A general procedure for protoplast recovery,callus and plant regeneration in plants

A portion of this work was supported by NSF grant DCB 8718314 and by the Missouri Agricultural Experiment Station. This research is a joint contribution from the Missouri Agricultural Experiment Station, Journal Series No. 10,789 and from the Massachusetts Agricultural Experiment Station, Amherst, MA 01003; Journal Paper No. MAES 2959. Ousama Zaghmout was supported by the Food for the 21st Century Program, College of Agriculture, University of Missouri, Columbia, MO.     

Population diversity of Cistopus indicus inferred from mitochondrial DNA (Cytochrome b) variation from China and Vietnam

ABSTRACT

The octopus Cistopus indicus is an important target of cephalopod fisheries in China. It is widely distributed in the South Pacific and tropical Indian Ocean, from the South China Sea, the Philippines, Malaysia, to Indian and Pakistan seas. We collected specimens from five sites in China and Vietnam (Zhoushan, Wenzhou, Shacheng, Zhanjiang and Mangjie). A fragment of 675bp of cytochrome b (Cytb) was amplified from 95 individuals. A total of 27 haplotypes and 78 variable nucleotide sites was observed. High haplotype diversity and low nucleotide diversity were observed in all populations. The phylogenetic analysis separated these populations into two clades; one was composed of three populations (Zhoushan, Wenzhou and Shacheng), the other of two (Zhanjiang, Mangjie). AMOVA analysis detected that 4.67% of the genetic variation occurred within populations and 95.33% occurred among populations. FST values ranged from 0.014 to 0.993, highlighting the high genetic variation among the populations. Assuming a molecular clock with a rate of 2.15–2.6%/Ma for the Cytb gene, the two clades may have diverged 2.88–3.49 million years ago (Pliocene). Neutral evolution tests and mismatch distribution analysis suggested recent population expansion. The present results revealed valuable information for genetic assessment, management and conservation of this species.     

Foliar application of ascorbic acid enhances salinity stress tolerance in barley (Hordeum vulgare L.) through modulation of morpho-physio-biochemical attributes,ions uptake,osmo-protectants and stress response genes expression

Barley (Hordeum vulgare L.) is a major cereal grain and is known as a halophyte (a halophyte is a salt-tolerant plant that grows in soil or waters of high salinity). We therefore conducted a pot experiment to explore plant growth and biomass, photosynthetic pigments, gas exchange attributes, stomatal properties, oxidative stress and antioxidant response and their associated gene expression and absorption of ions in H. Vulgare. The soil used for this analysis was artificially spiked at different salinity concentrations (0, 50, 100 and 150 mM) and different levels of ascorbic acid (AsA) were supplied to plants (0, 30 and 60 mM) shortly after germination of the seed. The results of the present study showed that plant growth and biomass, photosynthetic pigments, gas exchange parameters, stomatal properties and ion uptake were significantly (p < 0.05) reduced by salinity stress, whereas oxidative stress was induced in plants by generating the concentration of reactive oxygen species (ROS) in plant cells/tissues compared to plants grown in the control treatment. Initially, the activity of antioxidant enzymes and relative gene expression increased to a saline level of 100 mM, and then decreased significantly (P < 0.05) by increasing the saline level (150 mM) in the soil compared to plants grown at 0 mM of salinity. We also elucidated that negative impact of salt stress in H. vulgare plants can overcome by the exogenous application of AsA, which not only increased morpho-physiological traits but decreased oxidative stress in the plants by increasing activities of enzymatic antioxidants. We have also explained the negative effect of salt stress on H. vulgare can decrease by exogenous application of AsA, which not only improved morpho-physiological characteristics, ions accumulation in the roots and shoots of the plants, but decreased oxidative stress in plants by increasing antioxidant compounds (enzymatic and non-enzymatic). Taken together, recognizing AsA's role in nutrient uptake introduces new possibilities for agricultural use of this compound and provides a valuable basis for improving plant tolerance and adaptability to potential salinity stress adjustment.     

Chemical Polymorphism of Origanum compactum Grown in All Natural Habitats in Morocco

Origanum compactum L. (Lamiaceae) is one of the most important medicinal species in term of ethnobotany in Morocco. It is considered as a very threatened species as it is heavily exploited. Its domestication remains the most efficient way to safeguard it for future generations. For this purpose, wide evaluation of the existing variability in all over the Moroccan territory is required. The essential oils of 527 individual plants belonging to 88 populations collected from the whole distribution area of the species in Morocco were analyzed by GC/MS. The dominant constituents were carvacrol (0 – 96.3%), thymol (0 – 80.7%), p‐cymene (0.2 – 58.6%), γ‐terpinene (0 – 35.2%), carvacryl methyl ether (0 – 36.2%), and α‐terpineol (0 – 25.8%). While in the Middle Atlas region and the Central Morocco mainly carvacrol type samples were found, much higher chemotypic diversity was encountered within samples from the north part of Morocco (occidental and central Rif regions). The high chemical polymorphism of plants offers a wide range for selection of valuable chemotypes, as a part of breeding and domestication programs of this threatened species.     

The Effect of Thermophoresis on Unsteady Oldroyd-B Nanofluid Flow over Stretching Surface

There are currently only a few theoretical studies on convective heat transfer in polymer nanocomposites. In this paper, the unsteady incompressible flow of a polymer nanocomposite represented by an Oldroyd-B nanofluid along a stretching sheet is investigated. Recent studies have assumed that the nanoparticle fraction can be actively controlled on the boundary, similar to the temperature. However, in practice, such control presents significant challenges and in this study the nanoparticle flux at the boundary surface is assumed to be zero. We have used a relatively novel numerical scheme; the spectral relaxation method to solve the momentum, heat and mass transport equations. The accuracy of the solutions has been determined by benchmarking the results against the quasilinearisation method. We have conducted a parametric study to determine the influence of the fluid parameters on the heat and mass transfer coefficients.     

X-ray crystallography and isothermal titration calorimetry studies of the Salmonella zinc transporter ZntB

The ZntB Zn(2+) efflux system is important for maintenance of Zn(2+) homeostasis in Enterobacteria. We report crystal structures of ZntB cytoplasmic domains from Salmonella enterica serovar Typhimurium (StZntB) in dimeric and physiologically relevant homopentameric forms at 2.3 ? and 3.1 ? resolutions, respectively. The funnel-like structure is similar to that of the homologous Thermotoga maritima CorA Mg(2+) channel and a Vibrio parahaemolyticus ZntB (VpZntB) soluble domain structure. However, the central α7 helix forming the inner wall of the StZntB funnel is oriented perpendicular to the membrane instead of the marked angle seen in CorA or VpZntB. Consequently, the StZntB funnel pore is cylindrical, not tapered, which may represent an "open" form of the ZntB soluble domain. Our crystal structures and isothermal titration calorimetry data indicate that there are three Zn(2+) binding sites in the full-length ZntB, two of which could be involved in Zn(2+) transport.     

Activation of anti-tumor immune response and reduction of regulatory T cells with Mycobacterium indicus pranii (MIP) therapy in tumor bearing mice

Background

Role of immune system in protecting the host from cancer is well established. Growing cancer however subverts immune response towards Th2 type and escape from antitumor mechanism of the host. Activation of both innate and Th1 type response is crucial for host antitumor activity. In our previous study it was found, that Mycobacterium indicus pranii (MIP) also known as M. w induces Th1 type response and activates macrophages in animal model of tuberculosis. Hence, we studied the immunotherapeutic potential of MIP in mouse tumor model and the underlying mechanisms for its antitumor activity.

Methodology and Principal Findings

Tumors were implanted by injecting B16F10 melanoma cells subcutaneously into C57BL/6 mice. Using the optimized dose and treatment regimes, anti-tumor efficacy of heat killed MIP was evaluated. In MIP treated group, tumor appeared in only 50–60% of mice, tumor growth was delayed and tumor volume was less as compared to control. MIP mediated immune activation was analysed in the tumor microenvironment, tumor draining lymph node and spleen. Induction of Th1 response and higher infiltration of immune cells in the tumor microenvironment was observed in MIP treated mice. A large fraction of these immune cells were in activated state as confirmed by phenotypic and functional analysis. Interestingly, percentage of Treg cells in the tumor milieu of treated mice was less. We also evaluated efficacy of MIP along with chemotherapy and found a better response as compared to chemotherapy alone.

Conclusion

MIP therapy is effective in protecting mice from tumor. It activates the immune cells, increases their infiltration in tumor, and abrogates tumor mediated immune suppression.     

Selective Cu2+ binding, redox silencing, and cytoprotective effects of the small heat shock proteins alphaA- and alphaB-crystallin

Oxidative stress and Cu²⁺ have been implicated in several neurodegenerative diseases and in cataract. Oxidative stress, as well as Cu²⁺, is also known to induce the expression of the small heat shock proteins α-crystallins. However, the role of α-crystallins in oxidative stress and in Cu²⁺-mediated processes is not clearly understood. We demonstrate using fluorescence and isothermal titration calorimetry that α-crystallins (αA- and αB-crystallin and its phosphorylation mimic, 3DαB-crystallin) bind Cu²⁺ with close to picomolar range affinity. The presence of other tested divalent cations such as Zn²⁺, Mg²⁺, and Ca²⁺ does not affect Cu²⁺ binding, indicating selectivity of the Cu²⁺-binding site(s) in α-crystallins. Cu²⁺ binding induces structural changes and increase in the hydrodynamic radii of α-crystallins. Cu²⁺ binding increases the stability of α-crystallins towards guanidinium chloride-induced unfolding. Chaperone activity of αA-crystallin increases significantly upon Cu²⁺ binding. α-Crystallins rescue amyloid beta peptide, Aβ_1-40, from Cu²⁺-induced aggregation in vitro. α-Crystallins inhibit Cu²⁺-induced oxidation of ascorbate and, hence, prevent the generation of reactive oxygen species. Interestingly, α-synuclein, a Cu²⁺-binding protein, does not inhibit this oxidation process significantly. We find that the Cu²⁺-sequestering (or redox-silencing) property of α-crystallins confers cytoprotection. To the best of our knowledge, this is the first study to reveal high affinity (close to picomolar) for Cu²⁺ binding and redox silencing of Cu²⁺ by any heat shock protein. Thus, our study ascribes a novel functional role to α-crystallins in Cu²⁺ homeostasis and helps in understanding their protective role in neurodegenerative diseases and cataract.     

GIS and Remote Sensing for Detecting Yield Loss in Cranberry Culture

The primary goal of our research is to develop key elements of a precision agriculture program applicable to high-value woody perennial crops, such as cranberries. These crop systems exhibit tremendous variability in crop yields and quality as imposed by variations in soil properties (water availability and nutrient deficiency) that lead to crop stress (disease development and weed competition). Some of the variability present in the growing environment results in persistent yield losses as well as crop-quality reductions. We are using state-of-the-art methodologies (GIS, GPS, remote sensing) to identify and map spatial variations of the crop. Through image-processing methods (NDVI and unsupervised classification), approximately 65% of the variation in yield was described using 4-m multispectral satellite data as a base image.     

Vasoplegia after implantation of a continuous flow left ventricular assist device: incidence,outcomes and predictors

Background

Vasoplegia after routine cardiac surgery is associated with severe postoperative complications and increased mortality. It is also prevalent in patients undergoing implantation of pulsatile flow left ventricular assist devices (LVAD). However, less is known regarding vasoplegia after implantation of newer generations of continuous flow LVADs (cfLVAD). We aim to report the incidence, impact on outcome and predictors of vasoplegia in these patients.

Methods

Adult patients scheduled for primary cfLVAD implantation were enrolled into a derivation cohort (n?=?118, 2006–2013) and a temporal validation cohort (n?=?73, 2014–2016). Vasoplegia was defined taking into consideration low mean arterial pressure and/or low systemic vascular resistance, preserved cardiac index and high vasopressor support. Vasoplegia was considered after bypass and the first 48?h of ICU stay lasting at least three consecutive hours. This concept of vasoplegia was compared to older definitions reported in the literature in terms of the incidence of postoperative vasoplegia and its association with adverse outcomes. Logistic regression was used to identify independent predictors. Their ability to discriminate patients with vasoplegia was quantified by the area under the receiver operating characteristic curve (AUC).

Results

The incidence of vasoplegia was 33.1% using the unified definition of vasoplegia. Vasoplegia was associated with increased ICU length-of-stay (10.5 [6.9–20.8] vs 6.1 [4.6–10.4] p?=?0.002), increased ICU-mortality (OR 5.8, 95% CI 1.9–18.2) and one-year-mortality (OR 3.9, 95% CI 1.5–10.2), and a higher incidence of renal failure (OR 4.3, 95% CI 1.8–10.4). Multivariable analysis identified previous cardiothoracic surgery, preoperative dopamine administration, preoperative bilirubin levels and preoperative creatinine clearance as independent preoperative predictors of vasoplegia. The resultant prediction model exhibited a good discriminative ability (AUC 0.80, 95% CI 0.71–0.89, p?<? 0.01). Temporal validation resulted in an AUC of 0.74 (95% CI 0.61–0.87, p?<? 0.01).

Conclusions

In the era of the new generation of cfLVADs, vasoplegia remains a prevalent (33%) and critical condition with worse short-term outcomes and survival. We identified previous cardiothoracic surgery, preoperative treatment with dopamine, preoperative bilirubin levels and preoperative creatinine clearance as independent predictors.