Seguimiento a largo plazo de pacientes traqueostomizados post injuria cerebral adquirida grave / Long-term follow-up of tracheostomized patients post severe acquired brain injury

Resumen Introducción: Alrededor del 50% de los pacientes hos pitalizados por injuria cerebral adquirida grave requie ren traqueostomía y cuidados a largo plazo. El objetivo principal de este estudio fue describir la evolución de enfermos con injuria cerebral adquirida grave (ICAg) traqueostomizados que ingresaron a rehabilitación. Se cundariamente se estudió el fracaso de la decanulación y la supervivencia a los 12 meses del alta. Métodos: estudio cuantitativo observacional prospec tivo de centro único. Se incorporó al estudio, de forma prospectiva y consecutiva, usuarios mayores de 18 años, traqueostomizados posterior a ICAg ingresados a un cen tro de rehabilitación entre abril de 2018 y marzo de 2020. Resultados: se incluyeron para el análisis 50 pacien tes. La estancia en el centro fue de 203 (RIQ 93-320) días. Al alta de la institución, 32 (64%) pacientes pudieron ser decanulados exitosamente. El tiempo transcurrido desde el ingreso al centro hasta la decanulación fue de 49 (12-172) días. No se observó fracaso de la decanula ción. La mortalidad a los 12 meses de seguimiento fue de 32%, cinco (16%) de los 32 pacientes que pudieron ser decanulados y 11 (61%) de los 18 que no lograron la decanulación fallecieron dentro de los 12 meses de seguimiento. La relación entre la decanulación y la mortalidad a los 12 meses de seguimiento resultó esta dísticamente significativa (p = 0.002). Discusión: La supervivencia global fue relativamente elevada, el proceso de decanulación resulta relevante ya que puede tener impacto en la supervivencia a largo plazo.

Abstract Introduction: About 50% of patients hospitalized for severe acquired brain injury require tracheostomy, and many of them need long-term care. The main objective of this study was to describe the evolution of patients with severe acquired brain injury (sABI) tracheotomized who entered rehabilitation. Secondarily, mortality re lated to the success or failure of decannulation and survival at 12 months of discharge were studied. Methods: A single-center prospective observational quantitative study. Users over 18 years of age were recruited prospectively and consecutively, tracheosto mized after sABI, and admitted to a rehabilitation center between April 2018 and March 2020. Results: Fifty patients were included for analysis. The stay in the center was 203 (RIQ 93-320) days. At discharge to the institution, 32 (64%) patients managed to be successfully decannulated. The median number of days from admission to the center to decannulation was 49 (12-172). No decannulation failure was observed. Mortality at 12 months follow-up was 32%, five (16%) of the 32 patients who managed to be decannulated, and 11 (61%) of 18 who failed to achieve decannulation died within 12 months of follow-up. The relationship between decannulation success and mortality at 12 months of follow-up was statistically significant (p= 0.002). Discussion: Addressing the decannulation process early and properly guided is relevant as it may impact long-term survival.

[Long-term follow-up of tracheostomized patients post severe acquired brain injury]. / Seguimiento a largo plazo de pacientes traqueostomizados post injuria cerebral adquirida grave.

INTRODUCTION: About 50% of patients hospitalized for severe acquired brain injury require tracheostomy, and many of them need long-term care. The main objective of this study was to describe the evolution of patients with severe acquired brain injury (sABI) tracheotomized who entered rehabilitation. Secondarily, mortality related to the success or failure of decannulation and survival at 12 months of discharge were studied. METHODS: A single-center prospective observational quantitative study. Users over 18 years of age were recruited prospectively and consecutively, tracheostomized after sABI, and admitted to a rehabilitation center between April 2018 and March 2020. RESULTS: Fifty patients were included for analysis. The stay in the center was 203 (RIQ 93-320) days. At discharge to the institution, 32 (64%) patients managed to be successfully decannulated. The median number of days from admission to the center to decannulation was 49 (12-172). No decannulation failure was observed. Mortality at 12 months follow-up was 32%, five (16%) of the 32 patients who managed to be decannulated, and 11 (61%) of 18 who failed to achieve decannulation died within 12 months of follow-up. The relationship between decannulation success and mortality at 12 months of follow-up was statistically significant (p= 0.002). DISCUSSION: Addressing the decannulation process early and properly guided is relevant as it may impact long-term survival.

Introducción: Alrededor del 50% de los pacientes hospitalizados por injuria cerebral adquirida grave requieren traqueostomía y cuidados a largo plazo. El objetivo principal de este estudio fue describir la evolución de enfermos con injuria cerebral adquirida grave (ICAg) traqueostomizados que ingresaron a rehabilitación. Secundariamente se estudió el fracaso de la decanulación y la supervivencia a los 12 meses del alta. Métodos: estudio cuantitativo observacional prospectivo de centro único. Se incorporó al estudio, de forma prospectiva y consecutiva, usuarios mayores de 18 años, traqueostomizados posterior a ICAg ingresados a un centro de rehabilitación entre abril de 2018 y marzo de 2020. Resultados: se incluyeron para el análisis 50 pacientes. La estancia en el centro fue de 203 (RIQ 93-320) días. Al alta de la institución, 32 (64%) pacientes pudieron ser decanulados exitosamente. El tiempo transcurrido desde el ingreso al centro hasta la decanulación fue de 49 (12-172) días. No se observó fracaso de la decanulación. La mortalidad a los 12 meses de seguimiento fue de 32%, cinco (16%) de los 32 pacientes que pudieron ser decanulados y 11 (61%) de los 18 que no lograron la decanulación fallecieron dentro de los 12 meses de seguimiento. La relación entre la decanulación y la mortalidad a los 12 meses de seguimiento resultó estadísticamente significativa (p = 0.002). Discusión: La supervivencia global fue relativamente elevada, el proceso de decanulación resulta relevante ya que puede tener impacto en la supervivencia a largo plazo.

Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM.

BACKGROUND: Leishmaniasis is a vector-borne disease caused by a parasite protozoon from the genus Leishmania. Among the molecular techniques applied for detecting these parasites, real-time PCR with High Resolution Melting (PCR-HRM) proved advantageous since it simultaneously determines both the presence and species of the pathogen in one step, through amplification and later analysis of curves generated by melting temperature. METHODS: Based on this molecular technique, the goal of this study was to estimate the PCR-HRM sensitivity for Leishmania spp. detection in different canine tissues by evaluating biological samples obtained from popliteal, submandibular, and pre-scapular lymph nodes, from bone marrow and ear pinnae of 28 stray dogs captured in the metropolitan area of Asunción (Paraguay). RESULTS: The rk39 immunochromatographic test showed that 25/28 tested dogs (89%) presented antibodies against L. infantum. In 20/25 dogs that tested positive for rk39 (80%), it was possible to detect Leishmania spp. by PCR-HRM and determine that the species corresponded entirely to L. infantum. Regarding the analysis of different tissues, the parasite was detected in all popliteal lymph node samples, followed by high detection in submandibular (at 95%) and pre-scapular lymph nodes (at 90%), bone marrow (at 85%), and ear pinnae (at 85%). CONCLUSIONS: This study demonstrated that the use of real-time PCR-HRM using the molecular marker hsp70 was a highly sensitive method for simultaneously detecting and identifying Leishmania species in different tissues taken from infected dogs. In addition, the usefulness of ear pinnae as easily accessible tissue for molecular diagnosis was emphasized.

[Comparison between the first and second COVID-19 wave of patients who required invasive mechanical ventilation in an intensive care unit in Argentina]. / Comparación entre la primera y segunda ola de pacientes con COVID-19 que requirieron ventilación mecánica invasiva en una unidad de cuidados intensivos de Argentina.

INTRODUCTION: the information regarding characteristics and ventilatory results comparing the first (W1) and the second wave (W2) in Argentina are limited. The main objective of this study was to describe general characteristics and ventilatory variables in COVID-19 patients who required invasive mechanical ventilation (IMV) and compare differences between waves. Secondarily, factors associated with mortality in intensive care unit (ICU) were studied. METHODS: We conducted a prospective observational cohort study that included patients older than 18 years infected with SARS-CoV-2 consecutively admitted to ICU with IMV between August 1, 2020, and June 30, 2021. We included 412 patients. RESULTS: We found statistically significant differences (p < 0.001) in age [W1 64(55-72) vs W2 59 (50-66) years], presence of COPD [W1 n = 42 (19.8%) vs. W2 n = 13(6.3%)], plateau pressure [W1 27(25-30) cm H2O vsW2 24 (22-27) cmH2O], driving pressure (ΔP) [W1 15 (13-17) cmH2O vs. W2 12 (11-14) cm H2O] compliance [W1 40 mL/cmH2O (32-46) vs. W2 = 33 mL/cm H2O (27-40)]; reintubation [W1 30.4% (n = 63/207) vs. W2 13.7% (n = 28/205)]. We identified as independent factors associated with mortality the following variables: age [OR 1.07(95% CI 1.05-1.09)], the ΔP in the first 24 hours [OR 1.19(95% CI 1.10-1.28)] and W2 [OR 1.81 (95% IC1.12-2.93); p = 0.015. DISCUSSION: During W2 the patients were younger. It was possible to achieve ventilatory mechanics more adjusted to a protective ventilation strategy. In conclusion, in the patients studied, age and ΔP were independent predictors of mortality.

Introducción: la información sobre características y resultados ventilatorios que comparen la primera (O1) y segunda (O2) ola de COVID-19 en Argentina son limitados. El objetivo principal de este estudio fue describir las características generales y variables ventilatorias en pacientes COVID-19 que requirieron ventilación mecánica invasiva (VMI) y comparar las diferencias entre olas. Secundariamente se estudiaron factores asociados con mortalidad en la unidad de cuidados intensivos (UCI). Métodos: Realizamos un estudio observacional analítico de cohorte prospectiva que incluyó pacientes mayores de 18 años infectados por SARS-CoV-2 ingresados de forma consecutiva a la unidad de cuidados intensivos, con requerimiento de VMI, entre el 1 de agosto de 2020 y el 30 de junio de 2021. Se incluyeron 412 pacientes. Resultados: Encontramos diferencias estadísticamente significativas (p < 0.001) en: la edad [O1 64 (55-72) años versus O2 59 (50-66) años]; presencia de EPOC [O1 n = 41 (19.8%) versus O2 n = 13 (6.3%)]; Presión meseta [O1 27 (25-30) cmH2O versus O224 (22-27) cmH2O],presión de conducción (ΔP) [O1 15 (13-17) cmH2O versus O2 12 (11-14) cmH2O]; complacencia [O1 40 mL/cmH2O (32-46) versus O2 33 mL/cmH2O (27-40)]; reintubación [O1 30.4% (n=63/207) versus O2 13.7% (n=28/205)]. Se identificaron como factores independientes asociados a mortalidad las siguientes variables: edad [(OR 1.07 (IC 95% 1.05-1.09)], el ΔP en las primeras 24 h [(OR 1.19 (IC 95% 1.10-1.28)] y O2 [OR 1.81 (IC 95% 1.12-2.93); p = 0.015]. Discusión: Durante O2, los pacientes eran más jóvenes. Fue posible lograr una mecánica ventilatoria más ajustada a una estrategia de ventilación protectora. Como conclusión, en los pacientes estudiados la edad y ΔP fueron predictores independientes de mortalidad.

Evaluation of the In Vitro Antiparasitic Effect of the Essential Oil of Cymbopogon winterianus and Its Chemical Composition Analysis.

Cymbopogon winterianus, known as "citronella grass", is an important aromatic and medicinal tropical herbaceous plant. The essential oil of C. winterianus (EOCw) is popularly used to play an important role in improving human health due to its potential as a bioactive component. The present study aimed to identify the components of the essential oil of C. winterianus and verify its leishmanicidal and trypanocidal potential, as well as the cytotoxicity in mammalian cells, in vitro. The EOCw had geraniol (42.13%), citronellal (17.31%), and citronellol (16.91%) as major constituents. The essential oil only exhibited significant cytotoxicity in mammalian fibroblasts at concentrations greater than 250 µg/mL, while regarding antipromastigote and antiepimastigote activities, they presented values considered clinically relevant, since both had LC50 < 62.5 µg/mL. It can be concluded that this is a pioneer study on the potential of the essential oil of C. winterianus and its use against the parasites T. cruzi and L. brasiliensis, and its importance is also based on this fact. Additionally, according to the results, C. winterianus was effective in presenting values of clinical relevance and low toxicity and, therefore, an indicator of popular use.

Comparación entre la primera y segunda ola de pacientes con COVID-19 que requirieron ventilación mecánica invasiva en una unidad de cuidados intensivos de Argentina / Comparison between the first and second COVID-19 wave of patients who required invasive mechanical ventilation in an intensive care unit in Argentina

Resumen Introducción: la información sobre características y resultados ventilatorios que comparen la primera (O1) y segunda (O2) ola de COVID-19 en Argentina son limitados. El objetivo principal de este estudio fue describir las características generales y variables ventilatorias en pacientes COVID-19 que requirieron ventilación mecánica invasiva (VMI) y comparar las diferencias entre olas. Secundariamente se es tudiaron factores asociados con mortalidad en la unidad de cuidados intensivos (UCI). Métodos: Realizamos un estudio observacional analítico de cohorte prospectiva que incluyó pacientes mayores de 18 años infectados por SARS-CoV-2 ingresados de forma consecutiva a la unidad de cuidados intensivos, con requerimiento de VMI, entre el 1 de agosto de 2020 y el 30 de junio de 2021. Se incluyeron 412 pacientes. Resultados: Encontramos diferencias estadísticamente significativas (p < 0.001) en: la edad [O1 64 (55-72) años versus O2 59 (50-66) años]; presencia de EPOC [O1 n = 41 (19.8%) versus O2 n = 13 (6.3%)]; Presión meseta [O1 27 (25-30) cmH2O versus O224 (22-27) cmH2O],presión de conducción (ΔP) [O1 15 (13-17) cmH2O versus O2 12 (11-14) cmH2O]; complacencia [O1 40 mL/cmH2O (32-46) versus O2 33 mL/cmH2O (27-40)]; reintubación [O1 30.4% (n=63/207) versus O2 13.7% (n=28/205)]. Se identificaron como factores independientes asociados a mortalidad las siguientes variables: edad [(OR 1.07 (IC 95% 1.05-1.09)], el ΔP en las primeras 24 h [(OR 1.19 (IC 95% 1.10-1.28)] y O2 [OR 1.81 (IC 95% 1.12-2.93); p = 0.015]. Discusión: Durante O2, los pacientes eran más jóvenes. Fue posible lograr una mecánica ventilatoria más ajustada a una estrategia de ventilación protectora. Como conclusión, en los pacientes estudiados la edad y ΔP fueron predictores independientes de mortalidad.

Abstract Introduction: the information regarding characteristics and ventilatory results comparing the first (W1) and the second wave (W2) in Argentina are limited. The main objective of this study was to describe general charac teristics and ventilatory variables in COVID-19 patients who required invasive mechanical ventilation (IMV) and compare differences between waves. Secondarily, factors associated with mortality in intensive care unit (ICU) were studied. Methods: We conducted a prospective observational cohort study that included patients older than 18 years infected with SARS-CoV-2 consecutively admitted to ICU with IMV between August 1, 2020, and June 30, 2021. We included 412 patients. Results: We found statistically significant differences (p < 0.001) in age [W1 64(55-72) vs W2 59 (50-66) years], presence of COPD [W1 n = 42 (19.8%) vs. W2 n = 13(6.3%)], plateau pressure [W1 27(25-30) cm H2O vsW2 24 (22-27) cmH2O], driving pressure (ΔP) [W1 15 (13-17) cmH2O vs. W2 12 (11-14) cm H2O] compliance [W1 40 mL/cmH2O (32-46) vs. W2 = 33 mL/cm H2O (27-40)]; reintubation [W1 30.4% (n = 63/207) vs. W2 13.7% (n = 28/205)]. We identified as independent factors associated with mortality the following variables: age [OR 1.07(95% CI 1.05-1.09)], the ΔP in the first 24 hours [OR 1.19(95% CI 1.10- 1.28)] and W2 [OR 1.81 (95% IC1.12-2.93); p = 0.015. Discussion: During W2 the patients were younger. It was possible to achieve ventilatory mechanics more adjusted to a protective ventilation strategy. In conclusion, in the patients studied, age and ΔP were independent predictors of mortality.

A Potential New Source of Therapeutic Agents for the Treatment of Mucocutaneous Leishmaniasis: The Essential Oil of Rhaphiodon echinus.

Weeds are an important source of natural products; with promising biological activity. This study investigated the anti-kinetoplastida potential (in vitro) to evaluate the cytotoxicity (in vitro) and antioxidant capacity of the essential oil of Rhaphiodon echinus (EORe), which is an infesting plant species. The essential oil was analyzed by GC/MS. The antioxidant capacity was evaluated by reduction of the DPPH radical and Fe3+ ion. The clone Trypanosoma cruzi CL-B5 was used to search for anti-epimastigote activity. Antileishmanial activity was determined using promastigotes of Leishmania braziliensis (MHOM/CW/88/UA301). NCTC 929 fibroblasts were used for the cytotoxicity test. The results showed that the main constituent of the essential oil was γ-elemene. No relevant effect was observed concerning the ability to reduce the DPPH radical; only at the concentration of 480 µg/mL did the essential oil demonstrate a high reduction of Fe3+ power. The oil was active against L. brasiliensis promastigotes; but not against the epimastigote form of T. cruzi. Cytotoxicity for mammalian cells was low at the active concentration capable of killing more than 70% of promastigote forms. The results revealed that the essential oil of R. echinus showed activity against L. brasiliensis; positioning itself as a promising agent for antileishmanial therapies.

Leishmanicidal and trypanocidal potential of the essential oil of Psidium myrsinites DC.

Neglected tropical diseases (NTDs) are a group of pathologies caused by infectious agents or parasites, including the protozoa Leishmania sp. and Trypanosoma cruzi, which cause leishmaniasis and Chagas disease, respectively. The complications of the treatment regimen indicate the urgent need to search for new strategies and therapeutic agents. Among these is the essential oil of Psidium myrsinites DC. The essential oil of the leaves (PMEO) was evaluated in vitro, and cytotoxic activity was analysed against promastigotes of Leishmania braziliensis and Leishmania infantum and epimastigotes of Trypanosoma cruzi, as well as mammalian cells. The results showed that the PMEO had relevant activity against L. braziliensis, low cytotoxicity and a high selectivity index SI = 6.6. These results suggest that PMEO has antiparasitic potential against L. braziliensis, making this species is a possible alternative therapeutic source, given its effectiveness in the in vitro tests performed, opening the possibility of new biological studies in vivo.

Sensitivity comparison for the Leishmania spp. detection in different canine tissues using PCR-HRM

ABSTRACT Background: Leishmaniasis is a vector-borne disease caused by a parasite protozoon from the genus Leishmania. Among the molecular techniques applied for detecting these parasites, real-time PCR with High Resolution Melting (PCR-HRM) proved advantageous since it simultaneously determines both the presence and species of the pathogen in one step, through amplification and later analysis of curves generated by melting temperature. Methods: Based on this molecular technique, the goal of this study was to estimate the PCR-HRM sensitivity for Leishmania spp. detection in different canine tissues by evaluating biological samples obtained from popliteal, submandibular, and pre-scapular lymph nodes, from bone marrow and ear pinnae of 28 stray dogs captured in the metropolitan area of Asunción (Paraguay). Results: The rk39 immunochromatographic test showed that 25/28 tested dogs (89%) presented antibodies against L. infantum. In 20/25 dogs that tested positive for rk39 (80%), it was possible to detect Leishmania spp. by PCR-HRM and determine that the species corresponded entirely to L. infantum. Regarding the analysis of different tissues, the parasite was detected in all popliteal lymph node samples, followed by high detection in submandibular (at 95%) and pre-scapular lymph nodes (at 90%), bone marrow (at 85%), and ear pinnae (at 85%). Conclusions: This study demonstrated that the use of real-time PCR-HRM using the molecular marker hsp70 was a highly sensitive method for simultaneously detecting and identifying Leishmania species in different tissues taken from infected dogs. In addition, the usefulness of ear pinnae as easily accessible tissue for molecular diagnosis was emphasized.

Chemical Constituents and Biological Activities of Croton heliotropiifolius Kunth.

Croton heliotropiifolius Kunth (Euphorbiaceae), whose occurrence has already been registered in the most varied Brazilian biomes, is commonly found in the Chapada do Araripe, Ceará. The species is traditionally used to treat fungal, parasitic, and degenerative diseases. This study investigated the chemical composition and pharmacological potential (antioxidant, antifungal, antiparasitic, and cytotoxic) of an aqueous extract obtained from the roots of C. heliotropiifolius. Following a qualitative phytochemical screening, the chemical constituents were identified by ultra-efficiency liquid chromatography coupled witha quadrupole/time-of-flight system (UPLC-QTOF). The antioxidant potential was verified by thin-layer chromatography (TLC). The direct and combined antifungal activity of the extract against opportunistic Candida strains was investigated using the microdilution method. The minimal fungicidal concentration (MFC) was determined by subculture, while the modulation of the morphological transition (fungal virulence) was evaluated by light microscopy. The in vitro antiparasitic activity was analyzed using epimastigotes of Trypanosoma cruzi and promastigotes of Leishmania braziliensis and Leishmania infantum, while cytotoxicity was determined in cultures of mouse fibroblasts. The phytochemical analysis identified the presence of acids, terpenes, flavonoids, lignans, and alkaloids. Among these constituents, the presence of polar and non-polar phenolic compounds with known antioxidant action was highlighted. While the extract showed clinically ineffective antifungal effects, it could enhance the effectiveness of fluconazole, in addition to inhibiting the morphological transition associated with increased virulence in Candida strains. Although the extract showed low cytotoxicity against fibroblasts, it also had weak antiparasitic effects. In conclusion, Croton heliotropiifolius is a source of natural products with antifungal and antioxidant potential.

Triazole-Resistance in Environmental Aspergillus fumigatus in Latin American and African Countries.

Triazole-resistance has been reported increasingly in Aspergillus fumigatus. An international expert team proposed to avoid triazole monotherapy for the initial treatment of invasive aspergillosis in regions with >10% environmental-resistance, but this prevalence is largely unknown for most American and African countries. Here, we screened 584 environmental samples (soil) from urban and rural locations in Mexico, Paraguay, and Peru in Latin America and Benin and Nigeria in Africa for triazole-resistant A. fumigatus. Samples were screened using triazole-containing agars and confirmed as triazole-resistant by the European Committee on Antimicrobial Susceptibility Testing (EUCAST) broth dilution reference method. Isolates were further characterized by cyp51A sequencing and short-tandem repeat typing. Fungicide presence in samples was likewise determined. Among A. fumigatus positive samples, triazole-resistance was detected in 6.9% (7/102) of samples in Mexico, 8.3% (3/36) in Paraguay, 9.8% (6/61) in Peru, 2.2% (1/46) in Nigeria, and none in Benin. Cyp51A gene mutations were present in most of the triazole-resistant isolates (88%; 15/17). The environmentally-associated mutations TR34/L98H and TR46/Y121F/T289A were prevalent in Mexico and Peru, and isolates harboring these mutations were closely related. For the first time, triazole-resistant A. fumigatus was found in environmental samples in Mexico, Paraguay, Peru, and Nigeria with a prevalence of 7-10% in the Latin American countries. Our findings emphasize the need to establish triazole-resistance surveillance programs in these countries.

Chemical composition, antioxidant and antiprotozoal activity of Eugenia gracillima Kiaersk. leaves essential oil.

This work evaluated the volatile composition, antioxidant and antiprotozoal activities of the essential oil obtained from leaves of Eugenia gracillima Kiaersk. (EGEO) grown in Brazilian Northeast area (Araripe, Brazil). The volatile compounds of EGEO were analyzed by GC and GC-MS and its chemical composition is mainly composed of sesquiterpene hydrocarbons (91.22%), oxygenated sesquiterpenes (7.45%) and monoterpene (1.01%). The most abundant volatile constituents of the EGEO were germacrene D (16.10%), γ-muurolene (15.60%), bicyclogermacrene (8.53%), germacrene B (7.43%), and Δ-elemene (6.06%). The oil showed weak to moderate antioxidant activity. EGEO was highly selective to Leishmania braziliensis and Leishmania infantum promastigotes with selective indexes of 73.66 and 71.41, respectively. EGEO did not inhibit Trypanosoma cruzi. These data suggest that the E. gracillima essential oil is a relevant source of lead compounds for development of anti-Leishmania drugs.

Isolation, Biochemical Characterization and Antiparasitic Activity of BmatTX-IV, A Basic Lys49-Phospholipase A2 from the Venom of Bothrops mattogrossensis from Paraguay.

BACKGROUND: Functional and structural diversity of proteins of snake venoms is coupled with a wide repertoire of pharmacological effects. Snake venoms are targets of studies linked to searching molecules with biotechnological potential. METHODS: A homologue phospholipase A2 (BmatTX-IV) was obtained using two chromatographic techniques. Mass spectrometry and two-dimensional gel electrophoresis were used to determine the molecular mass and isoelectric point, respectively. By means of Edman degradation chemistry, it was possible to obtain the partial sequence of amino acids that comprise the isolated toxin. Trypanocidal, leishmanicidal and cytoxic activity against Trypanosoma cruzi, Leishmania infantum and murine fibrobasts was determinated. RESULTS: Combination of both chromatographic steps used in this study demonstrated efficacy to obtain the PLA2-Lys49. BmatTX-IV showed molecular mass and isoelectric point of 13.55 kDa and 9.3, respectively. Amino acid sequence of N-terminal region (51 residues) shows the presence of Lys49 residue at position 49, a distinctive trait of enzymatically inactive PLA2. Bothrops mattogrossensis snake venom showed IC50 values of 11.9 µg/mL against Leishmania infantum promastigotes and of 13.8 µg/mL against Trypanosoma cruzi epimastigotes, respectively. On the other hand, the venom showed a high cytotoxic activity (IC50 value of 16.7 µg/mL) against murine fibroblasts, whereas the BmatTX-IV showed IC50 value of 81.2 µg/mL. CONCLUSION: Physicochemical and biological characterization of snake venoms components is critically important, since these complex mixtures provide a source of molecules with antiparasitic potential, making further studies necessary to identify and characterize components with higher efficacy and selectivity.

Antiprotozoal action of synthetic cinnamic acid analogs.

INTRODUCTION: Leishmaniasis, Chagas disease, and malaria cause morbidity globally. The drugs currently used for treatment have limitations. Activity of cinnamic acid analogs against Leishmania spp., Trypanosoma cruzi, and Plasmodium falciparum was evaluated in the interest of identifying new antiprotozoal compounds. METHODS: In vitro effects of analogs against L. braziliensis, L. infantum chagasi, T. cruzi, and P. falciparum, and hemolytic and cytotoxic activities on NCTC 929 were determined. RESULTS: Three analogs showed leishmanicidal and tripanocidal activity. No antiplasmodial, hemolytic, or cytotoxic activity was observed. CONCLUSIONS: Antiprotozoal activity of analogs against L. infantum braziliensis, L. infantum chagasi, and T. cruzi was demonstrated.

Antiprotozoal action of synthetic cinnamic acid analogs

Abstract INTRODUCTION Leishmaniasis, Chagas disease, and malaria cause morbidity globally. The drugs currently used for treatment have limitations. Activity of cinnamic acid analogs against Leishmania spp., Trypanosoma cruzi, and Plasmodium falciparum was evaluated in the interest of identifying new antiprotozoal compounds. METHODS In vitro effects of analogs against L. braziliensis, L. infantum chagasi, T. cruzi, and P. falciparum, and hemolytic and cytotoxic activities on NCTC 929 were determined. RESULTS Three analogs showed leishmanicidal and tripanocidal activity. No antiplasmodial, hemolytic, or cytotoxic activity was observed. CONCLUSIONS Antiprotozoal activity of analogs against L. infantum braziliensis, L. infantum chagasi, and T. cruzi was demonstrated.

Antiparasitic Activity and Essential Oil Chemical Analysis of the Piper Tuberculatum Jacq Fruit.

With the increase of neglected diseases such as leishmaniasis and Chagas disease, there was a need for the search for new therapeutic alternatives that reduce the harm caused by medicine available for treatment. Thus, this study was performed to investigate the antiparasitic activity of the essential oil from the fruits of Piper tuberculatum Jacq, against lines of Leishmania braziliensis (MHOM/CO/88/UA301), Leishmania infantum (MHOM/ES/92/BCN83) and Trypanosoma cruzi (LC-B5 clone). Before running protocols, an analysis of the chemical composition of essential oil was conducted, which presented monoterpenes and sesquiterpenes. As major constituents, ß-pinene and α-pinene were identified. Regarding to antiparasitic activity, the essential oil had an EC50 values of 133.97 µg/mL and 143.59 µg/mL against variations promastigotes of L. infantum and L. braziliensis, respectively. As for trypanocidal activity, the oil showed EC50 value of 140.31 µg/mL against epimastigote form of T. cruzi. Moreover, it showed moderate cytotoxicity in fibroblasts with LC50 value of 204.71 µg/mL. The observed effect may be related to the presence of terpenes contained in the essential oil, since it has its antiparasitic activity proven in the literature.

Anti-Trypanosoma, anti-Leishmania and cytotoxic activities of natural products from Psidium brownianum Mart. ex DC. and Psidium guajava var. Pomifera analysed by LC-MS.

Neglected diseases are those that are prevalent in developing countries, even with a rich biodiversity. These diseases still persist because of the lack of scientific studies, government negligence or failures of the public health system. This study aims to identify the composition of extracts and fractions from Psidium brownianum and Psidium guajava through LC-MS, to evaluate its in vitro anti-parasitic and cytotoxic activity against Trypanosoma cruzi, Leishmania brasiliensis and L. infantum epismastigote and promastigote forms, as well as mammalian cells. The results showed the presence of chemical constituents in the two Psidium species as quercetin, myricetin and gallic acid derivatives. The P. brownianum extract and fractions showed low toxicity at all tested concentrations and all samples were effective at the concentration of 1000µg/mL against the parasites, with the extract being the most efficient against the L. infantum promastigote form. The ethanolic extract, and the flavonoid and tannic fractions, from P. guajava showed low toxicity for the fibroblasts. All samples showed effectiveness at the highest concentration tested and the extract was more effective against the promastigote forms tested. The results showed that the species Psidium brownianum and Psidium guajava demonstrated an anti-parasitic activity against the T. cruzi, L. brasiliensis and L. infantum parasite cell lines indicating these species as an alternative therapy given their efficacy in the in vitro assays performed, opening the possibility for new biological studies to further this knowledge through in vivo assays.

Morphometric Wings Similarity among Sylvatic and Domestic Populations of Triatoma infestans (Hemiptera: Reduviidae) from the Gran Chaco Region of Paraguay.

Despite sustained efforts for eliminating Triatoma infestans, reinfestation still persists in large part of the endemic area of Chagas disease from the Gran Chaco region. Sylvatic T. infestans populations seem to threat success of control programs of domestic T. infestans. In this study, we analyze whether T. infestans collected after a community-wide spraying were survivors or were immigrants from elsewhere using geometric morphometric tools. We used 101 right wings of female T. infestans captured before and after intervention program carried out in 12 de Junio and Casuarina, villages from Paraguayan Chaco, and in Puerto Casado during presprayed collection. There were no significant differences in wing size of domestic T. infestans between pre- and postspraying populations, and between domestic and sylvatic ones. When shape variables originating from postintervention individuals from 12 de Junio were introduced one by one into a discriminant analysis, the greatest weight (53%) was allocated to the sylvatic group. Furthermore, from the prespraying population, 25% were reallocated as postintervention individuals. Only 11% of the insects were reassigned to other groups Puerto Casado and Casuarina. These results suggest that postspraying individuals appear to have different origins. Half of the postspraying individuals from 12 de Junio were similar to the sylvatic ones and 25% of these were similar to those captured in the prespraying period. This remarkable morphometric wings similarity between sylvatic and domestic populations is new evidence suggesting that they could be highly related to each other in the Paraguayan Chaco; human-fed bugs from sylvatic area also support this.

Safety assessment and antioxidant activity of Lantana montevidensis leaves: Contribution to its phytochemical and pharmacological activity.

Lantana camara, the widely studied species, and L. montevidensis, the less studied species of the genus Lantana are both used in traditional medicine for the same purpose (anti-asthma, anti-ulcer, anti-tumor, etc). However, little is known about the toxicity of L. montevidensis and there is limited information on its chemical constituents. Here, we investigated for the first time the genotoxicity and cytotoxicity of the ethanolic (EtOH) and aqueous extracts from the leaves of Lantana montevidensis in human leukocytes, as well as their possible interaction with human erythrocyte membranes in vitro. The antioxidant activities of both extracts were also investigated in chemical and biological models. Treatment of leukocytes with EtOH or aqueous extracts (1-480 µg/mL) did not affect DNA damage index, but promoted cytotoxicity at higher concentrations (240-480 µg/mL). Both extracts did not modify the osmotic fragility of human erythrocytes. The extracts scavenged DPPH radical and prevented Fe2+-induced lipid peroxidation in rat's brain and liver homogenates, and this was likely not attributed to Fe (II) chelation. The HPLC analysis of the extracts showed different amounts of polyphenolic compounds (isoquercitrin, gallic acid, catechin, ellagic acid, apigenin, kaempferol, caffeic acid, rutin, quercitrin, quercetin, chlorogenic acid, luteolin) that may have contributed to these effects. These results supported information on the functional use of L. montevidensis in folk medicine.

A citotoxicidade e a avaliação da atividade antiprotozoária de Melissa officcinalis L (CIDRO-MELISA) / The cytotoxicity activity and evaluation of antiprotozoa Melissa officcinalis L (CIDRO-MELISA)

Introdução: as Leishmanioses correspondem a um complexo de doenças tropicais causada por mais de vinte diferentes espécies de protozoários intracelulares pertencentes ao gênero Leishmania. A Doença de Chagas é uma doença que representa um dos principais problemas de saúde pública em vários países, afetando sobretudo a população carente e rural. Melissa officinalis L., é uma erva perene, uma planta exótica, cuja família é Lamiaceae, que tem sido amplamente utilizada na medicina popular com finalidades terapêuticas. O seu óleo essencial tem sido usado como antioxidante, antimicótica, antifúngica, sedativa e antivirótica. Objetivo: avaliar a citotoxicidade, composição química e atividade antiparasitária do óleo essencial da espécie M. officinalis. Métodos: M. officcinalis foi coletada no município do Crato, CE, Brasil, e foi depositada no herbário da Universidade Regional do Cariri URCA. A análise da composição química do óleo essencial de M. officcinalis, foi realizada através de Cromatografia Gasosa acoplada à Espectrometria de Massas. No estudo da atividade leishmanicida in vitro, utilizou-se formas promastigotas de L. braziliensis e epimastigotas de Trypanosoma cruzi. Para a atividade citotóxica utilizou-se a linhagem de fibroblastos de mamífero NCTC clone 929. Resultados: a través da análise do óleo essencial de M. officcinalis feita por CG/EM, foi possível identificar 12 constituintes que totalizavam 99,83 por cento, dentre os quais alguns foram majoritários: Geranial, Z Citral, 1- Limonene e Carvone. Foi verificado baixa toxicidade contra os fibroblastos. M. officcinalis exibiu maior capacidade para impedir a replicação contra a forma promastigota do protozoário, nas concentrações testadas. Conclusão: a espécie pode ser uma fonte importante na busca de agentes novos e seletivos para o tratamento de doenças tropicais causadas por protozoários, cujas propriedades necessitam ser investigadas quanto aso seus mecanismos(AU)

Introducción: Leishmaniasis corresponde a un complejo de enfermedades tropicales causadas por más de veinte especies diferentes de protozoos intracelulares que pertenece al género Leishmania. La enfermedad de Chagas es una enfermedad que es uno de los principales problemas de salud pública en muchos países, que afectan especialmente a la población pobre y rural. Melissa officinalis L., es una hierba perenne, una planta exótica, cuya familia Lamiaceae, ha sido ampliamente utilizado en la medicina popular para fines terapéuticos. Su aceite esencial se ha usado como un antioxidante, antimicótico, antifúngica, antiviral y sedante. Objetivo: evaluar la citotoxicidad, la composición química y la actividad antiparasitaria del aceite esencial de la especie M. officinalis. Métodos: M. officinalis se recogió en el municipio de Crato, CE, Brasil, y fue depositado en el herbario de la Universidad Regional del Cariri URCA. El análisis de la composición química del aceite esencial de M. officcinalis se realizó por espectrometría de masa de cromatografía de gas. En el estudio de la actividad invitro leishmanicida se utilizó promastigotes L. braziliensis y Trypanosoma cruzi. Para la actividad citotóxica se utilizó la cepa de fibroblastos de mamífero NCTC clon 929. Resultados: a través de análisis de aceite esencial de M. officinalis hizo por GC/ EM se identificó un total de 12 constituyentes 99,83 por ciento, entre los cuales algunos eran mayoría: geranial, Z citral, limoneno y carvona 1. Se comprobó una baja toxicidad contra fibroblastos. M. officcinalis exhibió mayor capacidad para evitar la replicación contra la forma promastigote de parásito, a las concentraciones ensayadas. Conclusión: la especie puede ser una fuente importante en la búsqueda de nuevos agentes y selectivos para el tratamiento de enfermedades tropicales causadas por protozoos, cuyas propiedades deben ser investigados con respecto a sus mecanismos(AU)

Introduction: Leishmaniasis is a complex of tropical diseases caused by more than twenty different species of intracellular protozoa from the genus Leishmania. Chagas disease is one of the main public health problems in many countries. It mostly affects the poor, rural population. Melissa officinalis L. is a perennial herb, an exotic plant from the family Lamiaceae which has been widely used in folk medicine for therapeutic purposes. Its essential oil has been used as antioxidant, antifungal, antiviral and sedative. Objective: Evaluate the cytotoxicity, chemical composition and antiparasitic activity of essential oil of the species M. officinalis. Methods: M. officinalis specimens were collected from the municipality of Crato, CE, Brazil, and deposited in the herbarium at the Regional University of Cariri (URCA). Analysis of the chemical composition of essential oil of M. officinalis was performed by gas chromatography / mass spectrometry. L. braziliensis and Trypanosoma cruzi promastigotes were used for the study of in vitro leishmanicidal activity. Mammalian fibroblast strain NCTC clone 929 was used to evaluate cytotoxic activity. Results: Analysis of essential oil from M. officinalis by GC / MS led to identification of a total 12 constituents 99.83 percent, among which some were a majority: geranial, Z citral, limonene and 1-carvone. Toxicity against fibroblasts was found to be low. M. officinalis displayed greater capacity to avoid replication against the promastigote form of the parasite at the concentrations assayed. Conclusion: The species may be an important source of new agents and selective media for the treatment of tropical diseases caused by protozoa, whose properties should be studied with respect to their mechanisms(AU)