Mature seed-derived callus of the model indica rice variety Kasalath is highly competent in Agrobacterium-mediated transformation

We previously established an efficient Agrobacterium-mediated transformation system using primary calli derived from mature seeds of the model japonica rice variety Nipponbare. We expected that the shortened tissue culture period would reduce callus browning—a common problem with the indica transformation system during prolonged tissue culture in the undifferentiated state. In this study, we successfully applied our efficient transformation system to Kasalath—a model variety of indica rice. The Luc reporter system is sensitive enough to allow quantitative analysis of the competency of rice callus for Agrobacterium-mediated transformation. We unexpectedly discovered that primary callus of Kasalath exhibits a remarkably high competency for Agrobacterium-mediated transformation compared to Nipponbare. Southern blot analysis and Luc luminescence showed that independent transformation events in primary callus of Kasalath occurred successfully at ca. tenfold higher frequency than in Nipponbare, and single copy T-DNA integration was observed in ~40% of these events. We also compared the competency of secondary callus of Nipponbare and Kasalath and again found superior competency in Kasalath, although the identification and subsequent observation of independent transformation events in secondary callus is difficult due to the vigorous growth of both transformed and non-transformed cells. An efficient transformation system in Kasalath could facilitate the identification of QTL genes, since many QTL genes are analyzed in a Nipponbare × Kasalath genetic background. The higher transformation competency of Kasalath could be a useful trait in the establishment of highly efficient systems involving new transformation technologies such as gene targeting

Extracorporeal Membrane Oxygenation for Acute Pediatric Respiratory Failure

This article is made available for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.The use of extracorporeal membrane oxygenation (ECMO) to support children with acute respiratory failure has steadily increased over the past several decades, with major advancements having been made in the care of these children. There are, however, many controversies regarding indications for initiating ECMO in this setting and the appropriate management strategies thereafter. Broad indications for ECMO include hypoxia, hypercarbia, and severe air leak syndrome, with hypoxia being the most common. There are many disease-specific considerations when evaluating children for ECMO, but there are currently very few, if any, absolute contraindications. Venovenous rather than veno-arterial ECMO cannulation is the preferred configuration for ECMO support of acute respiratory failure due to its superior side-effect profile. The approach to lung management on ECMO is variable and should be individualized to the patient, with the main goal of reducing the risk of VILI. ECMO is a relatively rare intervention, and there are likely a minimum number of cases per year at a given center to maintain competency. Patients who have prolonged ECMO runs (i.e., greater than 21 days) are less likely to survive, though no absolute duration of ECMO that would mandate withdrawal of ECMO support can be currently recommended

Establishment of an efficient callus induction method from leaf and stem in kinnow mandarin (Citrus reticulata Blanco.) and citron (Citrus medica L.)

Kinnow mandarin (Citrus reticulata Blanco.) is a highly adaptable variety among citrus cultivars in the climate of the region of Panjab, Pakistan and citron (Citrus medica L.) was the first of the citrus fruits known to become visible in the Mediterranean Basin. In Sylhet region of Bangladesh, it is commonly known as Zara lemon. The establishment of in vitro technique for seed germination and callus induction has been done for kinnow and citron in this experiment. In case of seed germination, ½ MS media supplemented with 6-benzylaminopurine (BAP) (0.5 mg/L)+ 1-naphthaleneacetic acid (NAA) (2.0 mg/L)+KIN (1.0 mg/L) shows best seed germination response (90%) for kinnow and BAP (1.0 mg/L)+NAA (0.5 mg/L) shows best (92%) for citron. For callus induction, 5 weeks old plantlets were used as a source of leaf and stem explants for both. For callus induction from leaf and stem of kinnow mandarin, 2,4-D (1.0 mg/L) shows best result (90%) for leaf while 2,4-dichlorophenoxyacetic acid (2,4-D) (1.0 mg/L)+BAP (0.5 mg/L)+NAA (0.25 mg/L) shows best callus response (95%) for stem explants. In case of citron, 2,4-D (1.0 mg/L) shows best callus response (80%) for both leaf and stem explants. Key words:  In vitro, kinnow mandarin, citron, seed germination, callus induction