Poly(allylamine)-tripolyphosphate Ionic Assemblies as Nanocarriers: Friend or Foe?

Designing effective drug nanocarriers that are easy to synthesize, robust, and nontoxic is a significant challenge in nanomedicine. Polyamine-multivalent molecule nanocomplexes are promising drug carriers due to their simple and all-aqueous manufacturing process. However, these systems can present issues of colloidal instability over time and cellular toxicity due to the cationic polymer. In this study, we finely modulate the formation parameters of poly(allylamine-tripolyphosphate) complexes to jointly optimize the robustness and safety. Polyallylamine was ionically assembled with tripolyphosphate anions to form liquid-like nanocomplexes with a size of around 200 nm and a zeta potential of -30 mV. We found that nanocomplexes exhibit tremendous long-term stability (9 months of storage) in colloidal dispersion and that they are suitable as protein-loading agents. Moreover, the formation of nanocomplexes induced by tripolyphosphate anions produces a switch-off in the toxicity of the system by altering the overall charge from positive to negative. In addition, we demonstrate that nanocomplexes can be internalized by bone-marrow-derived macrophage cells. Altogether, these nanocomplexes have attractive and promising properties as delivery nanoplatforms for potential therapies based on the immune system activation.

Cytotoxic Screening and Enhanced Anticancer Activity of Lippia alba and Clinopodium nepeta Essential Oils-Loaded Biocompatible Lipid Nanoparticles against Lung and Colon Cancer Cells.

Plant and herbal essential oils (EOs) offer a wide range of pharmacological actions that include anticancer effects. Here, we evaluated the cytotoxic activity of EO from Lippia alba (chemotype linalool), L. alba (chemotype dihydrocarvone, LaDEO), Clinopodium nepeta (L.) Kuntze (CnEO), Eucalyptus globulus, Origanum × paniculatum, Mentha × piperita, Mentha arvensis L., and Rosmarinus officinalis L. against human lung (A549) and colon (HCT-116) cancer cells. The cells were treated with increasing EO concentrations (0-500 µL/L) for 24 h, and cytotoxic activity was assessed. LaDEO and CnEO were the most potent EOs evaluated (IC50 range, 145-275 µL/L). The gas chromatography-mass spectrometry method was used to determine their composition. Considering EO limitations as therapeutic agents (poor water solubility, volatilization, and oxidation), we evaluated whether LaDEO and CnEO encapsulation into solid lipid nanoparticles (SLN/EO) enhanced their anticancer activity. Highly stable spherical SLN/LaDEO and SLN/CnEO SLN/EO were obtained, with a mean diameter of 140-150 nm, narrow size dispersion, and Z potential around -5mV. EO encapsulation strongly increased their anticancer activity, particularly in A549 cells exposed to SLN/CnEO (IC50 = 66 µL/L CnEO). The physicochemical characterization, biosafety, and anticancer mechanisms of SLN/CnEO were also evaluated in A549 cells. SLN/CnEO containing 97 ± 1% CnEO was highly stable for up to 6 months. An increased in vitro CnEO release from SLN at an acidic pH (endolysosomal compartment) was observed. SLN/CnEO proved to be safe against blood components and non-toxic for normal WI-38 cells at therapeutic concentrations. SLN/CnEO substantially enhanced A549 cell death and cell migration inhibition compared with free CnEO.

Efectividad de la administración de hierro en forma semanal y diaria para la prevención de anemia por deficiencia de hierro en lactantes / Effectiveness of weekly and daily iron administration for the prevention of iron deficiency anemia in infants

Introducción. La deficiencia de hierro (DH) es la carencia nutricional más prevalente y la principal causa de anemia en lactantes. Existe consenso en la suplementación diaria con hierro como estrategia de prevención; también se demostró que la suplementación semanal es eficaz, pero la evidencia en lactantes es escasa. El objetivo fue comparar la efectividad de la administración diaria de hierro frente a la semanal para la prevención de la anemia por DH del lactante. Población y métodos. Ensayo clínico controlado y aleatorizado. Lactantes atendidos en un centro de salud público, sin anemia a los 3 meses de edad, aleatorizados en tres grupos: suplementación diaria (1 mg/kg/día), semanal (4 mg/kg/semana) o sin suplementación (grupo control con lactancia materna exclusiva [LME]). Se evaluó anemia y DH a los 3 y 6 meses. Se registró grado de adherencia y efectos adversos. Los datos se analizaron con el software R versión 4.0.3. Resultados. Participaron 227 lactantes. A los 6 meses el grupo de lactantes con LME sin suplementación (control) presentó prevalencias de DH y anemia por DH (ADH) mayores que los grupos intervenidos (diario y semanal). DH: 40,5 % vs. 13,5 % y 16,7 % (p = 0,002); ADH: 33,3 % vs. 7,8 % y 10 % (p < 0,001). No hubo diferencias entre los grupos diario y semanal. Tampoco hubo diferencias en el porcentaje de alta adherencia a la suplementación (50,6 % diaria vs. 57,1 % semanal), ni en los efectos adversos. Conclusiones. No se hallaron diferencias significativas en la efectividad entre la administración diaria y semanal para la prevención de ADH del lactante.

Introduction. Iron deficiency (ID) is the most prevalent nutritional deficiency and the main cause of anemia in infants. There is consensus on daily iron supplementation as a preventive strategy; and weekly iron supplementation has also been shown to be effective, but evidence in infants is scarce. The objective of this study was to compare the effectiveness of daily versus weekly iron administration for the prevention of ID anemia (IDA) in infants. Population and methods. Randomized, controlled clinical trial. Infants seen at a public health center, without anemia at 3 months of age, were randomized into 3 groups: daily supplementation (1 mg/kg/ day), weekly supplementation (4 mg/kg/week), or no supplementation (control group with exclusive breastfeeding [EB]). Anemia and ID were assessed at 3 and 6 months old. Adherence and adverse events were recorded. Data were analyzed using the R software, version 4.0.3. Results. A total of 227 infants participated. At 6 months, the group of infants with EB without supplementation(control) had a higher prevalence of ID and IDA than the intervention groups (daily and weekly). ID: 40.5% versus 13.5% and 16.7% (p = 0.002); IDA: 33.3% versus 7.8% and 10% (p < 0.001). There were no differences between the daily and weekly supplementation groups. There were also no differences in the percentage of high adherence to supplementation (50.6% daily versus 57.1% weekly) or adverse events. Conclusions. No significant differences in effectiveness were observed between daily and weekly administration for the prevention of infant IDA.

Preliminary transcriptomic analysis of peripheral blood from German Shepherd dogs with degenerative joint disease for the identification of diagnostic biomarkers.

OBJECTIVE: Degenerative joint disease (DJD) includes a group of disorders characterised by the deterioration of the articular cartilage. In this study, we investigated the transcriptomic profile of peripheral blood in German Shepherd dogs with DJD to identify putative diagnostic biomarkers. METHODS: Differential gene expression (DGE) and gene ontology (GO) analyses of the bulk RNA-seq experiment were performed in a cohort of 12 adult dogs (five cases and seven controls, classified by clinical and radiographic analyses). RESULTS: Radiographs of cases revealed severe signs of progressive DJD. Two up-regulated (LOC106559672 and THBS4) and one down-regulated (LOC106559235) differentially expressed genes (adjusted p value < 0.05) were identified. The DGE with log2 fold change < -1.5 and > 1.5 and non-adjusted p < 0.01 were selected for GO analysis. No significant enrichment terms were observed in the selected threshold. CONCLUSION: The gene-encoding protein THBS4 is correlated with DJD severity and long noncoding RNA LOC106559235 is probably involved in the DJD process. The THBS4 gene should be considered a good biomarker for DJD in dogs. Future studies using independent cohorts will be necessary to validate the present results.

Effectiveness of weekly and daily iron administration for the prevention of iron deficiency anemia in infants. / Efectividad de la administración de hierro en forma semanal y diaria para la prevención de anemia por deficiencia de hierro en lactantes.

Introduction. Iron deficiency (ID) is the most prevalent nutritional deficiency and the main cause of anemia in infants. There is consensus on daily iron supplementation as a preventive strategy; and weekly iron supplementation has also been shown to be effective, but evidence in infants is scarce. The objective of this study was to compare the effectiveness of daily versus weekly iron administration for the prevention of ID anemia (IDA) in infants. Population and methods. Randomized, controlled clinical trial. Infants seen at a public health center, without anemia at 3 months of age, were randomized into 3 groups: daily supplementation (1 mg/kg/day), weekly supplementation (4 mg/kg/week), or no supplementation (control group with exclusive breastfeeding [EB]). Anemia and ID were assessed at 3 and 6 months old. Adherence and adverse events were recorded. Data were analyzed using the R software, version 4.0.3. Results. A total of 227 infants participated. At 6 months, the group of infants with EB without supplementation (control) had a higher prevalence of ID and IDA than the intervention groups (daily and weekly). ID: 40.5% versus 13.5% and 16.7% (p = 0.002); IDA: 33.3% versus 7.8% and 10% (p < 0.001). There were no differences between the daily and weekly supplementation groups. There were also no differences in the percentage of high adherence to supplementation (50.6% daily versus 57.1% weekly) or adverse events. Conclusions. No significant differences in effectiveness were observed between daily and weekly administration for the prevention of infant IDA.

Introducción. La deficiencia de hierro (DH) es la carencia nutricional más prevalente y la principal causa de anemia en lactantes. Existe consenso en la suplementación diaria con hierro como estrategia de prevención; también se demostró que la suplementación semanal es eficaz, pero la evidencia en lactantes es escasa. El objetivo fue comparar la efectividad de la administración diaria de hierro frente a la semanal para la prevención de la anemia por DH del lactante. Población y métodos. Ensayo clínico controlado y aleatorizado. Lactantes atendidos en un centro de salud público, sin anemia a los 3 meses de edad, aleatorizados en tres grupos: suplementación diaria (1 mg/kg/día), semanal (4 mg/kg/semana) o sin suplementación (grupo control con lactancia materna exclusiva [LME]). Se evaluó anemia y DH a los 3 y 6 meses. Se registró grado de adherencia y efectos adversos. Los datos se analizaron con el software R versión 4.0.3. Resultados. Participaron 227 lactantes. A los 6 meses el grupo de lactantes con LME sin suplementación (control) presentó prevalencias de DH y anemia por DH (ADH) mayores que los grupos intervenidos (diario y semanal). DH: 40,5 % vs. 13,5 % y 16,7 % (p = 0,002); ADH: 33,3 % vs. 7,8 % y 10 % (p < 0,001). No hubo diferencias entre los grupos diario y semanal. Tampoco hubo diferencias en el porcentaje de alta adherencia a la suplementación (50,6 % diaria vs. 57,1 % semanal), ni en los efectos adversos. Conclusiones. No se hallaron diferencias significativas en la efectividad entre la administración diaria y semanal para la prevención de ADH del lactante.

Oxidative Stress and Genomic Damage Induced In Vitro in Human Peripheral Blood by Two Preventive Treatments of Iron Deficiency Anemia.

Iron deficiency is the most prevalent nutritional deficiency and the main cause of anemia worldwide. Since children aged 6-24 months are among the most vulnerable groups at risk, daily supplementation with ferrous sulfate is recommended by the Argentine Society of Pediatrics as preventive treatment of anemia. However, a single weekly dose would have fewer adverse side effects and has been therefore proposed as an alternative treatment. Ferrous sulfate is known by its pro-oxidative properties, which may lead to increased oxidative stress as well as lipid, protein, and DNA damage. We analyzed the effect of daily and weekly preventive treatment of iron deficiency anemia (IDA) on cell viability, oxidative stress, chromosome, and cytomolecular damage in peripheral blood cultured in vitro. The study protocol included the following: untreated negative control; bleomycin, hydrogen peroxide, or ethanol-treated positive control; daily 0.14 mg ferrous sulfate-supplemented group; and weekly 0.55 mg ferrous sulfate-supplemented group. We assessed cell viability (methyl-thiazolyl-tetrazolium and neutral red assays), lipid peroxidation (thiobarbituric acid reactive substances assay), antioxidant response (superoxide dismutase and catalase enzyme analysis), chromosome damage (cytokinesis-blocked micronucleus cytome assay), and cytomolecular damage (comet assay). Lipid peroxidation, antioxidant response, and chromosome and cytomolecular damage decreased after weekly ferrous sulfate supplementation (p < 0.05), suggesting less oxygen free radical production and decreased oxidative stress and genomic damage. Such a decrease in oxidative stress and genomic damage in vitro positions weekly supplementation as a better alternative for IDA treatment. Further studies in vivo would be necessary to corroborate whether weekly supplementation could improve IDA preventive treatment compliance in children.

Evaluation of resveratrol radiomodifying potential for radiotherapy treatment.

Radiotherapy is widely used for cancer treatment. However, its adverse effects that may develop during the course of treatment have forced to search agents to protect biological systems against the deleterious effects of radiation. Resveratrol (3,4,5-trihydroxy-trans-stilbene; RSV) is a natural polyphenol currently promoted for its beneficial pleiotropic effects on health, which has been shown to exhibit antioxidant properties while inhibiting the growth of tumor cells. In radioresistant tumors, RSV could contribute to reduce recurrence and treatment failure. We evaluated the radiomodulatory and genotoxic effects of RSV in CHO-k1 and A549 cell lines and in peripheral human blood lymphocytes through both conventional and hypofractionated protocols, due to the widespread use of hypofractionation in recent years. RSV genotoxic and cytotoxic action was assessed at 15 and 60 µM concentrations with the comet and the MTT assay and in cell proliferation experiments. Our results show that RSV administration to tumor cells at a dose of 60 µM exerted a genotoxic effect and that this concentration also had the capacity of modulating the cytomolecular damage induced by 4 and 16 Gy. These doses are delivered in conventional and hypofractionated radiotherapy, respectively. In both treatments, a radiosensitizing effect was evidenced by the decrease in cell viability that was exacerbated over time. These effects were not found in peripheral blood, suggesting that RSV had a dual response. Although the results obtained in CHO-k1 transformed cells corroborated the genotoxic effect of the 60 µM dose of RSV observed in the tumor system, they also showed a radio-protective effect at the lowest dose (15 µM). While more studies are necessary, our results together with the good systemic tolerance of RSV and the lack of toxicity position the compound as a potential candidate for the prevention and treatment of cancer as well as for the optimization of the radiotherapeutic ratio.

Possible radioprotective effect of folic acid supplementation on low dose ionizing radiation-induced genomic instability in vitro.

Ionizing radiation (IR) induces DNA damage through production of single and double-strand breaks and reactive oxygen species (ROS). Folic acid (FA) prevents radiation-induced DNA damage by modification of DNA synthesis and/or repair and as a radical scavenger. We hypothesized that in vitro supplementation with FA will decrease the sensitivity of cells to genetic damage induced by low dose of ionizing radiation. Annexin V, comet and micronucleus assays were performed in cultured CHO cells. After 7 days of pre-treatment with 0, 100, 200 or 300 nM FA, cultures were exposed to radiation (100 mSv). Two un-irradiated controls were executed (0 and 100 nM FA). Data were statistically analyzed with X2-test and linear regression analysis (P 0.05). We observed a significantly decreased frequency of apoptotic cells with the increasing FA concentration (P <0.05). The same trend was observed when analyzing DNA damage and chromosomal instability (P <0.05 for 300 nM). Only micronuclei frequencies showed significant differences for linear regression analysis (R2=94.04; P <0.01). Our results have demonstrated the radioprotective effect of folic acid supplementation on low dose ionizing radiation-induced genomic instability in vitro; folate status should be taken into account when studying the effect of low dose radiation in environmental or occupational exposure.

Assessment of the adverse effects of the acaricide amitraz: in vitro evaluation of genotoxicity.

Amitraz is a formamidine widely used in Veterinary Medicine for the treatment of ectoparasites. It is a highly liposoluble compound that is quickly absorbed through the skin and mucous membranes, thus making exposure potentially dangerous for humans and animals. The aim of this study was to compare the genotoxic potential of the active constituent of the insecticide amitraz and a commercial product containing amitraz in vitro in hamster cells. The induction of primary DNA damage was evaluated by alkaline single-cell gel electrophoresis (comet assay) and the apoptotic ability was examined by the Annexin V/propidium iodide staining assay. The commercial formulation significantly increased the index of DNA damage at concentrations of 2.50-3.75 µg/mL compared to the control. The active constituent only induced significant DNA damage with the highest concentration (3.75 µg/mL). Although both tested products increased the frequency of cell death, neither of them induced significant differences. Genotoxic potential is a primary risk factor for long-term effects such as carcinogenic and reproductive toxicology. Results presented here highlight the need for further investigation of the potential health risk of this veterinary medicine.

Comparación entre referencias de las prevalencias de sobrepeso y obesidad, estimadas a través del índice de masa corporal, en niños de Argentina / Comparison between references of the overweight and obesity prevalence, through the body mass index, in Argentinean children

La evaluación de la situación nutricional de los niños, depende en gran medida de las curvas de crecimiento utilizadas. El objetivo de esta investigación es comparar las prevalencias de sobrepeso y obesidad estimadas con tres referencias internacionales en una población de niños, a través del Índice de Masa Corporal. Se incluyeron 737 niños/as nacidos de término sanos, de 2 a 5 años (estudio transversal). La Participación fue voluntaria y consentida. Se estableció el Índice de Masa Corporal (kg/m²). Las técnicas de medición se basaron en guías nacionales. Las referencias utilizadas fueron: 1. Centers for Disease Control and Prevention (CDC) (>Pc85: sobrepeso; > Pc95: obesidad); 2. International Obesity Task Force (IOTF) (valores límites sexo-edad específicos) y 3. Organización Mundial de la Salud (OMS) (+2 desvíos estándar: sobrepeso; >+3: obesidad). Los resultados fueron evaluados estadísticamente mediante el programa Epi Info 6.0 (χ², p < 0,05 por ciento). La prevalencia de niños con sobrepeso estimada con la referencia del CDC es 1,1 veces mayor que la de IOTF y 2,33 veces la de OMS y la de IOTF es 2,1 veces la de OMS (p= 0,00001). La prevalencia de niños con obesidad estimada con la referencia del CDC es 5,4 veces la de IOTF y 23,9 veces la de OMS, mientras que la de IOTF es 4,4 veces la de OMS (p= 0,0000001). Las prevalencias de sobrepeso y obesidad, difieren substancialmente de acuerdo a la referencia y valores límite utilizados. No existiendo aún un criterio único, cada una de las referencias debe utilizarse con precaución.

The evaluation of child nutritional status is highly dependent on the growth charts used. The aim of this study was to compare different references assessing overweight and obesity in children population, through the Body Mass Index. A total of 737 healthy children born at term, aged 2-5 years, were included (cross-sectional study). The participation was voluntary and consent. Body Mass Index (kg/m²) was estimated. The measurements techniques were based on national guidelines. We compared three references: (1) Centers for Disease Control and Prevention (CDC) (>Pc85: overweight; >Pc95: obesity; (2) International Obesity Task Force (IOTF) (sex-age-specific body mass index cut-offs); (3) World Health Organization (WHO) (+2 standard deviation: overweight; +3: obesity). The Epi Info 6.0 software was used to the statistical evaluation ((χ2, p≤ , 05 percent). The prevalence of overweight was 1,1 and 2,33 times higher in CDC application respect to the IOTF and the WHO references respectively. The prevalence of overweight was 2,1 times higher in IOTF application respect to the WHO references (p=, 00001). The prevalence of obesity was 5,4 and 23,9 times higher in CDC application respect to the IOTF and the WHO references respectively. The prevalence of obesity was 4,4 times higher in IOTF application respect to the WHO references (p=, 0000001). The prevalence of overweight and obesity, calculated through the BMI, differs substantially according to the reference and cut-off points used. In the absence of even a single criterion, each of the references should be used with care.

[Comparison between references of the overweight and obesity prevalence, through the Body Mass Index, in Argentinean children]. / Comparación entre referencias de las prevalencias de sobrepeso y obesidad, estimadas a través del Indice de Masa Corporal, en niños de Argentina.

The evaluation of child nutritional status is highly dependent on the growth charts used. The aim of this study was to compare different references assessing overweight and obesity in children population, through the Body Mass Index. A total of 737 healthy children born at term, aged 2-5 years, were included (cross-sectional study). The participation was voluntary and consent. Body Mass Index (kg/m2) was estimated. The measurements techniques were based on national guidelines. We compared three references: (1) Centers for Disease Control and Prevention (CDC) (> Pc85: overweight; > Pc95: obesity; (2) International Obesity Task Force (IOTF) (sex-age-specific body mass index cut-offs); (3) World Health Organization (WHO) (+2 standard deviation: overweight; +3: obesity). The Epi Info 6.0 software was used to the statistical evaluation (chi2, p < or = .05%). The prevalence of overweight was 1.1 and 2.33 times higher in CDC application respect to the IOTF and the WHO references respectively. The prevalence of overweight was 2.1 times higher in IOTF application respect to the WHO references (p = .00001). The prevalence of obesity was 5.4 and 23.9 times higher in CDC application respect to the IOTF and the WHO references respectively. The prevalence of obesity was 4.4 times higher in IOTF application respect to the WHO references (p = .0000001). The prevalence of overweight and obesity, calculated through the BMI, differs substantially according to the reference and cut-off points used. In the absence of even a single criterion, each of the references should be used with care.

Malnutrición infantil: evaluación antropométrica y consecuencias cromosómicas

Contenido: Crecimiento y nutrición. Malnutrición. Malnutrición proteico-energética (MPE). Evaluación antropométrica del estado nutricional. Aberraciones cromosómicas estructurales (ACE). Malnutrición proteico-energética y ACE. Efectos de la malnutrición: ACE. Antropometría. Desarrollo histórico. De las muestras. De la obtención de los datos. Antropometría. Muestra total. De los indicadores antropométricos. De los indicadores citogenéticos. Perspectivas futuras...

Alteraciones cromosómicas en niños con malnutrición proteico-energética: resultados preliminares / Chromosomic alterations in child by malnutrition energetics-proteine: results preliminary

Objetivos: algunos autores encontraron una relación entre la Malnutrición Proteico Energética (MPE) y elaumento de la frecuencia de Aberraciones Cromosómicas Estructurales (ACE) en niños menores de 5 años.El objetivo del siguiente trabajo es analizar la incidencia de ACE en una población de niños del partido de La Plata que presentan MPE. Material y Métodos: se tomó una muestra de 30 niños de 1 a 60 meses de edad cronológica, 15 controles sanos y 15 diagnosticados como MPE primarios a través de una evaluación antropométrica y clínica. Se aplicó una encuesta semiestructurada en la que se tuvieron en cuenta peso al nacimiento, variables antropométricas, alimentación, enfermedades virales, y/o infecciosas y exposición a agentes potencialmente genotóxicos. Se realizaron cultivos de sangre periférica y se analizó la frecuencia de ACE en 100 células por individuo. Los resultados obtenidos fueron sometidos a evaluación estadística utilizando el programa Epi Info 6.0. Resultados: la diferencia en la frecuencia global de ACE entre ambas muestras resultó significativa. Al tratar cada tipo de aberración en forma independiente se encontraron diferencias significativas para gaps y fracturas de mono e isocromátidas, cromosomas dicéntricos, fragmentos y asociaciones telométricas. No se encontraron diferencias significativas respecto a la exposición de los niños a agentes potencialmente genotóxicos tales como medicamentos, radiaciones, pesticidas y desechos industriales. Conclusiones: la MPE es capaz de inducir daño genotóxico, este efecto podria explicarse por la distorsión del ambiente químico interno o bien por la disminución de la acción de los mecanismos de reparación del ADN como consecuencia del déficit proteico