Effect of Saccharomyces boulardii and Lactobacillus acidophilus fermentation on little millet (Panicum sumatrense)

The fermentation of little millet (Panicum sumatrense) carried out by probiotic yeast (Saccharomyces boulardii) and lactic acid bacteria (Lactobacillus acidophilus). The germinated little millets grains were fermented for 5 days grains were dried and grounded in a grinder and sieved. The sieved flours were analyzed for proximate composition, minerals and phytic acid content. The results showed that the little millet fermented by combined inoculation of Saccharomyces boulardii and L. acidophilus significantly enhanced protein content (10.95%) and decreases the fat (2.61%) and carbohydrate (82.01%) and single inoculation with L. acidophilus decreased the fibre (1.05%) and ash (2.30%). Single inoculation of L. acidophilus enhanced the minerals content Ca (41.44mg), Mg (141.08 mg), phosphorus (238.43mg), Fe (7.98 mg) and Zn (4.69mg) and over dual inoculation and control treatments. Similarly, Single inoculationof Lactobacillus acidophilus showed significant reduction in phytic acid (114.76 mg) content over dual inoculation (126.49 mg) and control (188.95 mg) treatments. The results indicated that fermentation of little millet by L. acidophilus enhanced mineral contents and reduction in phytic acid content

Nucleic acids delivery methods for genome editing in zygotes and embryos: the old, the new, and the old-new

In the recent years, sequence-specific nucleases such as ZFNs, TALENs, and CRISPR/Cas9 have revolutionzed the fields of animal genome editing and transgenesis. However, these new techniques require microinjection to deliver nucleic acids into embryos to generate gene-modified animals. Microinjection is a delicate procedure that requires sophisticated equipment and highly trained and experienced technicians. Though over a dozen alternate approaches for nucleic acid delivery into embryos were attempted during the pre-CRISPR era, none of them became routinely used as microinjection. The addition of CRISPR/Cas9 to the genome editing toolbox has propelled the search for novel delivery approaches that can obviate the need for microinjection. Indeed, some groups have recently developed electroporation-based methods that have the potential to radically change animal transgenesis. This review provides an overview of the old and new delivery methods, and discusses various strategies that were attempted during the last three decades. In addition, several of the methods are re-evaluated with respect to their suitability to deliver genome editing components, particularly CRISPR/Cas9, to embryos. Reviewers: Drs. Eugene Koonin and Haruhiko Siomi

Defect Investigation as a Tool for Quality Improvement

Defect investigation is an important tool for quality improvement as it identifies the root cause of errors and steps to be taken to eliminate these. In this paper, four cases of defect investigation are presented. The case studies cover link elevator, wing-fuselage attachment bolt, and stiff nut for Lakshya unmanned aerial vehicle. It was observed that the link elevator was discolourised even though the material used was the stainless steel. Investigations revealed that the stainless steel was heat-treated. In the case of leading edge, it was found that there was no provision for the location of a component in the tool, resulting in mismatch. Thread damage was noticed in wing-fuselage attachment bolts because of over tightening. All these studies resulted in improvement in the raw material bonding, inspection procedures, and fabrication methods, thus resulting in quality improvement

Easi-CRISPR: a robust method for one-step generation of mice carrying conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins

Background Conditional knockout mice and transgenic mice expressing recombinases, reporters, and inducible transcriptional activators are key for many genetic studies and comprise over 90% of mouse models created. Conditional knockout mice are generated using labor-intensive methods of homologous recombination in embryonic stem cells and are available for only ~25% of all mouse genes. Transgenic mice generated by random genomic insertion approaches pose problems of unreliable expression, and thus there is a need for targeted-insertion models. Although CRISPR-based strategies were reported to create conditional and targeted-insertion alleles via one-step delivery of targeting components directly to zygotes, these strategies are quite inefficient. Results Here we describe Easi-CRISPR (Efficient additions with ssDNA inserts-CRISPR), a targeting strategy in which long single-stranded DNA donors are injected with pre-assembled crRNA + tracrRNA + Cas9 ribonucleoprotein (ctRNP) complexes into mouse zygotes. We show for over a dozen loci that Easi-CRISPR generates correctly targeted conditional and insertion alleles in 8.5–100% of the resulting live offspring. Conclusions Easi-CRISPR solves the major problem of animal genome engineering, namely the inefficiency of targeted DNA cassette insertion. The approach is robust, succeeding for all tested loci. It is versatile, generating both conditional and targeted insertion alleles. Finally, it is highly efficient, as treating an average of only 50 zygotes is sufficient to produce a correctly targeted allele in up to 100% of live offspring. Thus, Easi-CRISPR offers a comprehensive means of building large-scale Cre-LoxP animal resources

Multi-modality Typical Imaging features in Superior Pulmonary Sulcus (Pancoast) tumour: A case report

A 53 years old male patient came with complaints of pain and tingling sensation of left shoulder joint & left upper limb since 3 months. Patient is a chronic smoker with past history pulmonary tuberculosis. The patient had completed ATT 15 years bac

Reprogramming human T cell function and specificity with non-viral genome targeting.

Decades of work have aimed to genetically reprogram T cells for therapeutic purposes1,2 using recombinant viral vectors, which do not target transgenes to specific genomic sites3,4. The need for viral vectors has slowed down research and clinical use as their manufacturing and testing is lengthy and expensive. Genome editing brought the promise of specific and efficient insertion of large transgenes into target cells using homology-directed repair5,6. Here we developed a CRISPR-Cas9 genome-targeting system that does not require viral vectors, allowing rapid and efficient insertion of large DNA sequences (greater than one kilobase) at specific sites in the genomes of primary human T cells, while preserving cell viability and function. This permits individual or multiplexed modification of endogenous genes. First, we applied this strategy to correct a pathogenic IL2RA mutation in cells from patients with monogenic autoimmune disease, and demonstrate improved signalling function. Second, we replaced the endogenous T cell receptor (TCR) locus with a new TCR that redirected T cells to a cancer antigen. The resulting TCR-engineered T cells specifically recognized tumour antigens and mounted productive anti-tumour cell responses in vitro and in vivo. Together, these studies provide preclinical evidence that non-viral genome targeting can enable rapid and flexible experimental manipulation and therapeutic engineering of primary human immune cells

Post total hip arthroplasty pseudo- tumour – A rare case report.

A 62-year-old female came with complaints of slow growing mass in the left hip region for 4-5 months, associated with dull aching pain. Range of movements were restricted due to pain and mass. She underwent Total Hip arthroplasty (THA) 11 months ago