Desiccation induced physiological and biochemical changes of Gymnacranthera canarica (King.) Warb. seeds in the Myristica swamp forests, Southern Western Ghats, India

Gymnacranthera canarica (King.) Warb. is an endemic tree species that dominates the Myristica swamp ecosystem of southern Western Ghats. This tropical tree species has become more threatened due to limited natural seed germination and habitat loss. Mature seeds were collected from the myristica swamp ecosystem subjected to desiccation study. This research evaluated the physiological (moisture content, tetrazolium reduction, lipid peroxidation, electrolyte leakage) and biochemical response of seeds during different desiccation treatments. Results showed that G. canarica seeds are highly sensitive to desiccation and total viability loss was seen within 15 days following harvest indicating the active seed metabolism of mature seeds showing absence of metabolic arrest. Desiccation enhanced malondialdehyde and electrolyte leakage while reducing formazan formation. Seed desiccation increases protease activity, which peaks when viability is lost. Desiccation reduced the quantity of phenol and starch, whereas proline, fat, sucrose and total soluble carbohydrates increased. The early viability loss in G. canarica seeds could be due to loss of membrane integrity, which was linked to ROS formation and associated lipid peroxidation products indicating seeds are truly recalcitrant

Embryo maturation, dormancy and seed storage behaviour of Gymnacranthera canarica (King) Warb., a threatened endemic tree species of Southern Western Ghats

Gymnacranthera canarica is a severely endangered endemic tree species found in the Myristica swamps of the Southern Western Ghats. Seed storage behaviour is an essential factor to consider when developing effective conservation methods for plant genetic resources whose ex-situ preservation is unclear. The seed storage behaviour, seed dormancy state, dormancy breaking treatments, germination, and phytohormonal analysis of G. canarica were explored in this work. G. canarica seed moisture at shedding was 28.86%, germinated to 34% at 25ºC, and had a low germination rate in natural conditions. Germination was considerably aided by Gibberellic acid pretreatment and drying stratification. G. canarica seeds may have non-deep simple morphophysiological dormancy, as evidenced by the fact that their embryos grew at temperatures between 20 and 25ºC (MPD). Seeds can be stored for up to 60 days at 20ºC after being desiccated to 19.37% suggest that G. canarica seeds are recalcitrant, with non-deep simple morphophysiological dormancy, and that seed pretreatment with Gibberellic acid prior to germination could be a viable approach for mass propagation and long term ex-situ conservation could be the potential storage of this critically endangered species

DESIGN AND DEVELOPMENT OF NANOSTRUCTURED LIPID CARRIER CONTAINING TRIAMCINOLONE ACETONIDE

Objective: The aim of present study was to prepare nanostructured lipid carriers (NLCs) based Triamcinolone acetonide (TA). Methods: Nanostructured lipid carriers (NLCs) consisted of solid lipid and liquid lipid are a new type of lipid nanoparticles, prepared by using solvent diffusion and high pressure homogenization methods, which offer the advantage of improved drug loading capacity and release properties. Glyceryl monostearate selected as the solid lipid, capmul MCM C8 as the liquid lipid, polyvinyl Alcohol (PVA) as the surfactant. NLCs dispersion was characterized by particle size analysis, zeta potential, scanning electron microscopy (SEM), differential scanning calorimetry, and an in vitro release study. Results: Optimized NLCs loaded with TA were exhibited spherical shape with particle size 286.1 nm, polydispersity index 0.317, zeta potential-21.9 mV and entrapment efficiency 86.19% respectively. The result of differential scanning calorimetry (DSC) showed that drug was dispersed in NLCs in a crystalline state. In vitro release studies revealed that drug release of optimized batch was 8.34 % and 88.84% at 1h and 8h respectively. The release kinetics of the optimized NLCs best fitted the peppas-korsmeyer model. Furthermore, morphological investigations by SEM showed that optimized batch exhibit a spherical shape and a smooth surface. Conclusion: Thus, the results indicated that successfully prepared TA-loaded NLCs and could potentially be exploited as a carrier with improved drug loading capacity and sustained drug release. The present results demonstrated that these systems could be a promising platform for inflammatory diseases, in particular for psoriasis topical therapy

Impact of fast and slow desiccation on Garcinia imberti seed and seedling vigour

Garcinia imberti Bourd. endemic to the southern Western Ghats is classified as endangered by the IUCN (2018). The seeds as the sole means of propagation with initial moisture content (MC) of 62.8% are sensitive to desiccation. Studies on the responses of the seed to drying require ascertaining of the critical moisture content (CMC) as a basic requisite for germplasm conservation. Responses of G. imberti seeds to fast drying with activated silica gel (25 ± 2 °C, 6 ± 1% RH) and to slow drying under laboratory conditions (28 ± 2°C, 60 ± 2% RH) were evaluated for seed and seedling vigour. When the MC was reduced to 56% by 48 hours of slow drying or 6 hours of fast drying, 75 to 90% normal seedlings were produced respectively. In the case of fast dried seed (6 hours), seed associated parameters except mean germination time showed peak values with maximum germination and enhanced root length. Below the CMCs of 16.4 and 26.3% (fast and slow drying respectively), half of the tested seeds become non viable. Thus for germplasm conservation the present study proposes 6 hour’s of fast drying to retain viability and normal seedling development of G. imberti

Bifurcation of rotating liquid drops: Results from USML-1 experiments in space

Experiments on rotational bifurcation of liquid drops, in which the drops were levitated and spun using acoustic fields in a low-gravity environment, were conducted during the first United States Microgravity Laboratory (USML-1) Space Shuttle flight. The experiments have successfully resolved the discrepancies existing between the previous experimental results and the theoretical predictions. In the case of a spherical drop, for which theory exists, the results agree well with the predictions. In the case of flattened drops, the experiments have extablished a family of curves, with the spherical drop as the limiting case

Effect of cryopreservation on germination of seeds and zygotic embryos of Calamus shendurunii an endemic rattan of Western Ghats

Calamus shendurunii is an endemic rattan of Western Ghats having restricted distribution and limited population. As a prerequisite to device an appropriate method for ex situ conservation of the species, desiccation and cryopreservation of seeds and zygotic embryo has been studied. Seeds extracted from ripened fruits possessed 35 per cent moisture content and exhibited 97 per cent germination. Desiccation to 28 per cent moisture content reduced the germination to 77 per cent. Desiccation below 14 per cent moisture content caused complete loss of seed germinability. Seeds stored under ambient conditions (28±2oC/60% RH) for more than seven days reduced germination to less than 40 per cent. Thus, conventional storage is not effective for their ex situ conservation. As an alternative method, excised zygotic embryos were subjected to desiccation and storage in liquid nitrogen. The embryos tolerated desiccation down to 5 per cent exhibiting 60 to 90 per cent germination upon culture into MS medium. Desiccated embryos subjected to liquid nitrogen exposure showed post freeze recovery and germination (80-90%) equal to that of desiccated control samples. Thus the study proved the extreme recalcitrance of C. shendurunii seeds and embryo cryopreservation as an alternative method of their ex situ conservation in gene banks

Population genetic structure of Garcinia imberti Bourd. an endangered endemic tree of southern Western Ghats, India

Assessing the genetic diversity of endemic plants is of great importance in future conservation programmes. The genetic diversity in Garcinia imberti from Agasthyamala Biosphere Reserve of southern Western Ghats was assessed through ISSR markers by molecular characterization with 15 primers. A total of 157 accessions from six populations were used for the study. They generated 102 amplified products, out of which 89 were polymorphic (87.25%). It produced an average of 6.8 bands per primer and 5.93% polymorphism per primer. The present study revealed that G. imberti has moderate level of genetic diversity at species level but differs at population level. The dendrogram constructed following UPGMA exhibited that all accessions were clustered together except Poonkulam population. The genetic diversity analysis of G. imberti showed that even though the populations are closely associated, every population have their own characteristic diversity and should be conserved. Among the populations, Chemunji is the largest one with more genetic diversity and may conserve as the potential source of gene pool of this species

Role of POTE-2 in hepatocellular carcinoma progression.

Background: Hepatocellular carcinoma (HCC) accounts for 85-90% of primary liver cancers. The Hispanic population had an incidence of 21.2 per 100,000 in Texas. Particularly, the Rio Grande Valley (RGV) is an underserved area facing disparities that increase risk factors of HCC and thus, yielding higher incidence and mortality. Therefore, early, faster, and inexpensive diagnostic biomarkers and methods are crucial to under-resourced areas such as the RGV. Recently, we have identified an extracellular cancer antigen, POTE-2. Preliminary data indicates high POTE-2 expression in HCC tumors. In this study, we will discuss the role of POTE-2 in HCC progression and its associated regulatory pathways. Methods: The Cancer Genome Atlas (TCGA) database of HCC patients (n=371 tumor; n=50 normal) was analyzed. Liver cancer cells were procured from ATCC. POTE-2 mRNA and protein expression analyzed via RT-PCR and western blot. Absolute copy number was determined using Digital Droplet PCR. Lentiviral-based plasmids were used for overexpression and knockdown studies. Signaling pathways were analyzed using Proteome Profiler array. Results: Comprehensive analysis of TCGA database revealed high POTE-2 expression tumors with upregulation in all stages of HCC. POTE-2 expression increases with nodal metastatic status leading to poor survival. The protein expression for POTE-2 was significantly higher in SK-HEP1 compared to C3A cells. Lentiviral transduction showed significant overexpression and knockdown of the POTE-2 protein. Modulation of POTE-2 expression led to changes in lncRNA and kinase pathways. Conclusion: These studies will help discover novel mechanisms of POTE-2 protein function, signaling pathways and roles in liver cancer progression