Development of a scalable single process for producing SARS-CoV-2 RBD monomer and dimer vaccine antigens.

We have developed a single process for producing two key COVID-19 vaccine antigens: SARS-CoV-2 receptor binding domain (RBD) monomer and dimer. These antigens are featured in various COVID-19 vaccine formats, including SOBERANA 01 and the licensed SOBERANA 02, and SOBERANA Plus. Our approach involves expressing RBD (319-541)-His6 in Chinese hamster ovary (CHO)-K1 cells, generating and characterizing oligoclones, and selecting the best RBD-producing clones. Critical parameters such as copper supplementation in the culture medium and cell viability influenced the yield of RBD dimer. The purification of RBD involved standard immobilized metal ion affinity chromatography (IMAC), ion exchange chromatography, and size exclusion chromatography. Our findings suggest that copper can improve IMAC performance. Efficient RBD production was achieved using small-scale bioreactor cell culture (2 L). The two RBD forms - monomeric and dimeric RBD - were also produced on a large scale (500 L). This study represents the first large-scale application of perfusion culture for the production of RBD antigens. We conducted a thorough analysis of the purified RBD antigens, which encompassed primary structure, protein integrity, N-glycosylation, size, purity, secondary and tertiary structures, isoform composition, hydrophobicity, and long-term stability. Additionally, we investigated RBD-ACE2 interactions, in vitro ACE2 recognition of RBD, and the immunogenicity of RBD antigens in mice. We have determined that both the monomeric and dimeric RBD antigens possess the necessary quality attributes for vaccine production. By enabling the customizable production of both RBD forms, this unified manufacturing process provides the required flexibility to adapt rapidly to the ever-changing demands of emerging SARS-CoV-2 variants and different COVID-19 vaccine platforms.

PEGylation Strategy for Improving the Pharmacokinetic and Antitumoral Activity of the IL-2 No-alpha Mutein.

BACKGROUND: In a previous work, an IL-2Rßγ biased mutant derived from human IL-2 and called IL-2noα, was designed and developed. Greater antitumor effects and lower toxicity were observed compared to native IL-2. Nevertheless, mutein has some disadvantages, such as a very short half-life of about 9-12 min, propensity for aggregation, and solubility problems. OBJECTIVE: In this study, PEGylation was employed to improve the pharmacokinetic and antitumoral properties of the novel protein. METHODS: Pegylated IL-2noα was characterized by polyacrylamide gel electrophoresis, size exclusion chromatography, in vitro cell proliferation and in vivo cell expansion bioassays, and pharmacokinetic and antitumor studies. RESULTS: IL-2noα-conjugates with polyethylene glycol (PEG) of 1.2 kDa, 20 kDa, and 40 kDa were obtained by classical acylation. No significant changes in the secondary and tertiary structures of the modified protein were detected. A decrease in biological activity in vitro and a significant improvement in half-life were observed, especially for IL-2noα-PEG20K. PEGylation of IL-2noα with PEG20K did not affect the capacity of the mutant to induce preferential expansion of T effector cells over Treg cells. This pegylated IL-2noα exhibited a higher antimetastatic effect compared to unmodified IL-2noα in the B16F0 experimental metastases model, even when administered at lower doses and less frequently. CONCLUSION: PEG20K was selected as the best modification strategy, to improve the blood circulation time of the IL-2noα with a superior antimetastatic effect achieved with lower doses.

Efficient capture of recombinant SARS-CoV-2 receptor-binding domain (RBD) with citrate-coated magnetic iron oxide nanoparticles.

Several vaccines against COVID-19 use a recombinant SARS-CoV-2 receptor-binding domain (RBD) as antigen, making the purification of this protein a key step in their production. In this work, citrate-coated magnetic iron oxide nanoparticles were evaluated as nano adsorbents in the first step (capture) of the purification of recombinant RBD. The nanoparticles were isolated through coprecipitation and subsequently coated with sodium citrate. The citrate-coated nanoparticles exhibited a diameter of 10 ± 2 nm, a hydrodynamic diameter of 160 ± 3 nm, and contained 1.9 wt% of citrate. The presence of citrate on the nanoparticles' surface was confirmed through FT-IR spectra and thermogravimetric analysis. The crystallite size (10.1 nm) and the lattice parameter (8.3646 Å) were determined by X-ray diffraction. In parallel, RBD-containing supernatant extracted from cell culture was exchanged through ultrafiltration and diafiltration into the adsorption buffer. The magnetic capture was then optimized using different concentrations of nanoparticles in the purified supernatant, and we found 40 mg mL-1 to be optimal. The ideal amount of nanoparticles was assessed by varying the RBD concentration in the supernatant (between 0.113 mg mL-1 and 0.98 mg mL-1), which resulted in good capture yields (between 83 ± 5% and 94 ± 4%). The improvement of RBD purity after desorption was demonstrated by SDS-PAGE and RP-HPLC. Furthermore, the magnetic capture was scaled up 100 times, and the desorption was subjected to chromatographic purifications. The obtained products recognized anti-RBD antibodies and bound the ACE2 receptor, proving their functionality after the developed procedure.

Rootstock increases the physiological defence of tomato plants against Pseudomonas syringae pv. tomato infection.

Climate change has intensified the infection of tomato plants by pathogens such as Pseudomonas syringae pv. tomato (Pst). Rootstocks may increase plant tolerance to leaf phytopathogens. The aim of this study was to evaluate the effects of the tolerant Poncho Negro (R) tomato rootstock on physiological defence and the role of hydrogen sulfide (H2S) in susceptible Limachino (L) tomato plant responses to Pst attack. Ungrafted (L), self-grafted (L/L), and grafted (L/R) plants were infected with Pst. Rootstock increased the concentration of antioxidant compounds including ascorbate in the scion. Tolerant rootstock induced an increase of H2S in the scion, which correlated with enhanced expression of the SlAPX2 gene. A high accumulation of salicylic acid was observed in Pst-inoculated grafted L/L and L/R plants, but this was higher in L/R plants. The increase of H2S during Pst infection was associated with a reduction of ethylene in L/R plants. Our study indicates that the Poncho Negro rootstock reduced the symptoms of bacterial speck disease in the Limachino tomato plants, conferring tolerance to Pst infection. This study provides new knowledge about the impact of rootstock in the defence of tomato plants against leaf pathogens that could be used in sustainable management of tomato cultivation.

Production of an anti-TNFα antibody in murine myeloma cells by perfusion culture.

Infliximab is a mouse/human chimeric IgG1 monoclonal antibody which recognizes the proinflammatory cytokine, tumor necrosis factor α (TNFα), and inhibits receptor interactions, thereby decreasing inflammation and autoimmune response in patients. This monoclonal antibody has been successfully used to treat rheumatoid arthritis, ankylosing spondylitis, and psoriatic arthritis. However, the high treatment cost limits patient access to this biotherapy. One alternative to this problem is the use of biosimilars. In this work, we describe the stable expression and physicochemical characterization of an anti-TNFα antibody. While infliximab is produced in recombinant murine SP2/0 cells, our anti-TNFα IgG antibody was expressed in recombinant murine NS0 myeloma cells. The best anti-TNFα antibody-expressing clone was selected from three clone candidates based on the stability of IgG expression levels, specific productivity as well as TNFα-binding activity compared to commercial infliximab. Our results indicate that the selected cell clone, culture medium, and fermentation mode allowed for the production of an anti-TNFα antibody with similar characteristics to the reference commercially available product. An optimization of the selected culture medium by metabolomics may increase the volumetric productivity of the process to satisfy the demand for this product. Further experiments should be performed to evaluate the biological properties of this anti-TNFα antibody. KEY POINTS: • An anti-TNFα antibody was produced in NS0 cells using perfusion culture. • A proprietary chemically defined culture medium was used to replace commercially available protein-free medium. • The purified anti-TNFα antibody was comparable to the reference marketed product.

Vacunación por elección contra COVID-19 por la comunidad mexicana / Vaccination by election against COVID-19 by the mexican community

La pandemia por COVID-19 es la crisis de salud mundial que define nuestro tiempo, es el mayor desafío que se ha enfrentado en mucho tiempo. Es por esto que las organizaciones de salud trabajan duro para encontrar una vacuna que reduzca el impacto de este virus. Objetivo: Describir la opinión del público sobre la vacuna COVID-19 para generar un panorama del conocimiento de la comunidad, del impacto que esta tendrá, así como de las áreas de salud preventiva que servirán de apoyo para que la comunidad esté convencida de aplicarse la vacuna. Método: Se realizó un estudio observacional descriptivo de carácter transversal mediante la aplicación de una encuesta en Google forms a hombres y mujeres de los estados de Hidalgo, Guerrero, Puebla y México a 150 personas con un rango de edad de 15-80 años. Resultados: El 29.33% no se se aplicaría la vacuna contra COVID-19 principalmente por el miedo a consecuencias adversas y la falta de conocimiento, en cambio un 70.67% indicó que si se la aplicaría debido a que confían en los avances de la ciencia en el área de la salud. Conclusión: Es necesario implementar más conocimiento en la población mexicana para generar interés sobre los avances de la salud, así como lo es la vacuna para el covid-19, de lo contrario los resultados serán negativos.(AU)

The COVID-19 pandemic is the defining global health crisis of our time, it is the greatest challenge that has been faced in a long time. This is why health organizations work hard to find a vaccine that reduces the impact of this virus.Objective: Describe the opinion of the public about the COVID-19 vaccine to generate an overview of the community's knowledge, the impact it will have, as well as the preventive health areas that will support the community to be convinced of applying the vaccine. Method: A descriptive, cross-sectional observational study was carried out by applying a survey in Google forms to men and women from the states of Hidalgo, Guerrero, Puebla and Mexico to 150 people with an age range of 15-80 years. Results: 29.33% would not apply the vaccine against COVID-19 mainly due to fear of adverse consequences and lack of knowledge, on the other hand 70.67% indicated that they would apply it because they trust the advances of science in the area Of the health. Conclusion: It is necessary to implement more knowledge in the Mexican population to generate interest in health advances, as is the vaccine for covid-19, otherwise the results will be negative.(AU)

Microbial Diversity of Psychrotolerant Bacteria Isolated from Wild Flora of Andes Mountains and Patagonia of Chile towards the Selection of Plant Growth-Promoting Bacterial Consortia to Alleviate Cold Stress in Plants.

Cold stress decreases the growth and productivity of agricultural crops. Psychrotolerant plant growth-promoting bacteria (PGPB) may protect and promote plant growth at low temperatures. The aims of this study were to isolate and characterize psychrotolerant PGPB from wild flora of Andes Mountains and Patagonia of Chile and to formulate PGPB consortia. Psychrotolerant strains were isolated from 11 wild plants (rhizosphere and phyllosphere) during winter of 2015. For the first time, bacteria associated with Calycera, Orites, and Chusquea plant genera were reported. More than 50% of the 130 isolates showed ≥33% bacterial cell survival at temperatures below zero. Seventy strains of Pseudomonas, Curtobacterium, Janthinobacterium, Stenotrophomonas, Serratia, Brevundimonas, Xanthomonas, Frondihabitans, Arthrobacter, Pseudarthrobacter, Paenarthrobacter, Brachybacterium, Clavibacter, Sporosarcina, Bacillus, Solibacillus, Flavobacterium, and Pedobacter genera were identified by 16S rRNA gene sequence analyses. Ten strains were selected based on psychrotolerance, auxin production, phosphate solubilization, presence of nifH (nitrogenase reductase) and acdS (1-aminocyclopropane-1-carboxylate (ACC) deaminase) genes, and anti-phytopathogenic activities. Two of the three bacterial consortia formulated promoted tomato plant growth under normal and cold stress conditions. The bacterial consortium composed of Pseudomonas sp. TmR5a & Curtobacterium sp. BmP22c that possesses ACC deaminase and ice recrystallization inhibition activities is a promising candidate for future cold stress studies.

Efecto de una Intervención Educativa en la presencia de ulceras vasculares en pacientes con Diabetes Mellitus / Effect of an Educational Intervention on the presence of Vascular Ulcers in patients with Diabetes Mellitus

Introducción: La diabetes mellitus se está convirtiendo en un problema de magnitud global, afectando la vida de las personas que la padecen, en específico cuando no se logra un control adecuado de la misma, bajo esta premisa, se ha considerado que la educación en diabetes es una buena herramienta para favorecer el cambio de conducta de las personas que la padecen y con esto establecer estilos de vida saludables.Objetivo: Identificar el efecto de una intervención educativa en el autocuidado de la diabetes mellitus y sus complicaciones, a través de la revisión de evidencia bibliográfica.Material y métodos: Se realizó una revisión sistemática de la literatura actual sobre las intervenciones en educación tanto en México como en las Américas, que han logrado tener efecto en el conocimiento de la misma enfermedad, en las complicaciones de la misma, en el autocontrol y automonitoreo, así como en el cambio de estilos de vida y la adherencia al tratamiento. Se eligieron estudios en buscadores como Crossref Metadata, revistas CONACYT, PubMed, Latindex y Elsevier.Resultados: Se comprobó que existe un efecto en los participantes de las intervenciones educativas, porque han logrado disminuir sus niveles de glucosa plasmática, así como conocer las características de la enfermedad y con ello conocen sobre el tratamiento y las condiciones físicas para evitar complicaciones de la enfermedad como lo son las ulceras vasculares, por lo que modifican sus estilos de vida.Conclusiones: Una intervención educativa es necesaria porque con ella se logró mayor conocimiento sobre los factores de riesgo, un mejor control metabólico, lo que conllevara a un efecto directo en la calidad de vida de las personas que padecen la enfermedad, esto significa que los resultados no son negativos.(AU)

Introduction: Diabetes mellitus is becoming into a global magnitude problem, affecting the life of people who suffer it, especially when a suitable control is not achieved, diabetes education has been considered a good tool to change persons conduct and establish healthy life styles.Objective: Identify the effect of an educational intervention about self-care in diabetes mellitus and its complications, through the review of bibliographic evidence.Material and methods: A bibliographic review of the current literature on educational interventions in Mexico and the Americas was carried out which provide information on the effect that they achieved in the management , knowledge effect in people with the disease, complications, self-control and self-monitoring impact of the disease, the people lifestyles changes and adherence to treatment. Search engine studies such as Crossref Metadata, CONACyT magazines, PubMed, Latindex and Elsevier were chosen.Results: a change has been demonstrated in people who participated in educational interventions, because they have been reduce their plasma glucose levels, get more information about the characteristics of the disease, treatment and physical conditions to avoid complications of the disease such as vascular ulcers, so they modify their lifestyles.Conclusions: A diabetes educational intervention is necessary because increase knowledge about risk factors of the disease and favors a better metabolic control, which lead a direct effect on the people with diabetes lifestyles. This means that the results are not negative.(AU)

Consenso Colombiano para la prevención, diagnóstico y manejo de condiciones trombóticas en pacientes adultos con COVID-19: aplicando el Marco de la Evidencia a la Decisión (EtD) GRADE / Colombian consensus for the prevention, diagnosis and treatment of thrombotic conditions in adult patients with COVID-19: applying GRADE Evidence to Decision (EtD) Frameworks

resumen está disponible en el texto completo

Abstract Recent studies have reported the occurrence of thrombotic phenomena or coagulopathy in patients with COVID-19. There are divergent positions regarding the prevention, diagnosis, and treatment of these phenomena, and current clinical practice is based solely on deductions by extension from retrospective studies, case series, observational studies, and international guidelines developed prior to the pandemic. In this context, the aim was to generate a group of recommendations on the prevention, diagnosis and management of thrombotic complications associated with COVID-19. Methods: A rapid guidance was carried out applying the GRADE Evidence to Decision (EtD) frameworks and an iterative participation system, with statistical and qualitative analysis. Results: 31 clinical recommendations were generated focused on: a) Coagulation tests in symptomatic adults with suspected infection or confirmed SARS CoV-2 infection; b) Thromboprophylaxis in adults diagnosed with COVID-19 (Risk scales, thromboprophylaxis for outpatient, in-hospital management, and duration of thromboprophylaxis after discharge from hospitalization), c) Diagnosis and treatment of thrombotic complications, and d) Management of people with previous indication of anticoagulant agents. Conclusions: Recommendations of this consensus guide clinical decision-making regarding the prevention, diagnosis, and treatment of thrombotic phenomena in patients with COVID-19, and represent an agreement that will help decrease the dispersion in clinical practices according to the challenge imposed by the pandemic.

Chemical Composition, Antioxidant and Anticancer Activities of Leptocarpha rivularis DC Flower Extracts.

An evaluation of antioxidant and anticancer activity was screened in Leptocarpha rivularis DC flower extracts using four solvents (n-hexane (Hex), dichloromethane (DCM), ethyl acetate (AcOEt), and ethanol (EtOH)). Extracts were compared for total extract flavonoids and phenol contents, antioxidant activity (2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), ferric reducing antioxidant potential (FRAP), total reactive antioxidant properties (TRAP) and oxygen radical absorbance capacity (ORAC)) across a determined value of reduced/oxidized glutathione (GSH/GSSG), and cell viability (the sulforhodamine B (SRB) assay). The most active extracts were analyzed by chromatographic analysis (GC/MS) and tested for apoptotic pathways. Extracts from Hex, DCM and AcOEt reduced cell viability, caused changes in cell morphology, affected mitochondrial membrane permeability, and induced caspase activation in tumor cell lines HT-29, PC-3, and MCF-7. These effects were generally less pronounced in the HEK-293 cell line (nontumor cells), indicating clear selectivity towards tumor cell lines. We attribute likely extract activity to the presence of sesquiterpene lactones, in combination with other components like steroids and flavonoids.

Effect of high temperatures on teeth fixed with an orthodontic bracket. An in vitro study.

In cases of incineration, where the identification of victims by conventional methods is impossible, teeth and dental materials become an available source of evidence - thanks to their ability to resist high temperatures. Currently, there is no experimental research to determine the behavior of the bracket and the dental structure when it is attached to the tooth after being exposed to high temperatures. An experimental in vitro study was carried out on 60 upper and lower premolar teeth (30 with a cemented bracket and 30 teeth without a bracket) that were exposed to six temperature ranges (200 °C, 400 °C, 600 °C, 800 °C, 1000 °C and 1200 °C). With the help of an Optiks® binocular stereomicroscope, macroscopic changes were described for each temperature; in addition, the mesio-distal diameter of the crown of the teeth and the bracket were measured before and after the incineration. A comparison of the diameters between the groups [the experimental group and the control group] and between the different temperatures was made. Both teeth, with brackets and without brackets showed macroscopic changes, such as fissures, cracks, fragmentation, enamel bursting and enamel and dentin separation according to the temperature range exposed. The detachment of the bracket was presented at 600 °C - leaving a mark on the dental surface. At above 1000 °C, the crown of the tooth with bracket acquired a pink color and the surface of the bracket became rough and opaque. At above 400 °C, both groups experienced an expansion in the crown dimension without significant differences. The results of this research suggest that teeth with attached brackets are equally resistant as the teeth without brackets, and that the adhesion of these attachments to the dental structure leave characteristics that allow them to be differentiated from teeth without braces -as well as suggesting the orientation to the temperature exposure.

Afecto negativo como mediador entre intolerancia a la incertidumbre, ansiedad y depresión / Negative affect as a mediator between intolerance to uncertainty, anxiety and depression

El modelo transdiagnóstico ha demostrado su relevancia en la comprensión y tratamiento de los trastornos emocionales que comparten procesos cognitivos y conductuales. El objetivo de esta investigación es analizar el papel mediador del afecto negativo (AN) en la relación entre la intolerancia a la incertidumbre (II) y los síntomas ansiosos y depresivos siguiendo un modelo transdiagnóstico. Se contó con una muestra de 506 participantes de edades entre 18 y 64 años (M = 31.9, DE = 11.26); los niveles de AN como estado y rasgo mediante la escala de afecto positivo y negativo (PANAS), la II con la escala de intolerancia a la incertidumbre, y los síntomas ansiosos y depresivos con los inventarios BAI y BDI-II. Los resultados señalan que las medidas afectivas de AN estado y rasgo, y cognitivas como la incertidumbre de inhibición y desconcierto correlacionaron con síntomas de ansiedad y depresión. Además, las medidas cognitivas y sus dimensiones, al asociarse con las medidas de depresión y ansiedad, disminuyeron significativamente controlando el AN, obteniendo un modelo mediador para las medidas afectivas y cognitivas en ansiedad y depresión en un modelo de senderos ajustado según los análisis de mediación y efectos indirectos en un modelo de ecuaciones estructurales. Se concluye que el AN estado y rasgo se considera un factor mediador entre la II y los síntomas de ansiedad de forma total y parcial con la depresión. Se genera nueva evidencia del papel del afecto como rasgo y estado en el planteamiento de un modelo transdiagnóstico en el área de la intervención clínica

The transdiagnostic model has demonstrated its relevance in the understanding and treatment of emotional disorders that share cognitive and behavioral processes. The objective of this research is to analyze the mediating role of negative affect (NA) in the relationship between intolerance to uncertainty (IU) and anxious and depressive symptoms following a transdiagnostic model. The sample consisted of 506 participants aged between 18 and 64 years (M = 31.9, SD = 11.26); the levels of NA as state and trait was obtained using the scale of positive and negative affect (PANAS), the IU with the scale of intolerance to uncertainty, and the anxious and depressive symptoms with the BAI and BDI-II. The results indicate that affective measures of state and trait NA, and cognitive measures such as uncertainty of inhibition and bewilderment correlated with symptoms of anxiety and depression. In addition, the cognitive measures and their dimensions when associated with measures of depression and anxiety decreased significantly controlling the AN, obtaining a mediating model for the affective and cognitive measures in anxiety and depression in a path model adjusted according to the mediation and indirect effects analyses in a model of structural equations. It is concluded that the state and trait NA is considered a mediating factor between IU and anxiety symptoms in a total and partial way with depression. New evidence of the role of affect is generated as a trait and state in the approach of a transdiagnostic model in the area of clinical intervention

Prevalence of breast and ovarian cancer subtypes in Hispanic populations from Puerto Rico.

BACKGROUND: Previous epidemiological studies aimed at describing characteristics of breast (BC) and ovarian cancer (OC) patients tend to examine Hispanic populations using a mix of individuals that come from ethnically different Hispanic backgrounds. Since most USA cancer statistics do not include cancer data from Puerto Rico (PR), there is a lack of historical and descriptive data analysis for Hispanic women in the island that suffer from these diseases. Therefore, the aim of our study is to provide a comprehensive clinicopathological characterization of BC and OC cases in PR. METHODS: Our study consisted of a longitudinal retrospective review of archived pathology reports at Southern Pathology Services (SPS), which mostly serves southwestern PR, from years 2000-2015. After filtering SPS records with pre-established criteria, tumor samples from 3451 BC and 170 OC cases were used for descriptive statistics and analysis using R program. RESULTS: In our cohort, the mean age of diagnosis for BC was 60.5 years and 60.3 years for OC. Available data for subtype characterization from BC cases, exhibited an expected subtype distribution that remained stable over time (Luminal A = 68.8%, Luminal B = 9.7%, HER-2 = 6.1% and Triple negative = 15.4%). Additionally, tumor grades distribution varied within different BC subtypes in which the majority of Luminal A tumors were G2 and most Triple negative tumors were G3. For OC cases, available subtype and tumor grade information identified serous histology in 64.71% of all cases and G3 as being the most prevalent tumor grade. Pathology reports revealed that 39.42% of all OC cases were described as late stage, while 50.5% as early stage (by pathological staging). CONCLUSION: Our data suggests that OC and BC subtypes distribution in Hispanic populations from PR are in-line with national averages. In a significant number of BC cases, subtype could not be determined due to study limitations, health insurance coverage, or other reasons described here and may constitute a health disparity. Altogether, and despite these gaps, this study represents one of the most complete reviews of BC and OC in PR and provides an opportunity to further study this population separate from other US Hispanic populations.

Biopesticide Activity from Drimanic Compounds to Control Tomato Pathogens.

Tomato crops can be affected by several infectious diseases produced by bacteria, fungi, and oomycetes. Four phytopathogens are of special concern because of the major economic losses they generate worldwide in tomato production; Clavibacter michiganensis subsp. michiganensis and Pseudomonas syringae pv. tomato, causative agents behind two highly destructive diseases, bacterial canker and bacterial speck, respectively; fungus Fusarium oxysporum f. sp. lycopersici that causes Fusarium Wilt, which strongly affects tomato crops; and finally, Phytophthora spp., which affect both potato and tomato crops. Polygodial (1), drimenol (2), isonordrimenone (3), and nordrimenone (4) were studied against these four phytopathogenic microorganisms. Among them, compound 1, obtained from Drimys winteri Forst, and synthetic compound 4 are shown here to have potent activity. Most promisingly, the results showed that compounds 1 and 4 affect Clavibacter michiganensis growth at minimal inhibitory concentrations (MIC) values of 16 and 32 µg/mL, respectively, and high antimycotic activity against Fusarium oxysporum and Phytophthora spp. with MIC of 64 µg/mL. The results of the present study suggest novel treatment alternatives with drimane compounds against bacterial and fungal plant pathogens.

Structure-Activity Relationship of Dialkoxychalcones to Combat Fish Pathogen Saprolegnia australis.

To investigate the anti-Saprolegnia activities of chalconic compounds, nine dialkoxychalcones 2⁻10, along with their key building block 2',4'-dihydroxychalcone 1, were evaluated for their potential oomycide activities against Saprolegnia australis strains. The synthesis afforded a series of O-alkylated derivatives with typical chalcone skeletons. Compounds 4⁻10 were reported for the first time. Interestingly, analogue 8 with the new scaffold demonstrated remarkable in vitro growth-inhibitory activities against Saprolegnia strains, displaying greater anti-oomycete potency than the standard drugs used in the assay, namely fluconazole and bronopol. In contrast, a dramatic loss of activity was observed for O-alkylated derivatives 2, 3, 6, and 7. These findings have highlighted the therapeutic potential of the natural compound 1 scaffold to be exploitable as a drug lead with specific activity against various Saprolegnia strains.

Sensitivity of ovarian cancer cells to acetaminophen reveals biological pathways that affect patient survival.

Experimental and epidemiological data support the potential activity of acetaminophen against ovarian cancer (OVCA). In this study, we sought to confirm the activity of acetaminophen in OVCA cell lines and to investigate the molecular basis of response. A total of 16 OVCA cell lines underwent pretreatment (baseline) genome-wide expression measurements and were then treated with and analyzed for acetaminophen sensitivity. Pearson's correlation analysis was performed to identify genes that were associated with OVCA acetaminophen response. The identified genes were subjected to pathway analysis, and the expression of each represented pathway was summarized using principal component analysis. OVCA acetaminophen response pathways were analyzed in 4 external clinico-genomic datasets from 820 women for associations with overall survival from OVCA. Acetaminophen exhibited antiproliferative activity against all tested OVCA cell lines, with half maximal inhibitory concentration values ranging from 63.2 to 403 µM. Pearson's correlation followed by biological pathway analysis identified 13 pathways to be associated with acetaminophen sensitivity (P<0.01). Associations were observed between patient survival from OVCA and expression of the following pathways: Development/angiotensin signaling via ß-arrestin (P=0.04), protein folding and maturation/angiotensin system maturation (P=0.02), signal transduction/c-Jun N-terminal kinase (JNK) pathway (P=0.03) and androstenedione and testosterone biosynthesis and metabolism (P=0.02). We confirmed that acetaminophen was active against OVCA cells in vitro. Furthermore, we identified 4 molecular signaling pathways associated with acetaminophen response that may also affect overall survival in women with OVCA, including the JNK pathway, which has been previously implicated in the mechanism of action of acetaminophen and is predictive of decreased survival in women with OVCA.

Micro-RNAs associated with the evolution of ovarian cancer cisplatin resistance.

OBJECTIVES: Ovarian cancer (OVCA) is the leading cause of mortality among women with gynecologic malignancy, in part due to the development of chemoresistance. We sought to identify micro-RNAs (miRNAs) associated with in vitro development of OVCA chemoresistance that may also represent potential targets for therapy. METHODS: In this study, four OVCA cell lines (A2780CP, A2780S, IGROV1, and OVCAR5) were serially treated with cisplatin in parallel with measurements of miRNA expression changes. RESULTS: Nine miRNAs were found to be associated with increasing cisplatin resistance (IC50) (p<0.01); however, only 5 of these miRNAs have publically available information. Pathway analysis identified 15 molecular signaling pathways that were represented by genes predicted to be targets of the 5 miRNAs (false discovery rate<0.05), 11 of which are associated with the epithelial-mesenchymal transition (EMT). Further analysis identified 2 of those pathways as being associated with overall survival in 218 patients with OVCA. CONCLUSIONS: Collectively, this panel of miRNAs associated with in vitro evolution of OVCA cisplatin resistance and the pathways identified to be associated with EMT and overall patient survival provide a framework for further investigations into EMT as a therapeutic target in patients with OVCA.

Hemorragia alveolar oculta en paciente con sarcoma de Kaposi broncopulmonar / Occult alveolar hemorrhage in patient with bronchopulmonary Kaposi sarcoma

No disponible

The Chinese herb polyphyllin D sensitizes ovarian cancer cells to cisplatin-induced growth arrest.

PURPOSE: We evaluated the effects of polyphyllin D (PD), a natural compound with anti-neoplastic activity and a major component of the Chinese herb Paris polyphylla, on ovarian cancer (OVCA) cell line proliferation and platinum sensitivity. METHODS: A panel of 20 OVCA cell lines was subjected to PD treatment, MTS proliferation assays, and determination of IC50. Pre-treatment, baseline genome-wide Affymetrix expression analysis was performed on each cell line, and Pearson's correlation was performed to identify genes associated with OVCA PD sensitivity. Twelve cell lines were treated with PD with and without cisplatin, and the effects of PD on cisplatin IC50 were quantified. Genes associated with OVCA PD sensitivity were evaluated for associations with survival in a publically available clinico-genomic dataset of 218 patients with OVCA. RESULTS: Our results showed that PD exhibited anti-proliferative effects against all OVCA cell lines tested, with IC50 values ranging from 0.2 to 1.4 µm. Furthermore, in all cell lines, PD treatment significantly decreased cisplatin IC50 (mean IC50 reduction of 2.1 µm; P < 0.02). Pearson's correlation test identified 25 probe sets, representing 18 unique genes to be associated with PD sensitivity (FDR = 0). We found that one of these genes was associated with overall survival in women with OVCA: CLDN4 (P = 0.014). CONCLUSION: Our findings highlight the value of PD as a natural product with anti-cancer properties, which may also enhance the activity of existing therapeutic agents.