as a competent advisor and provider of reliable expert information. Citation Format: • Scharwächter C, Röser A, Schwartz CA et al. Prenatal Radiation Exposure: Dose Calculation. Fortschr Röntgenstr 2015; 187: 338 - 346., (© Georg Thieme Verlag KG Stuttgart · New York.)

Differences in pre-harvest stress measurements and carcass characteristics between kosher and not-qualified-as-kosher cattle were evaluated. Finished heifers (n=157) were slaughtered by a shochet while held in an upright position using Glatt slaughter procedures. Stress measurements were collected prior to slaughter. Carcass data were collected, and 3.8-cm thick samples were taken from the loin at the 13th rib. Steaks from each sample were evaluated for mechanical tenderness and simulated retail display. Cattle with shorter times from gate to exsanguination and lower vocalization scores were more likely (P < 0.01) to qualify as kosher. Kosher carcasses had larger (P = 0.02) ribeye areas and higher (P < 0.0001) Warner-Bratzler shear values. At each day of simulated retail display, kosher steaks had lower (P < 0.05) L*, a*, and b* values. These data suggest that body composition and pre-harvest stress affect the likelihood of a beef animal qualifying as kosher and quality differences exist between kosher and non-kosher steaks.

The objective was to use digital color image texture features to predict troponin-T degradation in beef. Image texture features, including 88 gray level co-occurrence texture features, 81 two-dimension fast Fourier transformation texture features, and 48 Gabor wavelet filter texture features, were extracted from color images of beef strip steaks (longissimus dorsi, n = 102) aged for 10d obtained using a digital camera and additional lighting. Steaks were designated degraded or not-degraded based on troponin-T degradation determined on d 3 and d 10 postmortem by immunoblotting. Statistical analysis (STEPWISE regression model) and artificial neural network (support vector machine model, SVM) methods were designed to classify protein degradation. The d 3 and d 10 STEPWISE models were 94% and 86% accurate, respectively, while the d 3 and d 10 SVM models were 63% and 71%, respectively, in predicting protein degradation in aged meat. STEPWISE and SVM models based on image texture features show potential to predict troponin-T degradation in meat., (© 2013.)

History: A 77-year-old woman presented with occasional vomiting of hematin but no further complaints. 50 years ago a partial gastrectomy after Billroth II had been performed due to a gastric ulcer., Investigations: The physical examination revealed no pathologic findings. All laboratory values, including hemoglobin (HB), were in normal range. The gastroscopic examination showed traces of hematin in the stomach and in the efferent and afferent loop.  The Braun anastomosis could not be delineated. No other pathologic findings were noted., Treatment and Course: At the request of the patient additional regular controls of hemoglobin were planned on an outpatient basis. In case of hemoglobin decrease, further treatment should be arranged. Two days after discharge the patient was presented with sudden massive abdominal pain and vomiting. Clinical examination confirmed the assumption of mechanical ileus. An emergency abdominal CT showed an intussusception of a small bowel loop into the Braun anastomosis as the ileus trigger. A laparotomy was carried out immediately after the CT. Because of severe ischemic damage of the intussuscepted loop, an "en bloc" resection of the loop including the Braun anastomosis including reanastomosis had to be carried out., Conclusion: Intussusception is primarily a childhood disease, occurring in adults only in about 5% of cases. Postoperative intussusception is a rare complication after gastric surgery.Especially because of its exceptionality the diagnosis of intussusception in adulthood can be difficult. Crucial herein is to have intussusception in mind in the differential diagnostic pathway of a post gastric surgery ileus., (© Georg Thieme Verlag KG Stuttgart · New York.)

Opisthacanthus cayaporum belongs to the Liochelidae family, and the scorpions from this genus occur in southern Africa, Central America and South America and, therefore, can be considered a true Gondwana heritage. In this communication, the isolation, primary structure characterization, and K⁺-channel blocking activity of new peptide from this scorpion venom are reported. OcyKTx2 is a 34 amino acid long peptide with four disulfide bridges and molecular mass of 3807 Da. Electrophysiological assays conducted with pure OcyKTx2 showed that this toxin reversibly blocks Shaker B K⁺-channels with a Kd of 82 nM, and presents an even better affinity toward hKv1.3, blocking it with a Kd of ∼18 nM. OcyKTx2 shares high sequence identity with peptides belonging to subfamily 6 of α-KTxs that clustered very closely in the phylogenetic tree included here. Sequence comparison, chain length and number of disulfide bridges analysis classify OcyKTx2 into subfamily 6 of the α-KTx scorpion toxins (systematic name, α-KTx6.17)., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Three individuals of silver carp (Hypophthalmichthys molitrix) were collected biweekly from Paranoá Lake (DF, Brazil) for analysis of microcystin (MC) concentrations in their muscle and liver tissue. Analysis by high performance liquid chromatography and mass spectrometry revealed MC masses and fragmentation patterns that were identified as MC-LR (995.04 m/z), MC-LA (909.01 m/z) and an unknown MC (987.07 m/z). Concentrations were calculated as MC-LR equivalents using a calibration curve prepared with a standard of MC-LR. May/06 was the month with the highest MC-LR equivalent concentrations in muscle and liver (3.83 ± 2.78, and 12.94 ± 10.51 μg g(-1), respectively). Our results show that during the drought months (April-September), consumption of fish with these MC concentrations would result in exposure to MCs that greatly exceed the World Health Organization's recommended tolerable daily intake limit of 0.04 μg MC kg(-1) body weight.

In the present study we conducted proteomic and pharmacological characterizations of the venom extracted from the Brazilian tarantula Acanthoscurria paulensis, and evaluated the cardiotoxicity of its two main fractions. The molecular masses of the venom components were identified by mass spectrometry (MALDI-TOF-MS) after chromatographic separation (HPLC). The lethal dose (LD(50)) was determined in mice. Nociceptive behavior was evaluated by intradermal injection in mice and the edematogenic activity by the rat hind-paw assay. Cardiotoxic activity was evaluated on in situ frog heart and on isolated frog ventricle strip. From 60 chromatographic fractions, 97 distinct components were identified, with molecular masses between 601.4 and 21,932.3 Da. A trimodal molecular mass distribution was observed: 30% of the components within 500-1999 Da, 38% within 3500-5999 Da and 21% within 6500-7999 Da. The LD(50) in mice was 25.4 ± 2.4 μg/g and the effects observed were hypoactivity, anuria, constipation, dyspnea and prostration until death, which occurred at higher doses. Despite presenting a dose-dependent edematogenic activity in the rat hind-paw assay, the venom had no nociceptive activity in mice. Additionally, the venom induced a rapid blockage of electrical activity and subsequent diastolic arrest on in situ frog heart preparation, which was inhibited by pretreatment with atropine. In the electrically driven frog ventricle strip, the whole venom and its low molecular mass fraction, but not the proteic one, induced a negative inotropic effect that was also inhibited by atropine. These results suggest that despite low toxicity, A. paulensis venom can induce severe physiological disturbances in mice., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The objective of this study was to investigate the usefulness of raw meat surface characteristics (texture) in predicting cooked beef tenderness. Color and multispectral texture features, including 4 different wavelengths and 217 image texture features, were extracted from 2 laboratory-based multispectral camera imaging systems. Steaks were segregated into tough and tender classification groups based on Warner-Bratzler shear force. The texture features were submitted to STEPWISE multiple regression and support vector machine (SVM) analyses to establish prediction models for beef tenderness. A subsample (80%) of tender or tough classified steaks were used to train models which were then validated on the remaining (20%) test steaks. For color images, the SVM model correctly identified tender steaks with 100% accurately while the STEPWISE equation identified 94.9% of the tender steaks correctly. For multispectral images, the SVM model predicted 91% and STEPWISE predicted 87% average accuracy of beef tender., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Background: Arthropod venoms have attracted interest because they represent a source of neuroactive compounds that can be useful tools in neuroscience and pharmacological investigations., Objective: The purpose of the present work was to evaluate the anticonvulsant, anxiolytic, and behavioral effects of the peptide fraction separated from venom of the social wasp., Materials and Methods: The low- molecular-weight compounds of the venom were separated by ultrafiltration and the bioassays were performed to test anticonvulsant and anxiolytic effects, as well as alterations in the spontaneous behavior of the animals., Results: Intracerebroventricular injections of the compounds induced dose-dependent anticonvulsant effects and a potent anxiolytic activity. Regarding behavioral effects, no significant differences were observed in relation to the saline control group., Conclusion: The low-molecular-weight compounds of the venom of Polybia paulista include neuroactive peptides that can be used as pharmacological resources for anticonvulsant and anxiolytic drug research.

Purpose: To evaluate the feasibility of diagnosing in statu nascendi mesenteric ischemia using diffusion-weighted imaging (DWI) in a porcine model. All experiments were approved by the animal care committee at the district administration., Materials and Methods: Mesenteric blood supply was studied in ten healthy female pigs (weight ~50 kg) with artificially induced mesenteric ischemia. In the DSA technique a branch of the superior mesenteric artery was embolized with tissue glue or small particles. DWI was performed using a 1.5 T MR scanner before embolization and 30 and 60 minutes after embolization using a 32-channel receiver coil. ADC maps were calculated for all animals. The findings were correlated to angiographic and autoptic findings. The image quality of DWI was assessed by means of SNR measurements and diagnostic impact by CNR measurements., Results: Embolization of the mesenteric branches was technically successful in all cases. DWI of the bowel was feasible with the applied sequences. In all animals, DWI displayed distinct cytotoxic edema as the earliest sign of ischemia thirty minutes after induction of ischemia. Furthermore, DWI yielded a distinct reduction in the water diffusion coefficient in all animals. Variance analysis showed good correlation between CNR measurements and infarction areas. Autoptic findings could confirm the detected infarction areas by DWI., Conclusion: DWI using parallel imaging techniques is feasible for the early detection of acute mesenteric ischemia. The presented DWI results offer encouraging prospects regarding more rapid disease diagnosis., (© Georg Thieme Verlag KG Stuttgart · New York.)

The venom of social wasps has been poorly studied so far, despite the high number of accidents in humans and assessment of the use of these wasps as a biological control of pests. The study of the pharmacological effects of the venom is of great importance since the poisoning is dangerous causing serious systemic effects, including death in the case of multiple attacks. In this study, the pharmacological activities of venom from the social wasp Synoeca cyanea were evaluated by the following assays: LD50 in mice, the behavioural effects and the hemorrhagic activity induced by the venom in mice, the oedematogenic activity in rat, the haemolysis in human blood, the stimulating effect on guinea-pig smooth muscle, and the antimicrobial activity. The aim was to determine the toxic effects of venom and to perform a comparative study with earlier work conducted with venom from other wasp species. Results showed that S. cyanea venom produced a potent dose-dependent oedema, as well as antibacterial and haemolytic activities, suggesting the presence of histamine, serotonin, kinins and other molecules related to increased vascular permeability and cytolytic activity in this venom. Despite previous studies with wasp venoms, S. cyanea venom presented a slight hemorrhagic effect. Data obtained in the smooth muscle assay also suggest the presence of BK or analogues in S. cyanea whole venom. The knowledge of symptoms and effects produced by S. cyanea venom is critical for health organizations, in order to improve clinical treatment in accidents caused by wasp stings., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The kappa-KTx family of peptides, which is the newest K⁺-channel blocker family from scorpion venom, is present in scorpions from the families Scorpionidae and Liochelidae. Differently from the other scorpion KTx families, the three-dimensional structure of the known kappa-KTxs toxins is formed by two parallel α-helices linked by two disulfide bridges. Here, the characterization of a new kappa-KTx peptide, designated kappa-KTx 2.5, derived from the Liochelidae scorpion Opisthacanthus cayaporum, is described. This peptide was purified by HPLC and found to be identical to OcyC8, a predicted mature sequence precursor (UniProtKB C5J89) previously described by our group. The peptide was chemically synthesized and the circular dichroism (CD) spectra of both, native and synthetic, conducted at different temperatures in water and water/trifluoroethanol (TFE), showed a predominance of α-helices. The kappa-KTx 2.5 is heat stable and was shown to be a blocker of K⁺-currents on hKv1.1, and hKv1.4, with higher affinity for Kv1.4 channels (IC₅₀= 71 μM). Similarly to the other kappa-KTxs, the blockade of K⁺-channels occurred at micromolar concentrations, leading to uncertainness about their proper molecular target, and consequently their pharmacologic effect. In order to test other targets, kappa-KTx2.5 was tested on other K⁺-channels, on Na⁺-channels, on bacterial growth and on smooth muscle tissue, a known assay to identify possible bradykinin-potentiating peptides, due to the presence of two contiguous prolines at the C-terminal sequence. It has no effect on the targets used except on hKv1.1, and hKv1.4 expressed in Chinese hamster ovary cells. Since the only plausible function found for kappa-KTx2.5 seems to be the blockade of K⁺-channels, a discussion regarding the analysis of structure-function relationships is included in this communication, based on sequence alignments of members of the kappa-KTx toxin family, and on computational simulation of a structural model of the kappa-KTx2.5-Kv1.2 complex., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Background: Percutaneous cardiac interventions are currently performed under x-ray guidance. Magnetic resonance imaging (MRI) has been used to guide intravascular interventions in the past, but mainly in animals. Translation of MR-guided interventions into humans has been limited by the lack of MR-compatible and safe equipment, such as MR guide wires with mechanical characteristics similar to standard guide wires. The aim of the present study was to evaluate the safety and efficacy of a newly developed MR-safe and compatible passive guide wire in aiding MR-guided cardiac interventions in a swine model and describe the 2 first-in-man solely MR-guided interventions., Methods and Results: In the preclinical trial, the new MR-compatible wire aided the performance of 20 interventions in 5 swine. These consisted of balloon dilation of nondiseased pulmonary and aortic valves, aortic arch, and branch pulmonary arteries. After ethics and regulatory authority approval, the 2 first-in-man MR-guided interventions were performed in a child and an adult, both with elements of valvar pulmonary stenosis. Catheter manipulations were monitored with real-time MRI sequence with interactive modification of imaging plane and slice position. Temporal resolution was 11 to 12 frames/s. Catheterization procedure times were 110 and 80 minutes, respectively. Both patients had successful relief of the valvar stenosis and no procedural complications., Conclusions: The described preclinical study and case reports are encouraging that with the availability of the new MR-compatible and safe guide wire, certain percutaneous cardiac interventions will become feasible to perform solely under MR guidance in the future. A clinical trial is underway in our institution.

We report 7 cases of patients bitten by giant water bugs, large predatory insects belonging to the Belostomatidae family (Hemiptera, Heteroptera). These insects have toxic saliva capable of provoking intense pain and paralysis in vertebrates. Victims experienced intense, excruciating pain and 1 manifested hypoesthesia in the forearm. Bites by Belostomatidae are often reported by clinicians working in areas where these insects live, but there are no detailed case reports in the medical literature. There are no specific treatment modalities known to be effective, making prevention an important strategy., (Copyright (c) 2010 Wilderness Medical Society. Published by Elsevier Inc. All rights reserved.)

This communication reports the separation of 80 fractions from the venom of the Ischnuridae scorpion Opisthacanthus cayaporum by high-performance liquid chromatography (HPLC). From these, 93 distinct components were identified by liquid chromatography/electrospray mass spectrometry (LC/ESI-MS) analysis, with molecular weights varying from 229.2 to 61,144.0 atomic mass units. Additionally, the HPLC fractions were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) which resulted in 221 distinct components, among which were 52 of the 93 obtained by LC/ESI-MS. The entire set of different molecular species found (total of 262 molecular masses) has a trimodal molecular weight distribution, with 42% of the components possessing 229.2-2985.3Da, 37% within the range of 3045.0-7258.6Da and 12% within the range 7458.4-9429Da. Seventeen peptides/proteins were isolated and were sequenced by Edman degradation, among which were a scorpine-like peptide (8315Da), presenting antimicrobial activity, and two phospholipase A2 with a molecular weight around 14kDa. The pharmacological effects of the venom were tested on isolated rat and insect (cockroach) nerves using the single sucrose-gap assay. The ED50 of the venom was 1.1mg/ml in insect nerves. Venom concentrations in the order of 3mg/ml causes only 9% reduction of compound action potentials (APs) of rat nerves, suggesting that this venom is rather specific for insects. Comparative analysis of venom from male and female O. cayaporum was performed by HPLC and MALDI-TOF-MS showing no qualitative variations, but rather quantitative differences among both samples.

In Brazil, the center of cassava origin, cassava landraces have acquired through their domestication a large diversity in relation to many economic traits such as high content of carotenoids and excellent palatability among other characters. One of these clones, which has been grown by indigenous Brazilian farmers and is now being maintained in the University of Brasília gene bank, showed a high level of lycopene content (5 mg/kg viz. a viz. zero in common cultivars, and 12-20 mg/kg in tomato, a lycopene-rich vegetable). A second landrace called UnB 400 had a high content of beta-carotene, which reached 4 mg/kg.

A novel toxin was identified, purified and characterized from the venom of the Mexican scorpion Hadrurus gertschi (abbreviated HgeTx1). It has a molecular mass of 3950 atomic mass units (a.m.u.) and contains 36 amino acids with four disulfide bridges established between Cys1-Cys5, Cys2-Cys6, Cys3-Cys7 and Cys4-Cys8. It blocks reversibly the Shaker B K(+)-channels with a Kd of 52nM. HgeTx1 shares 60%, 45% and 40% sequence identity, respectively, with Heterometrus spinnifer toxin1 (HsTX1), Scorpio maurus K(+)-toxin (maurotoxin) and Pandinus imperator toxin1 (Pi1), all four-disulfide bridged toxins. It is 57-58% identical with the other scorpion K(+)-channel toxins that contain only three disulfide bridges. Sequence comparison, chain length and number of disulfide bridges analysis classify HgeTx1 into subfamily 6 of the alpha-KTx scorpion toxins (systematic name: alpha-KTx 6.14).

Toxicity and toxin identification in Colomesus asellus, an Amazonian (Brazil) freshwater puffer fish. By using four different techniques--mouse bioassay, ELISA, HPLC and mass spectrometry-we evaluated the toxicity in the extracts of C. asellus, a freshwater puffer fish from the rivers of the Amazon, and identified for the first time the components responsible for its toxicity. The T20G10 monoclonal antibody raised against TTX, and employed in an indirect competitive enzyme immunoassay, showed very low affinity for the C. asellus extracts, indicating that TTX and its analogs are not the main toxic components of the extracts. This antibody was efficient in detecting presence of TTX in a total extract of Sphoeroides spengleri, which is one of the most toxic puffer fish found in the Atlantic coast. Extracts of C. asellus were toxic when administered intraperitonially into mice with an average toxicity of 38.6+/-12 mouse unit (MU)/g, while HPLC analysis indicated a lower toxin content (7.6+/-0 5MU/g). The HPLC profile showed no traces of TTX, but only the presence of PSPs (STX, GTX 2 and GTX 3). These toxins were also confirmed by electrospray ionization mass spectrometry.

The increase in the emergence of antibiotic-resistant microorganisms and difficult to treat infections caused by these pathogens stimulate research aiming the identification of novel antimicrobials. Skin secretion of amphibian contains a large number of biologically active compounds, including compounds that performance defense mechanisms against microorganisms. In the present work, two antimicrobial bufadienolides, telocinobufagin (402.1609 Da) and marinobufagin (400.1515 Da), were isolated from skin secretions of the Brazilian toad Bufo rubescens. The specimens were collected in Brasilia (Distrito Federal, Brazil), the skin secretions extracted by electric stimulation, and submitted to purification by RP-HPLC. The molecular structure and mass determination were done by (1)H and (13)C NMR and mass spectrometry data, respectively. The antimicrobial activity was performed by liquid growth inhibition against Staphylococcus aureus and Escherichia coli. The minimum inhibitory concentrations of telocinobufagin and marinobufagin were, respectively, 64.0 and 16.0 microg/mL for E. coli and both 128 microg/mL for S. aureus. Besides the antimicrobial activity both bufadienolides promoted an increase of the contraction force in isolated frog ventricle strips.

Tetrodotoxin (TTX) is one of the most potent toxin already isolated, which occurs in a wide range of marine as well as terrestrial animals such as in newts and anurans. In this work, the occurrence of TTX and analogues was examined in three brachycephalid species: Brachycephalus ephippium, B. nodoterga and B. pernix using LC-FLD and LC-MS/MS. In toxicity assay (intra-peritonial injection in mice) B. nodoterga extracts were non-toxic, while B. pernix extract exhibit the highest toxicity among the studied species. Skin showed the highest toxic, followed by the liver. Retention time data in the LC-FLD system indicated the presence of TTX, 4-epiTTX, 4,9-anhydroTTX and TDA, SIM data confirmed the presence of these compounds and revealed other analogs such as 11-norTTX-6(S)-ol, 5-deoxyTTX, 11-deoxyTTX, 11-oxoTTX, 6-epiTTX. Two new components were also identified by mass spectrometry (348 and 330Da). These unknown compounds have daughter ions similar to TTX, suggesting new putative TTX analogues.

The emergence, in recent years, of microbial resistance to commonly used antibiotics has aroused a search for new naturally occurring bactericidal and fungicidal agents that may have clinical utility. In the present study, three new antimicrobial peptides were purified from the electrical-stimulated skin secretion of the South American frog Leptodactylus ocellatus by reversed-phase chromatographic procedures. Ocellatin 1 (1GVVDILKGAGKDLLAHLVGKISEKV25-CONH2), ocellatin 2 (1GVLDIFKDAAKQILAHAAEKQI25-CONH2) and ocellatin 3 (1GVLDILKNAAKNILAHAAEQI21-CONH2) are structurally related peptides. These peptides present hemolytic activity against human erythrocytes and are also active against Escherichia coli. Ocellatins exhibit significant sequence similarity to other amphibian antimicrobial peptides, mainly to brevinin 2ED from Rana esculenta.

Epipedobates flavopictus, Dendrobatidae, is a small aposematic frog found in Brazilian Cerrado bioma. In the present work, we isolated and characterized chemically the most abundant alkaloids present in the cutaneous extract of E. flavopictus. The specimens were collected in Pirenópolis (Goiás, Brazil), their skins were removed and extracted with methanol, and submitted to purification by HPLC and identification by gas chromatography and mass spectrometry. Pumiliotoxin 251D, histrionicotoxin 285Da and two decahydroquinolines, 219A and 243A, were identified. The pumiliotoxin 251D was tested on isolated frog sciatic nerve and on isolated guinea pig ileum muscle. The pumiliotoxin 251D slightly reduced the action potentials amplitude of frog sciatic nerve. The crude skin extract of E. flavopictus and the pumiliotoxin 251D produced rhythmic contractions and increased the muscular tension on isolated guinea pig ileum.

11-oxoTTX is an analogue 4-5 times more toxic than TTX itself, been rare even in marine animals. Two ions at m/z 320 and 336 corresponding to TTX and 11-oxoTTX (M+H(+)), respectively, were detected in the Brachycephalidae frog Brachycephalus ephippium extracts. The fragment ion pattern of 11-oxoTTX is similar to that TTX, although its possible to verify some specific fragments.

The effect of the skin secretion of the amphibian Siphonops paulensis was investigated by monitoring the changes in conductance of an artificial planar lipid bilayer. Skin secretion was obtained by exposure of the animals to ether-saturated air, and then rinsing the animals with distilled water. Artificial lipid bilayers were obtained by spreading a solution of azolectin over an aperture of a Delrin cup inserted into a cut-away polyvinyl chloride block. In 9 of 12 experiments, the addition of the skin secretion to lipid bilayers displayed voltage-dependent channels with average unitary conductance of 258 +/- 41.67 pS, rather than nonspecific changes in bilayer conductance. These channels were not sensitive to 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid or tetraethylammonium ion, but the experimental protocol used does not permit us to specify their characteristics.

In the present study Tityus fasciolatus crude venom toxicity was evaluated and we also report the purification and characterization of a 6.6 kDa neurotoxin isolated from T. fasciolatus venom. This new toxin, named Tf4, has a molecular mass of 6614Da and its primary structure is homologous to TbIT-I from T. bahiensis and TsTX-VI and TsNTxP from T. serrulatus. Tf4 delays frog sodium channel inactivation reversibly, but it is non-toxic to mammals or crustaceans. An attempt to identify the residues responsible for the partial loss of toxicity in Tf4 was carried out based on homology modeling and sequence comparison.

The composition of indolealkylamines of Bufo rubescens cutaneous secretions was compared to those from six other Brazilian bufonids. Skin, parotoid and tibial gland secretions were obtained for analysis by thin-layer chromatography. A triple-quadrupole mass spectrometer was used to confirm the indolealkylamines standards (serotonin, 5-HT; bufotenin, BTN; dehydrobufotenin, DHB and bufotenidin, BTD). We observed clear variation in the composition of indolealkylamines of the cutaneous secretions studied and also between those found in the skin and parotoid gland secretions of the same species. We discuss the utility of indolealkylamines to the phylogeny of this group of toads.

Brachycephalus ephippium is a diurnal frog, that shows aposematic colouration and inhabits Atlantic forest leaf litter in south-eastern Brazil. The presence of tetrodotoxin (TTX) in the skin, liver and ovaries of B. ephippium was demonstrated. The skin (260 M.U./g) exhibited the highest toxicity followed by liver (177 M.U./g). TTX and its analogues, tetrodonic acid, 4-epitetrodotoxin and 4,9 anhydrotetrodotoxin were isolated and identified by HPLC followed by fluorimetric analysis. TTX and 11-nortetrodotoxin-6(S)-ol had their presence confirmed by mass spectrometry (MALDI-TOF). The results confirm Brachycephalidae as a fourth family of anurans containing TTX., (Copright 2002 Elsevier Science Ltd.)

Cutaneous T-cell lymphomas (CTCL) are extranodal non-Hodgkin's T-cell lymphomas that present in the skin, the most common form being mycosis fungoides. The progression of disease is associated with acquired immunodeficiency and increased susceptibility to infections. Ten CTCL patients presented with dark brown to black eschars overlying ulcerated tumors that cultured positive for Enterococcus and healed with appropriate antibiotic therapy. Enterococcal infections in CTCL tumors may be recognized as a distinct clinical entity requiring specific intervention.

The skin secretion of the caecilian S. paulensis has an indirect cardiotoxic effect. It induces a rapid blockage of electrical activity and subsequent diastolic arrest on in situ toad heart preparations. This action is not blocked by atropine and is therefore not mediated through a cholinergic mechanism. In addition, S. paulensis skin secretion is ineffective in inducing any response in isolated perfused toad heart assay, in the spontaneously beating isolated toad atria or in the electrically driven toad ventricle strip. These results, and the observation that S. paulensis skin secretion exhibits an hemolytic activity ([Schwartz, E. F., Schwartz, C. A., Sebben, A., Mendes E. G. (1997) Cardiotoxic and hemolytic activities on the caecilian Siphonops paulensis skin secretion. J. Venom. Anim. Toxins 3(1), 190]), suggest that the skin secretion cardiotoxicity could be mediated through an endogenous agent. The cardiotoxic action of S. paulensis skin secretion was investigated in isolated toad cardiac tissues in the presence of toad red blood cells (TRBC). In both atria and ventricle it evoked the same responses observed in the in situ heart. The potassium concentrations of the suspending media, as determined by atomic absorption spectroscopy were 9.5 and 11.7 times greater, respectively, in the presence of TRBC than in its absence. Given that increased levels of potassium also showed a negative inotropic effect in the isolated ventricle strip, the action of the skin secretion was attributed to the hemolytic activity, which causesan increase in K+ concentration of the medium.

The skin secretion of the caecilian Siphonops paulensis (SpSS) induces a time-and dose-dependent hemolytic response on red blood cells (RBC). When RBC from various animals species were subjected to the action of SpSS, a range of sensitivities was evident, sheep erythrocytes being the most susceptible, human, mouse and rabbit having moderate susceptibility, cow, snake and toad erythrocytes being more resistant, while S. paulensis RBC were entirely resistant. The hemolytic activity of SpSS was inhibited at temperatures higher than 60 degrees C. Both trypsin- and chymotrypsin-treated SpSS were ineffective in inducing RBC lysis. The treatment of SpSS with sheep RBC ghosts reduced its activity. There is no phospholipase activity in the SpSS., (Copyright 1998 John Wiley & Sons, Ltd.)

An expansion of the utilisation of o-phthalaldehyde in sulphuric acid medium as spray reagent was carried out when tryptophan and some tryptophan-derived indole alkylamines such as tryptamine, serotonin, bufotenine, dehydrobufotenine and bufotenidine were examined by thin-layer chromatography. Rf-values and limits of detection ranging from 20 (serotonin) to 100 (dehydrobufotenine) ng per spot were found. Application of this reagent for the detection of some of these compounds was carried out, using either methanolic extracts or column chromatographic fractions of the skin secretion of the toads Bufo ictericus and Odontophrynus cultripes.

Water content of the various body compartments was estimated serially during the first postnatal month in six term baboon neonates. In absolute volumes (mL), total body water, intracellular water, and plasma volume all increased linearly with body weight and postnatal age, although the effect of weight was predominant. In proportion to body weight (mL/kg), total body water, intracellular water, and plasma volume increased linearly with postnatal age whereas extracellular water and red cell volume decreased. There was no linear relation between the proportions of any of the water volumes and body weight.

Water content of the various body compartments were estimated immediately after beginning extracorporeal membrane oxygenation (ECMO), and daily thereafter for 3 days, in seven healthy 2-month-old lambs. Total body water, extracellular water, and plasma volume were estimated simultaneously by 18O, bromide, and T-1824 dilution, respectively. Volumes of intracellular water, interstitial water, blood, and red cells were calculated from the experimental estimates. No statistically significant changes occurred in the water content of the various body compartments in relation to duration of ECMO. The data suggest that water retention, clinically noted in human neonates treated with ECMO for persistent pulmonary hypertension, may be related to the primary disease process and/or its medical management, rather than to ECMO.

We demonstrate the feasibility of quantifying the abundance of 2H in plasma by nuclear magnetic resonance (NMR) spectroscopy. After adding internal standard (tert-butyl-d9 alcohol) to deproteinized plasma samples containing 2H2O, we determined the ratio of NMR peak areas for 2H2O and tert-butyl-d9 alcohol. This peak-area ratio was directly proportional to the exogenous 2H enrichment of plasma (difference between measured and naturally occurring 2H) between 0 and 0.272 atom % (r = 0.999). The coefficient of variation was 1.34% at an exogenous enrichment of 0.136 atom %. We applied this method to a study of the dilution kinetics of 2H2O to determine the optimal time and method of blood sampling for estimation of total body water content. The 2H enrichment of plasma stabilized by 4 h after intravenous injection of 2H2O, 1 g/kg of body weight, and fluctuated within 2-4% of the 4- to 8-h mean thereafter.

Total body water was estimated simultaneously by dilution of antipyrine (antipyrine space; APS) and H2(18)O (18O-space; 18OS) in 5 baboon neonates and 2 two-month-old lambs (total of 14 studies). Calculations of 18OS were made either from a single plasma delta 18O measured 4 h after injection of the marker (18OS4) or by extrapolation to time zero of several plasma delta 18O obtained at precisely timed interval (18OS0). Mean (+/- SD) 18OS4 was on the average 23% lower than mean 18OS0 (796 +/- 70 ml/kg vs. 650 +/- 67 ml/kg, p less than 0.001). Mean APS was not statistically different from mean 18OS0 (664 +/- 127 ml/kg vs. 658 +/- 57 ml/kg), but APS ranged from 41% lower to 30% higher than 18OS0. These data and information from the literature suggest that antipyrine is of doubtful reliability and should be abandoned as a marker for total body water.

A major polypeptide component of bovine lens beta-crystallins has been found to be heat stable. This property is utilized for purification of this polypeptide on a preparative scale. A homogenate of lens crystallins buffered at pH of 7.3 with 0.95 M Tris containing 1mM DTT and 1mM EDTA is heated to 97-99 degrees C for 3-5 min. Denatured proteins are removed and the clear supernatant is lyophilized. The lyophilized material mostly consists of the major polypeptide component common to the beta-crystallins with minor quantities of some low molecular weight polypeptides. These contaminants can be removed by preparative polyacrylamide gel electrophoresis. Amino acid analysis, electrophoretic mobility and chromatographic behavior of the purified polypeptide indicate that it is similar to the beta Bp polypeptide. Antibodies raised against this polypeptide react with all beta-crystallins but not with the alpha-or gamma crystallins.