Divergent environmental preferences and areas of sympatry of tick species in the Amblyomma cajennense complex (Ixodidae)

Four species of Neotropical ticks, Amblyomma mixtum, Amblyomma cajennense, Amblyomma tonelliae and Amblyomma sculptum (formerly included in the catch-all name A. cajennense), have an allopatric distribution in much of their range, with areas of parapatry for at least two of them. We inferred the abiotic niches of these organisms using coefficients of a harmonic regression of the temperature and the Normalized Difference Vegetation Index (NDVI, reflecting plant stress) from remotely sensed data from MODIS satellites with 0.05° spatial resolution. Combinations of coefficients describing the phenology of these two variables pointed to divergent niche preferences, compatible with previous events of vicariance among the species. Amblyomma cajennense has been recorded in areas with small variations in temperature and NDVI. The remaining species were recorded in areas with large variations. The maximum environmental niche overlap was ∼73.6% between A. mixtum and A. cajennense and 73.5% between A. tonelliae and A. sculptum. Projecting these inferences on the geographical space revealed probable areas of sympatry or parapatry between A. mixtum and A. cajennense or between A. tonelliae and A. sculptum, the latter of which was confirmed with field collections. The A. sculptum distribution overlaps with that of A. tonelliae in northern Argentina and Paraguay; parapatry occurs at one extreme of the conditions occupied by both species. Compared with areas of allopatry, sites with both species had consistently lower temperatures, except for 10–12 weeks during the summer, and higher NDVI values throughout the year. We hypothesise that the overlap between A. tonelliae and A. sculptum resulted from secondary contact between populations, with A. sculptum adapting to sites with high water availability to balance high summer temperatures. Additional surveys of the areas of spatial overlap among these species are necessary to elucidate the forces driving their evolution and their adaptation to the environment.Fil: Estrada Peña, Agustín. Universidad de Zaragoza; EspañaFil: Tarragona, Evelina Luisa. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Vesco, Umberto. Università di Torino; ItaliaFil: De Meneghi, Daniele. Università di Torino; ItaliaFil: Mastropaolo, Mariano. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Mangold, Atilio Jose. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Guglielmone, Alberto Alejandro. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; ArgentinaFil: Nava, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentin

Comparative pharmacokinetics of a long-acting amoxicillin formulation following its subcutaneous and intramuscular administration to dogs

The influence of the route of administration on the amoxicillin pharmacokinetics in dogs was\ninvestigated following the subcutaneous or intramuscular administration of 15 mg/kg amoxicillin\nformulated as a long-acting aqueous suspension. Serial blood samples were collected at predetermined\ntimes. Pharmacokinetic parameters were calculated from the disposition curves for both routes for\neach animal. Significant differences were observed for the parameters area under the curve (73.8\n± 13.1 mg.h/ml versus 88.3 ± 17.0 mg.h/ml) and mean residence time (6.9 ± 2.8 h versus 10.3 ±\n6.1 h) following the intramuscular and subcutaneous injections, respectively. The subcutaneous\nadministration exhibited higher depot effect than the intramuscular administration. No significant\ndifferences were observed for the time above the minimun inhibitory concentrations for bacteria\nof low (4 mg/ml) and high (0.250 mg/ml) amoxicillin sensitivity. Our data suggest that the tested\nformulation can be used with a prolonged interval of 48 h for high sensitivity bacteria, and that\nboth routes of administration provide similar kinetic profiles and thus, similar clinical outcomes.Fil: Porta, N. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Farmacología; ArgentinaFil: Prados, A.P. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Farmacología; ArgentinaFil: Kreil, V. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Farmacología; ArgentinaFil: Tarragona, L. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Farmacología; ArgentinaFil: Monfrinotti, A. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Farmacología; ArgentinaFil: Rebuelto, M. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Farmacología; ArgentinaSe investigó la influencia de la vía de administración en la farmacocinética de la amoxicilina en caninos,\nluego de la administración por las vías subcutánea e intramuscular de 15 mg/kg de amoxicilina\nformulada como una suspensión acuosa de larga acción. Se tomaron muestras de sangre en tiempos\npredeterminados, y se calcularon los parámetros farmacocinéticos de las curvas de disposición para\ncada animal y para cada vía de administración. Se observaron diferencias significativas para los\nparámetros área bajo la curva (73.8 ± 13.1 mg.h/ml versus 88.3 ± 17.0 mg.h/ml) y tiempo medio\nde residencia (6.9 ± 2.8 h versus 10.3 ± 6.1 h) luego de las inyecciones intramuscular y subcutánea,\nrespectivamente. La administración subcutánea mostró un mayor efecto depot que la intramuscular.\nNo se observaron diferencias significativas para el tiempo sobre la concentración inhibitoria mínima\npara bacterias de baja (4 mg/ml) y alta (0.250 mg/ml) susceptibilidad a la amoxicilina. Nuestros datos\nsugieren que la formulación probada puede ser utilizada con un intervalo posológico prolongado de\n48 h para bacterias muy susceptibles, y que ambas vías de administración proveen perfiles cinéticos\nsimilares, y por ende, similares resultados clínicos. \

Equivalence of ELISpot Assays Demonstrated between Major HIV Network Laboratories

The Comprehensive T Cell Vaccine Immune Monitoring Consortium (CTC-VIMC) was created to provide standardized immunogenicity monitoring services for HIV vaccine trials. The ex vivo interferon-gamma (IFN-γ) ELISpot is used extensively as a primary immunogenicity assay to assess T cell-based vaccine candidates in trials for infectious diseases and cancer. Two independent, GCLP-accredited central laboratories of CTC-VIMC routinely use their own standard operating procedures (SOPs) for ELISpot within two major networks of HIV vaccine trials. Studies are imperatively needed to assess the comparability of ELISpot measurements across laboratories to benefit optimal advancement of vaccine candidates.We describe an equivalence study of the two independently qualified IFN-g ELISpot SOPs. The study design, data collection and subsequent analysis were managed by independent statisticians to avoid subjectivity. The equivalence of both response rates and positivity calls to a given stimulus was assessed based on pre-specified acceptance criteria derived from a separate pilot study.Detection of positive responses was found to be equivalent between both laboratories. The 95% C.I. on the difference in response rates, for CMV (-1.5%, 1.5%) and CEF (-0.4%, 7.8%) responses, were both contained in the pre-specified equivalence margin of interval [-15%, 15%]. The lower bound of the 95% C.I. on the proportion of concordant positivity calls for CMV (97.2%) and CEF (89.5%) were both greater than the pre-specified margin of 70%. A third CTC-VIMC central laboratory already using one of the two SOPs also showed comparability when tested in a smaller sub-study.The described study procedure provides a prototypical example for the comparison of bioanalytical methods in HIV vaccine and other disease fields. This study also provides valuable and unprecedented information for future vaccine candidate evaluations on the comparison and pooling of ELISpot results generated by the CTC-VIMC central core laboratories

NLRP3 inflammasome activation and symptom burden in KRAS-mutated CMML patients is reverted by IL-1 blocking therapy

Chronic myelomonocytic leukemia (CMML) is frequently associated with mutations in the rat sarcoma gene (RAS), leading to worse prognosis. RAS mutations result in active RAS-GTP proteins, favoring myeloid cell proliferation and survival and inducing the NLRP3 inflammasome together with the apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), which promote caspase-1 activation and interleukin (IL)-1(3 release. Here, we report, in a cohort of CMML patients with mutations in KRAS, a constitutive activation of the NLRP3 inflammasome in monocytes, evidenced by ASC oligomerization and IL-1(3 release, as well as a specific inflammatory cytokine signature. Treatment of a CMML patient with a KRASG12D mutation using the IL-1 receptor blocker anakinra inhibits NLRP3 inflammasome activation, reduces monocyte count, and improves the patient's clinical status, enabling a stem cell transplant. This reveals a basal inflammasome activation in RAS-mutated CMML patients and suggests potential therapeutic applications of NLRP3 and IL-1 blockers

One Health Approach to Identify Research Needs on Rhipicephalus microplus Ticks in the Americas

We aim to provide a harmonized view of the factors that affect the survival and promote the spread of R. microplus in the Neotropics, approaching its different facets of biology, ecology, distribution, and control. We review the interactions among environmental niche, landscape fragmentation, vegetal coverage (abiotic traits), and the biotic aspects of its ecology (abundance of domesticated or wild competent hosts), proposing emerging areas of research. We emphasize a holistic view integrating an economically and ecologically sustainable control of infestations and transmitted pathogens by R. microplus in the Neotropics. Examples of research link the trends of climate, the composition of the community of hosts, the landscape features, and a tailored management based on ecological grounds. Our view is that factors driving the spread of R. microplus are complex and deeply interrelated, something that has been seldom considered in control strategies. The effects of climate may affect the dynamics of wildlife or the landscape composition, promoting new patterns of seasonal activity of the tick, or its spread into currently free areas. In this paper we encourage a One Health approach highlighting the main aspects governing the components of the tick’s life cycle and its interactions with livestock and wild animals.Nuestro objetivo es ofrecer una visión armonizada de los factores que afectan a la supervivencia y promueven la propagación de R. microplus en el Neotrópico, abordando sus diferentes facetas de biología, ecología, distribución y control. Revisamos las interacciones entre el nicho ambiental, la fragmentación del paisaje la cobertura vegetal (rasgos abióticos), y los aspectos bióticos de su ecología (abundancia de hospedadores domesticados domésticos o silvestres competentes), proponiendo áreas emergentes de investigación. Hacemos hincapié en una visión holística integrando un control económica y ecológicamente sostenible de las infestaciones y los patógenos transmitidos patógenos transmitidos por R. microplus en el Neotrópico. Los ejemplos de investigación vinculan las tendencias del clima, la composición de la comunidad de hospedadores, las características del paisaje y una gestión adaptada basada en motivos ecológicos. Nuestra opinión es que los factores que impulsan la propagación de R. microplus son complejos y profundamente interrelacionados, algo que rara vez se ha tenido en cuenta en las estrategias de control. Los efectos del Los efectos del clima pueden afectar a la dinámica de la vida silvestre o a la composición del paisaje, promoviendo nuevos patrones de actividad estacional de la garrapata, o su propagación en zonas actualmente libres. En este artículo se fomenta un enfoque de En este documento fomentamos un enfoque de "salud", destacando los principales aspectos que rigen los componentes del ciclo de vida de la garrapata y sus interacciones con el ganado y los animales salvajes. sus interacciones con el ganado y los animales salvajes

Detection of kinase domain mutations in BCR::ABL1 leukemia by ultra-deep sequencing of genomic DNA

The screening of the BCR::ABL1 kinase domain (KD) mutation has become a routine analysis in case of warning/failure for chronic myeloid leukemia (CML) and B-cell precursor acute lymphoblastic leukemia (ALL) Philadelphia (Ph)-positive patients. In this study, we present a novel DNA-based next-generation sequencing (NGS) methodology for KD ABL1 mutation detection and monitoring with a 1.0E−4 sensitivity. This approach was validated with a well-stablished RNA-based nested NGS method. The correlation of both techniques for the quantification of ABL1 mutations was high (Pearson r = 0.858, p < 0.001), offering DNA-DeepNGS a sensitivity of 92% and specificity of 82%. The clinical impact was studied in a cohort of 129 patients (n = 67 for CML and n = 62 for B-ALL patients). A total of 162 samples (n = 86 CML and n = 76 B-ALL) were studied. Of them, 27 out of 86 harbored mutations (6 in warning and 21 in failure) for CML, and 13 out of 76 (2 diagnostic and 11 relapse samples) did in B-ALL patients. In addition, in four cases were detected mutation despite BCR::ABL1 < 1%. In conclusion, we were able to detect KD ABL1 mutations with a 1.0E−4 sensitivity by NGS using DNA as starting material even in patients with low levels of disease.Tis project was funded in part by CRIS CANCER FOUNDATION