In vitro conservation of Vanilla planifolia hybrids in minimal growth conditions

ABSTRACT Objective: In vitro conservation with minimal growth in interspecific hybrids of Vanilla planifolia. Design / Methodology / Approach: Explants from seven interspecific V. planifolia hybrids were used with parents of different origins. The treatments were different doses of mannitol and sucrose in the culture medium that varied from 0, 5, 10, 15, 20, 25 and 30 g l-1. The number of nodes, shoots and roots was recorded every 30 days for six months. Results: The results indicated that 30 g l-1 of mannitol and without sucrose in the culture medium allowed minimal growth for most hybrids. It was found that with a higher content of mannitol and less sucrose, the length, number of between nodes, shoots and roots of the explants were shorter, results that were different between treatments (P?0.05). Limitations of the study / implications: There is a differential behavior between the biological material and the culture media used, especially when it comes to hybrids due to the new genetic combinations that are generated, so for their conservation the components of the culture medium must be adjusted. Findings/conclusions: It is concluded that the mannitol at 30 g l-1 and without sucrose in the in vitro culture medium of vanilla hybrids is able to reduce its growth significantly during 180 days.Key words: Ex situ conservation, in vitro culture, Vanilla planifolia, mannitol.Objective: To maintain minimal growth in in vitro Vanilla planifolia hybrids. Design/Methodology/Approach: Explants of seven interspecific hybrids of V. planifolia with different origin parents were used. The treatments consisted of different doses of mannitol and sucrose in the culture medium which varied from 0, 5, 10, 15, 20, 25 and 30 g L-1.The number of nodes, shoots and roots was recorded every 30 d for six months. Results: 30 g L-1 mannitol and no sucrose in the culture medium allowed minimal growth in most of the hybrids. The higher the mannitol and lower the sucrose content, the length, number of between nodes, shoots and roots of the explants was lower (P?0.05). Limitations of the study/implications: There is a differential behavior between the biological material and the used culture medium, particularly in hybrids, due to their new genetic combinations. Therefore, for their conservation, the culture medium components must be adjusted. Conclusions: 30 g L-1 mannitol without sucrose in in vitro culture medium significantly reduces growth during 180 d in vanilla hybrids

SOMACLONAL VARIATION OF Phanaenopsis sp. var. Dudu PLANTS REGENERATED BY DIRECT SOMATIC EMBRIOGENESIS

Actualmente existe una creciente demanda de híbridos de Phalaenopsis sp., por lo cual es necesario desarrollar protocolos de propagación masiva que aseguren altos porcentajes de regeneración clonal tal como la técnica de embriogénesis somática. Así mismo, estudiar la variación genética dentro de las plantas regeneradas ofrece un mayor conocimiento de los alcances del protocolo de micropropagación en relación a la estabilidad genética de los materiales empleados. Objetivo: En el presente trabajo de investigación se evaluó la combinación de tres concentraciones y dos tipos de reguladores de crecimiento vegetal (RCV), la citocinina bencilaminopurina (BA) (1.0, 2.0 y 3.0 mgL-1) en combinación con tres concentraciones de ácido 2,4-diclorofenoxiacético (2,4-D) (3.0, 4.0 y 5.0 mgL-1), para la inducción de embriones somáticos, además, se analizó la estabilidad genética de las plántulas regeneradas mediante marcadores moleculares tipo RAPD (Random Amplified Polymorphic DNA, por sus siglas en inglés). Metodología: La embriogénesis somática se indujo a partir de explantes de hoja con distintos grados de desarrollo: primera hoja se consideró como explante maduro (PH) y la tercera hoja como explante joven (TH), obtenidas de una plántula de Phalaenopsis sp. var. Dudú cultivadas in vitro, de 15-20 cm de altura. Resultados: El mayor número de plantas regeneradas fue de 29.8 a los 135 días después de iniciado el cultivo (ddic) con 2.0 y 5.0 mgL-1 de BA y 2,4-D, respectivamente, utilizando como explante la TH en la respuesta morfogenética de los explantes regenerados, se observó una correlación entre la edad del explante y la concentración de RCV. El análisis de la variación genética mostró cambios en los patrones de bandeo, observando bandas polimórficas con los cuatro iniciadores utilizados, aludiendo variación somaclonal en las plantas regeneradas. Implicaciones: Los resultados obtenidos aportan una alternativa de regeneración, además de ofrecer una metodología para iniciar programas de mejoramiento genético en Phalaenopsis sp. var. Dudú. Conclusiones: Se logró la regeneración in vitro de Phalaenopsis sp. var. Dudú mediante embriogénesis somática, así como la detección de cambios genéticos en el material regenerado.Nowadays, a growing demand for hybrids of Phalaenopsis sp. exists to satisfy this demand it is necessary to develop protocols for massive propagation that ensure high percentages of clonal regeneration, such as somatic embryogenesis. Besides, studying genetic variation within regenerated plants offers a greater understanding of the suitability of the micropropagation protocol in relation with genetic stability of the materials used. Objective: The present research work aimed to evaluate three concentrations of two types of plant growth regulators (RCV). 6- benzylaminopurine cytokinin (BA) (1.0, 2.0 y 3.0 mgL-1) in combination with three concentrations of 2,4- diclorofenoxiacetic acid (2,4-D) (3.0, 4.0, 5.0 mgL-1), for the induction of somatic embryos. In addition, the genetic stability of the regenerated plants was analyzed using molecular markers type RAPD (Random Amplified Polymorphic DNA). Methodology: The induction of somatic embryogenesis was induced from two leaf explants with different stages of develop from 15-20 cm in height Phalaenopsis sp. var. Dudú seedlings, cultivated in vitro; first leaf as mature explant (PH) and third leaf as young explant (TH). Results: The highest number of regenerated plants was 29.8 at 135 days after the start of the culture (ddic) with 2.0 and 5.0 mgL-1 of BA and 2,4-D, respectively, using TH as explant. In the morphogenetic response of the regenerated explants, a correlation was observed between the age of the explant and the RCV concentration. Polymorphic bands were observed with the four primers used, indicating somaclonal variation in regenerated plants. Implications: The results obtained provide an alternative for regeneration, as well as offering a methodology to initiate genetic improvement programs in Phalaenopsis sp. var. Dudú. Conclusions: In vitro regeneration of Phalaenopsis sp. var. Dudú by somatic embryogenesis was achieved, as well as the analysis of the genetic integrity of the regenerated material.UNIVERSIDAD AUTONOMA DEL ESTADO DE MEXIC

Ciencia Odontológica 2.0

Libro que muestra avances de la Investigación Odontológica en MéxicoEs para los integrantes de la Red de Investigación en Estomatología (RIE) una enorme alegría presentar el segundo de una serie de 6 libros sobre casos clínicos, revisiones de la literatura e investigaciones. La RIE está integrada por cuerpos académicos de la UAEH, UAEM, UAC y UdeG

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Ten-year clinical and echocardiographic follow-up of patients undergoing percutaneous mitral commissurotomy with Inoue balloon

Percutaneous mitral commissurotomy (PMC) has emerged as an effective nonsurgical technique for the treatment of patients with symptomatic mitral stenosis. This report highlights the immediate and long-term follow-up results of this procedure in an unselected cohort of patients with rheumatic mitral stenosis from a single center. PMC with Inoue balloon was performed in 70 patients in a 2-year period (1993-1994). Age of patients ranged from 18 to 67 years (mean 38 ± 11). Atrial fibrillation was present in 18 (30%) patients. A detailed clinical and echocardiographic (two-dimensional, continuous-wave Doppler and color-flow imaging) assessment was done at follow-up. The procedure was technically successful in 61 (85%) patients with an increase in mitral valve area (MVA) from 0.96 ± 0.2 to 1.7 ± 0.28 cm² (P < .001) and a reduction in mean trans-mitral gradient from 14.3 ± 4.8 to 6.0 ± 2.8 mmHg (P < .01). Mitral regurgitation appeared or worsened in 25 (30%) patients, of which 3 (4%) developed severe mitral regurgitation. Urgent mitral valve replacement was performed in these 3 patients. Data of 52 patients followed over a period of 105 ± 10 months revealed MVA of 1.4 ± 0.4 cm. Elective mitral valve replacement was done in 14 (23%) patients. Mitral restenosis diagnosed with echocardiography was seen in 24 (50%) patients, of which 14 were having recurrence of class III or more symptoms and were treated with surgery. Thus, percutaneous mitral commissurotomy is an effective and safe procedure and over 2/3 of the patients were event-free at the end of follow-up. The benefits are sustained in most of these patients on long-term follow-up

Paclitaxel coated-stent for early-onset thrombosis after liver transplantation

Hepatic artery thrombosis (HAT) is the most common vascular complication of orthotopic liver transplantation (OLT) and constitutes a potential emergency during the postoperative period. Surgical revascularization and retransplantation are the treatments of choice for this condition. The aim of this report is to present long-term follow-up on survival and graft function of three patients with paclitaxel-coated hepatic artery stents placed percutaneously after early-onset HAT. Three patients developed early onset HAT after cadaveric-donor OLT in a tertiary care center in Mexico. These patients were treated percutaneously with balloon angioplasty and paclitaxel-coated stents. After 24 months or more of follow-up, 2 patients present total occlusion of the stent and one patient, intra-stent stenosis; interestingly, all patients have normal graft function and excellent quality of life. In conclusion, although balloon angioplasty and stent placement may be a therapeutic option for suitable patients with early-onset HAT after OLT, long-term patency is unlikely even with the use of paclitax-el-coated materials

Efecto de dos citoquininas, ácido ascórbico y sacarosa en la obtención de plantas in vitro de sorghum bicolor para la formación de callos

Título en ingles: Effect of two cytokinin, ascorbic acid and sucrose to obtain in vitro shoots of sorghum for calli formation Resumen: El trabajo se realizó con el objetivo de obtener brotes in vitro vía organogénesis directa en sorgo variedad CIAP 2E-95, para la formación de callos con estructuras embriogénicas. Se tomaron semillas maduras de plantas que crecían en condiciones controladas, las que se desinfectaron y fueron colocadas a germinación en un medio de cultivo con las sales MS, mioinositol 100 mg L-1, sacarosa 3%, pH 5,7, Fitagel 2,5 g L-1. Para la multiplicación de los brotes, se estudiaron diferentes concentraciones de 6 Bencilaminopurina (6-BAP) y kinetina. El empleo en el medio de cultivo de multiplicación con las sales MS y 6- BAP 0,22 mg L-1, incrementó el número de brotes por explante y se observó la presencia de oxidación fenólica. La oxidación fenólica se contrarrestó con la adición al medio de cultivo de 50 mg L-1 de ácido ascórbico y se incrementó significativamente el número de brotes, altura y número de hojas. En la obtención de brotes in vitro de calidad para la formación de callos, estos fueron engrosados con las sales MS, sin reguladores de crecimiento, ácido ascórbico 50 mg L-1, Fitagel 2,5 g L-1, pH 5,7 y 40 g L-1 de sacarosa. La formación de callos con estructuras embriogénicas a partir del cilindro central de las hojas enrolladas del segmento más próximo a la base de los brotes engrosados se alcanzó a los 45 días de cultivo. Los segmentos del cilindro central de las hojas enrolladas constituyen una fuente segura y promisoria de explantes para la formación de callos embriogénicos. Palabras clave: 6 BAP; brote in vitro; callos con estrucutras embriogénicos; kinetina. Abstract: The current work aimed to obtain in vitro shoots by direct organogenesis from sorghum variety CIAP 2E-95, for the formation of calluses with embryogenic structures. Mature seeds were collected from plants grown under controlled conditions, which were disinfected and were placed in a germination medium with MS salts, myo-inositol 100 mg L-1, 3% sucrose, pH 5.7, phytagel 2.5 g L-1. Different concentrations of 6-benzylaminopurine (6-BAP) and kinetin were studied for shoot multiplication. Using 6 - BAP 0.22 mg L-1 in the multiplication culture medium increased the number of shoots per explant, showing phenolic oxidation. Adding 50 mg L-1 of ascorbic acid to the culture medium resisted phenolic oxidation and the number of sprouts (5.0), height and number of leaves increased significantly. Quality in vitro buds for callus formation were obtained  stimulating the thickening by using MS salts, without growth regulators, ascorbic acid 50 mg L-1, phytagel 2.5 g L-1 and pH 5.7 and 40 g L-1 of sucrose. The largest diameter of shoots, height, number of leaves and roots of in vitro plants, was achieved after 21 days of cultured in MS salts with 40 g L-1 sucrose. Calluses formation with embryogenic structures from the central cylinder of curled leaves of the nearest segment to the base of thickened shoots was achieved after 45 days of culture. Segments from the central cylinder of the curled leaves are a safe source for the formation of embryogenic calli. Key words: 6-BAP; in vitro shoots; callus with embryogenic structures; kineti

Efecto de dos citoquininas, ácido ascórbico y sacarosa en la obtención de plantas in vitro de Sorghum bicolor para la formación de callos

The aim of the current work was to obtain in vitro shoots by direct organogenesis from sorghum variety CIAP 2E-95, for the formation of calluses with embryogenic structures. Mature seeds were collected from plants grown under controlled conditions, which were disinfected and were placed in a germination medium with MS salts, myo-inositol 100 mg L-1, 3% sucrose, pH 5.7, phytagel 2.5 g L-1. Different concentrations of 6-benzylaminopurine (6-BAP) and kinetin were studied for shoot multiplication. Using 6 - BAP 0.22 mg L-1 in the multiplication culture medium increased the number of shoots per explant, showing phenolic oxidation. Adding 50 mg L-1 of ascorbic acid to the culture medium resisted phenolic oxidation and the number of sprouts (5.0), height and number of leaves increased significantly. Quality in vitro buds for callus formation were obtained stimulating the thickening by using MS salts, without growth regulators, ascorbic acid 50 mg L-1, phytagel 2.5 g L-1 and pH 5.7 and 40 g L-1 of sucrose. The largest diameter of shoots, height, number of leaves and roots of in vitro plants, was achieved after 21 days of cultured in MS salts with 40 g L-1 sucrose. Calluses formation with embryogenic structures from the central cylinder of curled leaves of the nearest segment to the base of thickened shoots was achieved after 45 days of culture. Segments from the central cylinder of the curled leaves are a safe source for the formation of embryogenic callus.El trabajo se realizó con el objetivo de obtener brotes in vitro vía organogénesis directa en sorgo variedad CIAP 2E-95, para la formación de callos con estructuras embriogénicas. Se tomaron semillas maduras de plantas que crecían en condiciones controladas, las que se desinfectaron y fueron colocadas a germinación en un medio de cultivo con las sales MS, mioinositol 100 mg L-1, sacarosa 3%, pH 5,7, Fitagel 2,5 g L-1. Para la multiplicación de los brotes, se estudiaron diferentes concentraciones de 6 Bencilaminopurina (6-BAP) y kinetina. El empleo en el medio de cultivo de multiplicación con las sales MS y 6- BAP 0,22 mg L-1, incrementó el número de brotes por explante y se observó la presencia de oxidación fenólica. La oxidación fenólica se contrarrestó con la adición al medio de cultivo de 50 mg L-1 de ácido ascórbico y se incrementó significativamente el número de brotes, altura y número de hojas. En la obtención de brotes in vitro de calidad para la formación de callos, estos fueron engrosados con las sales MS, sin reguladores de crecimiento, ácido ascórbico 50 mg L-1, Fitagel 2,5 g L-1, pH 5,7 y 40 g L-1 de sacarosa. La formación de callos con estructuras embriogénicas a partir del cilindro central de las hojas enrolladas del segmento más próximo a la base de los brotes engrosados se alcanzó a los 45 días de cultivo. Los segmentos del cilindro central de las hojas enrolladas constituyen una fuente segura y promisoria de explantes para la formación de callos embriogénicos