Coffee Cell Suspensions as a Platform for Transient Gene Expression Analysis.

Coffea arabica L. is a crucial crop globally, but its genetic homogeneity leads to its susceptibility to diseases and pests like the coffee berry borer (CBB). Chemical and cultural control methods are difficult due to the majority of the CBB life cycle taking place inside coffee beans. One potential solution is the use of the gene cyt1Aa from Bacillus thuringiensis as a biological insecticide. To validate candidate genes against CBB, a simple, rapid, and efficient transient expression system is necessary. This study uses cell suspensions as a platform for expressing the cyt1Aa gene in the coffee genome (C. arabica L. var. Catuaí) to control CBB. The Agrobacterium tumefaciens strain GV3101::pMP90 containing the bar and cyt1Aa genes are used to genetically transform embryogenic cell suspensions. PCR amplification of the cyt1Aa gene is observed 2, 5, and 7 weeks after infection. This chapter describes a protocol that can be used for the development of resistant varieties against biotic and abiotic stresses and CRISPR/Cas9-mediated genome editing.

Using historical habitat loss to predict contemporary mammal extirpations in Neotropical forests.

Understanding which species will be extirpated in the aftermath of large-scale human disturbance is critical to mitigating biodiversity loss, particularly in hyperdiverse tropical biomes. Deforestation is the strongest driver of contemporary local extinctions in tropical forests but may occur at different tempos. The 2 most extensive tropical forest biomes in South America-the Atlantic Forest and the Amazon-have experienced historically divergent pathways of habitat loss and biodiversity decay, providing a unique case study to investigate rates of local species persistence on a single continent. We quantified medium- to large-bodied mammal species persistence across these biomes to elucidate how landscape configuration affects their persistence and associated ecological functions. We collected occurrence data for 617 assemblages of medium- to large-bodied mammal species (>1 kg) in the Atlantic Forest and the Amazon. Analyzing natural habitat cover based on satellite data (1985-2022), we employed descriptive statistics and generalized linear models (GLMs) to investigate ecospecies occurrence patterns in relation to habitat cover across the landscapes. The subregional erosion of Amazonian mammal assemblage diversity since the 1970s mirrors that observed since the colonial conquest of the Atlantic Forest, given that 52.8% of all Amazonian mammals are now on a similar trajectory. Four out of 5 large mammals in the Atlantic Forest were prone to extirpation, whereas 53% of Amazonian mammals were vulnerable to extirpation. Greater natural habitat cover increased the persistence likelihood of ecospecies in both biomes. These trends reflected a median local species loss 63.9% higher in the Atlantic Forest than in the Amazon, which appears to be moving toward a turning point of forest habitat loss and degradation. The contrasting trajectories of species persistence in the Amazon and Atlantic Forest domains underscore the importance of considering historical habitat loss pathways and regional biodiversity erosion in conservation strategies. By focusing on landscape configuration and identifying essential ecological functions associated with large vertebrate species, conservation planning and management practices can be better informed.

Uso de la pérdida histórica de hábitat para predecir la desaparición de mamíferos contemporáneos en los bosques neotropicales Resumen Tener conocimiento de cuáles especies desaparecerán después de una perturbación humana es de suma importancia para mitigar la pérdida de la biodiversidad, particularmente en los biomas híper diversos. La deforestación es la principal causante de las extinciones locales contemporáneas en los bosques tropicales, aunque puede ocurrir en diferentes tiempos. Los dos bosques tropicales más extensos de América del Sur ­ el Bosque Atlántico y la Amazonia ­ han experimentado formas históricamente divergentes de pérdida de hábitat y decadencia de biodiversidad, lo que proporciona un caso único de estudio para investigar las tasas de persistencia de las especies locales en un solo continente. Cuantificamos la persistencia de las especies de mamíferos de talla mediana a grande en estos dos bosques para aclarar cómo la configuración del paisaje afecta su persistencia y las funciones ecológicas asociadas. Recolectamos datos de presencia de 617 ensambles de especies de mamíferos de talla mediana a grande (>1 kg) en el Bosque Atlántico y en la Amazonia. Analizamos la cobertura natural del hábitat con base en datos satelitales (1985­2022) y empleamos estadística descriptiva y modelos lineales generalizados (MLG) para investigar los patrones de presencia de las eco especies en relación con la cobertura del hábitat en los distintos paisajes. La erosión subregional de la diversidad de ensambles de mamíferos en la Amazonia desde los 70s es igual a la observada en el Bosque Atlántico desde la conquista colonial, dado que 52.8% de todos los mamíferos amazónicos se encuentran en una trayectoria similar. Cuatro de los cinco grandes mamíferos en el Bosque Atlántico estaban propensos a desaparecer, mientras que el 53% de los mamíferos amazónicos estaban vulnerables a desaparecer. Una mayor cobertura natural del hábitat incrementó la probabilidad de persistencia de las eco especies en ambos bosques. Estas tendencias reflejaron una pérdida mediana de especies locales 63.9% mayor en el Bosque Atlántico que en la Amazonia, lo cual parece dirigirse hacia un momento decisivo para la degradación y pérdida del hábitat del bosque. Las trayectorias contrastantes de la persistencia de especies en el Bosque Atlántico y la Amazonia destacan la importancia de considerar dentro de las estrategias de conservación las maneras en las que se ha perdido históricamente el hábitat y la erosión de la biodiversidad regional. Si nos enfocamos en la configuración del paisaje y en la identificación de las funciones ecológicas esenciales asociadas con las especies grandes de vertebrados, podemos informar de mejor manera a la planeación de la conservación y las prácticas de manejo.

A Capsaicin-Based Phytogenic Solution Improves Performance and Thermal Tolerance of Heat-Stressed Growing Pigs.

Exposure to heat stress (HS) detrimentally affects pig performance. This study explored whether a dietary phytogenic solution based on Capsicum spp. (PHY) could enhance the thermal tolerance of heat-stressed growing pigs. Forty-two individually housed pigs were randomly assigned to three treatments: thermoneutral pigs on a control diet (TN-C) and pigs subjected to HS fed the control diet either without (HS-C) or with supplemental PHY (HS-PHY). The TN-C group exhibited increased average daily gain (ADG) and feed intake (FI) compared to both HS-C (p < 0.01) and HS-PHY pigs (p < 0.05) and better feed efficiency compared to HS-C pigs only (p < 0.01). However, the HS-PHY pigs showed significantly higher FI (p < 0.01) and ADG (p < 0.05) compared to HS-C pigs. HS pigs displayed higher body temperatures (BTs) than TN pigs (p < 0.01), yet HS-PHY pigs experienced a lesser increase in BT compared to HS-C pigs (p < 0.05). Supplementation with PHY mitigated some effects of HS, increasing serum superoxide dismutase and catalase activity, reducing HSP90 expression in longissimus dorsi muscle, and elevating jejunal villus height compared to HS-C pigs (p < 0.05), reaching levels akin to TN-C pigs. Additionally, PHY supplementation resulted in lower serum urea levels than HS-C pigs (p < 0.01) and similar myosin gene expression to TN-C pigs (p > 0.1), suggesting enhanced amino acid post-absorptive utilization for lean tissue growth. In conclusion, dietary PHY supplementation partially offset the adverse effects of HS on pig performance by improving thermal tolerance.

Multiprotease improves amino acid release in vitro, energy, and nutrient utilization in broilers fed diets varying in crude protein levels.

Low crude protein (CP) diets can reduce nitrogen (N) excretion and costs by increasing N utilization efficiency. Exogenous proteases may further improve protein digestibility in low CP diets. This study first evaluated in vitro the efficacy of a multiprotease on amino acid (AA) release from feedstuffs and broiler feed. Later, a broiler study evaluated the effect of feeding corn-soybean meal diets containing 3 CP levels (17, 19, and 21% CP) with supplementation on top of 0 or 2,400 U/kg multiprotease on chicken growth performance, total tract CP, and ileal AA digestibilities, and energy utilization. Ross 708 male chickens were placed in 42 cages and assigned to 6 treatments resulting from a 3 × 2 factorial arrangement. Three isocaloric basal diets were formulated to reduce CP, but all diets maintained digestible Lys:CP in 5.47% and the same ideal protein profile. Data were analyzed in a completely randomized design. On average, the multiprotease increased (P < 0.05) in vitro free AA release by 27.81% in most feedstuffs evaluated compared to the control. For broiler feed, 1,200 U/g multiprotease addition improved (P < 0.001) in vitro free AA release by 18.90%. This multiprotease showed interaction effects (P < 0.05) on chicken FCR, energy, and CP digestibility. As expected, BW at 24 d, BW gain, and FCR (8-24 d) worsened (P < 0.001) as dietary CP reduced from 21 to 17%, and multiprotease addition did not improve (P > 0.05) these parameters. BW gain decreased by 12.9% when N intake was reduced from 49.32 to 38.49 g/bird. Multiprotease supplementation improved (P < 0.01) AMEn by 71 kcal/kg, CP digestibility from 59.45 to 63.51%, ileal AA digestibility, and DM digestibility from 67.08 to 73.49%, but only in the 21% CP diet. No differences in ileal AA digestibility due to CP level (P > 0.05) were detected, except for Cys digestibility (P < 0.01). In conclusion, low CP diets reduced growth performance and improved N utilization but negatively affected energy utilization efficiency. Exogenous multiprotease supplementation improved AME, AMEn, protein, ileal AA, and DM digestibility in the 21% CP diet without significantly affecting growth performance.

Ecologically mediated differences in electric organ discharge drive evolution in a sodium channel gene in South American electric fishes.

Active electroreception-the ability to detect objects and communicate with conspecifics via the detection and generation of electric organ discharges (EODs)-has evolved convergently in several fish lineages. South American electric fishes (Gymnotiformes) are a highly species-rich group, possibly in part due to evolution of an electric organ (EO) that can produce diverse EODs. Neofunctionalization of a voltage-gated sodium channel gene accompanied the evolution of electrogenic tissue from muscle and resulted in a novel gene (scn4aa) uniquely expressed in the EO. Here, we investigate the link between variation in scn4aa and differences in EOD waveform. We combine gymnotiform scn4aa sequences encoding the C-terminus of the Nav1.4a protein, with biogeographic data and EOD recordings to test whether physiological transitions among EOD types accompany differential selection pressures on scn4aa. We found positive selection on scn4aa coincided with shifts in EOD types. Species that evolved in the absence of predators, which likely selected for reduced EOD complexity, exhibited increased scn4aa evolutionary rates. We model mutations in the protein that may underlie changes in protein function and discuss our findings in the context of gymnotiform signalling ecology. Together, this work sheds light on the selective forces underpinning major evolutionary transitions in electric signal production.

Biochemical and microbiological characterization of a thermotolerant bacterial consortium involved in the corrosion of Aluminum Alloy 7075.

Microorganisms can play a significant role in material corrosion, with bacterial biofilms as major participants in microbially influenced corrosion (MIC). The exact mechanisms by which this takes place are poorly understood, resulting in a scarcity of information regarding MIC detection and prevention. In this work, a consortium of moderately thermophilic bacteria isolated from a biofilm growing over aluminum alloy 7075 was characterized. Its effect over the alloy was evaluated on a 40-day period using Electron Microscopy, demonstrating acceleration of corrosion in comparison to the abiotic control. The bacterial consortium was biochemically and microbiologically characterized as an attempt to elucidate factors contributing to corrosion. Molecular analysis revealed that the consortium consisted mainly of members of the Bacillus genus, with lower abundance of other genera such as Thermoanaerobacterium, Anoxybacillus and Paenibacillus. The EPS polysaccharide presented mainly mannose, galactose, rhamnose and ribose. Our observations suggest that the acidification of the culture media resulting from bacterial metabolism acted as the main contributor to corrosion, hinting at an unspecific mechanism. The consortium was not sulfate-reducing, but it was found to produce hydrogen, which could also be a compounding factor for corrosion.

Initial validation of the International Trauma Questionnaire (ITQ) in a sample of Chilean adults.

BACKGROUND: ICD-11 Posttraumatic Stress Disorder (PTSD) and Complex PTSD (CPTSD) are stress-related disorders. The International Trauma Questionnaire (ITQ) is a widely used instrument to assess PTSD and CPTSD. To date, there is no evidence of the psychometric characteristics of the ITQ in Latin American countries. OBJECTIVE: The aim of this study was to assess the construct and concurrent validity of the Latin American Spanish adaptation of the ITQ in a sample of Chilean adults. METHODS: A sample of 275 Chilean young adults completed the ITQ, a traumatic life events checklist, the Adverse Childhood Experiences Questionnaire, the Depression Anxiety Stress Scales-21, and the Columbia-Suicide Severity Rating Scale short version. Four alternative confirmatory factor analysis models were tested. Correlation analyses were performed to determine concurrent validity with associated measures (number of reported traumatic events, number of adverse childhood experiences, anxiety, depression, and suicidal risk). RESULTS: The second-order two-factor (PTSD and DSO) and the correlated first-order six-factor model provided acceptable fit; however, the first model showed a better fit based on the BIC difference. The PTSD and DSO dimensions, as well as the six ITQ clusters showed positive correlations with reported number of traumatic life-events, reported number of adverse childhood experiences, levels of anxiety, depression, and suicidal risk. CONCLUSIONS: The ITQ Latin American Spanish adaptation provides acceptable psychometric evidence to assess PTSD and CPTSD in accordance with the ICD-11.

This study is an initial validation of the Latin American Spanish adaptation of the ITQ with a Chilean young adults sample.The latent structure of the Latin American Spanish ITQ was better supported by a two-factor second-order model (PTSD/DSO); a six-factor correlated model was also acceptable.The six ITQ symptom clusters, as well as the PTSD/CPTSD dimensions were significantly positively correlated with three criterion variables: anxiety, depression, and suicidal risk.The number of potentially traumatic experiences, as well as number of exposure to adverse childhood experiences, was significantly associated with PTSD/CPTSD symptoms.

Pharmacotherapeutic monitoring of dipyrone in northeastern Brazilian donkeys (Equus asinus).

This study aimed to monitor the effects of dipyrone following multiple administrations in northeastern donkeys. Ten castrated male donkeys, aged 6.4 ± 3 years and weighing 130.6 ± 9.8 kg, were administered dipyrone (25 mg/kg IV) every 12 h, resulting in six administrations (D1 to D6) per animal. Blood samples were collected over a 72 h monitoring period. A validated UHPLC-MS/MS method was employed to determine the plasma concentrations of the 4- methylaminoantipyrine (4-MAA) and 4-aminoantipyrine (4-AA). The calculated pharmacokinetic variables of 4-MAA after D1 and D6 were, respectively: Cmax (µg/mL) = 163.60 ± 179.72 and 178.79 ± 196.94; T1/2beta (h) = 2.65 ± 0.65 and 3.37 ± 1.03; and AUC0-t (µg/mL × h) = 240.38 ± 130.87 and 373.52 ± 78.85. The same variables for 4-AA were: Cmax, (µg/mL) = 0.44 ± 0.27 and 0.90 ± 0.31, T1/2beta (h) = 14.77 ± 13.13 and 35.97 and AUC0-t (µg/mL × h) = 3.20 ± 0.43 and 27.73 ± 11.99. Concentrations of 4-MAA exceeded the minimum concentration required for 50% inhibition of cyclooxygenases 1 and 2. However, an accumulation of 4-AA, was observed. Further clinical studies are necessary to ascertain the implications of these findings on the pharmacodynamic response to dipyrone in northeastern donkeys.

Alterations of Plasmatic Biomarkers of Neurodegeneration in Mucopolysaccharidosis Type II Patients Under Enzyme Replacement Therapy.

Mucopolysaccharidosis type II (MPS II) is a disorder caused by a deficient activity of iduronate-2-sulfatase, a lysosomal enzyme responsible for degrading glycosaminoglycans (GAGs). The abnormal storage of GAGs within lysosomes disrupts cellular homeostasis and leads to a severe symptomatology. Patients present neuropsychiatric impairment characterized by mental retardation and impaired cognition. The aim of this study was to quantify four neurodegeneration biomarkers in plasma: brain-derived neurotrophic factor (BDNF), platelet-derived growth factor (PDGF-AA), neural cell adhesion molecule (NCAM) and cathepsin-D, as well as to identify possible correlations with urinary GAGs in seven patients undergoing treatment with ERT (Elaprase® 0.5 mg/kg of body weight). Patients with both severe and attenuated forms of MPS II showed signs of neurodegeneration in neuroimaging exams. Patients have a decrease in BDNF and PDGF-AA concentrations, and an increase in NCAM level compared to controls. No alterations in cathepsin-D concentration were seen. GAGs levels were higher in patients than in controls, but no significant correlations between GAGs and biomarkers were observed. These results evidence that patients have neurodegeneration and that monitoring these biomarkers might be useful for assessing this process. To this date, this is the first work to analyze these plasmatic markers of neurodegeneration in patients.

Community and domestic violence are associated with DNA methylation GrimAge acceleration and heart rate variability in adolescents.

Background: Cumulative exposure to violence can change the regulation of epigenetic and physiological markers. Although violence has been associated with accelerated cellular aging, little is known about associations with cardiac autonomic activity.Objective: The current study aimed to investigate the relationship of exposure to community and domestic violence (CDV) with vagal activity and epigenetic aging acceleration.Methods: A total of 86 adolescents (57% female) were evaluated and interviewed at two time-points in São Gonçalo (2014-2019), a Brazilian city with high levels of violence. Exposure to CDV was assessed in both time-points. GrimAge acceleration was calculated from saliva DNA methylation using Infinium HumanMethylation450K (Illumina) collected in the first assessment. Heart rate variability (HRV) was collected during two stress tasks at the second assessment.Results: The exposure to violence witnessed or directly experienced at home and in the community increased significantly (t = 4.87, p < .01) across two-time points, and males had reported higher violence exposure (t = 2.06, p = .043). Violence at 1st assessment was significantly associated with GrimAge acceleration (B = .039, p value = .043). Violence at both assessments were associated with HRV measured during the narration of the worst trauma (traumaHRV) (B = .009, p value = .039, and B = .007, p value = .024, 1st and 2nd assessment respectively). GrimAge acceleration was significantly associated with traumaHRV (B = .043, p value = .049), and HRV measured during a 3D roller coaster video (B = .061, p value = .024).Conclusions: We found relevant evidence that experiencing violence during adolescence is associated with epigenetic aging and stress-related vagal activity. Understanding these factors during this period could contribute to the development of early interventions for health promotion.HIGHLIGHTS Higher exposure to Community and domestic violence is associated with increased GrimAge acceleration.Higher GrimAge acceleration is associated with increased stress-related vagal activity.Exposure to community and domestic violence increased significantly over time.

Devouring and Asphyxia: Symptoms of a Cultural Complex in Present Times.

The article discusses dominance and oppression in society due to cultural complexes filled with collective memories of destructiveness and perpetration, implicit memories which have remained repressed. Individual personal complexes and traumas are intertwined with traumatizing historical circumstances, setting up pairs of perpetrator and victim. The metaphors of devouring and asphyxiation are used to denote interpersonal and group relationships in which feelings of imprisonment, suffocating anxiety and expulsion are predominant, all of which leads to painful projections and introjections, dissociation and suffering. Asphyxiating death symbolizes not only environmental devastation by fire, the pandemic and the plague, but is also seen as a symptom of acute anxiety in modern times. Devouring stands for the annulment of the objectified Other in a patriarchal society best revealed in fratricidal struggles, the oppression of women and, ultimately, wars.

L'article traite de la domination et de l'oppression dans la société, du fait de complexes culturels chargés de souvenirs de destruction et de culpabilité, de mémoires implicites qui sont restées refoulées. Les complexes personnels et les traumatismes des individus sont entremêlés avec les circonstances historiques traumatisantes. Ainsi se forment des couples coupable-victime. Les métaphores de dévoration et d'asphyxie sont utilisées pour désigner des relations interpersonnelles et de groupe dans lesquelles les sentiments d'emprisonnement, l'angoisse d'étouffement et l'expulsion sont prédominants, tout cela menant à des projections et des introjections douloureuses, à de la dissociation et de la souffrance. La mort par asphyxie symbolise non seulement la dévastation de l'environnement par l'incendie, la pandémie et la peste, mais est aussi comprise comme un symptôme d'anxiété aigüe à notre époque. La dévoration signifie l'annulation d'un Autre traité comme un objet dans une société patriarcale qui se dévoile dans des batailles fratricides, l'oppression des femmes et au final, les guerres.

El artículo analiza la dominación y la opresión en la sociedad debido a complejos culturales llenos de memorias colectivas de destructividad y perpetración, memorias implícitas que han permanecido reprimidas. Los complejos y traumas personales individuales se entrelazan con circunstancias históricas traumatizantes, estableciendo pares de perpetrador y víctima. Las metáforas de la devoración y la asfixia se utilizan para denotar relaciones interpersonales y grupales en las que predominan sentimientos de aprisionamiento, ansiedad sofocante y expulsión, todo lo cual conduce a proyecciones e introyecciones dolorosas, disociación y sufrimiento. La muerte asfixiante simboliza no sólo la devastación medioambiental por el fuego, la pandemia y la peste, sino que también se considera un síntoma de ansiedad aguda en los tiempos modernos. Devorar representa la anulación del Otro cosificado en una sociedad patriarcal que sobre todo se revela en las luchas fratricidas, la opresión de la mujer y, en última instancia, las guerras.

A comprehensive analysis of immunogenic cell death and its key gene HSP90AA1 in bladder cancer.

BACKGROUND: Bladder cancer (BLCA) is defined as a type of urinary cancer with high incidence and lack of specific biomarkers and drug targets. Immunogenic cell death (ICD) has been classified as a regulated type of cell death. Growing evidence suggested that ICD can reshape the tumor immune microenvironment, which may contribute to the development of immunotherapy strategies. The aim of this study was to reveal the specific mechanism of ICD in bladder cancer and to further predict the prognostic immunotherapy outcomes. METHODS: By consensus clustering analysis, bladder cancer patients in TCGA database were divided into different ICD subtypes. Additionally, we developed an ICD-scoring system and constructed the ICD score-based risk signature and nomogram to better characterize patients. Furthermore, we carried out a series of experiments to verify the relevant findings. RESULTS: Based on the transcriptome expression levels of ICD-related genes, a total of 403 BLCA patients in the TCGA database were divided into two subgroups with different ICD molecular patterns by consensus cluster analysis. These subgroups showed different clinicopathological features, survival outcomes, tumor microenvironment (TME) characteristics, immune-related scores, and treatment response. Moreover, the established prediction model and ICD score can effectively distinguish high risk/score patients from low risk/score patients, which has excellent predictive value. Finally, we found that the key gene HSP90AA1 was highly expressed in the high-ICD score group and in bladder cancer tissues, and was confirmed to be associated with the proliferation of bladder cancer cells. CONCLUSION: To sum up, we established a new classification system for BLCA based on ICD-related genes. This stratification has significant predictive power for clinical outcomes and can effectively evaluate the prognosis and immunotherapy of BLCA patients. Finally, it was proved that HSP90AA1 was highly expressed in BLCA and would be a promising therapeutic target for BLCA.

Facilitating biodiversity conservation through partnerships to achieve transformative outcomes.

Conservation biology is a mission-driven discipline that must navigate a new relationship between conservation and science. Because conservation is a social and political as well as an ecological project, conservation biologists must practice interdisciplinarity and collaboration. In a comparative study of 7 cases (Jaguars in the Chaco, Grevy's zebra in Kenya, Beekeeping in Tanzania, Andean cats in Argentina, Jaguars in Mexico, Lobster fishing, and Black bears in Mexico), we examined motivations for collaboration in conservation, who can collaborate in conservation, and how conservation professionals can work well together. In 5 case studies, successful conservation outcomes were prioritized over livelihood benefits. In the other 2 cases, livelihoods were prioritized. All case studies employed participatory approaches. There were multiple external actors, including local and Indigenous communities, nongovernmental organizations, agencies, regional and national governments, and international organizations, which enhanced conservation and wider sustainability outcomes. Key collaboration aspects considered across the case studies were time (mismatch between relationship building and project schedules), trust required for meaningful partnerships, tools employed, and transformative potential for people, nature, and the discipline of conservation biology. We developed guidelines for successful collaboration, including long-term commitment, knowledge integration, multiscalar and plural approaches, cultivation of trust, appropriate engagement, evaluation, supporting students, and efforts for transformation.

Facilitación de la conservación de la biodiversidad a través de colaboraciones para lograr resultados transformadores Resumen La biología de la conservación es una disciplina impulsada por sus objetivos que debe navegar una nueva relación entre la conservación y la ciencia. Dado que la conservación es un proyecto social y político así como ecológico, los biólogos de la conservación deben practicar la interdisciplinariedad y la colaboración. En un estudio comparativo de siete casos (jaguares en el Chaco, cebras de Grevy en Kenia, apicultura en Tanzania, gatos andinos en Argentina, jaguares en México, pesca de langosta, osos negros en México), analizamos las motivaciones para colaborar en la conservación, quién puede colaborar en la conservación y cómo los profesionales de la conservación pueden trabajar bien juntos. En cinco estudios de caso, se priorizaron los resultados satisfactorios de la conservación sobre los beneficios para los medios de subsistencia. En los otros 2 casos, se dio prioridad a los medios de sustento. Todos los estudios de caso emplearon enfoques participativos. Hubo múltiples actores externos, como comunidades locales e indígenas, organizaciones no gubernamentales, agencias, gobiernos regionales y nacionales y organizaciones internacionales, que mejoraron los resultados de conservación y sostenibilidad en general. Los aspectos clave de la colaboración considerados en los estudios de caso fueron el tiempo (desajuste entre el establecimiento de relaciones y los calendarios de los proyectos), la confianza necesaria para establecer colaboraciones significativas, las herramientas empleadas y el potencial transformador para las personas, la naturaleza y la disciplina de la biología de la conservación. Se elaboraron directrices para el éxito de la colaboración, como el compromiso a largo plazo, la integración de conocimientos, los enfoques multiescalares y plurales, el fomento de la confianza, la participación adecuada, la evaluación, el apoyo a los estudiantes y los esfuerzos de transformación. Facilitación de la conservación de la biodiversidad a través de colaboraciones para lograr resultados transformadores.

Mpp23Aa/Xpp37Aa Insecticidal Proteins from Bacillus thuringiensis (Bacillales: Bacillaceae) Are Highly Toxic to Anthonomus grandis (Coleoptera: Curculionidae) Larvae.

The beetle Anthonomus grandis Boheman, 1843, is the main cotton pest, causing enormous losses in cotton. The breeding of genetically modified plants with A. grandis resistance is seen as an important control strategy. However, the identification of molecules with high toxicity to this insect remains a challenge. The susceptibility of A. grandis larvae to proteins (Cry1Ba, Cry7Ab, and Mpp23Aa/Xpp37Aa) from Bacillus thuringiensis Berliner, 1915, with toxicity reported against Coleopteran, has been evaluated. The ingestion of different protein concentrations (which were incorporated into an artificial diet) by the larvae was tested in the laboratory, and mortality was evaluated after one week. All Cry proteins tested exhibited higher toxicity than that the untreated artificial diet. These Cry proteins showed similar results to the control Cry1Ac, with low toxicity to A. grandis, since it killed less than 50% of larvae, even at the highest concentration applied (100 µg·g-1). Mpp/Xpp proteins provided the highest toxicity with a 0.18 µg·g-1 value for the 50% lethal concentration. Importantly, this parameter is the lowest ever reported for this insect species tested with B. thuringiensis proteins. This result highlights the potential of Mpp23Aa/Xpp37Aa for the development of a biotechnological tool aiming at the field control of A. grandis.

Alopecia Areata in Latin America: Where are We and Where are We Going?

Alopecia areata (AA) represents an underrecognized burden in Latin America (LA), severely impacting quality of life (QoL). This impact is exacerbated by limited access to specialized dermatologic care and therapies for AA within and among nations. Many of the unmet needs for AA globally also exist in LA. The region has geographic, ethnic, cultural, and economic conditions. With new AA medicines targeting immunologic pathways on the horizon, LA must prepare regarding regulatory issues, reimbursement, awareness, and education to give adequate and timely treatment for patients with AA. To address these issues, the Americas Health Foundation convened a panel of six dermatologists from Argentina, Brazil, Colombia, and Mexico who are experts in AA and its comorbidities for a 3-day virtual meeting to discuss AA diagnosis and treatment in LA and create a manuscript offering recommendations to address discussed barriers. This publication examines unmet AA needs in LA, treatment, and innovative therapies and recommends improving AA care. Access constraints to conventional and novel medicines hinder appropriate treatments for patients. Therapy initiation delays can affect QoL, mental health, and disease progression. People with AA face stigmas, discrimination, and misconceptions owing to a lack of disease awareness. With promising new treatments for AA on the horizon, all stakeholders must coordinate efforts to enhance LA's AA management landscape and improve patient outcomes.

Molecular diagnosis of abdominal angiostrongyliasis by PCR using serum samples.

Abdominal angiostrongyliasis (AA) is a zoonotic disease caused by the nematode Angiostrongylus costaricensis, which is endemic in southern Brazil. Humans become infected by ingesting third-stage (L3) larvae and are considered accidental hosts since neither eggs nor first-stage (L1) larvae are found in feces. The definitive diagnosis can be made by histopathologic examination of surgical specimens or intestinal biopsies. The present study assessed the use of PCR to carry out the molecular detection of AA from serum samples. A total of 62 human serum samples were divided into three groups: (i) 28 serum samples from human patients with presumptive histopathological diagnosis of AA; (ii) 23 serum samples from individuals with unknown serology for AA; (iii) 11 serum samples from patients that suffered from different parasitosis were included. The serum samples were initially tested by in-house indirect ELISA and then by PCR. A total of 14 samples were positive by ELISA, and 6 were positive by PCR. Six samples that were negative by ELISA were positive by PCR. Amplicons were sequenced, and Angiostrongylus DNA was confirmed. We conclude that PCR amplification can be used to confirm Angiostrongylus DNA in serum, which is especially important in cases where antibody levels are too low to be detected. It may also serve as a useful target for survey studies.

A critical review of the precautionary approach of the IUCN impact classification for non-native taxa.

The International Union for the Conservation of Nature (IUCN) proposes the use of the Environmental Impact Classification for Alien Taxa to standardize the classification of introduced species (IS) based on their environmental impact. The IUCN invoked the precautionary principle (PP) via 2 rules: the impact assigned to a taxon must be the maximum recorded impact across different impact assessments, and when the main driver of environmental damage is unclear, it must be assumed to be caused by the IS. The validity of PP is conditioned on the degree of emergency that warrants urgent decisions and on the scientific evidence demonstrating the advantages of applying a preventive measure. The application of an impact classification system does not arise in the context of an emergency that requires management; it occurs before the decision-making phase. Thus, PP should not be used in early steps of the risk analysis process. The IUCN also did not provide enough scientific basis to justify the use of PP. Instead, the PP rules appear to be rooted primarily in the ethical value system underlying conservation science. Conservationists assign intrinsic value to native species by virtue of their roles and relationships within ecological and evolutionary systems and processes; thus, individuals introduced in new environments not only cease to have value because they are no longer part of that natural diversity and lack those links with the rest of the ecosystem, but they become a threat to what conservationists value most. The consequence of this belief is that all introduced taxa will have an impact at some level, suggesting that values justify the PP rules.

La Unión Internacional para la Conservación de la Naturaleza (UICN) propone usar la Clasificación del Impacto Ambiental para Taxones Exóticos (CIATE) para estandarizar la clasificación de las especies introducidas con base en su impacto ambiental. La UICN invocó el principio de precaución (PP) mediante dos reglas: el impacto asignado a un taxón debe ser el mayor impacto registrado en diferentes evaluaciones de impacto y cuando no sea claro el principal causante del daño ambiental, se debe asumir que lo causa la especie introducida. La validez del PP está condicionada por el grado de emergencia que amerite decisiones urgentes y por la evidencia científica que demuestre las ventajas de aplicar una medida preventiva. La aplicación de un sistema de clasificación de impactos no surge del contexto de una emergencia que requiere gestión, sino que ocurre antes de la fase de toma de decisiones. Por esto, el PP no debería usarse durante los primeros pasos del proceso de análisis de riesgo. La UICN tampoco proporcionó suficientes fundamentos científicos para justificar el uso del PP. En su lugar, las reglas del PP parecen estar ancladas principalmente en el sistema de valor ético subyacente a las ciencias de la conservación. Los conservacionistas les asignan un valor intrínseco a las especies nativas por su papel y sus relaciones dentro de los sistemas y procesos ecológicos y evolutivos; los individuos introducidos en ambientes nuevos no sólo dejan de tener valor porque ya no forman parte de la diversidad natural y carecen de esas conexiones con el resto del ecosistema, sino también se vuelven una amenaza para lo que más valoran los conservacionistas. Esta creencia tiene como consecuencia que todos los taxones introducidos tendrán un impacto a cierto nivel, lo que sugiere que el valor justifica las reglas del PP.

Systemic amyloidosis journey from diagnosis to outcomes: a twelve-year real-world experience of a single center in a middle-income country.

BACKGROUND: Systemic amyloidosis is caused by the deposition of misfolded protein aggregates in tissues, leading to progressive organ dysfunction and death. Epidemiological studies originate predominantly from high-income countries, with few data from Latin America. Due to the non-specific clinical manifestations, diagnosing amyloidosis is often challenging and patients experience a long journey and delay in diagnosis. This study aimed to assess clinical and laboratory characteristics, the diagnostic journey, and outcomes of patients with biopsy-proven systemic amyloidosis diagnosed between 2009 and 2020 at a university referral center in a middle-income Latin American country. Patients´ medical records were retrospectively reviewed. RESULTS: One hundred and forty-three patients were included. The median age at diagnosis was 60 years and 54% were male. Until the diagnosis, most of the patients (52%) were seen by at least 3 specialists, the main ones being: general practitioners (57%), nephrologists (45%), and cardiologists (38%). The most common manifestations were renal (54%) and cardiac (41%) disorders, and cachexia was seen in 36% of patients. In 72% of the cases, ≥ 2 biopsies were required until the final diagnosis. The median time from symptoms onset to diagnosis was 10.9 months, and most patients (75%) had ≥ 2 organs involved. The following subtypes were identified: AL (68%), ATTR (13%), AA (8%), AFib (4%), and inconclusive (7%). Median OS was 74.3 months in the non-AL subgroup and 18.5 months in AL. Among AL patients, those with advanced cardiac stage had the worst outcome [median OS 8.6 months versus 52.3 for stage III versus I-II, respectively (p < 0.001)]. AL subtype, cardiac involvement, and ECOG ≥ 2 were identified as independent risk factors for reduced survival. CONCLUSIONS: Systemic amyloidosis is still an underdiagnosed condition and the delay in its recognition leads to poor outcomes. Medical education, better diagnostic tools, improvement in access to therapies, and establishment of referral centers may improve patient outcomes in middle-income countries.

Sinus Node Artery Occlusion During Cardiac Denervation Procedures.

Cardioneural ablation is a novel treatment for functional bradycardia. However, the risk of acute complications is still unknown. The aim of this case report is to describe acute occlusion of the sinus node artery after cardiac denervation procedures in 2 patients and to encourage measures to prevent it, such as evaluating the aortic angulation in older patients before the procedure and by monitoring signs of sinus failure during ablation in patients with electroanatomical maps showing a constricted aspect of the right atrium. (Level of Difficulty: Advanced.).

Oleuropein-Enriched Extract From Olive Mill Leaves by Homogenizer-Assisted Extraction and Its Antioxidant and Antiglycating Activities.

Olive oil consumption has increased in the last two decades and consequently, its wastes have increased, which generates a tremendous environmental impact. Among the by-products are the olive mill leaves, which are easier and inexpensive to treat than other olive by-products. However, little research has been done on their chemical composition and potential bioactivity. Hence, in this study, olive mill leaves were used to obtain Oleuropein-Enriched Extracts (OLEU-EE) using Conventional Extraction, Ultrasound-Assisted Extraction, and Homogenization-Assisted Extraction. These three techniques were evaluated using a Factorial Design to determine the parameters to obtain an OLEU-EE with high contents of Total Phenolic Compounds (TPC), Antioxidant Activity (AA), and Oleuropein concentration (OLEU). From the results, the Homogenizer-Assisted Extraction (HAE) technique was selected at 18,000 rpm, solid:liquid ratio 1:10, and 30 s of homogenization with 70% ethanol, due to its high TPC (5,196 mg GA/100 g), AA (57,867 µmol of TE/100 g), and OLEU (4,345 mg of OLEU/100 g). In addition, the antiglycating effect of OLEU-EE on the levels of (1) fluorescent Advanced Glycation End Products (AGEs) were IC50 of 0.1899 and 0.1697 mg/mL for 1λEXC 325/λEM 440 and 2λEXC 389/λEM 443, respectively; (2) protein oxidative damage markers such as dityrosine (DiTyr), N-formylkynurenine (N-formyl Kyn), and kynurenine (Kyn) were IC50 of 0.1852, 0.2044, and 0.1720 mg/mL, respectively. In conclusion, OLEU-EE from olive mill leaves has different capacities to inhibit AGEs evidenced by the IC50 of fluorescent AGEs and protein oxidation products, together with the scavenging free radical evidenced by the concentration of Trolox Equivalent. Therefore, OLEU-EE could be potential functional ingredients that prevent oxidative damage caused by free radicals and AGEs accumulation.