Circulating Organ-Specific MicroRNAs Serve as Biomarkers in Organ-Specific Diseases: Implications for Organ Allo- and Xeno-Transplantation

Different cell types possess different miRNA expression profiles, and cell/tissue/organ-specific miRNAs (or profiles) indicate different diseases. Circulating miRNA is either actively secreted by living cells or passively released during cell death. Circulating cell/tissue/organ-specific miRNA may serve as a non-invasive biomarker for allo- or xeno-transplantation to monitor organ survival and immune rejection. In this review, we summarize the proof of concept that circulating organ-specific miRNAs serve as non-invasive biomarkers for a wide spectrum of clinical organ-specific manifestations such as liver-related disease, heart-related disease, kidney-related disease, and lung-related disease. Furthermore, we summarize how circulating organ-specific miRNAs may have advantages over conventional methods for monitoring immune rejection in organ transplantation. Finally, we discuss the implications and challenges of applying miRNA to monitor organ survival and immune rejection in allo- or xeno-transplantation.     

Pheromonal Discrimination of Sex by Male and Female Leopard Geckos (Eublepharis macularius)

The ability of male and female Eublepharis macularius to discriminate among pheromones of males and females and a blank control was investigated. Stimuli were presented on ceramic tiles in the animal's home cages. Males tongue-flicked at significantly lower rates in response to male stimuli than to female and control stimuli. Males also performed aggressive behaviors toward male, but not female or control, stimuli, and tail vibrations toward female, but not male or control, stimuli. Mean tongue-flick rates by females did not differ significantly among conditions. Discrimination of male pheromones by females was demonstrated by greater performance of labial-licking and chin-rubbing in response to male stimuli than to either female or control stimuli.     

Soil nutrient maps of Sub-Saharan Africa: assessment of soil nutrient content at 250 m spatial resolution using machine learning

Spatial predictions of soil macro and micro-nutrient content across Sub-Saharan Africa at 250 m spatial resolution and for 0–30 cm depth interval are presented. Predictions were produced for 15 target nutrients: organic carbon (C) and total (organic) nitrogen (N), total phosphorus (P), and extractable—phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), sulfur (S), sodium (Na), iron (Fe), manganese (Mn), zinc (Zn), copper (Cu), aluminum (Al) and boron (B). Model training was performed using soil samples from ca. 59,000 locations (a compilation of soil samples from the AfSIS, EthioSIS, One Acre Fund, VitalSigns and legacy soil data) and an extensive stack of remote sensing covariates in addition to landform, lithologic and land cover maps. An ensemble model was then created for each nutrient from two machine learning algorithms—random forest and gradient boosting, as implemented in R packages ranger and xgboost—and then used to generate predictions in a fully-optimized computing system. Cross-validation revealed that apart from S, P and B, significant models can be produced for most targeted nutrients (R-square between 40–85%). Further comparison with OFRA field trial database shows that soil nutrients are indeed critical for agricultural development, with Mn, Zn, Al, B and Na, appearing as the most important nutrients for predicting crop yield. A limiting factor for mapping nutrients using the existing point data in Africa appears to be (1) the high spatial clustering of sampling locations, and (2) missing more detailed parent material/geological maps. Logical steps towards improving prediction accuracies include: further collection of input (training) point samples, further harmonization of measurement methods, addition of more detailed covariates specific to Africa, and implementation of a full spatio-temporal statistical modeling framework.     

Retardation of Crystallization through the Addition of Dairy Phospholipids

The objective of this work was to identify the effects that milk phospholipids (PL) have on crystallization of anhydrous milk fat (AMF). Three mixtures were prepared by adding 0%, 0.01%, and 0.1% PL to AMF. Each mixture was crystallized for 90 min at 24, 26, and 28 °C. The solid fat content was measured as a function of time and fitted to the Avrami equation. Melting point, thermal behavior, viscoelastic properties, and crystal morphology were all measured at 90 min. All assays were repeated, as well as hardness, after being stored at 5 °C for 48 hours. Samples containing PL showed slower crystallization as concentration increased especially at higher temperatures (26 and 28 °C). The addition of PL caused a difference in crystal morphology resulting in visibly larger crystals at 90 min. The elasticity and hardness at 90 min were influenced by the addition of PL at 24 °C with lower values obtained in samples with PL compared to the AMF alone. No differences in hardness nor in elasticity was observed for samples crystallized at 26 and 28 °C. A decrease in melting enthalpy was observed in samples with PL indicating a reduction in crystallization at all temperatures, which was supported by crystal morphology.     

Exterior wood coatings. Part-2: Modeling correlation between coating properties and their weathering performance

Understanding how a coating’s properties are related to its performance characteristics and how measuring its properties can be used for predicting the performance of coating in service was the main focus of this study. A number of exterior penetrating wood stains were characterized in terms of glass transition temperature (T _g), solid content, viscosity, and surface tension. The contact angles of liquid coatings were measured on wood treated with chromated copper arsenate, alkaline copper quat, and copper azole as well as untreated wood. Also, the film thicknesses of the cured coatings on wood surfaces were measured by back-scattered electron imaging of osmium-treated samples. Partial least square-regression (PLS-R) modeling was used to correlate measured coating properties with their weathering performance on the preservative-treated and untreated wood substrates, based on water repellency, color retention, and visual ratings after 3 years of natural weathering. A PLS model was developed with 77% fitting accuracy and 69% prediction ability. This model indicated that preservative treatments significantly affected coating performance, and among coating properties, film thickness, viscosity, and glass transition temperature had the greatest effects on the coatings’ performance in exterior.     

Responses to major categories of food chemicals by the lizard Podarcis lilfordi

Many lizards are capable of identifying food using only chemical cues from food surfaces, but almost nothing is known about the types of compounds that are effective stimuli. We experimentally studied lingual and biting responses by a lacertid lizard, Podarcis lilfordi, to single representatives of three major categories of food chemicals, sucrose as a carbohydrate, pure pork fat as a mixture of lipids, and bovine gamma globulin as a protein. In 60-sec trials in which stimuli were presented on cotton swabs, the lizards detected all three stimuli, exhibiting more tongue-flicks, licks, or bites, or a greater tongue-flick attack score (TFAS; overall measure of response strength to prey stimuli) than to deionized water. The initial response to all stimuli was tongue-flicking, but the lizards discriminated among the types of chemical stimuli. After preliminary tongue-flicks, the lizards responded to sucrose solutions by licking at high rates, to pure pork fat by biting, and to protein by a combination of additional tongue-flicks and biting. Biting is a feeding response to prey or solid plant material. Licking is a feeding response to sugars in nectar or ripe fruit. Its frequency increased with sucrose concentration. Our data suggest that lizards can identify several types of chemicals associated with food and direct feeding attempts to sources of such chemicals in the absence of visual cues.     

Enhanced biosurfactant production by <Emphasis Type="Italic">Corynebacterium alkanolyticum</Emphasis> ATCC 21511 using self-cycling fermentation

Enhanced biosurfactant production by Corynebacterium alkanolyticum ATCC 21511 was accomplished in a self-cycling fermenter (SCF) on a hexadecane substrate. The phospholipid biosurfactant produced during each cycle could be monitored rapidly using fluorescence spectroscopy. By optimizing the cycling pattern of the SCF, significantly better yields of biosurfactant were obtained than previously reported for this microorganism. It was also possible to virtually eliminate the hydrocarbon residue in the product. Harvest concentrations of 1.9 g L⁻¹ were obtained by using a two-stage fermentation. The first step was the growth of C. alkanolyticum in an SCF to yield a harvest of synchronous cells. These cells were transferred to a second vessel for the production stage. The concentration of biosurfactant could be further increased to 2.7 g L⁻¹ by the addition of more hexadecane at the beginning of the second stage.     

Inhibition of cholesterol synthesis by cyclopropylamine derivatives of squalene in human hepatoblastoma cells in culture

Two squalene derivatives, trisnorsqualene cyclopropylamine and trisnorsqualeneN-methylcyclopropylamine, were synthesized and tested for inhibition of lanosterol and squalene epoxide formation from squalene in rat hepatic microsomes, and for the inhibition of cholesterol syntheses in human cultured hepatoblastoma (HepG2) cells. Trisnorsqualene cyclopropylamine inhibited [³H]-squalene conversion to [³H]squalene epoxide in microsomes (IC₅₀=5.0 μM), indicating that this derivative inhibited squalene mono-oxygenase. Trisnorsqualenen-methylcyclopropylamine inhibited [³H]squalene conversion to [³H]lanosterol (IC₅₀=12.0 μM) and caused [³H]-squalene epoxide to accumulate in microsomes, indicating that this derivative inhibited 2,3-oxidosqualene cyclase. Cholesterol biosynthesis from [¹⁴C]acetate in HepG2 cells was inhibited by both derivatives (IC₅₀=1.0 μM for trisnorsqualene cyclopropylamine; IC₅₀=0.5 μM for trisnorsqualeneN-methylcyclopropylamine). Cells incubated with trisnorsqualene cyclopropylamine accumulated [¹⁴C]squalene, while cells incubated with trisnorsqualeneN-methylcyclopropylamine accumulated [¹⁴C]squalene epoxide and [¹⁴C]squalene diepoxide. The concentration range of inhibitor which caused these intermediates to accumulate coincided with that which inhibited cholesterol synthesis. The results indicate that cyclopropylamine derivatives of squalene are effective inhibitors of cholesterol synthesis, and that substitutions at the nitrogen affect enzyme selectivity and thus the mechanism of action of the compounds.     

Lipase G-Catalyzed synthesis of monoglycerides in organic solvent and analysis by HPLC

Monoglycerides were synthesized with Lipase G fromPenicillium sp. as biocatalyst. This enzyme successfully catalyzed the esterification of glycerol with oleic acid (18:1 n-9) or eicosapentaenoic acid, EPA (20:5 n-3) in hexane. Esterification at 40°C for 24 hr resulted in 86.3 mol% and 64.3 mol% incorporation of 18:1 n-9 and 20:5 n-3, respectively. Lipase GC fromGeotrichum candidum was not effective in esterifying the fatty acids under the present experimental conditions. Lipase G was able to incorporate 98.5 mol% and 98.1 mol% of 18:1 n-9 onto glycerol in 24 hr or less at 25°C and 37°C, respectively, in the absence of molecular sieve 4A. The product formed was analyzed by high performance liquid chromatography (HPLC) with a combined evaporative light scattering mass detector (ELSD) and ultraviolet detection at 206 nm. The result of this study demonstrates that Lipase G is capable of incorporating long-chain fatty acids of potential health benefit onto glycerol.