Optically transparent polymer devices for in situ assessment of cell electroporation

In order to study cell electroporation in situ, polymer devices have been fabricated from poly-dimethyl siloxane with transparent indium tin oxide parallel plate electrodes in horizontal geometry. This geometry with cells located on a single focal plane at the interface of the bottom electrode allows a longer observation time in both transmitted bright-field and reflected fluorescence microscopy modes. Using propidium iodide (PI) as a marker dye, the number of electroporated cells in a typical culture volume of 10-100 mu l was quantified in situ as a function of applied voltage from 10 to 90 V in a series of 2-ms pulses across 0.5-mm electrode spacing. The electric field at the interface and device current was calculated using a model that takes into account bulk screening of the transient pulse. The voltage dependence of the number of electroporated cells could be explained using a stochastic model for the electroporation kinetics, and the free energy for pore formation was found to be kT at room temperature. With this device, the optimum electroporation conditions can be quickly determined by monitoring the uptake of PI marker dye in situ under the application of millisecond voltage pulses. The electroporation efficiency was also quantified using an ex situ fluorescence-assisted cell sorter, and the morphology of cultured cells was evaluated after the pulsing experiment. Importantly, the efficacy of the developed device was tested independently using two cell lines (C2C12 mouse myoblast cells and yeast cells) as well as in three different electroporation buffers (phosphate buffer saline, electroporation buffer and 10 % glycerol)

Comparative Evaluation of Serum Lithium Estimation Using Plain Glass Vial and Serum Clot Activator Vacutainer by Reflectance Photometry

Background The collection of blood samples in different vacutainers can affect the result of serum lithium estimation due to the presence of distinct additives in the blood collection vacutainer for enhancing the clot formation process. Due to the low therapeutic index and threat of toxicity of lithium, it is imperative to correctly report the test result. Thus, it has become a challenge for the laboratory physician to estimate lithium in any clinical laboratory setup. Materials and Methods Sample of 100 patients were collected and paired into clot activator vacutainers and plain glass vials. After centrifugation, samples from the paired collection tubes were processed immediately for serum lithium estimation by VITROS 4600 analyzer working on the principle of reflectance photometry. Both the paired tubes were stored at 2 to 8°C and were further analyzed, at 24 and 48 hours, respectively, from the time of their collection. The statistical analysis was done in IBM SPSS software version 23. Results There was a statistically significant differences between the mean of lithium values when processed within 1st hour of collection, obtained from clot activator vacutainers in comparison to glass vials. However, within tube comparison, there was no statistical difference in the lithium values estimated at 1st hour, 24 hours, and 48 hours of collection. Conclusion In this study, lithium values measured by clot-activated vacutainers are found to be lower as compared with values measured through glass vials

Estimation of activation energy for electroporation and pore growth rate in liquid crystalline and gel phases of lipid bilayers using molecular dynamics simulations

Molecular dynamics simulations of electroporation in POPC and DPPC lipid bilayers have been carried out at different temperatures ranging from 230 K to 350 K for varying electric fields. The dynamics of pore formation, including threshold field, pore initiation time, pore growth rate, and pore closure rate after the field is switched off, was studied in both the gel and liquid crystalline (L-alpha) phases of the bilayers. Using an Arrhenius model of pore initiation kinetics, the activation energy for pore opening was estimated to be 25.6 kJ mol(-1) and 32.6 kJ mol(-1) in the L-alpha phase of POPC and DPPC lipids respectively at a field strength of 0.32 V nm(-1). The activation energy decreases to 24.2 kJ mol(-1) and 23.7 kJ mol(-1) respectively at a higher field strength of 1.1 V nm(-1). At temperatures below the melting point, the activation energy in the gel phase of POPC and DPPC increases to 28.8 kJ mol(-1) and 34.4 kJ mol(-1) respectively at the same field of 1.1 V nm(-1). The pore closing time was found to be higher in the gel than in the L-alpha phase. The pore growth rate increases linearly with temperature and quadratically with field, consistent with viscosity limited growth models

Mapping of Soil Fertility Status in a Coastal Village of Odisha Using Geospatial Technology

The most fundamental decision-making tool for sustainable soil nutrient management is the assessment of soil fertility in a region. During 2021, a soil fertility status inventory was prepared through National Innovations on Climate Resilient Agriculture (NICRA) project in Achyutadaspur village of Jagatsinghpur district, Odisha. A total of 125 surface soil samples were randomly collected from a depth of 0-15 cm with GPS locations and soil parameters like pH, electrical conductivity, organic carbon, available N, P, K, S, Zn, and B were evaluated using standard analytical procedures. GIS maps have been prepared using ArcGIS software based on the analysis report. The majority of the soil was found acidic around 62.4% area of the village, followed by neutral soil pH in 33.6% area. The organic carbon status of the soil was found low in 79.2% area and medium in 20.8% area, available N was found low in 96% area and medium in 4% area, available P was found medium in 82.4% area and high in 17.6% area, available K was low in 27.2% area, medium in 64% area, and high in 8.8% area, and available S was found low in 37.6% area and medium in 62.4% area. Among the available micronutrients, Zn was found low in 79.2% area and medium in 20.8% area. Similarly, hot water extractable boron was found low in 92.6% area of the region and medium in 7.4% area. By adopting soil map based fertilizer recommendation with improved practices in the Achyutadaspur village, 17.92% yield was increased over farmers’ practices. The benefit cost ratio (B:C ratio) of rice was found to be 1.89 whereas 1.65 in farmers practices

Thirteen Year Long Term Fertilization Effect on Soil Phosphorus Fractions of an Acid Inceptisol and Their Contribution to Phosphorus Uptake by a Double Crop of Rice under Sub-Tropical Climate

A 13 year old long term fertilizer experiment conducted on an acid soil with rice-rice system was used to study the impact of continuous application of fertilizer nutrients and amendments on changes in soil phosphorus fractions and their relative contribution to P nutrition. There was accumulation of total P on surface soil in all P fertilized treatments and depletion in P minus treatments. Depending on the P balance the treatments differed significantly with respect to P accumulation. 150%NPK treatment had highest P accumulation (841.92kgha-1) in surface soil. The high yielding treatments viz. 100%NPK+FYM and 100% NPK +FYM+Lime had relatively less P built up than 100%NPK. The total P comprised of inorganic P, 64.43-89.60% and organic P, 10.40– 35.57%. The inorganic fraction in terms of their abundance were in the order: Sal-P (42.13 kgha-1) <Occl-P (52.16 kgha-1) < Al-P (52.98 kgha-1) < Ca-P (55.44 kgha-1) < Red–P (134.22 kgha-1) < Fe-P (218.70 kgha-1). Olsen P which was significantly influenced by the fertilizer treatments had a very strong positive correlation (p<0.01) with Sal-P(r= 0.469***), Occl-P(r=0.639***) and Ca-P (r=0.739***). Among the inorganic P fractions, Ca-P contributed highest of 42.57% to P uptake, followed by Occl-P (28.59%), Sal-P (18.56%) and Al-P (7.63%). The fractions in order of their relative importance for P uptake under submerged rice-rice situation are Ca-P > Occl-P > Sal-P > Al-P > Red-P > Fe-P. The study indicated that 100%NPK+FYM and 100% NPK+FYM+Lime are the best treatments for maintaining more active fractions of P required for P nutrition of rice-rice system under the tropical acid situation

Underutilized legumes: nutrient status and advanced breeding approaches for qualitative and quantitative enhancement

Underutilized/orphan legumes provide food and nutritional security to resource-poor rural populations during periods of drought and extreme hunger, thus, saving millions of lives. The Leguminaceae, which is the third largest flowering plant family, has approximately 650 genera and 20,000 species and are distributed globally. There are various protein-rich accessible and edible legumes, such as soybean, cowpea, and others; nevertheless, their consumption rate is far higher than production, owing to ever-increasing demand. The growing global urge to switch from an animal-based protein diet to a vegetarian-based protein diet has also accelerated their demand. In this context, underutilized legumes offer significant potential for food security, nutritional requirements, and agricultural development. Many of the known legumes like Mucuna spp., Canavalia spp., Sesbania spp., Phaseolus spp., and others are reported to contain comparable amounts of protein, essential amino acids, polyunsaturated fatty acids (PUFAs), dietary fiber, essential minerals and vitamins along with other bioactive compounds. Keeping this in mind, the current review focuses on the potential of discovering underutilized legumes as a source of food, feed and pharmaceutically valuable chemicals, in order to provide baseline data for addressing malnutrition-related problems and sustaining pulse needs across the globe. There is a scarcity of information about underutilized legumes and is restricted to specific geographical zones with local or traditional significance. Around 700 genera and 20,000 species remain for domestication, improvement, and mainstreaming. Significant efforts in research, breeding, and development are required to transform existing local landraces of carefully selected, promising crops into types with broad adaptability and economic viability. Different breeding efforts and the use of biotechnological methods such as micro-propagation, molecular markers research and genetic transformation for the development of underutilized crops are offered to popularize lesser-known legume crops and help farmers diversify their agricultural systems and boost their profitability