Synthesis, characterization, and preliminary application of new biocompatible nanogels useful as anticancer drug delivery systems

253 p.

Inherited Photoreceptor Degeneration Causes the Death of Melanopsin-Positive Retinal Ganglion Cells and Increases Their Coexpression of Brn3a

Purpose: To study the population of intrinsically photosensitive retinal ganglion cells (melanopsin-expressing RGCs, m+RGCs) in P23H-1 rats, a rat model of inherited photoreceptor degeneration. Methods: At postnatal (P) times P30, P365, and P540, retinas from P23H dystrophic rats (line 1, rapid degeneration; and line 3, slow degeneration) and Sprague Dawley (SD) rats (control) were dissected as whole-mounts and immunodetected for melanopsin and/or Brn3a. The dendritic arborization of m+RGCs and the numbers of Brn3a+RGCs and m+RGCs were quantified and their retinal distribution and coexpression analyzed. Results: In SD rats, aging did not affect the population of Brn3a+RGCs or m+RGCs or the percentage that showed coexpression (0.27%). Young P23H-1 rats had a significantly lower number of Brn3a+RGCs and showed a further decline with age. The population of m+RGCs in young P23H-1 rats was similar to that found in SD rats and decreased by 22.6% and 28.2% at P365 and P540, respectively, similarly to the decrease of the Brn3a+RGCs. At these ages the m+RGCs showed a decrease of their dendritic arborization parameters, which was similar in both the P23H-1 and P23H-3 lines. The percentage of coexpression of Brn3a was, however, already significantly higher at P30 (3.31%) and increased significantly with age (10.65% at P540). Conclusions: Inherited photoreceptor degeneration was followed by secondary loss of Brn3a+RGCs and m+RGCs. Surviving m+RGCs showed decreased dendritic arborization parameters and increased coexpression of Brn3a and melanopsin, phenotypic and molecular changes that may represent an effort to resist degeneration and/or preferential survival of m+RGCs capable of synthesizing Brn3a.

The interaction between Sertoli cells and luekemia inhibitory factor on the propagation and differentiation of spermatogonial stem cells in vitro

Background: Sertoli cells play a pivotal role in creating microenvironments essential for spermatogonial stem cells (SSCs) self-renewal and commitment to differentiation. Maintenance of SSCs and or induction of in vitro spermiogenesis may provide a therapeutic strategy to treat male infertility. Objective: This study investigated the role of luekemia inhibitory factor (LIF) on the propagation of SSCs and both functions of Sertoli cells on the proliferation and differentiation of these cells. Materials and Methods: SSCs were sorted from the testes of adult male mice by magnetic activated cell sorting and thymus cell antigen 1 antibody. On the other hand, isolated Sertoli cells were enriched using lectin coated plates. SSCs were cultured on Sertoli cells for 7 days in the absence or presence of LIF. The effects of these conditions were evaluated by microscopy and expression of meiotic and post meiotic transcripts by reverse transcriptase polymerase chain reaction. Results: Our data showed that SSCs co-cultured with Sertoli cells in the presence of LIF formed colonies on top of the Sertoli cells. These colonies had alkaline phosphatesase activity and expressed SSCs specific genes. SSCs were enjoyed limited development after the mere removal of LIF, and exhibiting expression of meiotic and postmeiotic transcript and loss of SSCs specific gene expression (p< 0.05). Conclusion: Our findings represent co-culture of SSCs with Sertoli cells provides conditions that may allow efficient proliferation and differentiation of SSCs for male infertility treatment.

Measuring the Impact of Melaleuca quinquenervia Biochar Application on Soil Quality, Plant Growth, and Microbial Gas Flux

Biochar has been heralded a mechanism for carbon sequestration and an ideal amendment for improving soil quality. Melaleuca quinquenervia is an aggressive and wide-spread invasive species in Florida. The purpose of this research was to convert M. quinquenervia biomass into biochar and measure how application at two rates (2% or 5% wt/wt) impacts soil quality, plant growth, and microbial gas flux in a greenhouse experiment using Phaseolus vulgaris L. and local soil. Plant growth was measured using height, biomass weight, specific leaf area, and root-shoot ratio. Soil quality was evaluated according to nutrient content and water holding capacity. Microbial respiration, as carbon dioxide (CO2), was measured using gas chromatography. Biochar addition at 5% significantly reduced available soil nutrients, while 2% biochar application increased almost all nutrients. Plant biomass was highest in the control group, p2 flux decreased significantly in both biochar groups, but reductions were not long term.

Effects of plant density and the number of emitters on plant growth and nitrate concentration in spinach cultivated in substrate

The effects of plant density and the number of emitters per Styrofoam box on plant growth and nitrate (NO3-) concentration were evaluated in spinach (Spinacia oleracea L. cv. Tapir). Spinach seedlings were transplanted at 45 days after emergence into Styrofoam boxes filled with the substrate and were grown during winter in an unheated greenhouse with no supplemental lighting. The experiment was carried out with four treatments, including two plant densities (160 and 280 plants/m2) and two number of emitters per Styrofoam box (4 and 8 emitters). Each planting box was irrigated daily and fertigated with a complete nutrient solution. Shoot dry weight was not affected by plant density. However, yield increased with plant density and emitter number. Leaf-blade NO3- concentration was not affected by the interaction between plant density and number of emitters, but petioles NO3- concentration was greater in treatment with 160 plants/m2 and 8 emitters. Although leaf-blade NO3- concentration was not affected by plant density, it decreased with the number of emitters. On the other hand, petiole NO3- concentration was not affected by plant density or number of emitters. Leaf-blade NO3- concentration ranged from 3.2 to 4.1 mg/g fresh weight, occurring the highest value in the treatment with 280 plants/m2 and 4 emitters. Petiole NO3- concentration ranged from 3.5 to 5.3 mg/g fresh weight, values that were higher than allowed by EU regulation.

Quantification of free auxins in semi-hardwood plant cuttings and microshoots by dispersive liquid–liquid microextraction/microwave derivatization and GC/MS analysis

Several studies have suggested that differences in the natural rooting ability of plant cuttings could be attributed to differences in endogenous auxin levels. Hence, during rooting experiments, it is important to be able to routinely monitor the evolution of endogenous levels of plant hormones. This work reports the development of a new method for the quantification of free auxins in auxin-treated Olea europaea (L.) explants, using dispersive liquid–liquid microextraction (DLLME) and microwave assisted derivatization (MAD) followed by gas chromatography/mass spectrometry (GC/MS) analysis. Linear ranges of 0.5–500 ng mL 1 and 1–500 mg mL 1 were used for the quantification of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), respectively. Determined by serial dilutions, the limits of detection (LOD) and quantification (LOQ) were 0.05 ng mL 1 and 0.25 ng mL 1, respectively for both compounds. When using the calibration curve for determination, the LOQ corresponded to 0.5 ng mL 1 (IAA) and 0.5 mg mL 1 (IBA). The proposed method proved to be substantially faster than other alternatives, and allowed free auxin quantification in real samples of semi-hardwood cuttings and microshoots of two olive cultivars. The concentrations found in the analyzed samples are in the range of 0.131–0.342 mg g 1 (IAA) and 20–264 mg g 1 (IBA).

Effects of substrate type on plant growth and nitrogen and nitrate concentration in spinach

Abstract The effects of three commercial substrates (a mixture of forest residues, composted grape husks, and white peat, black peat and coir) on plant growth and nitrogen (N) and nitrate (NO3) concentration and content were evaluated in spinach (Spinacia oleracea L. cv. Tapir). Spinach seedlings were transplanted at 45 days after emergence into Styrofoam boxes filled with the substrates and were grown during winter and early spring in an unheated greenhouse with no supplemental lighting. Each planting box was irrigated daily by drip and fertilized with a complete nutrient solution. The NO3 content of the drainage water was lower in coir than in the other substrates. However, shoot NO3 concentration was not affected by substrate type, while yield and total shoot N and NO3 content were greater when plants were grown in peat than in the mixed substrate or the coir. Leaf chlorophyll meter readings provided a good indication of the amount of N in the plants and increased linearly with total shoot N. Keywords Spinacia oleracea; chlorophyll meter; coir; peat; soilless culture systems

Effects of substrate type on plant growth and nitrogen and nitrate concentration in spinach.

The effects of three commercial substrates (a mixture of forest residues, composted grape husks, and white peat, black peat and coir) on plant growth and nitrogen (N) and nitrate (NO3) concentration and content were evaluated in spinach (Spinacia oleracea L. cv. Tapir). Spinach seedlings were transplanted at 45 days after emergence into Styrofoam boxes filled with the substrates and were grown during winter and early spring in an unheated greenhouse with no supplemental lighting. Each planting box was irrigated daily by drip and fertilized with a complete nutrient solution. The NO3 content of the drainage water was lower in coir than in the other substrates. However, shoot NO3 concentration was not affected by substrate type, while yield and total shoot N and NO3 content were greater when plants were grown in peat than in the mixed substrate or the coir. Leaf chlorophyll meter readings provided a good indication of the amount of N in the plants and increased linearly with total shoot N.

Dedifferentiation of Leaf Cells and Growth Pattern of Calluses of Capsicum annuumcv. Etna.

Background:In vitrocell suspension cultivation systems have been largely reported assafe and standardized methods for production of secondary metabolites with medicinaland agricultural interest.Capsicum annuumis one of the most widely grown vegetablein the world and its biological activities have been demonstrated against insects, fungi,bacteria and other groups of organisms. The determination of procedures for thededifferentiation of cells into callus cells and the subsequent study of the callus growthpattern are necessary for the establishment of cellsuspensions and also to subsidizestudies regarding the bioactivity of its secondarymetabolites. To date, no study hasdescribed the development of protocols for callus induction inC. annuumL. cv. Etna. Objective:The objective of this study was to establish a protocol for dedifferentiationof leaf cells of the cultivarC. annuumcv. Etna and to determine the growth pattern ofthe calluses with a focus on the deceleration phase, when the callus cells must besubcultured into a liquid medium in order to establish cell suspension cultivationsaiming at the production of secondary metabolites.Results:The treatment that resultedin the highest %CI, ACCC and callus weight was thecombination of 4.52 μ M 2,4-D +0.44 μ M BA. The calluses produced were friable andwhitish and their growth patternfollowed a sigmoid shape. The deceleration phase started on the 23rdday of cultivation.Conclusion:Callus induction in leaf explants ofC. annuumcv. Etnacan be achieved inMS medium supplemented with 4.52 μ M 2,4-D + 0.44 μ MBA, which results in highcellular proliferation; in order to start a cell suspension culture, callus cells on the 23rdday of culture should be used.

Evolution and challenges of dynamic global vegetation models for some aspects of plant physiology and elevated atmospheric CO2.

Dynamic global vegetation models (DGVMs) simulate surface processes such as the transfer of energy, water, CO2, and momentum between the terrestrial surface and the atmosphere, biogeochemical cycles, carbon assimilation by vegetation, phenology, and land use change in scenarios of varying atmospheric CO2 concentrations. DGVMs increase the complexity and the Earth system representation when they are coupled with atmospheric global circulation models (AGCMs) or climate models. However, plant physiological processes are still a major source of uncertainty in DGVMs. The maximum velocity of carboxylation (Vcmax), for example, has a direct impact over productivity in the models. This parameter is often underestimated or imprecisely defined for the various plant functional types (PFTs) and ecosystems. Vcmax is directly related to photosynthesis acclimation (loss of response to elevated CO2), a widely known phenomenon that usually occurs when plants are subjected to elevated atmospheric CO2 and might affect productivity estimation in DGVMs. Despite this, current models have improved substantially, compared to earlier models which had a rudimentary and very simple representation of vegetation?atmosphere interactions. In this paper, we describe this evolution through generations of models and the main events that contributed to their improvements until the current state-of-the-art class of models. Also, we describe some main challenges for further improvements to DGVMs.

Plant density and mineral nitrogen fertilization influencing yield, yield components and concentration of oil and protein in soybean grains.

There are few studies on the interaction between soybean plant density and nitrogen fertilization. This research aimed to assess the effect of mineral nitrogen associated to different plant densities on yield, yield components and oil and protein concentrations of soybean grains. Two experiments were conducted in the 2013/2014 and 2014/2015 growing seasons, with randomized complete block design, in a split plots scheme, with six replications. Four sowing densities (150, 300, 440 and 560 thousand viable seeds; ha-1) were allocated in the plots, and two nitrogen levels (0 and 45 kg N; ha-1, applied at V2, using ammonium sulfate) were allocated in the subplots. There was no interaction between soybean plant density and the application of mineral nitrogen on yield, yield components and oil and protein concentrations in soybean grains. Higher plant population reduced the number of pods per plant and the contribution of branch sinks to the grain yield, but the effects on yield differed among the growing seasons. The mineral nitrogen fertilization did not increase yield and protein and oil concentrations in the grains, thus it was unnecessary.

Histology challenge : an interactive program for 1st year students

Our students come from diverse backgrounds. They need flexibility in their learning, and opportunities to review aspects of curriculum they are less confident with. An online teaching and learning programme called the Histology Challenge has been developed to supplement learning experiences offered in several first year anatomy and anatomy & physiology units at QUT. The programme is designed to be integrated with the existing Blackboard sites. The Histology Challenge emphasises the foundation concept that a complex system, such as the human body, can be better understood by examining its simpler components. The tutorial allows students to examine the cells and tissues which ultimately determine structural and functional properties of body organs. The program is interactive, asking students to make decisions and choices, demonstrating an integrated understanding of systemic and cellular aspects. It provides users with the ability to progress at their own pace and to test their understanding and knowledge. For the developer the learning activity can be easily controlled and modified via the use of text files. There are several key elements of this programme, designed to promote specific aspects of student learning. Minimum text is used, instead there is a strong emphasis on instructive artwork and original, high quality histology images presented within a framework that reinforces learning and promotes problem solving skills.

Plasmas meet nanoparticles—where synergies can advance the frontier of medicine

Nanoparticles and low-temperature plasmas have been developed, independently and often along different routes, to tackle the same set of challenges in biomedicine. There are intriguing similarities and contrasts in their interactions with cells and living tissues, and these are reflected directly in the characteristics and scope of their intended therapeutic solutions, in particular their chemical reactivity, selectivity against pathogens and cancer cells, safety to healthy cells and tissues and targeted delivery to diseased tissues. Time has come to ask the inevitable question of possible plasma–nanoparticle synergy and the related benefits to the development of effective, selective and safe therapies for modern medicine. This perspective paper offers a detailed review of the strengths and weakenesses of nanomedicine and plasma medicine as a stand-alone technology, and then provides a critical analysis of some of the major opportunities enabled by synergizing nanotechnology and plasma technology. It is shown that the plasma–nanoparticle synergy is best captured through plasma nanotechnology and its benefits for medicine are highly promising.

Clinical impacts of mammalian target of rapamycin expression in human colorectal cancers

This study investigated the clinicopathologic roles of mammalian target of rapamycin (mTOR) expression and its relationship to carcinogenesis and tumor progression in a colorectal adenoma-adenocarcinoma model. Two colon cancer cell lines with different pathologic stages (SW480 and SW48) and 1 normal colonic epithelial cell line (FHC) were used, in addition to 119 colorectal adenocarcinomas and 32 adenomas. mTOR expression profiles at messenger RNA (mRNA) and protein levels were investigated in the cells and tissues using real-time quantification polymerase chain reaction and immunohistochemistry. The findings were correlated with the clinicopathologic features of the tumors. The colon cell line from stage III cancer (SW48) showed higher expression of mTOR mRNA than that from stage II cancer (SW480). At the tissue level, mTOR showed higher mRNA and protein expression in colorectal carcinoma than in adenoma. The mRNA and protein expression was correlated with each other in approximately one-third of the carcinomas and adenomas. High levels of mTOR mRNA expression were noted more in carcinoma or adenoma arising from the distal portion of the large intestine (P = .025 and .019, respectively). Within the colorectal cancer population, a high level of expression of mTOR mRNA was related to the presence of lymph node metastases (P = .031), advanced pathologic stage (P = .05), and presence of persistent disease or tumor recurrence (P = .035). To conclude, the study has indicated that mTOR is likely to be involved in the development and progression of colorectal cancer and is linked to cancer initiation, invasiveness, and progression.