Somatic Embryogenesis and Agrobacterium-Mediated Gene Transfer Procedures in Chilean Temperate Japonica Rice Varieties for Precision Breeding.

Rice (Oryza sativa) varieties are generated through breeding programs focused on local requirements. In Chile, the southernmost rice producer, rice productivity relies on the use and generation of temperate japonica germplasms, which need to be adapted to the intensifying effects of climate change. Advanced biotechnological tools can contribute to these breeding programs; new technologies associated with precision breeding, including gene editing, rely on procedures such as regeneration and gene transfer. In this study, the local rice varieties Platino, Cuarzo, Esmeralda, and Zafiro were evaluated for somatic embryogenesis potential using a process that involved the combined use of auxins and cytokinins. An auxin-based (2,4-D) general medium (2N6) allowed for the induction of embryogenic masses in all the genotypes. After induction, masses required culturing either in N6R (kinetin; Platino) or N6RN (BAP, kinetin, IBA, and 2,4-D; Cuarzo, Esmeralda, and Zafiro) to yield whole plants using regeneration medium (N6F, no hormone). The sprouting rates indicated Platino as the most responsive genotype; for this reason, this variety was evaluated for gene transfer. Fifteen-day-old embryo masses were assayed for Agrobacterium-mediated transformation using the bacterial strain EHA105 harboring pFLC-Myb/HPT/GFP, a modified T-DNA vector harboring a geminivirus-derived replicon. The vector included the green fluorescent protein reporter gene, allowing for continuous traceability. Reporter mRNA was produced as early as 3 d after agroinfiltration, and stable expression of the protein was observed along the complete process. These achievements enable further biotechnological steps in these and other genotypes from our breeding program.

Platelet VPS16B is dependent on VPS33B expression, as determined in two siblings with arthrogryposis, renal dysfunction, and cholestasis syndrome.

BACKGROUND: Platelet α-granule biogenesis in precursor megakaryocytes is critically dependent on VPS33B and VPS16B, as demonstrated by the platelet α-granule deficiency seen in the rare multisystem disorder arthrogryposis, renal dysfunction, and cholestasis (ARC) syndrome associated with biallelic pathogenic variants in VPS33B and VIPAS39 (encoding VPS16B). VPS33B and VPS16B are ubiquitously expressed proteins that are known to interact and play key roles in protein sorting and trafficking between subcellular locations. However, there remain significant gaps in our knowledge of the nature of these interactions in primary cells from patients with ARC syndrome. OBJECTIVES: To use primary cells from patients with ARC syndrome to better understand the interactions and roles of VPS33B and VPS16B in platelets and precursor megakaryocytes. PATIENTS/METHODS: The proband and his male sibling were clinically suspected to have ARC syndrome. Confirmatory genetic testing and platelet phenotyping, including electron microscopy and protein expression analysis, was performed with consent in a research setting. RESULTS: We describe the first case of ARC syndrome identified in Costa Rica, associated with a novel homozygous nonsense VPS33B variant that is linked with loss of expression of both VPS33B and VPS16B in platelets. CONCLUSION: These results indicate that stable expression of VPS16B in platelets, their precursor megakaryocytes, and other cells is dependent on VPS33B. We suggest that systematic evaluation of primary cells from patients with a range of VPS33B and VIPAS39 variants would help to elucidate the interactions and functions of these proteins.

[Utility of prenatal screening for Congenital Chagas Disease in Chile]. / Utilidad del tamizaje prenatal de enfermedad de Chagas congénita en Chile.

Chagas Disease is a pathology caused by the parasite Trypanosoma cruzi. It is considered a public health issue, especially its congenital transmission, not only in endemic countries but also in non endemic ones, being the main form of transmission in Chile. Objectives: To describe a congenital Chagas Disease case and to report about the usefulness of prenatal screening for Chagas Disease. Cli nical Case: A 29-week preterm newborn of a multiparous mother with Chagas Disease detected in the perinatal screening of current pregnancy. Due to history of Trypanosoma cruzi, PCR was performed on the 4th day of life resulting undetectable. At the 6th and the 8th week of life, a macular erythema tous rash and fever and respiratory distress were observed, respectively, both self-limited. At the 8th week of life, as a routine prematurity follow-up, a fundus examination was performed which showed bilateral retinal granulation, therefore, a TORCH study was performed with a negative result. Also, in this same week, a second Trypanosoma cruzi PCR was performed, with a positive result. Lab tests reported visualization of trypomastigotes, confirming the Congenital Chagas Disease diagnosis. The patient was treated with Nifurtimox, presenting an adverse reaction at 35 days of treatment with neu tropenia, vomiting, and poor weight gain. The treating drug was replaced by Benznidazole, achieving 60 days of treatment. At 22 months of chronologic age, the patient has remained asymptomatic. Con clusions: It is necessary to increase efforts in prevention, early detection, and study of direct relatives. This report is an example of the usefulness of Chagas disease prenatal screening for the detection and early treatment of new cases.

First Report of Fruit Rot of Sweet Cultivars of Japanese Plum caused by Alternaria alternata, A. arborescens, and A. tenuissima in Chile.

During the last two seasons, an unusual fruit rot was observed in four orchards of sweet Japanese plum (Prunus salicina) cultivars located in the Chilean Central Valley (30°00'S, 70°42'W). The incidence was 5% in Black Majesty, 4% in Red Lyon, and 6% in Sweet Mary cultivars in 2020. Fruits in the field showed a firm, dehydrated, and slightly sunken rot on the blossom end, along with rough and irregular epidermis in the affected area. Internally, the fruit flesh appeared light to dark-brown or olive-green. Symptomatic fruits (n=119) were superficially disinfected (75% ethanol) and, pieces of the pericarp (3 x 3 mm) were removed and placed on potato dextrose agar (PDA). Isolates of Alternaria spp. were obtained and 9 of these were selected for identification. Colonies were dark olive to gray-brown with white margins, small, catenulate and muriform conidia, produced in single or branched conidiophores. Isolates produced brown to golden-brown, ovoid, ellipsoidal to obclavate conidia with dimensions of 19.7 to 26.7 × 10.0 to 11.9 µm with two to four transverse and zero to three longitudinal septa on 0.05× PDA (Pryor and Michailides 2002) after 7 d at 20°C under 10/14 h light/dark cycles. A molecular analysis was performed by sequencing the nuclear genes RNA polymerase II subunit (RPB2), plasma membrane ATPase (ATP), and the calmodulin (Cal) gene using primers RPB2-5F2/fRPB2-7cR, ATPDF1/ATPDR1, and CALDF1/CALDR1, respectively (Lawrence et al. 2013; Woudenberg et al. 2013). A BLAST search revealed the presence of Alternaria spp. with a 99% to 100% identity with the reference sequences of A. alternata (JQ905182, JQ671874, JQ646208), A. arborescens (JQ646487, JQ671880, JQ646214), and A. tenuissima (JQ811961, JQ811989, JQ646209). Maximum parsimony phylogenetic analysis confirmed the identifications. Sequences were deposited in GenBank as numbers MW514249 to MW514257, MT872324 to MT872332, and MT872314 to MT872322 for RPB2, ATP, and Cal sequences, respectively. All these Alternaria isolates were deposited in the Colección Chilena de Recursos Genéticos Microbianos - INIA, Chillán Chile (RGM3069 to RGM3077). Pathogenicity of A. alternata (n=4), A. arborescens (n=3) and, A. tenuissima (n=2) was tested in Red Lyon plum fruits. Plums were disinfected in 1% sodium hypochlorite for 2 min, rinsed in sterile distilled water for 1 min and dried on absorbent towels in a laminar flow hood. Then, the plums were wounded on the blossom end with a sterile needle (1 x 0.5 mm), inoculated with 10 µl of a conidial suspension (106 conidia/ml), wrapped with Parafilm and maintained in a humid chamber (>95% relative humidity). An equal number of fruits wounded and inoculated with sterile water were used as a control. After 7 days at 20°C, all inoculated fruits developed a dark-brown firm rot with lesion lengths of 24.4 (±3.0) mm, 19.6 (±0.7) mm, and 16.8 (±2.4) mm for A. alternata, A. arborescens and A. tenuissima, respectively. A. alternata was the most aggressive species (P < 0.001). Control fruits remained asymptomatic. Koch's postulates were fulfilled after the re-isolating the causal agent from the border of the lesions. Leaf spots and fruit rots caused by Alternaria isolates have been reported in stone fruits, including plums (Kim et al. 2005; Long et al. 2021; Moosa et al., 2019; Yang et al. 2014). To our knowledge, this is the first report of A. alternata, A. arborescens, and A. tenuissima associated with fruit rot in sweet Japanese plum cultivars in the field, in Chile.

Novel polyester-based thermoplastic elastomers for 3D-printed long-acting drug delivery applications.

To improve patient compliance and personalised drug delivery, long-acting drug delivery devices (LADDDs), such as implants and inserts, greatly benefit from a customisation in their shape through the emerging 3D-printing technology, since their production usually follows a one-size-fits-most approach. The use of 3D-printing for LADDDs, however, is mainly limited by the shortage of flawlessly 3D-printable, yet biocompatible materials. The present study tackles this issue by introducing a novel, non-biodegradable material, namely a polyester-based thermoplastic elastomer (TPC) - a multi-block copolymer containing alternating semi-crystalline polybutylene terephthalate hard segments and poly-ether-terephthalate amorphous soft segments. Next to a detailed description of the material's 3D-printability by mechanical, rheological and thermal analyses, which was found to be superior to that of conventional polymers (ethylene-vinyl acetates (EVA)), this study establishes the fundamental understandings of the interactions between progesterone (P4) and TPC and drug-releasing properties of TPC for the first time. P4-loaded LADDDs based on TPC, prepared via an elaborated solvent-immersion technique, enable the release of P4 at pharmacologically relevant rates, similar to those of marketed formulations based on EVA and silicones. Additionally, TPC demonstrated an exceptional 3D-printability for a wide selection of implant sizes and complex geometries.

Novel Cleaning-in-Place Strategies for Pharmaceutical Hot Melt Extrusion.

To avoid any type of cross-contamination, residue-free production equipment is of utmost importance in the pharmaceutical industry. The equipment cleaning for continuous processes such as hot melt extrusion (HME), which has recently gained popularity in pharmaceutical applications, necessitates extensive manual labour and costs. The present work tackles the HME cleaning issue by investigating two cleaning strategies following the extrusion of polymeric formulations of a hormonal drug and for a sustained release formulation of a poorly soluble drug. First, an in-line quantification by means of UV-Vis spectroscopy was successfully implemented to assess very low active pharmaceutical ingredient (API) concentrations in the extrudates during a cleaning procedure for the first time. Secondly, a novel in-situ solvent-based cleaning approach was developed and its usability was evaluated and compared to a polymer-based cleaning sequence. Comparing the in-line data to typical swab and rinse tests of the process equipment indicated that inaccessible parts of the equipment were still contaminated after the polymer-based cleaning procedure, although no API was detected in the extrudate. Nevertheless, the novel solvent-based cleaning approach proved to be suitable for removing API residue from the majority of problematic equipment parts and can potentially enable a full API cleaning-in-place of a pharmaceutical extruder for the first time.

Impacto de un programa de educación terapéutica en asma en un niño y sus cuidadores / Impact of a program of health education for the asthmatic child and their families

Las guías para el manejo del asma promueven la educación sanitaria como un escalón más en su terapéutica. Se creó el programa Educación Sanitaria del Niño Asmático y su Familia. El objetivo fue evaluar el impacto del Programa sobre el bienestar del niño asmático y su familia.Estudio retrospectivo, descriptivo, durante 3 años, que analizó características epidemiológicas, impacto y satisfacción de las familias. Se incluyeron 386 pacientes. Asma controlada al corte: el 78 %. Descendieron el uso de beta-2-adrenérgicos y las admisiones hospitalarias (p < 0,05), el uso de corticoides orales y consultas en Urgencias (p < 0,001). Evaluó el Programa como excelente el 98 %; comprendió y aceptó la enfermedad el 99 %; reconoció el inicio de crisis y comenzó el tratamiento el 96 %; aplicó bien la terapia inhalatoria el 92 %. El Programa tuvo alto impacto en el control del asma. Constituyó un modelo útil para replicar.

Guidelines for management establish health education as part of asthma care. A Health Education Program for Asthmatic Children and Families was started in our institution. This descriptive retrospective study was designed in order to evaluate the Program impact in children and family satisfaction; 386 children were enrolled. Their asthma was properly controlled in 78 %. There was a significant decrease (p < 0.05) of rapid action beta 2 agonists use and hospital admissions, and a highly significant decrease (p < 0.001) of corticosteroids use and emergency visits. Educational content was found to be excellent in 98 %; 99 % understood and accepted the condition; 96 % were able to recognize early symptoms and started home treatment; 92 % correctly managed inhalation technique. The Program had a high impact on children asthma control, with a significant decrease in emergency visits, admissions, and drug use, becoming a useful model

[Impact of a program of health education for the asthmatic child and their families]. / Impacto de un programa de educación terapéutica en asma en un niño y sus cuidadores.

Guidelines for management establish health education as part of asthma care. A Health Education Program for Asthmatic Children and Families was started in our institution. This descriptive retrospective study was designed in order to evaluate the Program impact in children and family satisfaction; 386 children were enrolled. Their asthma was properly controlled in 78 %. There was a significant decrease (p < 0.05) of rapid action beta 2 agonists use and hospital admissions, and a highly significant decrease (p < 0.001) of corticosteroids use and emergency visits. Educational content was found to be excellent in 98 %; 99 % understood and accepted the condition; 96 % were able to recognize early symptoms and started home treatment; 92 % correctly managed inhalation technique. The Program had a high impact on children asthma control, with a significant decrease in emergency visits, admissions, and drug use, becoming a useful model.

Las guías para el manejo del asma promueven la educación sanitaria como un escalón más en su terapéutica. Se creó el programa Educación Sanitaria del Niño Asmático y su Familia. El objetivo fue evaluar el impacto del Programa sobre el bienestar del niño asmático y su familia. Estudio retrospectivo, descriptivo, durante 3 años, que analizó características epidemiológicas, impacto y satisfacción de las familias. Se incluyeron 386 pacientes. Asma controlada al corte: el 78 %. Descendieron el uso de beta-2-adrenérgicos y las admisiones hospitalarias (p < 0,05), el uso de corticoides orales y consultas en Urgencias (p < 0,001). Evaluó el Programa como excelente el 98 %; comprendió y aceptó la enfermedad el 99 %; reconoció el inicio de crisis y comenzó el tratamiento el 96 %; aplicó bien la terapia inhalatoria el 92 %. El Programa tuvo alto impacto en el control del asma. Constituyó un modelo útil para replicar.

Fate of Spoilage and Pathogenic Microorganisms in Acidified Cold-Filled Hot Pepper Sauces.

Consumption of spicy foods and hot sauces is currently a popular trend worldwide. Shelf-stable acidified sauces are commonly hot-filled to ensure commercial sterility, but cold-fill-hold processes might also be suitable if microbial safety and stability are ensured. For this study, model acidified hot pepper sauces were developed and characterized. The effects of sauce pH and of two different organic acids on the survival of Pichia manshurica and Lactobacillus curvatus isolated from contaminated commercial hot sauces and on pathogenic Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes were assessed. Full factorial designs with three levels for pH (3.2, 3.5, and 3.9) and two for organic acid (citric and acetic) were used to determine the effects of these factors and their interactions on the survival of the microorganisms. Commercially sterile sauces were independently inoculated and kept at ambient temperature. Microbial counts were determined at different sampling times, depending on the treatment evaluated. Sauces acidified to pH 3.2 with citric or acetic acid were inoculated with cocktails of five strains or serotypes of the three pertinent pathogens, and inactivation curves were determined. Trials were performed in triplicate. A greater than 5-log reduction of P. manshurica and L. curvatus was achieved in less than 6 h in sauces adjusted to pH 3.2 with acetic acid. Greater than 5-log reductions of pathogenic bacteria were achieved 0.5 h after inoculation in sauces acidified to pH 3.2 with acetic acid. In contrast, at least 48 h was required to guarantee the same inactivation for the most tolerant pathogen when citric acid was used. Thus, a cold-fill-hold process may be a suitable alternative for acidified hot pepper sauces. Based on survival of the microorganisms evaluated in this study, microbial safety and stability can be achieved by adjusting the pH to 3.2 or less by the addition of acetic acid.

Leishmaniasis: Opciones terapéuticas en la población pediátrica / Leishmaniasis: Therapeutic options in the pediatric population

Resumen La leishmaniasis es una enfermedad parasitaria crónica endémica en muchas partes del mundo. La variabilidad de cepas, su clínica y respuesta a tratamiento ha hecho que se clasifique en dos grandes grupos: la leishmaniasis del Nuevo Mundo y la del Viejo Mundo. Según esto, varían las recomendaciones respecto a manejo y seguimiento. En esta revisión se hace énfasis a la leishmaniasis de nuestro medio, revisando opciones terapéuticas y posibilidades principalmente en la población pediátrica.

Abstract Leishmaniasis is a chronic parasitic disease endemic in many parts of the world. The variability of strains, their clinic and response to treatment has led to their classification into two major groups: New World leishmaniasis and Old World leishmaniasis. According to this, the recommendations regarding management and follow-up vary. In this review, emphasis is placed on leishmaniasis in our environment, reviewing therapeutic options and possibilities mainly in the pediatric population.

Lesiones graves y fatales en niños y adolescentes uruguayos secundarias a siniestros de tránsito por motos. Estudio epidemiológico / Serious and fatal injuries in Uruguayan children and adolescents secondary to traffic accidents by motorcycle. Epidemiological study

En países de ingresos medios, como Uruguay, los conductores de motos son parte importante de los lesionados o muertos en siniestros de tránsito. Objetivo: conocer la cantidad de niños y adolescentes que sufrieron siniestros con motos y que: a) fallecieron o, b) ingresaron a CTI/CI. De estos últimos describir las principales lesiones y características. Metodología: estudio descriptivo, retrospectivo, entre el 1 de junio y el 31 de diciembre de 2009. Fuentes de información: bases de datos del MSP, CTI/CI, certificados de defunción, partes policiales e historias clínicas. Resultados: en los 6 meses estudiados: a) fallecieron 20 menores de 19 años, 54% de los fallecidos por accidentes de tránsito del rango etario. 70% varones, todos conductores o pasajeros. 16/20 en vía pública, el resto en CTI. Al menos 45% no usaban casco; b) ingresaron a CTI/CI 69 menores de 19 años (2.7% de las causas de ingreso /edad), 26% menores de 15 años, 71% varones, la mayoría conductores o pasajeros. 91% sufrieron TEC y 87% politraumatismos de 3 a 6 sectores. Presentaron lesiones de: miembros 65%, tórax 60%, facial 54%, abdomen 40%, pelvis 25%; coma 40%, shock 25%. No usaba casco 68%. Se encontró carencias de datos en los documentos analizados. Conclusiones: es necesario: 1) mejorar la calidad de la información relativa a los siniestros de tránsito. 2) legislar la pertinencia o no de los niños a bordo de moto y si corresponde, las condiciones de traslado 3) realizar campañas educativas permanentes de seguridad vial, además de fiscalización y penalización en caso de no cumplimiento de la ley...

Mass spectrometry/gas chromatography-mass spectrometry approach for rapid screening/quantitative determination of perchloroethylene in air.

A new mass spectrometry/gas chromatography-mass spectrometry (MS/GC-MS) approach has been developed for the screening and quantitative determination of perchloroethylene (PERC) in workplace and outdoor air samples, which could be extended to the screening and analysis of other analytes and samples. This approach may be rapidly modified in order to be used directly as an MS detector for screening purposes or alternatively as a common GC-MS, for confirmation. The screening alternative by MS is approximately 20 times faster than the quantitative-confirmatory determination by GC-MS. Detection limits of both alternatives are sufficiently low to screen and determine PERC in the above-mentioned matrixes. The advantage of this approach over others previously described is that, in the present case, the sample passes through the chromatographic column only when the confirmatory GC-MS is used. For the MS screening method, the chromatographic column is bypassed by using an appropriate selection valve. In this way, the column lifetime is extended and screening time is considerably shortened.

¿Podría tener la cafeína algún efecto vasculoprotector? / Could caffeine have some vasoprotective effect?

El café es la bebida estimulante caliente de mayor consumo en el mundo y hace parte de la dieta corriente y cultural de nuestra sociedad. Existe un ®mito¼ alrededor del café, considerándolo nocivo con base en estudios clínicos, cuyos resultados han sido contradictorios y sin un sustento científico sólido. Sólo en los últimos años, se han venido describiendo las bondades del café a nivel cognoscitivo, digestivo y cardiovascular.Se presenta el efecto vasodilatador de la cafeína en un modelo experimental in-vitro que utiliza anillos de aorta procedentes de conejos sanos, colocados en un baño de órganos de 4 canales (Kent Scientific Corporation, Litchfield CT), en solución de Krebs, a 37°C, pH 7.40 +/- 0.05 y aireada con O2 95 porciento y CO2 5 porciento. Los anillos fueron pre-contraídos con norepinefrina (- 5.5 Log M) y relajados con nitroglicerina (relajación independiente de endotelio), acetilcolina (relajación dependiente de endotelio) y cafeína.

Manual de normas de atención integral a la niñez y los niños nos unieron / Manual of standards for comprehensive care for children and children joined us

El presente documento tiene cinco capítulos: el primero trata del marco conceptual de la Atención Integral a la Niñez; el segundo, define y describe la modalidad institucional, que es la forma de brindar atención integral a través de los Centros Infantiles Comunitarios (CICOs); el tercero define y describe la Modalidad Interante, que es una de las lecciones aprendidas para ofrecer y ampliar la cobertura de la atención cuando las comunidades están geográficamente dispersas; el cuarto capítulo, describe la atención a las mujeres embarazadas y madres lactantes, uno de los nuevos servicios que brindará el Programa para captar tempranamente a los/las niños(as) y promocionar la salud de las madres e hijos; y el quinto, describe la Participación Social a través de la Red Juvenil, El círculo de padres y la Comisión Municipal de la Niñez y la Adolescencia (CMNA), que es una de las estrategias fundamentales para la sostenibilidad del Proyecto Municipal y el empoderamiento de las comunidades en busca de su propio desarrollo. En consecuencia la normativa es el marco regulador que define los procedimientos y actividades a seguir para la ejecución del Proyecto, el cual responde a la realidad y contexto de los municipios; además, establece las funciones y responsabilidades de los actores involucrados...

Manual de normas de atención integral a la niñez... y los niños nos unieron / Manual of standards for comprehensive care for children... and children joined us

El presente documento tiene cinco capítulos: el primero trata del marco conceptual de la Atención Integral a la Niñez; el segundo, define y describe la modalidad institucional, que es la forma de brindar atención integral a través de los Centros Infantiles Comunitarios (CICOs); el tercero define y describe la Modalidad Interante, que es una de las lecciones aprendidas para ofrecer y ampliar la cobertura de la atención cuando las comunidades están geográficamente dispersas; el cuarto capítulo, describe la atención a las mujeres embarazadas y madres lactantes, uno de los nuevos servicios que brindará el Programa para captar tempranamente a los/las niños(as) y promocionar la salud de las madres e hijos; y el quinto, describe la Participación Social a través de la Red Juvenil, El círculo de padres y la Comisión Municipal de la Niñez y la Adolescencia (CMNA), que es una de las estrategias fundamentales para la sostenibilidad del Proyecto Municipal y el empoderamiento de las comunidades en busca de su propio desarrollo. En consecuencia la normativa es el marco regulador que define los procedimientos y actividades a seguir para la ejecución del Proyecto, el cual responde a la realidad y contexto de los municipios; además, establece las funciones y responsabilidades de los actores involucrados...