Drying kinetics, thermodynamic properties and physicochemical characteristics of Rue leaves.

Generally, medicinal plants are harvested with high amount of water, so it is essential to subject the product to drying as soon as possible to prevent degradation before application. Most compounds from medicinal plants are sensitive to drying processes, so it is important to adjust the drying conditions. The objective of this study was to describe the drying of Rue (Ruta chalepensis L.) leaves, select the models that best fit each drying condition, determine the activation energy and thermodynamic properties of the leaves, and evaluate their quality after drying. Leaves were harvested with moisture content of 3.55 ± 0.05 kg water kg-1dry matter and subjected to drying at temperatures of 40, 50, 60 and 70 °C. Valcam model showed the best fit to represent the drying kinetics of Rue leaves at temperatures of 40 and 70 °C, and Midilli model proved to be better for the temperatures of 50 and 60 °C. Effective diffusion coefficient increased linearly with the increase in drying air temperature, and the activation energy was 60.58 kJ mol-1. Enthalpy, entropy and Gibbs free energy values ranged from 57.973 to 57.723 kJ mol-1, from - 0.28538 to - 0.28614 kJ mol-1 K-1 and from 147.34 to 155.91 kJ mol-1, respectively, for the temperature range of 40-70 °C. Drying air temperature promoted darkening or tendency to loss of green color; increase in drying air temperature leads to greater discoloration, as well as a higher concentration of total phenolic compounds (about 221.10 mg GAE mL-1 g-1 dm), with a peak at temperature of 60 °C.

Multidisciplinary Clinical Care in the Management of Patients Receiving Anti-GD2 Immunotherapy for High-Risk Neuroblastoma.

The addition of anti-disialoganglioside-2 (GD2) monoclonal antibodies (mAbs) such as dinutuximab and naxitamab to standard therapies for high-risk (HR) neuroblastoma has significantly improved outcomes for children with this devastating disease. The care for these young patients receiving treatment for HR neuroblastoma is complex, with need for the involvement of a multidisciplinary team. Clinical implementation of anti-GD2 mAb treatment requires the same harmonized team approach. The authors share the development process of this coordinated team method and practical recommendations for administration of anti-GD2 mAbs and adverse event (AE) management. Successful collaboration between nurses and other team members ensures optimal treatment and comfort of patients and their families. The primary focus of this approach is to mitigate and manage AEs associated with anti-GD2 mAb treatments, such as pain, hypotension, allergic reactions, and hypertension, and to ensure safe and effective use of anti-GD2 mAbs. The two treatments approved for use in patients with neuroblastoma, dinutuximab for patients with HR disease following a partial response or better to frontline multimodal therapy and naxitamab for refractory or relapsed HR disease in the bone or bone marrow, were studied in different administration settings and follow different regimens and infusion schedules. Therefore, AE management requirements are specific to each treatment. The awareness of these differences and implementation of appropriate AE management strategies in clinical practice are important to ensure the best possible outcomes for patients with HR neuroblastoma.

Construction and validation of educational technology for family members of people with venous ulcers.

OBJECTIVE: To build and validate an educational technology in the form of a booklet, aimed at the family members of people with venous ulcers to assist them in their care. METHODS: A methodological study, which went through the stages of bibliographic survey and situational diagnosis for the construction of the booklet and validation of content, appearance, and adequacy with judges and the target audience. The Content Validity Index, the Suitability Assessment of Materials, and the Concordance Index were used. RESULTS: In the validation with the content and appearance judges, the booklet showed an excellent overall Content Validity Index (tCVI=0.92). The technical judges evaluated the booklet as "superior" (average of 91%). The booklet underwent adjustments, and validation was performed with the target population, reaching an agreement rate higher than 75%. CONCLUSION: The educational booklet developed was validated for content and appearance and considered suitable for use by family members of people with venous ulcers.

Construction and validation of educational technology for family members of people with venous ulcers / Elaboración y validación de tecnología educacional para parientes de personas con úlcera venosa / Construção e validação de tecnologia educacional para familiares de pessoas com úlcera venosa

ABSTRACT Objective: To build and validate an educational technology in the form of a booklet, aimed at the family members of people with venous ulcers to assist them in their care. Methods: A methodological study, which went through the stages of bibliographic survey and situational diagnosis for the construction of the booklet and validation of content, appearance, and adequacy with judges and the target audience. The Content Validity Index, the Suitability Assessment of Materials, and the Concordance Index were used. Results: In the validation with the content and appearance judges, the booklet showed an excellent overall Content Validity Index (tCVI=0.92). The technical judges evaluated the booklet as "superior" (average of 91%). The booklet underwent adjustments, and validation was performed with the target population, reaching an agreement rate higher than 75%. Conclusion: The educational booklet developed was validated for content and appearance and considered suitable for use by family members of people with venous ulcers.

RESUMEN Objetivo: Elaborar y validar tecnología educativa en forma de cartilla, vuelta a parientes de personas con úlceras venosas para auxiliarlos en el cuidado. Métodos: Estudio metodológico, que pasó por etapas de levantamiento bibliográfico y diagnóstico situacional para elaboración de la cartilla y validez de contenido, apariencia y adecuación con jueces y público objeto. Utilizado el Índice de Validez de Contenido, el Suitability Assessment of Materials e Índice de Concordancia. Resultados: En la validación con jueces de contenido y apariencia, la cartilla presentó excelente Índice de Validez de Contenido total (IVCt=0,92). Los jueces técnicos evaluaron la cartilla clasificándola con grado de recomendación "superior" (mediana de 91%). La cartilla pasó por adecuaciones, y fue realizada validación con la población objeto, alcanzando índice de concordancia superior a 75%. Conclusión: La cartilla educativa desarrollada fue validada cuanto al contenido y apariencia y considerada adecuada para ser utilizada por parientes de personas con úlcera venosa.

RESUMO Objetivo: Construir e validar uma tecnologia educativa em forma de cartilha, voltada aos familiares de pessoas com úlceras venosas para auxiliá-los no cuidado. Métodos: Estudo metodológico, que percorreu etapas de levantamento bibliográfico e diagnóstico situacional para construção da cartilha e validação de conteúdo, aparência e adequação com juízes e público-alvo. Utilizou-se o Índice de Validade de Conteúdo, o Suitability Assessment of Materials e Índice de Concordância. Resultados: Na validação com os juízes de conteúdo e aparência, a cartilha apresentou excelente Índice de Validade de Conteúdo total (IVCt=0,92). Os juízes técnicos avaliaram a cartilha classificando-a com grau de recomendação "superior" (média de 91%). A cartilha passou por adequações, e foi realizada validação com a população-alvo, alcançando índice de concordância superior a 75%. Conclusão: A cartilha educativa desenvolvida foi validada quanto ao conteúdo e aparência e considerada adequada para ser utilizada pelos familiares de pessoas com úlcera venosa.

Eleven High-Quality Reference Genome Sequences and 360 Draft Assemblies of Shiga Toxin-Producing Escherichia coli Isolates from Human, Food, Animal, and Environmental Sources in Canada.

We report high-quality closed reference genomes for 1 bovine strain and 10 human Shiga toxin (Stx)-producing Escherichia coli (STEC) strains from serogroups O26, O45, O91, O103, O104, O111, O113, O121, O145, and O157. We also report draft assemblies, with standardized metadata, for 360 STEC strains isolated from watersheds, animals, farms, food, and human infections.

SNVPhyl: a single nucleotide variant phylogenomics pipeline for microbial genomic epidemiology.

The recent widespread application of whole-genome sequencing (WGS) for microbial disease investigations has spurred the development of new bioinformatics tools, including a notable proliferation of phylogenomics pipelines designed for infectious disease surveillance and outbreak investigation. Transitioning the use of WGS data out of the research laboratory and into the front lines of surveillance and outbreak response requires user-friendly, reproducible and scalable pipelines that have been well validated. Single Nucleotide Variant Phylogenomics (SNVPhyl) is a bioinformatics pipeline for identifying high-quality single-nucleotide variants (SNVs) and constructing a whole-genome phylogeny from a collection of WGS reads and a reference genome. Individual pipeline components are integrated into the Galaxy bioinformatics framework, enabling data analysis in a user-friendly, reproducible and scalable environment. We show that SNVPhyl can detect SNVs with high sensitivity and specificity, and identify and remove regions of high SNV density (indicative of recombination). SNVPhyl is able to correctly distinguish outbreak from non-outbreak isolates across a range of variant-calling settings, sequencing-coverage thresholds or in the presence of contamination. SNVPhyl is available as a Galaxy workflow, Docker and virtual machine images, and a Unix-based command-line application. SNVPhyl is released under the Apache 2.0 license and available at http://snvphyl.readthedocs.io/ or at https://github.com/phac-nml/snvphyl-galaxy.

Effects of activin and TGFß on p21 in colon cancer.

Activin and TGFß share SMAD signaling and colon cancers can inactivate either pathway alone or simultaneously. The differential effects of activin and TGFß signaling in colon cancer have not been previously dissected. A key downstream target of TGFß signaling is the cdk2 inhibitor p21 (p21(cip1/waf1)). Here, we evaluate activin-specific effects on p21 regulation and resulting functions. We find that TGFß is a more potent inducer of growth suppression, while activin is a more potent inducer of apoptosis. Further, growth suppression and apoptosis by both ligands are dependent on SMAD4. However, activin downregulates p21 protein in a SMAD4-independent fashion in conjunction with increased ubiquitination and proteasomal degradation to enhance migration, while TGFß upregulates p21 in a SMAD4-dependent fashion to affect growth arrest. Activin-induced growth suppression and cell death are dependent on p21, while activin-induced migration is counteracted by p21. Further, primary colon cancers show differential p21 expression consistent with their ACVR2/TGFBR2 receptor status. In summary, we report p21 as a differentially affected activin/TGFß target and mediator of ligand-specific functions in colon cancer, which may be exploited for future risk stratification and therapeutic intervention.

Activin signaling in microsatellite stable colon cancers is disrupted by a combination of genetic and epigenetic mechanisms.

BACKGROUND: Activin receptor 2 (ACVR2) is commonly mutated in microsatellite unstable (MSI) colon cancers, leading to protein loss, signaling disruption, and larger tumors. Here, we examined activin signaling disruption in microsatellite stable (MSS) colon cancers. METHODS: Fifty-one population-based MSS colon cancers were assessed for ACVR1, ACVR2 and pSMAD2 protein. Consensus mutation-prone portions of ACVR2 were sequenced in primary cancers and all exons in colon cancer cell lines. Loss of heterozygosity (LOH) was evaluated for ACVR2 and ACVR1, and ACVR2 promoter methylation by methylation-specific PCR and bisulfite sequencing and chromosomal instability (CIN) phenotype via fluorescent LOH analysis of 3 duplicate markers. ACVR2 promoter methylation and ACVR2 expression were assessed in colon cancer cell lines via qPCR and IP-Western blots. Re-expression of ACVR2 after demethylation with 5-aza-2'-deoxycytidine (5-Aza) was determined. An additional 26 MSS colon cancers were assessed for ACVR2 loss and its mechanism, and ACVR2 loss in all tested cancers correlated with clinicopathological criteria. RESULTS: Of 51 MSS colon tumors, 7 (14%) lost ACVR2, 2 (4%) ACVR1, and 5 (10%) pSMAD2 expression. No somatic ACVR2 mutations were detected. Loss of ACVR2 expression was associated with LOH at ACVR2 (p<0.001) and ACVR2 promoter hypermethylation (p<0.05). ACVR2 LOH, but not promoter hypermethylation, correlated with CIN status. In colon cancer cell lines with fully methylated ACVR2 promoter, loss of ACVR2 mRNA and protein expression was restored with 5-Aza treatment. Loss of ACVR2 was associated with an increase in primary colon cancer volume (p<0.05). CONCLUSIONS: Only a small percentage of MSS colon cancers lose expression of activin signaling members. ACVR2 loss occurs through LOH and ACVR2 promoter hypermethylation, revealing distinct mechanisms for ACVR2 inactivation in both MSI and MSS subtypes of colon cancer.

TGFbeta modulates PTEN expression independently of SMAD signaling for growth proliferation in colon cancer cells.

Signaling pathways enabling transforming growth factor-beta (TGFbeta)'s conversion from a tumor suppressor to a tumor promoter are not well characterized. TGFbeta utilizes intracellular SMADs to mediate growth suppression; however, TGFbeta-induced proliferative pathways may become more apparent when SMAD signaling is abrogated. Here, we determined regulation of the tumor suppressor PTEN by TGFbeta utilizing SMAD4-null colon cancer cells. TGFbeta downregulated PTEN mRNA and simultaneously induced growth proliferation. TGFbeta also induced both SMAD2 and SMAD3 nuclear translocation, but only triggered SMAD2-specific transcriptional activity in the absence of SMAD4. Interference of SMAD2 with DN-SMAD2 enhanced TGFbeta-induced cell proliferation, but downregulation of PTEN expression by TGFbeta was unaffected. TGFbeta increased PI3K tyrosine phosphorylation, and inhibition of PI3K pharmacologically or by DN-p85 transfection reversed both TGFbeta-induced PTEN suppression and TGFbeta-induced cell proliferation. Thus, TGFbeta activates PI3K to downregulate PTEN for enhancement of cell proliferation that is independent of SMAD proteins.

RAS/ERK modulates TGFbeta-regulated PTEN expression in human pancreatic adenocarcinoma cells.

Phosphatase and tensin homolog deleted on chromosome 10 (PTEN) is rarely mutated in pancreatic cancers, but its regulation by transforming growth factor (TGF)-beta might mediate growth suppression and other oncogenic actions. Here, we examined the role of TGFbeta and the effects of oncogenic K-RAS/ERK upon PTEN expression in the absence of SMAD4. We utilized two SMAD4-null pancreatic cell lines, CAPAN-1 (K-RAS mutant) and BxPc-3 (WT-K-RAS), both of which express TGFbeta surface receptors. Cells were treated with TGFbeta1 and separated into cytosolic/nuclear fractions for western blotting with phospho-SMAD2, SMAD 2, 4 phospho-ATP-dependent tyrosine kinases (Akt), Akt and PTEN antibodies. PTEN mRNA levels were assessed by reverse transcriptase-polymerase chain reaction. The MEK1 inhibitor, PD98059, was used to block the downstream action of oncogenic K-RAS/ERK, as was a dominant-negative (DN) K-RAS construct. TGFbeta increased phospho-SMAD2 in both cytosolic and nuclear fractions. PD98059 treatment further increased phospho-SMAD2 in the nucleus of both pancreatic cell lines, and DN-K-RAS further improved SMAD translocation in K-RAS mutant CAPAN cells. TGFbeta treatment significantly suppressed PTEN protein levels concomitant with activation of Akt by 48 h through transcriptional reduction of PTEN mRNA that was evident by 6 h. TGFbeta-induced PTEN suppression was reversed by PD98059 and DN-K-RAS compared with treatments without TGFbeta. TGFbeta-induced PTEN expression was inversely related to cellular proliferation. Thus, oncogenic K-RAS/ERK in pancreatic adenocarcinoma facilitates TGFbeta-induced transcriptional down-regulation of the tumor suppressor PTEN in a SMAD4-independent manner and could constitute a signaling switch mechanism from growth suppression to growth promotion in pancreatic cancers.

Activin type 2 receptor restoration in MSI-H colon cancer suppresses growth and enhances migration with activin.

BACKGROUND & AIMS: Colon cancers with high-frequency microsatellite instability (MSI-H) develop frameshift mutations in tumor suppressors as part of their pathogenesis. ACVR2 is mutated at its exon 10 polyadenine tract in >80% of MSI-H colon cancers, coinciding with loss of protein. ACVR2 transmits the growth effects of activin via phosphorylation of SMAD proteins to affect gene transcription. The functional effect of activin in colon cancers has not been studied. We developed and characterized a cell model in which we studied how activin signaling affects growth. METHODS: hMLH1 and ACVR2 mutant HCT116 cells were previously stably transferred with chromosome 2 (HCT116+chr2), restoring a single regulated copy of wild-type ACVR2 but not hMLH1. Both HCT116+chr2 and parental HCT116 cells (as well as HEC59 and ACVR2 and hMSH2 complemented HEC59+chr2 cells) were assessed for genetic complementation and biologic function. RESULTS: HCT116+chr2 cells and HEC59+chr2 cells, but not ACVR2-mutant HCT116 or HEC59 cells, acquired wild-type ACVR2 as well as expression of ACVR2 wild-type messenger RNA. Complemented ACVR2 protein complexed with ACVR1 with activin treatment, generating nuclear phosphoSMAD2 and activin-specific gene transcription. ACVR2-restored cells showed decreased growth and reduced S phase but increased cellular migration following activin treatment. ACVR2 small interfering RNA reversed these effects in complemented cells. CONCLUSIONS: ACVR2-complemented MSI-H colon cancers restore activin-SMAD signaling, decrease growth, and slow their cell cycle following ligand stimulation but show increased cellular migration. Activin is growth suppressive and enhances migration similar to transforming growth factor beta in colon cancer, indicating that abrogation of the effects of activin contribute to the pathogenesis of MSI-H colon cancers.