A quantitative and semiautomated method for determining misaligned and lagging chromosomes during mitosis

Accurate chromosome alignment at metaphase facilitates the equal segregation of sister chromatids to each of the nascent daughter cells. Lack of proper metaphase alignment is an indicator of defective chromosome congression and aberrant kinetochore–microtubule attachments which in turn promotes chromosome missegregation and aneuploidy, hallmarks of cancer. Tools to sensitively, accurately, and quantitatively measure chromosome alignment at metaphase will facilitate understanding of the contribution of chromosome segregation errors to the development of aneuploidy. In this work, we have developed and validated a method based on analytical geometry to measure several indicators of chromosome misalignment. We generated semiautomated and flexible ImageJ2/Fiji pipelines to quantify kinetochore misalignment at metaphase plates as well as lagging chromosomes at anaphase. These tools will ultimately allow sensitive and systematic quantitation of these chromosome segregation defects in cells undergoing mitosis.     

Effect of the diverse combinations of useful microbes and chemical fertilizers on important traits of potato

The belowground soil environment is an active space for microbes, particularly Arbuscular Mycorrhizal Fungi (AMF) and P hosphate Solubilizing Bacteria (PSB) that can colonize with roots of higher plants. In the present experiment, we evaluated the combination of microbial inoculants with the different doses of urea and superphosphate in a complete randomized block design (CRBD). Three different doses of urea and superphosphate were tested, i.e., recommended dose, 75% of the recommended dose and 125% of the recommended dose, independently and in combination with three microbial groups viz. Glomus mosseae (AMF), Bacillus subtilis (PSB) and Nitrifying microorganisms (Nitrosomonas + Nitrobacter, NN). Overall, there were 16 treatment combinations used, and studied the number of tubers per plant, the weight of tubers, moisture content, and the number of nodes per tubers which were best in treatment comprising of AMF + PSB + NN + 75% of urea + superphosphate. From our results, it is suggested for the growers to use a lesser quantity of fertilizers from the recommended dose along with some bioinoculants to maintain the soil fertility and also to achieve the yield targets by decreasing the cost of chemical fertilizers.     

Effects of arbuscular mycorrhizal fungi and P-solubilizing Pseudomonas fluorescence (ATCC-17400) on morphological traits and mineral content of sesame

Sesame (Sesamum indicum L.) is an important staple crop of the family Pedaliaceae. The commercial production of sesame is still dependent on the applications of chemical fertilizers. Mycorrhiza inoculum resulted in better morphological and biochemical traits in vegetables. Thus, here the outcome of arbuscular mycorrhizal fungi (AMF) and Pseudomonas fluorescence (ATCC-17400) inoculation was studied in the pot culture experiment. Primarily, there seems to be a promising opportunity of AMF in sesame under pot and field trials because of enhanced morphological parameters, especially root weight, and disparities in nutrients and metabolites. The AMF appears to be an option to boost plant growth, mineral content, and sesame yield. The AMF treatment with Pseudomonas fluorescence strain (ATCC-17400) determined the maximum values for the morphological traits and mineral content. Overall, our study highlights mycorrhizal fungi and other microbes efficacy in achieving a successful sesame production.     

Osmotin-expressing transgenic tea plants have improved stress tolerance and are of higher quality

Drought is a major stress that affects the yield and quality of tea, a widely consumed beverage crop grown in more than 20 countries of the world. Therefore, osmotin gene-expressing transgenic tea plants produced using earlier optimized conditions were evaluated for their tolerance of drought stress and their quality. Improved tolerance of polyethylene glycol-induced water stress and faster recovery from stress were evident in transgenic lines compared with the normal phenotype. Significant improvements in growth under in-vitro conditions were also observed. Besides enhanced reactive oxygen species-scavenging enzyme activity, the transgenic lines contained significantly higher levels of flavan-3-ols and caffeine, key compounds that govern quality and commercial yield of the beverage. The selected transgenic lines have the potential to meet the demands of the tea industry for stress-tolerant plants with higher yield and quality. These traits of the transgenic lines can be effectively maintained for generations because tea is commercially cultivated through vegetative propagation only.     

Awareness and attitudes of final-year pharmacy students towards chronotherapy: a needs analysis

Chronotherapy involves the administration of medication in coordination with the body’s circadian rhythms to maximise effectiveness and/or minimise/avoid adverse effects. A deeper understanding of the concepts underpinning chronotherapy and emerging applications in pharmacy practice would be an important addition to the scope of education in future graduates. This study aimed to assess baseline awareness level and attitude towards the principles of chronotherapy among final-year pharmacy students at the University of Sydney and to uncover underlying ‘gaps’ in their knowledge of the evidence-based information on chronotherapy in practice. A cross-sectional survey instrument was utilised. A questionnaire was designed to establish the knowledge base and attitudes toward principles of chronotherapy. Out of 216 students, 212 students completed the questionnaire resulting in a response rate of 98 %. Mean total awareness and attitude scores were 6.6 ± 2.0 (score range 0–13) and 47.4 ± 6.9 (score range 12–60), respectively. The study indicated that the final-year pharmacy students have a positive attitude and willingness to apply principles of chronotherapy in future practice despite having a lack of awareness regarding circadian rhythms and evidence-based information on chronotherapy. The finding points to the need for an educational intervention based on the principles of chronotherapy in pharmacy schools.

     

Regulation of calcium ion and its effect on growth and developmental behavior in wild type and ntcA mutant of Anabaena sp. PCC 7120 under varied levels of CaCl2

A study of calcium ion regulation in Anabaena 7120 and its derivative mutant (CSE2) strain impaired in ntcA gene were investigated in terms of altered morphological and physiological responses against various levels of calcium stress (0–100 mM). Calcium concentration of 10 mM was found to be inhibitory while 100 mM proved lethal for both wild type and mutant strain. The involvement of Ca²⁺ in the regulation of cellular processes has been described in terms of an influx or efflux of Ca²⁺ from the cytosol. A biphasic calcium uptake with difference in calcium influx and efflux rate was responsible for differential amount of remaining calcium which followed a decreasing trend both for wild type and mutant. Low K _{s 0.5} and high V _max in mutant suggest heavy and less restricted influx of calcium ion. Further, the interactive effect of calcium influx/efflux rate, remaining Ca²⁺ and intracellular levels of Na⁺ and K⁺ may be attributed for the degree of membrane damage and growth sustenance during exogenous supply of calcium salt. Widening in heterocyst spacing pattern, decreased heterocyst frequency and formation of abnormal cell structures at higher concentration (100 mM CaCl₂) suggest that calcium mediated regulatory process modulate heterocyst frequency and maintenance of cell structure. Further, poor regulation of calcium ion homeostasis in ntcA suggests that the calcium level and ntcA gene expression are inter-related.     

Kinetic analysis of late steps of eukaryotic translation initiation

Little is known about the molecular mechanics of the late events of translation initiation in eukaryotes. We present a kinetic dissection of the transition from a preinitiation complex after start codon recognition to the final 80S initiation complex. The resulting framework reveals that eukaryotic initiation factor (eIF)5B actually accelerates the rate of ribosomal subunit joining, and this acceleration is influenced by the conformation of the GTPase active site of the factor mediated by the bound nucleotide. eIF1A accelerates joining through its C-terminal interaction with eIF5B, and eIF1A release from the initiating ribosome, which occurs only after subunit joining, is accelerated by GTP hydrolysis by eIF5B. Following subunit joining, GTP hydrolysis by eIF5B alters the conformation of the final initiation complex and clears a path to promote rapid release of eIF1A. Our data, coupled with previous work, indicate that eIF1A is present on the ribosome throughout the entire initiation process and plays key roles at every stage.     

S-nitrosoglutathione modulates CXCR4 and ICOS expression

The expression of CXCR4, a membrane protein which is involved in the entry of HIV-1, is down-modulated from the cell surface by Phorbol 12-myristate 13-acetate (PMA) and the Ca+ ionophore, Ionomycin. Inducible co-stimulator (ICOS), which contributes to lymphocyte proliferation, is up-regulated by PMA/Ionomycin. We examined the influence of S-nitrosoglutathione (SNG), an inhibitor of Vacuolar H+-ATPase (V-ATPase), on the expression of CXCR4 and ICOS in PMA/Ionomycin-treated peripheral mononuclear cells (PBMC), and of CXCR4 alone in lymphoid cell lines. In this report, we show that SNG interferes with both effects of PMA/Ionomycin, namely CXCR4 down-regulation and ICOS up-regulation. These studies imply opposing roles of V-ATPase in the regulation of CXCR4 and ICOS. The influence of SNG in modulating the susceptibility of T cells to HIV-1 and on their immune responses needs further investigation.     

Acid-induced unfolding of didecameric keyhole limpet hemocyanin: detection and characterizations of decameric and tetrameric intermediate states

Keyhole limpet hemocyanin (KLH) is widely used as an immune stimulant and hapten carrier derived from a marine mollusc Megathura crenulata. To provide details of the stability and equilibrium of KLH, different intermediate species were investigated with a series of biophysical techniques: circular dichroism, binding of hydrophobic dye, 1-anilino-8-naphthalene sulfonic acid, acrylamide-induced fluorescence quenching, thermal stability and dynamic light scattering. KLH in its native state at pH 7.4 exists in the stable didecameric form with hydrodynamic radii (R _h) of 28.22 nm, which is approximately equal to a molecular mass of 8.8 ± 0.6 MDa. The experimental results demonstrated the presence of two structurally distinct species in the conformational transition of KLH under acidic conditions. One species populates at pH 2.8, characterized as decameric (4.8 ± 0.2 MDa; R _h = 22.02 nm), molten globule-like state, while the other accumulates at pH 1.2 and is characterized as a tetramer (2.4 ± 0.8 MDa; R _h = 16.47 nm) with more organized secondary and tertiary structures. Our experimental manipulation of the oligomeric states of KLH has provided data that correlate well with the known oligomeric forms obtained from total KLH formed in vivo and extends our understanding of multimer formation by KLH. The results are of particular interest in light of the important role of the mechanistic pathway of pH-dependent structural changes of Hc stability in the biochemical and medical applications of these respiratory proteins.