Red blood cells imaging and antigen-antibody interaction measurement

In the present study the atomic force microscope (AFM) was used to image the surface morphology of red blood cells (RBC) for the first time. The AFM yielded very reproducible images without appreciable modifications of the sample surfaces. In addition to this topographical imaging, we have developed an experimental approach to measure the binding strength between antibody (anti-A), and the RBC antigen A, when reversible bonds between specific molecules such as antigen and antibody mediate the adhesion. The experimental results suggest that the procedure established here may be used for specific antibody detection. This study has also enhanced our understanding under physiological conditions of molecular interaction in particular antigen-antibody.     

Large, rapidly evolving intergenic spacers in the mitochondrial DNA of the salamander family Ambystomatidae (Amphibia: Caudata)

We report the presence, in the mitochondrial DNA (mtDNA) of all of the sexual species of the salamander family Ambystomatidae, of a shared 240- bp intergenic spacer between tRNAThr and tRNAPro. We place the intergenic spacer in context by presenting the sequence of 1,746 bp of mtDNA from Ambystoma tigrinum tigrinum, describe the nucleotide composition of the intergenic spacer in all of the species of Ambystomatidae, and compare it to other coding and noncoding regions of Ambystoma and several other vertebrate mtDNAs. The nucleotide substitution rate of the intergenic spacer is approximately three times faster than the substitution rate of the control region, as shown by comparisons among six Ambystoma macrodactylum sequences and eight members of the Ambystoma tigrinum complex. We also found additional inserts within the intergenic spacers of five species that varied from 87-444 bp in length. The presence of the intergenic spacer in all sexual species of Ambystomatidae suggests that it arose at least 20 MYA and has been a stable component of the ambystomatid mtDNA ever since. As such, it represents one of the few examples of a large and persistent intergenic spacer in the mtDNA of any vertebrate clade.      

Induced change in <Emphasis Type="Italic">Arthrospira</Emphasis> sp. (<Emphasis Type="Italic">Spirulina</Emphasis>) intracellular and extracellular metabolites using multifactor stress combination approach

The interactive effects of light intensity, NaCl, nitrogen, and phosphorus on intracellular biomass content and extracellular polymeric substance production were assessed for Arthrospira sp. (Spirulina) in a two-phase culture process using principal component analysis and central composite face design. Under high light intensity (120 μmol photons m^?2?s^?1) and low NaCl (1 gL^?1), NaNO₃, and K₂HPO₄ (0.5 g L^?1), the carbohydrate content was maximized to 26.61%. Interaction of both K₂HPO₄ (1.6 gL^?1) and NaCl (1.19 gL^?1) with low NaNO₃ (0.5 gL^?1) achieved the maximum content of lipids (15.62%), while high NaCl (40 gL^?1), K₂HPO₄, and NaNO₃ (4.5 gL^?1) enhanced mainly total carotenoids (0.85%). Conversely, under low light intensity of 10 μmol photons m^?2?s^?1 combined with 11.76 gL^?1 of NaCl, 0.5 gL^?1 of NaNO₃, and 2.68 gL^?1 of K₂HPO₄, the phycobiliprotein content reached its highest level (16.09%). The maximum extracellular polymeric substance (EPS) production (0.902 gg^?1?DW) was triggered under moderate light of 57.25 μmol photons m^?2?s^?1 and interaction of high NaCl (40 gL^?1) and K₂HPO₄ (4.5 gL^?1) with low NaNO₃ (0.5 gL^?1). The maximization ratios of intracellular biomass content in terms of carbohydrate, lipid, total carotenoid, phycobiliprotein, and EPS production were 3.55-, 1.73-, 9.55-, 2.92-, and 1.46-fold, respectively, greater than those obtained at optimal growth conditions. This study demonstrated that the multiple stress factors applied to the adopted two-phase culture process could be a promising strategy to produce biomass enriched in various high-value compound.     

Combined effect of temperature and light intensity on growth and extracellular polymeric substance production by the cyanobacterium Arthrospira platensis

The effects of light intensity and temperature on Arthrospira platensis growth and production of extracellular polymeric substances (EPS) in batch culture were evaluated using a three-level, full-factorial design and response surface methodology. Three levels were tested for each parameter (temperature: 30, 35, 40°C; light intensity: 50, 115, 180 μmol photons m⁻² s⁻¹). Both growth and EPS production are influenced mainly by the temperature factor but the interaction term temperature*light intensity also had a significant effect. In addition, conditions optimising EPS production are different from those optimising growth. The highest growth rate (0.414 ± 0.003 day⁻¹) was found at the lowest temperature (30°C) and highest light intensity (180 μmol photons m⁻² s⁻¹) tested, no optima were detectable within the given test range. Obviously, optima for growth must be at a temperature lower than 30°C and a light intensity higher than 180 μmol photons m⁻² s⁻¹. For EPS production, light intensity had a positive linear effect (optimum obviously higher than 180 μmol photons m⁻² s⁻¹), but for the temperature parameter a maximum effect was detectable at 35°C.     

Site localization of sialyl Lewis(x) antigen on alpha1-acid glycoprotein by high performance liquid chromatography-electrospray mass spectrometry

A simple, fast and sensitive method was developed to verify the presence of the sialyl Lewis(x) antigen on an N-linked glycoprotein. High performance liquid chromatography-electrospray mass spectrometry (HPLC-ESI/MS) was used to identify which of the five N-linked glycosylation sites of human plasma alpha1-acid-glycoprotein (orosomucoid, OMD) contain the sialyl Lewis(x) antigen. OMD was digested with proteolytic enzymes and analyzed by reversed phase chromatography coupled with on-line ESI/MS. A tandem mass spectrometry experiment was designed to detect the presence of the sialyl Lewis(x) antigen based on the observation of an 803 mass to charge ratio ( m/z ) ion produced in the intermediate pressure region of the ESI interface. The ESI/MS signal at m/z 803 is consistent with an oxonium ion for a glycan structure containing NeuAc, Gal, GlcNAc, and Fuc. The identity of the m/z 803 ion was confirmed by ESI/MS/MS analysis of the m/z 803 fragment ion and comparison with a sialyl Lewis(x) standard. The stereochemistry and linkage positions were assigned using previous NMR analysis but could be determined with permethylation analysis if necessary. The analysis of OMD gave a pattern showing signal for the sialyl Lewis(x) antigen coeluting with each of the five N-linked glycopeptides. The ability to monitor sialyl Lewis(x) expression at each of the five sites is of interest in the study of OMD's role in inflammatory diseases.      

Modeling growth and photosynthetic response in <Emphasis Type="Italic">Arthrospira platensis</Emphasis> as function of light intensity and glucose concentration using factorial design

Combined effect of light intensity and glucose concentration on Arthrospira platensis growth and photosynthetic response was evaluated using a 3² factorial design. This design was carried out with light levels of 50, 100, and 150 μmol photons m⁻² s⁻¹ and glucose concentrations of 0.5, 1.5, and 2.5 g L⁻¹. Results from the response surface methodology were that the highest level of light intensity and glucose concentration improved biomass (1.33 g L⁻¹), maximum specific growth rate (0.49 day⁻¹), and net photosynthetic rate (139.89 μmol O₂ mg Chl⁻¹ h⁻¹). Furthermore, the interaction of both factors showed that at low light, glucose had a low effect on maximum biomass and maximal net photosynthetic rate. However, at the highest light levels, the effect of glucose was more sensitive and the increase of glucose concentration increased the levels of all responses. The rates of the instantaneous relative growth, net photosynthesis, and dark respiration of growth cultures showed two different phases in mixotrophic condition. The first was distinguished by the preponderance of the photoautotrophic mode; the second was based mainly on photoheterotrophy.     

Preservation of fresh edible cactus stems (<i>Opuntia ficus indica</i> Mill.) by modified atmosphere packaging

Cactus stems, the cladodes of Opuntia spp. cacti, are consumed in Mexico and other countries due to their fresh and herbaceous flavor, and because of their widely known nutraceutical benefits. In order to extend the postharvest life of this vegetable, the effect of a modified atmosphere packaging (MAP) was studied in cactus stems of the cultivar Atlixco stored at 4 ± 1 °C for 20 days under three types of atmospheres: (1) air (passive atmosphere), (2) 5 kPa O₂ + 4 kPa CO₂, and (3) N₂. During storage, the titratable acidity decreased and the color of cladodes became darker and less green; however, the 5 kPa O₂ + 4 kPa CO₂ atmosphere was able to preserve both quality characteristics. All modified atmospheres reduced weight loss (from 8 to <2%) and the symptoms of chilling injury, and this physiological disorder appeared earlier in controls than in MAP-stored cladodes. The levels of fermentation metabolites were low in all three evaluated atmospheres. Because of this, only cladodes stored under the N₂ atmosphere were selected for furthersensory analysis of the MAP effect on odor perception as evaluated by a trained panel. Results indicated that there was no detrimental effect (atypical odors) of MAP on this sensory characteristic. We conclude that cultivar Atlixco is suitable for preservation using MAP technology.     

Hydraulic properties of layered soils influence survival of Rhodes grass (<Emphasis Type="Italic">Chloris gayana</Emphasis> Kunth.) during water stress

Survival of vegetation on soil-capped mining wastes is often impaired during dry seasons due to the limited amount of water stored in the shallow soil capping. Growth and survival of Rhodes grass (Chloris gayana) during soil drying on various layered capping sequences constructed of combinations of topsoil, subsoil, seawater-neutralised residue sand and low grade bauxite was determined in a glasshouse. The aim was to describe the survival of Rhodes grass in terms of plant and soil water relationships. The soil water characteristic curve and soil texture analysis was a good predictor of plant survival. The combination of soil with a high water holding capacity and low soil water diffusivity (e.g. subsoil with high clay contents) with soil having a high water holding capacity and high diffusivity (e.g. residue sand) gave best survival during drying down (up to 88 days without water), whereas topsoil and low grade bauxite were unsuitable (plants died within 18–39 days). Clayey soil improved plant survival by triggering a water stress response during peak evaporative water demand once residue sand dried down and its diffusivity fell below a critical range. Thus, for revegetation in seasonally dry climates, soil capping should combine one soil with low diffusivity and one or more soils with high total water holding capacity and high diffusivity.