Transformation Events Research Articles

Small molecule inhibitors of mammalian GSK-3β promote in vitro plant cell reprogramming and somatic embryogenesis in crop and forest species.

Plant in vitro regeneration systems, such as somatic embryogenesis, are essential in breeding; they permit propagation of elite genotypes, production of doubled-haploids, and regeneration of whole plants from gene editing or transformation events. However, in many crop and forest species, somatic embryogenesis is highly inefficient. We report a new strategy to improve in vitro embryogenesis using synthetic small molecule inhibitors of mammalian glycogen synthase kinase 3β (GSK-3β), never used in plants. These inhibitors increased in vitro embryo production in three different systems and species, microspore embryogenesis of Brassica napus and Hordeum vulgare, and somatic embryogenesis of Quercus suber. TDZD-8, a representative compound of the molecules tested, inhibited GSK-3 activity in microspore cultures, and increased expression of embryogenesis genes FUS3, LEC2, and AGL15. Plant GSK-3 kinase BIN2 is a master regulator of brassinosteroid (BR) signalling. During microspore embryogenesis, BR biosynthesis and signalling genes CPD, GSK-3-BIN2, BES1, and BZR1 were up-regulated and the BAS1 catabolic gene was repressed, indicating activation of the BR pathway. TDZD-8 increased expression of BR signalling elements, mimicking BR effects. The findings support that the small molecule inhibitors promoted somatic embryogenesis by activating the BR pathway, opening up the way for new strategies using GSK-3β inhibitors that could be extended to other species.

LATE–a novel sensitive cell-based assay for the study of CRISPR/Cas9-related long-term adverse treatment effects

Since the introduction of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9), genome editing has been broadly applied in basic research and applied biotechnology, whereas translation into clinical testing has raised safety concerns. Indeed, although frequencies and locations of off-target events have been widely addressed, little is known about their potential biological consequences in large-scale long-term settings. We have developed a long-term adverse treatment effect (LATE) in vitro assay that addresses potential toxicity of designer nucleases by assessing cell transformation events. In small-scale proof-of-principle experiments we reproducibly detected low-frequency (<0.5%) growth-promoting events in primary human newborn foreskin fibroblasts (NUFF cells) resulting from off-target cleavage in the TP53 gene. Importantly, the LATE assay detected not only off-target effects in TP53 not predicted by popular online tools but also growth-promoting mutations in other tumor suppressor genes, such as p21 and PLZF. It convincingly verified strongly reduced off-target activities of high fidelity compared with first-generation Cas9. Finally, the LATE assay was readily adapted to other cell types, namely clinically relevant human mesenchymal stromal cells (hMSCs) and retinal pigmented epithelial (RPE-1) cells. In conclusion, the LATE assay allows assessment of physiological adverse effects of the CRISPR/Cas system and might therefore be useful for preclinical safety studies.

Fertility Myth of Oldoinyo Lengai and Its Impacts to the Maasai Community of Northern Tanzania

The paper aims to explore the myth of fertility associated with Oldoinyo Lengaiand its impacts to the Maasai community in the Gregorian rift valley system innorthern Tanzania. It assumes that in African spirituality, myths are used toguide knowledge, values and practices in the community and Mother Nature.Fertility is among the major transformation event in human life circles that canbe reflected through community myth. Through in-depth interviews andobservation techniques primary data relating to culture, customs and traditionswere obtained. Documentary search was employed to supplement field data. Thestudy found out that the Maasai community depended on the Oldoinyo Lengai(Mountain of God) for the fertility of human and animals. The fertility mythamong the Maasai pressurized them to prefer many children. The myth alsoencouraged the dominance of promiscuousness; and the deterioration of healthdue to bearing many children with limited maternal health care and the spreadof sexually transmitted diseases. Thus the myth of Oldoinyo Lengai in relation to fertility has caused an incredibly adverse impact on the Maasai community, particularly in Tanzania. The study recommends that health education and services be extended to villages in the vicinity of Oldoinyo Lengai with service providers who are well-trained to integrate the nature of the myth and its side effects to the community in relation to the changing socio-economic landscape. Moreover, it is imperative to empower the community members economically and socially so that they can decide their fate and exercise their consent.

Prognostic role and biologic features of Musashi-2 expression in colon polyps and during colorectal cancer progression.

The RNA-binding protein Musashi-2 (MSI2) controls the translation of proteins that support stem cell identity and lineage determination and is associated with progression in some cancers. We assessed MSI2 as potential clinical biomarker in colorectal cancer (CRC) and tubulovillous adenoma (TA) of colon mucosa. We assessed 125 patients, of whom 20 had polyps of the colon (TAs), and 105 had CRC. Among 105 patients with CRC, 45 had stages I-III; among metastatic CRC (mCRC) patients, 31 had synchronous and 29 metachronous liver metastases. We used immunohistochemistry to measure MSI2 expression in matching specimens of normal tissue versus TAs, primary CRC tumors, and metastases, correlating expression to clinical outcomes. We analyzed the biological effects of depleting MSI2 expression in human CRC cells. MSI2 expression was significantly elevated in polyps versus primary tissue, and further significantly elevated in primary tumors and metastases. MSI2 expression correlated with decreased progression free survival (PFS) and overall survival (OS), higher tumor grade, and right-side localization (p = 0.004) of tumors. In metastases, high MSI2 expression correlated with E-cadherin expression. Knockdown of MSI2 in CRC cells suppressed proliferation, survival and clonogenic capacity, and decreased expression of TGFβ1, E-cadherin, and ZO1. Elevated expression of MSI2 is associated with pre-cancerous TAs in the colonic mucosa, suggesting it is an early event in transformation. MSI2 expression is further elevated during CRC progression, and associated with poor prognosis. Depletion of MSI2 reduces CRC cell growth. These data imply a causative role of MSI2 overexpression at multiple stages of CRC formation and progression.

Improving Adventitious Shoot Regeneration and Transient Agrobacterium-Mediated Transformation of Apricot (Prunus armeniaca L.) Hypocotyl Sections

The improvement of previously described protocols for the regeneration of shoots from ‘Canino’ mature seed hypocotyl slices has been accomplished. The effects of different factors such as the part of the hypocotyl used, vacuum-infiltration, 2,4-Dichlorophenoxyacetic acid pulse, vacuum-infiltration and sonication on regeneration and transient transformation were analyzed. When the three slices obtained from the hypocotyls were evaluated separately on regeneration medium, the highest percentages of regenerating explants were achieved in the part close to the epicotyl and in the central part. On the other hand, sonication of the explants for 30 s followed by vacuum-infiltration during Agrobacterium infection for 20 min allowed for an increase in the transformation events. The application of these modifications to the procedure increased the regeneration efficiencies, and transient transformation events and may reduce the frequency of failed experiments. An efficient regeneration/transformation protocol could facilitate its use as a biotechnological technique for apricot breeding.

Dasatinib Discontinuation in Patients (Pts) with Chronic-Phase Chronic Myeloid Leukemia (CML-CP) and Stable Deep Molecular Response (DASFREE)

Dasatinib Discontinuation in Patients (Pts) with Chronic-Phase Chronic Myeloid Leukemia (CML-CP) and Stable Deep Molecular Response (DASFREE)

Phenotypic and Prognostic Relevance of the Who Diagnostic Distinction between Polycythemia Vera and JAK2 -Mutated Essential Thrombocythemia: Combined Analysis of 785 Informative Cases

Background:The 2016 World Health Organization (WHO) classification system (Arber et al. Blood 2016;127:2391) distinguishes essential thrombocythemia (ET) from polycythemia vera (PV), based primarily on hemoglobin/hematocrit level. However, there is significant overlap between the two in terms of other clinical and laboratory parameters and the majority of patients with both ET and PV share the JAK2 V617F mutation. These observations have suggested the possibility that JAK2 -mutated ET and PV represent a clinic-pathologic disease continuum with similar phenotype and prognosis; we explored this possibility by a combined analysis of 785 consecutive cases of WHO-defined PV or JAK2 -mutated ET.Methods:Study patients were selected from our institutional database of myeloproliferative neoplasms (MPN). Diagnoses of ET, PV, blast phase disease, and post-ET/PV myelofibrosis (MF) were according to WHO (Arber et al. Blood 2016;127:2391) and international working group for MPN research and treatment (IWG-MRT; Barosi et al. Leukemia 2008;22:437) criteria. Statistical analyses considered clinical and laboratory data collected at the time of diagnosis. Overall (OS), leukemia-free (LFS), myelofibrosis-free (MFFS) and thrombosis-free (TFS) survivals were calculated from the date of diagnosis to the date of death, event or last contact.Results:The study population (n=785) was selected from a larger cohort of 1383 consecutive cases of PV and ET and included 456 cases with PV and 329 with JAK2 -mutated ET. 1.Phenotypic comparisons at diagnosis between PV and JAK2-mutated ETOutside of the expected differences in hemoglobin level and platelet count, the 329 patients with JAK2 -mutated ET, compared to the 456 with PV, displayed a preponderance of female gender (63% vs 48%; p<0.0001) and lower incidences of leukocytosis (28% vs 55%; p<0.0001), microcirculatory symptoms (20% vs 34%; p=0.0005) and palpable splenomegaly (20% vs 29%; p=0.005). On the other hand, notable similarities included the incidences of age ≥60 years (51% vs 53%; p=0.6), thrombosis history (24% vs 25%; p=0.70), arterial thrombosis history (18% vs 16%; p=0.66) and abnormal karyotype (14% vs 19%; p=0.1). Of note, when PV was compared to all ET cases regardless of JAK2 mutational status (n=927), significant differences were also noted for age, abnormal karyotype and venous thrombosis. 1.Rates of complications post-diagnosis in PV vs JAK2-mutated ETAt a median follow-up of 9 years, 323 (41%) deaths, 26 (3%) leukemic transformations, 89 (11%) fibrotic progressions, and 161 (20%) thrombotic complications, including 115 (15%) arterial events were recorded. Compared to that of JAK2 -mutated ET, mortality (44% vs 36%; p<0.0001) and fibrotic progression (14% vs 8%; p=0.02) rates were higher in PV; there was no significant difference in blast transformation (4% vs 3%; p=0.44) or post-diagnosis thrombotic events (21% vs 20%; p=0.8). 1.Combined analysis of OS, LFS, MFFS and TFS:When all 785 study patients were analyzed together, multivariable analysis identified diagnosis of “PV” vs “ JAK2 -mutated ET” (HR 1.5, 95% CI 1.03-2.1), age ≥60 years (HR 4.3, 95% CI 3.0-6.2), leukocyte count ≥11 x 10(9)/L (HR 1.4, 95% CI 1.03-2.0) and abnormal karyotype (HR 1.9, 95% CI 1.3-2.8) as significant risk factors for OS; figure 1 depicts survival comparisons of PV vs JAK2 -mutated ET vs JAK2- unmutated ET; the apparent difference in survival between the latter two was accounted for by the significant difference in age distribution. A similar analysis for TFS identified history of diabetes (HR 2.0, 95% CI 1.3-3.2) and thrombosis (HR 1.9, 95% CI 1.4-2.7), but not the specific diagnosis of PV vs ET (p=0.7) as significant risk factors. The number of events was too small to determine the relevance of PV vs ET diagnosis for LFS or MFFS.Conclusions:The current study validates the phenotypic and prognostic relevance of the current WHO-defined diagnostic distinction between JAK2 -mutated ET and PV; phenotypically, the two were primarily different in their gender distribution and leukocyte count while survival was worse in PV; thrombotic complication rates between the two were similar, implicating the JAK2 mutation as the common offender. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Phylogeny and Evolutionary History of Respiratory Complex I Proteins in Melainabacteria.

The evolution of oxygenic photosynthesis was one of the most transformative evolutionary events in Earth’s history, leading eventually to the oxygenation of Earth’s atmosphere and, consequently, the evolution of aerobic respiration. Previous work has shown that the terminal electron acceptors (complex IV) of aerobic respiration likely evolved after the evolution of oxygenic photosynthesis. However, complex I of the respiratory complex chain can be involved in anaerobic processes and, therefore, may have pre-dated the evolution of oxygenic photosynthesis. If so, aerobic respiration may have built upon respiratory chains that pre-date the rise of oxygen in Earth’s atmosphere. The Melainabacteria provide a unique opportunity to examine this hypothesis because they contain genes for aerobic respiration but likely diverged from the Cyanobacteria before the evolution of oxygenic photosynthesis. Here, we examine the phylogenies of translated complex I sequences from 44 recently published Melainabacteria metagenome assembled genomes and genomes from other Melainabacteria, Cyanobacteria, and other bacterial groups to examine the evolutionary history of complex I. We find that complex I appears to have been present in the common ancestor of Melainabacteria and Cyanobacteria, supporting the idea that aerobic respiration built upon respiratory chains that pre-date the evolution of oxygenic photosynthesis and the rise of oxygen.

Clinical outcomes of interferon therapy for polycythemia vera and essential thrombocythemia: a systematic review and meta-analysis

Interferon therapy has been used in clinical practice for more than three decades to treat polycythemia vera (PV) and essential thrombocythemia (ET). However, there has been no systematic investigation of its expected outcomes and potential risks. We performed a systematic review and single-arm meta-analysis to assess the clinical outcomes (hematological response, molecular response, vascular events, hematological transformation, and adverse events) after interferon therapy for patients with PV and ET. A systematic search identified 37 reports, including data from 1794 patients that were published before March 2021. The pooled overall hematological response (OHR) rate was 86%, with better OHR rates observed in studies using long-acting interferon (p < 0.001) and studies with younger patients (p = 0.038). The pooled overall molecular response rate was 48%, and inter-study heterogeneity was also related to patient age (p = 0.009). The overall incidence was 0.42/100 person-years for thrombosis, 0.01/100 person-years for hemorrhage, 0.21/100 person-years for myelofibrotic transformation, and 0.08/100 person-years for leukemic transformation. Compared with hydroxyurea, interferon produced a non-inferior hematological response and a superior molecular response. In conclusion, interferon therapy provided high rates of hematological and molecular response for patients with PV and ET and was associated with a favorable prognosis.

Upfront identification of high-risk follicular lymphoma.

Follicular lymphoma (FL) is a common disease with clinically indolent behavior, and a long natural history for the majority of patients. Despite excellent therapeutic strategies currently available for FL, approximately 10%-20% of patients will experience early disease progression, defined as occurring within two years of diagnosis. These patients have poor outcomes, with overall survival at 5 years ranging between 37% and 50%. Much of the biology driving early progression and inferior survival is attributed to early transformation events; however, transformation alone does not account for all the observed clinical heterogeneity and survival differences among patients. Several clinical, genetic, and molecular alterations in FL have been discovered that help define subsets of patients at risk for multiply relapses and refractory disease, and are slowly making their way into risk calculators to be used in daily practice. Additionally, the role of functional imaging with PET scan, as well as circulating and cell free tumor DNA are being evaluated as tools to define high-risk subsets of patients with FL. This review seeks to provide an over view of current and evolving biomarkers that define high-risk FL at diagnosis. The goal is for these tools to assist clinicians in integrating these rapidly evolving prognosis models into clinical practice, in the hopes of risk-stratifying treatments and improving outcomes for patients.

COVID-19 and dynamics of environmental awareness, sustainable consumption and social responsibility in Malaysia.

Despite a profound evidence of the human unsustainable behaviours’ impact on the environment, stark disparities prevail on this narrative especially in the context of the current epidemiological situation ushered by the COVID-19. The ongoing pandemic is a global public health concern due to its sagacious impacts on environmental sustainability, social responsibility and people’s quality of life. This study primarily focuses on analysing the impact of COVID-19 (COV) on the environmental awareness (EA), sustainable consumption (SC) and social responsibility (SR). Additionally, we aspire to investigate the impact of demographics of generations and religion on the proposed nexus in this study. The data was collected from 700 participants of different age groups and religious backgrounds in Malaysia, and structural equation modelling (SEM) was used to analyse this data and test the hypotheses. The findings indicate that COVID-19 has a significantly positive impact on EA, SC and SR, and the generations and religiosity moderate the relationship between COVID-19 and its impact on sustainable behaviours. This study contributes to analyse the difference in the perception of EA, SC and SR among the people that eventually will stimulate the scientific reasoning among the governments, policymakers and scientists to develop a holistic framework to combat unprecedented event such as COVID-19 and ensure the authentication of sustainable environment and exceptional quality of life. The policymakers in Malaysia may use the findings of this study to inspect the social and environmental aspects of the people during the transformation events.

Disruption of mitochondrial open reading frame 352 partially restores pollen development in cytoplasmic male sterile rice.

Plant mitochondrial genomes sometimes carry cytoplasmic male sterility (CMS)-associated genes. These genes have been harnessed in various crops to produce high-yielding F1 hybrid seeds. The gene open reading frame 352 (orf352) was reported to be an RT102-type CMS gene in rice (Oryza sativa), although the mechanism underlying its role in CMS is unknown. Here, we employed mitochondrion-targeted transcription activator-like effector nucleases (mitoTALENs) to knockout orf352 from the mitochondrial genome in the CMS rice RT102A. We isolated 18 independent transformation events in RT102A that resulted in genome editing of orf352, including its complete removal from the mitochondrial genome in several plants. Sequence analysis around the mitoTALEN target sites revealed their induced double-strand breaks were repaired via homologous recombination. Near the 5ʹ-target site, repair involved sequences identical to orf284, while repair of the 3ʹ-target site yielded various new sequences that generated chimeric genes consisting of orf352 fragments. Plants with a chimeric mitochondrial gene encoding amino acids 179–352 of ORF352 exhibited the same shrunken pollen grain phenotype as RT102A, whereas plants either lacking orf352 or harboring a chimeric gene encoding amino acids 211–352 of ORF352 exhibited partial rescue of pollen viability and germination, although these plants failed to set seed. These results demonstrated that disruption of orf352 partially restored pollen development, indicating that amino acids 179–210 from ORF352 may contribute to pollen abortion.

Remembering summerdale: A scandal that reformed policing

The following article contains never-before-published accounts from first-hand witnesses to the events surrounding the Summerdale Scandal, garnered by one of the authors in personal interviews with key stakeholders. These include interviews with former Evanston and Bartlett Police Chief William McHugh, former Illinois Police Superintendent James McGuire, former Cook County State’s Attorney Chief Investigator Paul Newey and renowned attorneys Gerald Getty, Julius Echels and Louis Galippo. Though all have now passed away, their words bring to life an event that forever changed policing in America. In 1960, a massive scandal rocked the Chicago Police Department. Rife with clandestine schemes, audacious exploits and a level of corruption that stunned the public, the Summerdale Scandal nearly brought down the entire Chicago police organization. Once the scandal unfurled, it would transform policing, not just in Chicago, but around the country. 2020 will mark the sixtieth anniversary of the Summerdale Scandal, and once again find ourselves recovering from police corruption. The scandal surrounding the events of the 2017 Chicago Police shooting of Laquan McDonald resulted in the firing of a police superintendent, affected the decision of Mayor Rahm Emmanuel in not running for re-election, and forced the Chicago Police Department into a Federal Oversight Consent Decree. Now, as we approach the 60th anniversary of the Summerdale Scandal, it bears revisiting this transformative event that dramatically overhauled policing four decades earlier.

High strain rate in situ micropillar compression of a Zr-based metallic glass

High strain rate micromechanical testing can assist researchers in elucidating complex deformation mechanisms in advanced material systems. In this work, the interactions of atomic-scale chemistry and strain rate in affecting the deformation response of a Zr-based metallic glass was studied by varying the concentration of oxygen dissolved into the local structure. Compression of micropillars over six decades of strain rate uncovered a remarkable reversal of the strain rate sensitivity from negative to positive above ~ 5 s−1 due to a delocalisation of shear transformation events within the pre-yield linear regime for both samples, while a higher oxygen content was found to generally decrease the strain rate sensitivity effect. It was also identified that the shear band propagation speed increases with the actuation speed, leading to a transition in the deformation behaviour from serrated to apparent non-serrated plastic flow at ~ 5 s−1.Graphic abstract

ASGE Past President's Address.

John J. Vargo, MD, MPH, MASGE (Author photo 1, Author photo 2, Author photo 3, Author photo 4, Author photo 5, Author photo 6), is an associate professor of medicine at the Cleveland Clinic Lerner College of Medicine at Case Western University. He is a member of the Department of Gastroenterology, Hepatology and Nutrition in the Digestive Disease and Surgery Institute at the Cleveland Clinic.Author photo 2Department of Gastroenterology and Hepatology at the Cleveland Clinic. I am blessed to be surrounded by one of the finest faculties and groups of fellows in the world.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Author photo 3World Cup of Endoscopy at Digestive Disease Week. The alter ego comes out. Now, what to do with the costumes?View Large Image Figure ViewerDownload Hi-res image Download (PPT)Author photo 4With my dear friends Seiichiro Abe, Yutaka Saito, and Amit Bhatt.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Author photo 5With my friend of many years, the incomparable G.S. Raju.View Large Image Figure ViewerDownload Hi-res image Download (PPT)Author photo 6My family: my wife Mary and our sons (left to right) Kevin, John, and Matthew.View Large Image Figure ViewerDownload Hi-res image Download (PPT) Dr Vargo received his medical degree from the University of Rochester School of Medicine and Dentistry and completed his internal medicine residency training at the University of Pittsburgh Medical Center. He then completed gastroenterology and hepatology fellowships at the Cleveland Clinic and received training in advanced endoscopy at the same institution. After fulfilling a 3-year commitment in the United States Navy while stationed at Bethesda National Naval Medical Center as director of endoscopy and consultant in gastroenterology for the United States Capitol, he returned to the Cleveland Clinic in the section of advanced endoscopy. During his tenure, Dr Vargo developed a keen interest in clinical research. After securing a career development award from the American Society for Gastrointestinal Endoscopy (ASGE) and the American Digestive Health Foundation, Dr Vargo obtained a Master of Public Health degree from Case Western University. This turned out to be a transformational event, which led to the development of numerous endoscopic research projects focusing on clinical outcomes, quality, and cost-effectiveness. This resulted in the publication of over 250 articles in the peer-reviewed literature. Dr Vargo served as chair of the Department of Gastroenterology and Hepatology and vice chair for the Digestive Disease and Surgery Institute for 8 years. He also served as president of the Cleveland Clinic’s medical staff and sat on the institution’s Board of Governors. He is currently the director of Enterprise Endoscopy Operations, director of Endoscopic Research and Innovation, and head of the Section of Advanced Endoscopy. He has lectured and performed endoscopic demonstrations around the globe and has been honored with fellowships in 3 international gastroenterology societies. He has been invited faculty for numerous courses for ASGE and has been co-director of ASGE’s annual postgraduate course twice. He has held numerous positions in ASGE, including chair of the Training Committee. During his tenure, the committee was able to develop an advanced endoscopy training directory and began development of the forerunner of the advanced endoscopy match program. Dr Vargo served as president of ASGE from 2019 to 2020. During this entropic time of the COVID-19 pandemic, he led the society through a transition to a virtual education and communications platform, which ensured that information on the COVID-19 pandemic was expediently distributed via weekly webinars, fast-tracked articles in GIE, and expedient practice guidelines that were co-authored with ASGE’s sister societies. During his presidency, the entire framework of ASGE’s educational mission was reframed and crystallized to meet the needs of gastroenterologists in the United States and around the world. Dr Vargo has trained a multitude of general gastroenterology and advanced endoscopy fellows over the years, who are now scattered around the United States and throughout the world. He is a perennial Top Doctors and Best Doctors in America selection. Dr Vargo received teacher-of-the-year awards from his gastroenterology fellows a total of 5 times over his career. Areas of interest in his practice include ERCP, EUS, the treatment of Barrett’s esophagus, artificial intelligence, and the development of endoscopic devices. All authors disclosed no financial relationships. https://www.videogie.org/cms/asset/776b53df-e503-4e22-94df-3f45da5ce4ab/mmc1.mp4Loading ... Download .mp4 (179.53 MB) Help with .mp4 files Video 1

Thymol production in hairy root culture of Sahendian savory (Satureja sahendica Bornm)

Sahendian savory (Satureja sahendica Bornm) is a medicinal plant species which produces valuable secondary metabolites including thymol, p-cymene and γ-terpinene. In this study, optimization of hairy roots culture in S. sahendica using Agrobacterium rhizogenes was efficiently established. High frequency of genetic transformation was obtained using node and internode explants and three bacterial strains (ATCC 15834, A4, and LBA 9402). However, leaf explants showed no susceptibility to induce hairy roots with the A. rhizogenes strains. The node explants and LBA 9402 strain were the best combination for root induction. Infection time of 20 min caused greater transformation occurrence compared to 10 and 30 min, respectively. Node explants infected with LBA 9402 were found to produce more transformants upon co-cultivation for 2 days. The maximum induction of hairy roots was obtained from bacterial infection at a density of OD600 = 0.8. Polymerase chain reaction by gene-specific primers from rol B gene confirmed the positive transformation events. S. sahendica efficiently provided and produced a large number of hairy roots which could be used for the production of secondary metabolites or other molecular and biotechnological applications. Production of thymol an important secondary metabolite of S. sahendica from hairy root cultures was confirmed by TLC and GC analysis. Elicitor-treated hairy root cultures with methyl jasmonate and ammonium nitrate produced more amount of thymol compared to untreated roots. These results demonstrate that secondary metabolite production such as thymol in hairy root culture of Sahendian savory could be used to increase low secondary metabolite production in this valuable medicinal plant.

The role of (dynamic) capabilities in the transformation of a multi-organizational setting

This paper analyzes the role of dynamic capabilities in the transformation of a multi-organizational setting. I conduct an embedded multi-case study in which the cases represent critical transformational events in the history of a shopping center. The five critical transformational events, which have exerted substantial influence on the organizations operating in the context, are concerned with the construction of new premises, modernization of existing premises, and the introduction of new services. I analyze and classify related (dynamic) capabilities of individual organizations, resulting in aggregated outcomes that determine the evolution of the entity. Applying an abductive case research strategy, I then generate five distinct theoretical results on the effects and the interconnectedness of the multi-organizational context, the individual organizations, and their dynamic capabilities.

Enacting the aesthetic: A model for raw cognitive dynamics

One challenge faced by aesthetics is the development of an account able to trace out the continuities and discontinuities between general experience and aesthetic experiences. Regarding this issue, in this paper, I present an enactive model of some raw cognitive dynamics that might drive the progressive emergence of aesthetic experiences from the stream of general experience. The framework is based on specific aspects of John Dewey’s pragmatist philosophy and embodied aesthetic theories, while also taking into account research in ecological psychology, cognitive sciences, and dynamic systems theory. The model focuses on dynamically relevant nodes at the pre-reflective and the reflective side of experience that would work as nested rhythmic constraints at different cognitive timescales with the potential to shunt experiences toward the aesthetic in everyday situations. My proposal constitutes a way to explore aesthetic experiences from an enactive perspective that regards them as transformative events in which cognitive processes entrain and are entrained by changes taking place in the environment, the brain, and the body.

Agrobacterium-mediated In-planta transformation of bread wheat (Triticum aestivum L.)

Agrobacterium-mediated in-planta transformation method allows efficient plant transformation without tissue culture. In the present study, a tissue culture independent protocol for floral transformation in bread wheat was developed. Three bread wheat genotypes namely, HD 2967, HD 3086 and Bobwhite were used for floral transformation. At pre-anthesis stage, clipped florets were sprayed with inoculation medium containing Agrobacterium tumefaciens strain EHA 105 culture harboring binary vector pCAMBIA1301-p35S:hptII-p35S:gus and allowed to grow till maturity. Positive transformants were confirmed through antibiotic selection, PCR analysis and southern blotting. The method led to generation of transformation events in Bobwhite and HD 2967 wheat genotypes. The transformation efficiency of T1 transgenic plants was higher in Bobwhite as compared to HD 2967. Floral spray method of transformation developed in the present study is efficient for development of transgenic wheat and can be used for improvement of wheat using transgenic approach.

Plastid transformation: Advances and challenges for its implementation in agricultural crops

Chloroplast biotechnology has emerged as a promissory platform for the development of modified plants to express products aimed mainly at the pharmaceutical, agricultural, and energy industries. This technology’s high value is due to its high capacity for the mass production of proteins. Moreover, the interest in chloroplasts has increased because of the possibility of expressing multiple genes in a single transformation event without the risk of epigenetic effects. Although this technology solves several problems caused by nuclear genetic engineering, such as turning plants into safe bio-factories, some issues must still be addressed in relation to the optimization of regulatory regions for efficient gene expression, cereal transformation, gene expression in non-green tissues, and low transformation efficiency. In this article, we provide information on the transformation of plastids and discuss the most recent achievements in chloroplast bioengineering and its impact on the biopharmaceutical and agricultural industries; we also discuss new tools that can be used to solve current challenges for their successful establishment in recalcitrant crops such as monocots. How to cite: Quintín Rascón-Cruz Q, González-Barriga CD, Iglesias-Figueroa BF, et al. Plastid transformation: Advances and challenges for its implementation in agricultural crops. Electron J Biotechnol 2021;51. https://doi.org/10.1016/j.ejbt.2021.03.005