Targeted insertion of transgenes at pre-determined plant genomic safe harbors provides a desirable alternative to insertions at random sites achieved through conventional methods. Most existing cases ...of targeted gene insertion in plants have either relied on the presence of a selectable marker gene in the insertion cassette or occurred at low frequency with relatively small DNA fragments (<1.8 kb). Here, we report the use of an optimized CRISPR-Cas9-based method to achieve the targeted insertion of a 5.2 kb carotenoid biosynthesis cassette at two genomic safe harbors in rice. We obtain marker-free rice plants with high carotenoid content in the seeds and no detectable penalty in morphology or yield. Whole-genome sequencing reveals the absence of off-target mutations by Cas9 in the engineered plants. These results demonstrate targeted gene insertion of marker-free DNA in rice using CRISPR-Cas9 genome editing, and offer a promising strategy for genetic improvement of rice and other crops.
The availability of a whole-genome sequenced mutant population and the cataloging of mutations of each line at a single-nucleotide resolution facilitate functional genomic analysis. To this end, we ...generated and sequenced a fast-neutron-induced mutant population in the model rice cultivar Kitaake (Oryza sativa ssp japonica), which completes its life cycle in 9 weeks. We sequenced 1504 mutant lines at 45-fold coverage and identified 91,513 mutations affecting 32,307 genes, i.e., 58% of all rice genes. We detected an average of 61 mutations per line. Mutation types include single-base substitutions, deletions, insertions, inversions, translocations, and tandem duplications. We observed a high proportion of loss-of-function mutations. We identified an inversion affecting a single gene as the causative mutation for the short-grain phenotype in one mutant line. This result reveals the usefulness of the resource for efficient, cost-effective identification of genes conferring specific phenotypes. To facilitate public access to this genetic resource, we established an open access database called KitBase that provides access to sequence data and seed stocks. This population complements other available mutant collections and gene-editing technologies. This work demonstrates how inexpensive next-generation sequencing can be applied to generate a high-density catalog of mutations.
Hydrogels based on protein and polysaccharides have ameliorated the arena of tissue regeneration, and other healthcare applications. We report for the first time a new class of conjugated hydrogel ...based on carbohydrate polymer consisting of biomass derived nanocellulose and collagen inspired complementary ionic peptides. Interestingly, these conjugate hydrogels were constructed via simple non-covalent interactions. The synthetic strategy utilized TEMPO oxidized cellulose nanofibers which presented the anionic surface charge, and thus offer the scope of electrostatic interactions with the oppositely charged moieties of peptides. Further, these biomolecular hydrogel constructs were explored towards cellular studies in order to develop a superior replica of native extracellular matrix. Interestingly, differential cellular response could be induced in such novel biopolymeric matrix by judicious tuning of the intermolecular interactions to fabricate suitable matrix with variable physical properties, like, porosity, mechanical stiffness etc. Tunable porous network of these conjugated biopolymeric hydrogels stabilized by combination of CH/π, hydrogen bonding interactions along with electrostatic interactions between the cellulose and collagen mimetic peptides present a scaffold that successfully mimic the merits of the native extracellular matrix by combing peptide and sugar leading to significant cellular adhesion, growth, and proliferation. Interestingly, these combined scaffolds supported cellular behaviour of both fibroblast as well as neural cells, highlighting the diversities of these conjugate hydrogel.
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Ezetimibe inhibits intestinal cholesterol absorption and lowers low‐density lipoprotein cholesterol. Uncontrolled studies have suggested that it reduces liver fat as estimated by ultrasound in ...nonalcoholic steatohepatitis (NASH). Therefore, we aimed to examine the efficacy of ezetimibe versus placebo in reducing liver fat by the magnetic resonance imaging‐derived proton density‐fat fraction (MRI‐PDFF) and liver histology in patients with biopsy‐proven NASH. In this randomized, double‐blind, placebo‐controlled trial, 50 patients with biopsy‐proven NASH were randomized to either ezetimibe 10 mg orally daily or placebo for 24 weeks. The primary outcome was a change in liver fat as measured by MRI‐PDFF in colocalized regions of interest within each of the nine liver segments. Novel assessment by two‐dimensional and three‐dimensional magnetic resonance elastography was also performed. Ezetimibe was not significantly better than placebo at reducing liver fat as measured by MRI‐PDFF (mean difference between the ezetimibe and placebo arms ‐1.3%, P = 0.4). Compared to baseline, however, end‐of‐treatment MRI‐PDFF was significantly lower in the ezetimibe arm (15%‐11.6%, P < 0.016) but not in the placebo arm (18.5%‐16.4%, P = 0.15). There were no significant differences in histologic response rates, serum alanine aminotransferase and aspartate aminotransferase levels, or longitudinal changes in two‐dimensional and three‐dimensional magnetic resonance elastography‐derived liver stiffness between the ezetimibe and placebo arms. Compared to histologic nonresponders (25/35), histologic responders (10/35) had a significantly greater reduction in MRI‐PDFF (‐4.35 ± 4.9% versus ‐0.30 ± 4.1%, P < 0.019). Conclusions: Ezetimibe did not significantly reduce liver fat in NASH. This trial demonstrates the application of colocalization of MRI‐PDFF‐derived fat maps and magnetic resonance elastography‐derived stiffness maps of the liver before and after treatment to noninvasively assess treatment response in NASH. (Hepatology 2015;61:1239–1250)
The coronavirus disease 2019 (COVID-19) pandemic is an issue of global significance that has taken the lives of many across the world. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is ...the virus responsible for its pathogenesis. The pulmonary manifestations of COVID-19 have been well described in the literature. Initially, it was thought to be limited to the respiratory system; however, we now recognize that COVID-19 also affects several other organs, including the nervous system. Two similar human coronaviruses (CoV) that cause severe acute respiratory syndrome (SARS-CoV-1) and Middle East respiratory syndrome (MERS-CoV) are also known to cause disease in the nervous system. The neurological manifestations of SARS-CoV-2 infection are growing rapidly, as evidenced by several reports. There are several mechanisms responsible for such manifestations in the nervous system. For instance, post-infectious immune-mediated processes, direct virus infection of the central nervous system (CNS), and virus-induced hyperinflammatory and hypercoagulable states are commonly involved. Guillain-Barré syndrome (GBS) and its variants, dysfunction of taste and smell, and muscle injury are numerous examples of COVID-19 PNS (peripheral nervous system) disease. Likewise, hemorrhagic and ischemic stroke, encephalitis, meningitis, encephalopathy acute disseminated encephalomyelitis, endothelialitis, and venous sinus thrombosis are some instances of COVID-19 CNS disease. Due to multifactorial and complicated pathogenic mechanisms, COVID-19 poses a large-scale threat to the whole nervous system. A complete understanding of SARS-CoV-2 neurological impairments is still lacking, but our knowledge base is rapidly expanding. Therefore, we anticipate that this comprehensive review will provide valuable insights and facilitate the work of neuroscientists in unfolding different neurological dimensions of COVID-19 and other CoV associated abnormalities.
Background: Cervical cancer is one of the leading causes of death among women facing gynecological problems. It requires early detection by various screening methods.
Aims and Objectives: The current ...research aimed to assess the diagnostic utility of pap smear cytology and visual inspection of acetic acid (VIA) as a screening test for various cervical lesions.
Materials and Methods: The current prospective study was conducted on 210 non-pregnant women. Cervical pap smears were taken and reported according to the revised Bethesda system, 2014. VIA examination was also done, and any dense opaque aceto-white lesion observation near the squamo-columnar junction was reported as VIA positive. The findings of pap smear cytology and VIA were correlated with findings of histopathology.
Results: Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy of pap smear cytology for the diagnosis of cervical lesions were found to be 90.16%, 85.22%, 89.43%, 86.2%, and 88.09%, respectively; while sensitivity, specificity, PPV, NPV, and diagnostic accuracy of VIA for the diagnosis of cervical lesions were found to be 88.37%, 55.55%, 76%, 75%, and 75.71%, respectively. Our study showed a substantial degree of agreement between pap smear cytology and histopathology for the diagnosis of cervical lesions with Cohen’s kappa value being 0.755.
Conclusion: The current study revealed that pap smear and VIA have high sensitivity with good diagnostic accuracy for the detection of neoplastic cervical lesions, which indicate that they can be used as an efficient screening tool for the detection of cervical lesions in their earlier stages.
This paper presents a grid connected solar photovoltaic (PV)-battery system using a double stage topology. The ESOGI (Enhanced Second Order Generalised Integrator)-based control estimates the ...positive sequence components (PSCs) of grid voltages, the amplitude of grid voltages, and unit templates. The VSC (voltage source converter) converts the DC power to AC power and providing constant power to the grid under a predefined power mode. VSC also performs multiple functions like balancing the grid currents, harmonics elimination, and providing reactive power compensation to the loads. Moreover, BDC (bidirectional converter) control maintains the voltage at the DC link and battery current. At low insolation levels, the battery supplies the required power to the load under a predefined power mode. Moreover, the battery supplies the predefined power to the grid under a predefined power mode condition. Under various dynamic conditions, simulated results and experimental results have shown a good performance.
The availability of thousands of complete rice genome sequences from diverse varieties and accessions has laid the foundation for in-depth exploration of the rice genome. One drawback to these ...collections is that most of these rice varieties have long life cycles, and/or low transformation efficiencies, which limits their usefulness as model organisms for functional genomics studies. In contrast, the rice variety Kitaake has a rapid life cycle (9 weeks seed to seed) and is easy to transform and propagate. For these reasons, Kitaake has emerged as a model for studies of diverse monocotyledonous species.
Here, we report the de novo genome sequencing and analysis of Oryza sativa ssp. japonica variety KitaakeX, a Kitaake plant carrying the rice XA21 immune receptor. Our KitaakeX sequence assembly contains 377.6 Mb, consisting of 33 scaffolds (476 contigs) with a contig N50 of 1.4 Mb. Complementing the assembly are detailed gene annotations of 35,594 protein coding genes. We identified 331,335 genomic variations between KitaakeX and Nipponbare (ssp. japonica), and 2,785,991 variations between KitaakeX and Zhenshan97 (ssp. indica). We also compared Kitaake resequencing reads to the KitaakeX assembly and identified 219 small variations. The high-quality genome of the model rice plant KitaakeX will accelerate rice functional genomics.
The high quality, de novo assembly of the KitaakeX genome will serve as a useful reference genome for rice and will accelerate functional genomics studies of rice and other species.
In this research, α − ψ − θ contraction has been defined to find the best proximity point in partially ordered metric spaces. Proper support for the result has been given in the form of a suitable ...example. The third part is fully devoted to the positive definite solution of matrix equations.
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