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Abstract Improved therapies are needed against snakebite envenoming, which kills and permanently disables thousands of people each year. Recently developed neutralizing monoclonal antibodies against several snake toxins have shown promise in preclinical rodent models. Here, we use phage display technology to discover a human monoclonal antibody and show that this antibody causes antibody-dependent enhancement of toxicity (ADET) of myotoxin II from the venomous pit viper, Bothrops asper, in a mouse model of envenoming that mimics a snakebite. While clinical ADET related to snake venom has not yet been reported in humans, this report of ADET of a toxin from the animal kingdom highlights the necessity of assessing even well-known antibody formats in representative preclinical models to evaluate their therapeutic utility against toxins or venoms. This is essential to avoid potential deleterious effects as exemplified in the present study.

Ananteris is a scorpion genus that inhabits dry and seasonal areas of South and Central America. It is located in a distinctive morpho-group of Buthids, the ‘Ananteris group’, which also includes species distributed in the Old World. Because of the lack of information on venom composition, the study of Ananteris species could have biological and medical relevance. We conducted a venomics analysis of Ananteris platnicki, a tiny scorpion that inhabits Panama and Costa Rica, which shows the presence of putative toxins targeting ion channels, as well as proteins with similarity to hyaluronidases, proteinases, phospholipases A2, members of the CAP-domain family, and hemocyanins, among others. Venom proteolytic and hyaluronidase activities were corroborated. The determination of the primary sequences carried out by mass spectrometry evidences that several peptides are similar to the toxins present in venoms from Old World scorpion genera such as Mesobuthus, Lychas, and Isometrus, but others present in Tityus and Centruroides toxins. Even when this venom displays the characteristic protein families found in all Buthids, with a predominance of putative Na+-channel toxins and proteinases, some identified partial sequences are not common in venoms of the New World species, suggesting its differentiation into a distinctive group separated from other Buthids.

Fabry disease is an invalidating multisystemic disorder affecting α-Galactosidase, a rate-limiting hydrolase dedicated to lipid catabolism. Non-metabolized substrates, such as Globotriaosylceramide and its derivatives trigger the direct or indirect activation of inflammatory events and endothelial dysfunction. In spite of the efficacy demonstrated by enzyme replacement therapy or pharmacological chaperones in delaying disease progression, few studies have analyzed whether these treatments can improve the pro-inflammatory state of FD patients. Therefore, the aim of this work was to assess cytokines and cardiovascular risk-related proteins detectable in plasma from FD patients, whether treated or not with ERT, to evaluate the reliability of these markers in monitoring disease stage and treatment effects. We identified inflammatory and endothelial dysfunction markers (ADAMTS-13, TNF-α, GDF-15, MIP-1β, VEGFA, MPO, and MIC-1) that cooperate in a common pathway and are increased in FD patients’ plasma samples. As shown by the assessment of these proteins over time, they can help to evaluate the risk of higher severity in FD, as well as ERT effects. Even though the analyzed proteins cannot be considered as proper biomarkers due to their non-specificity to FD, taken together they can provide a signature of reference molecules with prognostic value for early diagnosis, and evaluation of disease progression and treatment efficacy, using blood samples.

El síndrome conocido como “oreja caída” es un hallazgo clínico observado en los sistemas productivos bovinos. Varias son las etiologías descriptas como causa de esa patología. El objetivo del presente trabajo es describir tres casos de oreja caída en bovinos. Se estudiaron dos casos en engorde a corral y uno en guachera. Se realizaron exámenes clínicos, necropsias, histopatologías y diferentes técnicas entre ellas cultivo de hongos, de bacterias, de virus, etc. En todos los animales necropsiados se observó aumento de tamaño del ganglio de Gasser. En los estudios histopatológicos se encontró reacción granulomatosa y presencia de estructuras compatibles con criptococos que fueron positivas a la tinción de PAS. El aislamiento y tipificación de Cryptococcus neoformans de 1 animal, los resultados de los exámenes anatomo-histopatológicos, y las características estructurales y tintoriales observadas en todos los casos indican un diagnóstico compatible con criptococosis nerviosa.

Abstract Viperid snake venoms contain a unique family of cytotoxic proteins, the Lys49 PLA2 homologs, which are devoid of enzymatic activity but disrupt the integrity of cell membranes. They are known to induce skeletal muscle damage and are therefore named ‘myotoxins’. Single intact and skinned (devoid of membranes and cytoplasm but with intact sarcomeric proteins) rat cardiomyocytes were used to analyze the cytotoxic action of a myotoxin, from the venom of Bothrops asper. The toxin induced rapid hypercontraction of intact cardiomyocytes, associated with an increase in the cytosolic concentration of calcium and with cell membrane disruption. Hypercontraction of intact cardiomyocytes was abrogated by the myosin inhibitor para-aminoblebbistatin (AmBleb). No toxin-induced changes of key parameters of force development were observed in skinned cardiomyocytes. Thus, although myosin is a key effector of the observed hypercontraction, a direct effect of the toxin on the sarcomeric proteins -including the actomyosin complex- is not part of the mechanism of cytotoxicity. Owing to the sensitivity of intact cardiomyocytes to the cytotoxic action of myotoxin, this ex vivo model is a valuable tool to explore in further detail the mechanism of action of this group of snake venom toxins.

Fabry disease (FD) is an X-linked lysosomal storage disorder (LSD) secondary to mutations in the GLA gene that causes dysfunctional activity of lysosomal hydrolase α-galactosidase A and results in the accumulation of globotriaosylceramide (Gb3) and globotriaosylsphingosine (lyso-Gb3). The endothelial accumulation of these substrates results in injury to multiple organs, mainly the kidney, heart, brain and peripheral nervous system. The literature on FD and central nervous system involvement is scarce when focusing on alterations beyond cerebrovascular disease and is nearly absent in regard to synaptic dysfunction. In spite of that, reports have provided evidence for the CNS’ clinical implications in FD, including Parkinson’s disease, neuropsychiatric disorders and executive dysfunction. We aim to review these topics based on the current available scientific literature.

Coralsnakes belong to the family Elapidae and possess venoms which are lethal to humans and can be grouped based on the predominance of either three finger toxins (3FTxs) or phospholipases A2 (PLA2s). A proteomic and toxicological analysis of the venom of the coralsnake Micrurus yatesi was performed. This species, distributed in southeastern Costa Rica, was formerly considered a subspecies of M. alleni. Results showed that this venom is PLA2-rich, in contrast with the previously studied venom of Micrurus alleni. Toxicological evaluation of the venom, in accordance with proteomic data, revealed that it has a markedly higher in vitro PLA2 activity upon a synthetic substrate than M. alleni. The evaluation of in vivo myotoxicity in CD-1 mice using histological evaluation and plasma creatine kinase release also showed that M. yatesi venom caused muscle damage. A commercial equine antivenom prepared using the venom of Micrurus nigrocinctus displayed a similar recognition of the venoms of M. yatesi and M. nigrocinctus by enzyme immunoassay. This antivenom also immunorecognized the main fractions of the venom of M. yatesi and was able to neutralize its lethal effect in a murine model.

Peptaibols (Paib), are a class of biologically active peptides isolated from soil, fungi and molds, which have interesting properties as antimicrobial agents. Paib production was optimized in flasks by adding sucrose as a carbon source, 2-aminoisobutyric acid (Aib) as an additive amino acid, and F. oxysporum cell debris as an elicitor. Paib were purified, sequenced and identified by High-performance liquid chromatography (HPLC)coupled to mass spectrometry. Afterward, a Paib extract was obtained from the optimized fermentations. The biological activity of these extracts was evaluated using in vitro and in vivo methods. The extract inhibited the growth of specific plant pathogens, and it showed inhibition rates similar to those from commercially available fungicides. Growth inhibition rates were 92.2, 74.2, 58.4 and 36.2% against Colletotrichum gloeosporioides, Botrytis cinerea, Alternaria alternata and Fusarium oxysporum, respectively. Furthermore, the antifungal activity was tested in tomatoes inoculated with A. alternata, the incidence of the disease in tomatoes treated with the extract was 0%, while the untreated fruit showed a 92.5% incidence of infection Scanning electron microscopy images showed structural differences between the fungi treated with or without Paib. The most visual alterations were sunk and shriveled morphology in spores, while the hyphae appeared to be fractured, rough and dehydrated.

The exchange rate pass-through into the consumer price index on healthcare goods and services was measured by estimating a FAVAR model for Colombia. Results provide evidence of an incomplete and heterogeneous effect. There is no indication of transmission to the services or insurance indexes, but there is a significant effect on the medicines and devices indexes that have implications for out-of-pocket expenditure. Therefore, this indicates that the Colombian healthcare system effectively protects consumers from exchange rate volatility, but may need to design policies to protect consumers from price rises in medicines and goods that are not covered by the national benefits package.

Snake envenoming is a major but neglected human disease in tropical and subtropical regions. Among venomous snakes in the Americas, coral snakes of the genus Micrurus are particularly dangerous because they cause a peripheral neuroparalysis that can persist for many days or, in severe cases, progress to death. Ventilatory support and the use of snake species-specific antivenoms may prevent death from respiratory paralysis in most cases. However, there is a general consensus that additional and non-expensive treatments that can be delivered even long after the snake bite are needed. Neurotoxic degeneration of peripheral motor neurons activates pro-regenerative intercellular signaling programs, the greatest of which consist of the chemokine CXCL12α, produced by perisynaptic Schwann cells, which act on the CXCR4 receptor expressed on damaged neuronal axons. We recently found that the CXCR4 agonist NUCC-390 promotes axonal growth. Here, we show that the venom of the highly neurotoxic snake Micrurus nigrocinctus causes a complete degeneration of motor axon terminals of the soleus muscle, followed by functional regeneration whose time course is greatly accelerated by NUCC-390. These results suggest that NUCC-390 is a potential candidate for treating human patients envenomed by Micrurus nigrocinctus as well as other neurotoxic Micrurus spp. in order to improve the recovery of normal neuromuscular physiology, thus reducing the mortality and hospital costs of envenoming.

Snake species within the Bothrops complex (sensu lato) are of medical relevance in Latin America, but knowledge on their venom characteristics is limited, or even unavailable, for some taxa. Perú harbors 17 species of pit vipers, within the genera Bothrops, Bothriechis, Bothrocophias, Porthidium, Crotalus, and Lachesis. This study compared the venoms of twelve species, through chromatographic and electrophoretic profiles, as well as proteolytic and phospholipase A2 (PLA2) activities. Also, proteomic profiles were analyzed for nine of the venoms using a shotgun approach. Results unveiled conspicuous differences in the expression of venom PLA2s among species, six of them presenting scarce levels as judged by RP-HPLC profiles. Since most species within the bothropoid lineage possess venoms with high to intermediate abundances of this protein family, our findings suggest the existence of a phenotypic duality in the expression of venom PLA2s within the Bothrops (sensu lato) complex. Bothrops barnetti and Bothrocophias andianus venoms, very scarce in PLA2s, were shown to lack significant myotoxic activity, highlighting that the observed variability in PLA2 expression bears toxicological correlations with effects attributed to these proteins. Finally, an attempt to identify phylogenetic relationships of bothropoid species from Perú presenting low- or high-PLA2 venom phenotypes showed an interspersed pattern, thus precluding a simple phylogenetic interpretation of this venom compositional dichotomy.

Snakebite envenoming is a major neglected tropical disease that affects millions of people every year. The only effective treatment against snakebite envenoming consists of unspecified cocktails of polyclonal antibodies purified from the plasma of immunized production animals. Currently, little data exists on the molecular interactions between venom-toxin epitopes and antivenom-antibody paratopes. To address this issue, high-density peptide microarray (hdpm) technology has recently been adapted to the field of toxinology. However, analysis of such valuable datasets requires expert understanding and, thus, complicates its broad application within the field. In the present study, we developed a user-friendly, and high-throughput web application named "Snake Toxin and Antivenom Binding Profiles" (STAB Profiles), to allow straight-forward analysis of hdpm datasets. To test our tool and evaluate its performance with a large dataset, we conducted hdpm assays using all African snake toxin protein sequences available in the UniProt database at the time of study design, together with eight commercial antivenoms in clinical use in Africa, thus representing the largest venom-antivenom dataset to date. Furthermore, we introduced a novel method for evaluating raw signals from a peptide microarray experiment and a data normalization protocol enabling intra-microarray and even inter-microarray chip comparisons. Finally, these data, alongside all the data from previous similar studies by Engmark et al., were preprocessed according to our newly developed protocol and made publicly available for download through the STAB Profiles web application (http://tropicalpharmacology.com/tools/stab-profiles/). With these data and our tool, we were able to gain key insights into toxin-antivenom interactions and were able to differentiate the ability of different antivenoms to interact with certain toxins of interest. The data, as well as the web application, we present in this article should be of significant value to the venom-antivenom research community. Knowledge gained from our current and future analyses of this dataset carry the potential to guide the improvement and optimization of current antivenoms for maximum patient benefit, as well as aid the development of next-generation antivenoms.

Fabry disease is an X-linked multisystemic disorder caused by the impairment of lysosomal α-Galactosidase A, which leads to the progressive accumulation of glycosphingolipids and to defective lysosomal metabolism. Currently, Fabry disease is treated by enzyme replacement therapy or the orally administrated pharmacological chaperone Migalastat. Both therapeutic strategies present limitations, since enzyme replacement therapy has shown low half-life and bioavailability, while Migalastat is only approved for patients with specific mutations. The aim of this work was to assess the efficacy of PBX galactose analogues to stabilize α-Galactosidase A and therefore evaluate their potential use in Fabry patients with mutations that are not amenable to the treatment with Migalastat. We demonstrated that PBX compounds are safe and effective concerning stabilization of α-Galactosidase A in relevant cellular models of the disease, as assessed by enzymatic activity measurements, molecular modelling, and cell viability assays. This experimental evidence suggests that PBX compounds are promising candidates for the treatment of Fabry disease caused by mutations which affect the folding of α-Galactosidase A, even for GLA variants that are not amenable to the treatment with Migalastat.

Lysosomal Storage Diseases are multisystemic disorders determined by genetic variants, which affect the proteins involved in lysosomal function and cellular metabolism. Different therapeutic approaches, which are based on the physiologic mechanisms that regulate lysosomal function, have been proposed for these diseases. Currently, enzyme replacement therapy, gene therapy, or small molecules have been approved or are under clinical development to treat lysosomal storage disorders. The present article reviews the main therapeutic strategies that have been proposed so far, highlighting possible limitations and future perspectives.

Aspergillus species are widely distributed throughout the world and can develop parasitic and saprophytic ways of life, allowing Aspergillus to infect living hosts, including plants, insects, birds and mammals. The most common form of aspergillosis in poultry and other birds is respiratory infection. Clinical manifestations depend on the infective dose, pre-existing diseases, and the immune response of the host. The aim of the present research was to study aspergillosis in domestic and wild birds from Argentina. We carried out morphological and molecular identification, and determination of antifungal susceptibility against seven antifungal drugs. Six birds from different cities of Buenos Aires Province of Argentina were studied. Three of the samples belonged to broiler chicks, while the other three belonged to an eagle, a pheasant, and a kelp gull. Two isolates were identified as Aspergillus fumigatus by morphological characteristics and growth at 50 °C. Morphology and BenA sequencing enabled us to identify three isolates as Aspergillus flavus, and one as Aspergillus sydowii. All antifungal drugs tested showed low MIC values, ranging from 0.008 to 1 mg/L. Aspergillosis in birds causes high economic losses and could be controlled by sanitation, avoidance of moldy food, nest and litter and reducing stress factors.

Species of the genus Scedosporium (family Microascaceae, phylum Ascomycota) are responsible for a wide range of opportunistic human infections, and have a low susceptibility to most antifungal drugs. It is well known that the pattern of Scedosporium species distribution varies according to geographic region. To assess the diversity of Scedosporium species in Argentina involved in human infections, we carried out a retrospective study reviewing 49 strains from clinical samples sent for diagnosis to the National Clinical Mycology Reference Laboratory between 1985 and 2019. Then, a phenotypic characterization, a phylogenetic study and and in vitro susceptibility test to antifungals were carried out. An analysis of combined nucleotide sequences dataset of the internal transcribed spacer of the ribosomal DNA (ITS) and of a fragment of the β-tubulin gene (BT2) demonstrated that 92% of the strains belonged to the species S. boydii, S. apiospermum and S. angustum, all them pertaining to S. apiospermum species complex. However, two strains (4%) were identified as S. aurantiacum, a species never reported in clinical settings in the Americas’. Surprisingly, one of them displayed a polycytella-like conidiogenesis, up to date only reported for S. apiospermum. In addition, the strain DMic 165285 was phylogenetically located far away from the rest of the species, so is proposed as the novel species Scedosporium americanum. On the other hand, from all seven antifungals tested, voriconazole and posaconazole were the most active drugs against Scedosporium spp.

This article analyzes different international share price indices for the period 1995-2013, in order to test for the existence and date of appearance of asset price explosions in the world’s stock markets. A sign test is employed to construct different indices of bubbles in representative financial markets for each region, using dynamic factor models. These indices permit a characterization to be made of each region in terms of risk and, also, of the occurrence of financial bubbles. Evidence is found that indicates a certain degree of synchronization between episodes of financial bubbles in the markets analyzed and, generally, at international level.

The global proliferation of plug-in electric vehicles (PEVs) poses a major challenge for current and future distribution systems. If uncoordinated, their charging process may cause congestion on both network transformers and feeders, resulting in overheating, deterioration, protection triggering and eventual risk of failure, seriously compromising the stability and reliability of the grid. To mitigate such impacts and increase their hosting capacity in radial distribution systems, the present study compares the levels of effectiveness and performances of three alternative centralized thermal management formulations for a real-time agent-based charge control algorithm that aims to minimize the total impact upon car owners. A linear formulation and a convex formulation of the optimization problem are presented and solved respectively by means of integer linear programming and a genetic algorithm. The obtained results are then compared, in terms of their total impact on the end-users and overall performance, with those of the current heuristic implementation of the algorithm. All implementations were tested using a simulation environment considering multiple vehicle penetration and base load levels, and equipment modeled after commercially available charging stations and vehicles. Results show how faster resolution times are achieved by the heuristic implementation, but no significant differences between formulations exist in terms of network management and end-user impact. Every vehicle reached its maximum charge level while all thermal impacts were mitigated for all considered scenarios. The most demanding scenario showcased over a 30% reduction in the peak load for all thermal variants.

Se plantea el coeficiente de dependencia asintótica, basado en cópulas, como una medida para la administración del riesgo en portafolios de acciones. Se describen algunos aspectos de las estructuras macro y microeconómicas del mercado en Colombia, motivando la introducción de medidas como la propuesta. La aplicación a catorce de las acciones más líquidas en el período de estudio (2007-2012) permite dar algunas recomendaciones de diversificación en caso de materialización de eventos extremos. En consecuencia, con la hipótesis de partida, se encuentran indicios de una alta vulnerabilidad del mercado de acciones colombiano ante eventos de tipo sistémico.

Con el fin de poner a prueba hipótesis recientes en la literatura económica, según las cuales la formación de burbujas periódicas y sincronizadas en los mercados globales es consecuencia de la migración recurrente de capitales de portafolio, en este trabajo se estiman los periodos de burbujas financieras en los mercados de acciones latinoamericanos de mayor tamaño relativo; asimismo, la cronología de tales burbujas se compara con la estimada para el caso estadounidense. Estas burbujas se estiman haciendo uso de un estadístico de signo, con ajuste recursivo de mediana, el cual sirve para contrastar la hipótesis de caminata aleatoria versus alternativas explosivas. Se encuentra evidencia favorable para la hipótesis de partida y se exploran posibles ampliaciones de ésta.

Using variational arguments, we prove some nonexistence and multiplicity results for positive solutions of a system of p-Laplace equations of gradient form. Then we study a p-Laplace-type problem with nonlinear boundary conditions.

Many Massively Multiplayer Online Role-Playing Games (MMORPGs) use TCP flows for communication between the server and the game clients. The utilization of TCP, which was not initially designed for (soft) real-time services, has many implications for the competing traffic flows. In this paper we present a series of studies which explore the competition between MMORPG and other traffic flows. For that aim, we first extend a source-based traffic model, based on player’s activities during the day, to also incorporate the impact of the number of players sharing a server (server population) on network traffic. Based on real traffic traces, we statistically model the influence of the variation of the server’s player population on the network traffic, depending on the action categories (i.e., types of in-game player behaviour). Using the developed traffic model we prove that while server population only modifies specific action categories, this effect is significant enough to be observed on the overall traffic. We find that TCP Vegas is a good option for competing flows in order not to throttle the MMORPG flows and that TCP SACK is more respectful with game flows than other TCP variants, namely, Tahoe, Reno, and New Reno. Other tests show that MMORPG flows do not significantly reduce their sending window size when competing against UDP flows. Additionally, we study the effect of RTT unfairness between MMORPG flows, showing that it is less important than in the case of network-limited TCP flows.

Two subtypes of phospholipases A2 (PLA2s) with the ability to induce myonecrosis, 'Asp49' and 'Lys49' myotoxins, often coexist in viperid snake venoms. Since the latter lack catalytic activity, two different mechanisms are involved in their myotoxicity. A synergism between Asp49 and Lys49 myotoxins from Bothrops asper was previously observed in vitro, enhancing Ca2+ entry and cell death when acting together upon C2C12 myotubes. These observations are extended for the first time in vivo, by demonstrating a clear enhancement of myonecrosis by the combined action of these two toxins in mice. In addition, novel aspects of their synergism were revealed using myotubes. Proportions of Asp49 myotoxin as low as 0.1% of the Lys49 myotoxin are sufficient to enhance cytotoxicity of the latter, but not the opposite. Sublytic amounts of Asp49 myotoxin also enhanced cytotoxicity of a synthetic peptide encompassing the toxic region of Lys49 myotoxin. Asp49 myotoxin rendered myotubes more susceptible to osmotic lysis, whereas Lys49 myotoxin did not. In contrast to myotoxic Asp49 PLA2, an acidic non-toxic PLA2 from the same venom did not markedly synergize with Lys49 myotoxin, revealing a functional difference between basic and acidic PLA2 enzymes. It is suggested that Asp49 myotoxins synergize with Lys49 myotoxins by virtue of their PLA2 activity. In addition to the membrane-destabilizing effect of this activity, Asp49 myotoxins may generate anionic patches of hydrolytic reaction products, facilitating electrostatic interactions with Lys49 myotoxins. These data provide new evidence for the evolutionary adaptive value of the two subtypes of PLA2 myotoxins acting synergistically in viperid venoms.

This paper analyzes and compares two different video conference solutions, widely used in corporate and home environments, with a special focus on the mechanisms used for adapting the traffic to the network status. The results show how these mechanisms are able to provide a good quality in the hostile environment of the public Internet, a best effort network without delay or delivery guarantees. Both solutions are evaluated in a laboratory, where different network impairments (bandwidth limit, delay, and packet loss) are set, in both the uplink and the downlink, and the reaction of the applications is measured. The tests show how these solutions modify their packet size and interpacket time, in order to increase or reduce the sent data. One of the solutions also uses a scalable video codec, able to adapt the traffic to the network status and to the end devices.

The behaviour of the routers’ buffer may affect the quality of service (QoS) of network services under certain conditions, since it may modify some traffic characteristics, as delay or jitter, and may also drop packets. As a consequence, the characterization of the buffer is interesting, especially when multimedia flows are transmitted and even more if they transport information with real-time requirements. This work presents a new methodology with the aim of determining the technical and functional characteristics of real buffers (i.e., behaviour, size, limits, and input and output rate) of a network path. It permits the characterization of intermediate buffers of different devices in a network path across the Internet.