SONICATION AND VACUUM INFILTRATION ENHANCED AGROBACTERIUM RHIZOGENES MEDIATED TRANSFORMATION IN SOYBEAN

AbstractObjective: The present study involved the formulation of protocol of Agrobacterium rhizogenes-mediated transformation for the detailed study of isoflavones metabolism in soybean.Methods: Cotyledons were separated from 4-day-old soybean seedlings and infected with three different A. rhizogenes strains under various time durations of sonication, vacuum infiltration and co-cultivated on MS medium supplemented with various concentrations of acetosyringone. The induced hairy roots were established as a culture with the selection agent hygromycin B. Transgenes integrated in hairy roots were analysed at molecular level by PCR assay.Results: A. rhizogenes strain R1000 harbouring pCAMBIA1301 resulted in better transformation efficiency when compared with other strains. The optimum duration of sonication (2 min) and vacuum infiltration (2 min) enhanced the transformation efficiency up to 76.47 %.  PCR analyses revealed the integration of transgene in hairy roots lines.Conclusion:  From the present study, we could conclude that, sonication and vacuum infiltration techniques could be employed to produce genotype independent transgenic soybean hairy root lines and which could be used to study for the improved production of potent anti-cancer compounds, isoflavones in soybean.Keywords: Agrobacterium, isoflavones, soybean, sonication, vacuum infiltratio

An endophyte Paenibacillus dendritiformis strain APL3 promotes Amaranthus polygonoides L. sprout growth and their extract inhibits food-borne pathogens

Green leafy vegetables are rich sources of antioxidants and minerals, which prevent food-borne pathogen infections during our diet. This study was aimed to isolate and identify the plant growth-promoting endophytic bacterium from several plant species to enhance the growth of Amaranthus polygonoides L. and their antimicrobial potential against food-borne pathogens. Seven endophytic bacterial isolates were tested on two Amaranthus species to identify the suitable beneficial bacterium. The antioxidants capacity and antimicrobial activity of bacterial isolate (APL3) treated plants were analyzed. The bacterial isolate, APL3 showed a significantly higher growth of A. polygonoides L. than other isolates. It was identified as Paenibacillus dendritiformis strain APL3 by 16S rRNA gene sequencing and phylogenetic analysis. The endophyte (APL3) treated A. polygonoides L. sprouts had higher antioxidants potentials and significantly inhibited the growth of Escherichia coli, Salmonella sp., Staphylococcus sp. and Pseudomonas sp. The results of the present study suggest that utilization of P. dendritiformis strain APL3 triggers the growth of A. polygonoides L. and induces metabolic changes in plants to improve their antimicrobial properties to prevent foodborne pathogens

Mechanical properties of 3D printed concrete:a RILEM TC 304-ADC interlaboratory study — flexural and tensile strength

This paper discusses the flexural and tensile strength properties of 3D printed concrete, based on the results of a RILEM TC 304-ADC interlaboratory study on mechanical properties. These properties are determined using different testing techniques, including 3- and 4-point flexural tests, splitting tests, and uniaxial tension tests, on specimens extracted from large 3D printed elements in accordance with a prescribed study plan. The relationship between compressive and flexural or tensile strengths, cast or printed samples, different types of tests, and different loading orientations, are analysed to understand the influence of 3D printing. As expected, the strength can reduce significantly when the main tensile stress is acting perpendicular to the interface between layers. The role of deviations from the standard study procedure, in terms of the time interval between the placing of subsequent layers, or the adoption of a different curing strategy, are also assessed. While the increased time interval significantly impacts the strength in the critical direction, the use of variable curing conditions does not seem to have a clear-cut effect on the strength ratios of the printed to cast specimens. Additionally, the paper looks at the variability in the results for the printed specimens, in order to emphasize the need for multiple replicates for obtaining a proper result. An extensive insight into the aspects affecting the variability is presented in the paper. Finally, with the limited dataset available for specimens tested at a larger scale, it is difficult to arrive at a clear understanding of the role of specimen size (i.e., greater number of layers)

Mechanical properties of 3D printed concrete: a RILEM 304-ADC interlaboratory study – compressive strength and modulus of elasticity

Traditional construction techniques, such as in-situ casting and pre-cast concrete methods, have well-established testing protocols for assessing compressive strength and modulus of elasticity, including specific procedures for sample preparation and curing. In contrast, 3D concrete printing currently lacks standardized testing protocols, potentially contributing to the inconsistent results reported in previous studies. To address this issue, RILEM TC 304-ADC initiated a comprehensive interlaboratory study on the mechanical properties of 3D printed concrete. This study involves 30 laboratories worldwide, contributing 34 sets of data, with some laboratories testing more than one mix design. The compressive strength and modulus of elasticity were determined under three distinct conditions: Default, where each laboratory printed according to their standard procedure followed by water bath curing; Deviation 1, which involved creating a cold joint by increasing the time interval between printing layers; and Deviation 2, where the standard printing process was used, but the specimens were cured under conditions different from water bath. Some tests were conducted at two different scales based on specimen size—“mortar-scale” and “concrete-scale”—to investigate the size effect on compressive strength. Since the mix design remained identical for both scales, the only variable was the specimen size. This paper reports on the findings from the interlaboratory study, followed by a detailed investigation into the influencing parameters such as extraction location, cold joints, number of interlayers, and curing conditions on the mechanical properties of the printed concrete. As this study includes results from laboratories worldwide, its contribution to the development of relevant standardized testing protocols is critical

Zika Virus Encoding Nonglycosylated Envelope Protein Is Attenuated and Defective in Neuroinvasion

ABSTRACT Zika virus (ZIKV), a mosquito-transmitted flavivirus responsible for sporadic outbreaks of mild and febrile illness in Africa and Asia, reemerged in the last decade causing serious human diseases, including microcephaly, congenital malformations, and Guillain-Barré syndrome. Although genomic and phylogenetic analyses suggest that genetic evolution may have led to the enhanced virulence of ZIKV, experimental evidence supporting the role of specific genetic changes in virulence is currently lacking. One sequence motif, VNDT, containing an N-linked glycosylation site in the envelope (E) protein, is polymorphic; it is absent in many of the African isolates but present in all isolates from the recent outbreaks. In the present study, we investigated the roles of this sequence motif and glycosylation of the E protein in the pathogenicity of ZIKV. We first constructed a stable full-length cDNA clone of ZIKV in a novel linear vector from which infectious virus was recovered. The recombinant ZIKV generated from the infectious clone, which contains the VNDT motif, is highly pathogenic and causes lethality in a mouse model. In contrast, recombinant viruses from which the VNDT motif is deleted or in which the N-linked glycosylation site is mutated by single-amino-acid substitution are highly attenuated and nonlethal. The mutant viruses replicate poorly in the brains of infected mice when inoculated subcutaneously but replicate well following intracranial inoculation. Our findings provide the first evidence that N-linked glycosylation of the E protein is an important determinant of ZIKV virulence and neuroinvasion. IMPORTANCE The recent emergence of Zika virus (ZIKV) in the Americas has caused major worldwide public health concern. The virus appears to have gained significant pathogenicity, causing serious human diseases, including microcephaly and Guillain-Barré syndrome. The factors responsible for the emergence of pathogenic ZIKV are not understood at this time, although genetic changes have been shown to facilitate virus transmission. All isolates from the recent outbreaks contain an N-linked glycosylation site within the viral envelope (E) protein, whereas many isolates of the African lineage virus lack this site. To elucidate the functional significance of glycosylation in ZIKV pathogenicity, recombinant ZIKVs from infectious clones with or without the glycan on the E protein were generated. ZIKVs lacking the glycan were highly attenuated for the ability to cause mortality in a mouse model and were severely compromised for neuroinvasion. Our studies suggest glycosylation of the E protein is an important factor contributing to ZIKV pathogenicity

Plant derived secondary metabolites in cancer therapy: actions, applications, and future prospective of dietaryflavonol (quercetin)

Higher plants are prominent sources for several bioactive chemical constituents (secondary metabolites) who include photochemical, flavoring agents, fragrant molecules, and food additives. According to WHO estimates, it has been reported that more than 80% of population in developing countries prefer these natural bioactive active compounds for their primary health requirement. At present, conventional chemotherapy is constrained due to the nonselective toxicity to human organs and their usage is limited now a days. In a recent survey, more than 60% of cancer patients have been preferring adjuvant phototherapy along with chemotherapy. Thus, photochemical are being widely used as anticancer agents to target specific pathological pathways underlying cancer with low toxic profiles and side effects. These photochemical are cost-effective and easily accessible to the public to treat cancer diseases. These bioactive photochemical are meticulously belongs to secondary metabolites such as alkaloids, flavonoids, polyphenols.Among them, the flavonoids are polyphenolic substances, which are found in all parts of the plant such as flowers, fruits, leaves, roots, seeds, and bark. They possess high medicinal properties like being anti-cancer, anti-hypertensive, anti-inflammatory, anti-obesity, anti-malarial, antioxidant agents. Quercetin is major flavones associated with a profound antioxidant and medicinal property to prevent the oxidation of lipids in vitro and in vivo, and also exhibits direct proapoptotic effects on tumor cells. This compound has proven efficacy in targeting several cancer cells of breast, colon, prostate, ovarian, and lung tumor in vitro. The present review focuses on the effect of quercetin in cancer therapy.</jats:p

Synthesis and Characterization of Nickel Nanoparticles of Gloriosa Superba and Its Anti-Angiogenic Activity in Cervical Cancer

Abstract Cancer is a leading cause of death worldwide that arises from the transformation of normal cells into tumour cells in a multi-stage process. There is always a stable demand for new therapies to treat cancer. Research communities is showing interest towards nanoparticles along with naturally-derived compounds as they are considered as less toxic compared to chemotherapy and radiation. The present study is to develop a fast, eco-friendly synthesis of Gloriosa superba nickel nanoparticles (GSNiNPs) using methanolic extract of Gloriosa superba tuber which acts as a reducing agent and has an active principle against cancer activity. Nickel nanoparticles are considered as good adsorbents due to their chemical and magnetic properties. The synthesised GSNiNPs were characterized using UV Visible spectroscopy, which showed a prominent peak at 350 nm verified the formation of NiNps by the reduction of bioactive compounds of Gloriosa superba towards metal salts. FTIR confirmed that the GSNiNPs were functionalized with biomolecules. SEM and TEM analysis revealed that GSNiNPs are slightly spherical and agglomerated nanoparticles. The in-vitro cytotoxic activity of Gloriosa superba nickel nanoparticles in HeLa cancer cell lines was analyzed using MTT assay, wound healing assay, DAPI staining and double staining. In vitro anti-angiogenic efficacy of Gloriosa superba-nickel nanoparticles was evaluated via the expression pattern of caspase 3 and 9, VEGF A and VEGF B through reverse transcriptase-PCR. The results revealed that Gloriosa superba nickel nanoparticles possess apoptotic and anti-angiogenic effects in HeLa cell lines by inhibiting VEGF A and B and increasing the levels of caspase 3 and 9. GSNiNPs can act as an effective anti-cancer therapeutic agent for cervical cancer cells. In conclusion, the synthesized GSNiNPs can bring a new approach to improve the bioavailability of drug-responsive for the treatment of cervical cancer.</jats:p

Super Nutritive Marine Astaxanthin, an Effectual Dietary Carotenoid for Neurodegenerative Diseases

The red-pigmented astaxanthin (3,-3’-dihydroxy-?,?-carotene-4,4’-dione) were commonly found in marine algae and aquatic animals such as shrimp, lobster, and trout. These pigments are produced as secondary metabolites which fall in arytenoids under class xanthophylls. Synthetic astaxanthin has a wide range of commercial applications such as color additives, usage in cosmetics and immune-boosters. In aquaculture, supplementing synthetic astaxanthin as feed, enhances skin pigmentation which possesses commercial importance. However, synthetic astaxanthin is not highly efficient compared to naturally derived counter forms. On the other hand, humans should only depend on microbial and aquatic sources for their dietary intake of natural astaxanthin. Being a powerful antioxidant, natural astaxanthin is called as king of antioxidants which has scavenging activity 6000 times stronger than vitamin C and 50 times more powerful than vitamin E in protecting cell membranes. It also has a single oxygen quenching activity up to 800 times stronger than coenzyme Q, 550 times more powerful than green tea catechins, 4.9 times stronger than beta-carotene and three times stronger than lute in. Furthermore, researchers revealed that this carotenoid has the capacity to alleviate tumor activity, protecting against lipid per oxidation, free radicals, oxidative damage to LDL-cholesterol and UV light affects on cell membranes and tissues. Also, it is mainly recommended for curing the macular degeneration of cataracts. Anti-aging properties of astaxanthin improve skin health by reducing wrinkles and repairs the UV-induced DNA damage in human cells. Interestingly, the ability of astaxanthin in crossing the blood-brain barrier has brought this compound to limelight as a potential target in treating neurodegenerative diseases including Parkinson's and Alzheimer's disease. Hence, in this review, we are mainly focusing on the therapeutic usage of astaxanthin in neurodegenerative diseases.</jats:p

Role of Pyridoxine in Alleviating Cardiovascular Diseases: A Brief Review

Micronutrients are class of dietary components which are highly essential for the optimal health, growth, and development. These nutrients can overcome birth defects, maternal impairment, and increased risk of death. According to 2017 statistics, around 2 billion people in the developing countries are affected by micronutrient deficiency. There are around 13 essential vitamins which are found in nature. Vitamin B is more important as it plays a vital role in maintaining good health and well-being. Among several existing forms, the native and active form of vitamin B6 is PLP (pyridoxal 5- phosphate). PLP is a co-factor for several biochemical reactions and plays an imperative role in synthesis of amino acids, carbohydrates and fatty acids. Vitamin B6 supplementation in RDA showed to be highly health promising in controlling several clinical alignments such as cardiovascular disease, diabetes, neurological disease, premenstrual syndrome, stroke, peripheral vascular disease, coronary artery disease, pellagra skin disease, ataxia, hyperacuosis, suppression of colon tumor genesis and hyper irritability. Recently, it has been reported that 43.8% people are suffering from cardiovascular disease followed by stroke (16.8%) and is also expected to reach up to1.1 trillion by the end of 2035. It has been found that vitamin B6 lowers Homocystein in blood, which damages the inner linings of arteries causing blood clots and thereby reducing the symptoms of cardiovascular diseases. In addition to it humans have to depend on dietary sources of vitamin B6 which are produced by plants, fungi and bacteria. The present review currently focuses on the action of Vitamin B6 in the management of cardiovascular diseases.</jats:p