Stem cell factor supports migration in canine mesenchymal stem cells

Adult Mesenchymal Stem Cells (MSC) are cells that can be defined as multipotent cells able to differentiate into diverse lineages, under appropriate conditions. These cells have been widely used in regenerative medicine, both in preclinical and clinical settings. Initially discovered in bone marrow, MSC can now be isolated from a wide spectrum of adult and foetal tissues. Studies to evaluate the therapeutic potential of these cells are based on their ability to arrive to damaged tissues. In this paper we have done a comparative study analyzing proliferation, surface markers and OCT4, SOX9, RUNX2, PPARG genes expression in MSC cells from Bone marrow (BMMSC) and Adipose tissue (ASC). We also analyzed the role of Stem Cell Factor (SCF) on MSC proliferation and on ASCs metalloproteinases MMP-2, MMP-9 secretion. Healthy dogs were used as BMMSC donors, and ASC were collected from omentum during elective ovariohysterectomy surgery. Both cell types were cultured in IMDM medium with or without SCF, 10% Dog Serum (DS), and incubated at 38 °C with 5% CO2. Growth of BMMSCs and ASCs was exponential until 25–30 days. Flow citometry of MSCs revealed positive results for CD90 and negative for CD34, CD45 and MCH-II. Genes were evaluated by RT-PCR and metalloproteinases by zymografy. Our findings indicate morphological and immunological similarities as well as expression of genes from both origins on analyzed cells. Furthermore, SCF did not affect proliferation of MSCs, however it up-regulated MMP-2 and MMP-9 secretion in ASCs. These results suggest that metalloproteinases are possibly essential molecules pivoting migration.     

Natural hybridization between wheat (Triticum aestivum L.) and its wild relatives Aegilops geniculata Roth and Aegilops triuncialis L.

BACKGROUND

Cultivated bread wheat (Triticum aestivum L.) spontaneously hybridizes with wild/weedy related Aegilops populations, but little is known about the actual rates at which this hybridization occurs under field conditions. It is very important to provide reliable empirical data on this phenomenon in order to assess the potential crop–wild introgression, especially in the context of conducting risk assessments for the commercialization of genetically modified (GM) wheat, as gene flow from wheat to Aegilops species could transfer into the wild species genes coding for traits such as resistance to herbicides, insects, diseases or environmental stresses.

RESULTS

The spontaneous hybridization rates between wheat and A. geniculata and A. triuncialis, which are very abundant in the Mediterranean area, have been estimated for the first time in the northern part of the Meseta Central, the great central plateau which includes the largest area of wheat cultivation in Spain. Hybridization rates averaged 0.12% and 0.008% for A. geniculata and A. triuncialis, respectively. Hybrids were found in 26% of A. geniculata and 5% of A. triuncialis populations, at rates that can be ≤3.6% for A. geniculata and 0.24% for A. triuncialis.

CONCLUSION

The detection of Aegilops spp.–wheat hybrids in Aegilops populations indicates that gene flow can occur, although wheat is considered a crop with a low-to-medium risk for transgene escape. These data on field hybridization rates are essential for GM wheat risk assessment purposes. © 2023 Society of Chemical Industry.     

SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis

Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.     

Correlation of selected constituents with the total antioxidant capacity of coffee beverages: influence of the brewing procedure

Relationships between volatile and nonvolatile compounds and the antioxidant capacity of coffee brews prepared from commercial conventional and torrefacto roasted coffees, employing commonly used doses and prepared by four brewing procedures (filter, plunger, mocha, and espresso machine) were assessed. Significant correlations between volatile Maillard reaction products and antioxidant capacity (measured by both 2,2-diphenyl-1-picrylhydrazyl radical and redox potential methods) were not observed. Highly positive correlations between browned compounds and caffeine with both antioxidant capacity parameters were reported. Principal component analysis allowed coffee brews separation according to coffee roasting processes (PC1) and brewing procedures (PC2), showing that in all cases coffee brews from torrefacto roasted coffee were more antioxidant that those extracted from conventional ones; also, coffee brews extracted by an espresso machine were more antioxidant than those extracted by mocha, plunger, and filter machines.     

Fermentation as a bio-process to obtain functional soybean flours

The effect of fermentation on the antioxidant compounds [vitamins C and E, total phenolic compounds (TPC), and reduced glutathione (GSH)], and antioxidant capacity [superoxide anion scavenging activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), inhibition of phosphatidylcholine (PC) peroxidation, and Trolox equivalent antioxidant capacity (TEAC)] of soybean (Glycine max cv. Merit) was studied. Fermentation was carried out in solid state in cracked seeds inoculated with Aspergillus oryzae, Rhizopus oryzae, Bacillus subtilis, and Lactobacillus plantarum and in liquid state either in cracked seeds or milled soybean flours fermented naturally by only the microorganisms present in the seeds or by inoculation with L. plantarum. Vitamin C was not detected in the studied samples. Fermentation caused a decrease in vitamin E activity, except when cracked seed was fermented with A. oryzae, R. oryzae, or B. subtilis that increased 31, 30, and 89%, respectively. Fermentation produced an increase in TPC content and did not affect or reduce the GSH content. Fermentation decreased SOD-like activity drastically, while PRTC increased except when it was carried out naturally in cracked seed. TEAC values rose sharply when soybeans were fermented with B. subtilis. Processed soybean extracts inhibited PC peroxidation in comparison with the control assay. On the basis of the results obtained, the relative contributions of vitamin E, TPC, and GSH to antioxidant capacity were calculated and results showed a very high TPC contribution and a low contribution of GSH and vitamin E activity. Optimum results for functional soybean flours were achieved when fermentation was carried out with B. subtilis inoculum.     

Total chemically available (free and intrachain) lysine and furosine in pea, bean, and lentil sprouts

The effect of the germination of peas, beans, and lentils under differing conditions of illumination for different times on parameters linked to the Maillard reaction (chemically available free and intrachain lysine, lysine availability, and furosine) was evaluated. The chemically available free lysine content in the raw seeds of the three legumes was quite small compared to the chemically available intrachain lysine content, and furosine was detectable only in the beans and the lentils. The effect of germination was to increase lysine availability compared with levels in the raw seeds in all of the germinated samples, the smallest increase taking place in the lentils. In addition, furosine became detectable in all of the germinated samples. Quantities varied depending on the germination conditions but in all cases were higher than the quantities observed in the raw seeds. Linear correlations were observed to exist between some of the parameters considered in the three legumes tested.     

Changes in the Solubility of Corn Proteins through Interaction with the Arabinoxylans in Extruded Nixtamalized Corn Flour Treated with Xylanase

The extrusion process allows the production of nixtamalized corn flour rich in arabinoxylans, which help to prevent cardiovascular and intestinal diseases. During extrusion, physiochemical properties of nixtamalized corn flour are negatively modified. The use of enzymes such as xylanase in order to obtain nixtamalized corn flour using extrusion has been studied as an alternative to reduce these changes in corn flour tortilla. The aim of this research was to evaluate changes in protein solubility of extruded nixtamalized corn flour with and without different concentrations of xylanase enzyme (0.05, 0.075, and 0.1 %, w/w). Soluble proteins of each corn flour were extracted and analyzed by SE-HPLC, while insoluble proteins were determined by the combustion method. In addition, each corn flour was analyzed by scanning electron microscopy (SEM). Results showed that the extruded nixtamalized corn flour, with and without xylanase, increased the protein solubility, and this effect was lower in extruded nixtamalized corn flour with xylanase. Insoluble protein diminished in corn flours either with or without xylanase enzyme. The addition of xylanase reduces the effect that the extrusion process has on the solubility proteins of extruded nixtamalized corn flour.     

Effect of processing on the antioxidant vitamins and antioxidant capacity of Vigna sinensis Var. Carilla

Cowpea (Vigna sinensis L. var. Carilla) flours obtained by fermentation with inoculum Lactobacillus plantarum (PF) or with the natural microorganisms present in the flour (NF) and subsequent heat treatment in an autoclave were prepared to study the effect of fermentation on the antioxidant vitamin content and on the antioxidant capacity. Bacterial counts and pH values, vitamins C and E, carotenoids, glutathione (GSH), superoxide dismutase-like activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), lipid peroxidation in unilamillar liposomes, and Trolox equivalent antioxidant capacity (TEAC) were evaluated in raw and processed cowpea flours. gamma-Tocopherol and delta-tocopherol were found in raw cowpea, whereas vitamin C and carotenoids were not detected. An increase in the vitamin E activity was observed in PF, whereas vitamin C and carotenoids were not detected in fermented cowpea flours. Fermentation or heat treatment in an autoclave after fermentation produced processed cowpea flours with lower PRTC, glutathione content, and SOD-like activity than those of the raw seeds. However, those processes increased the capacity to inhibit the lipid peroxidation in unilamellar lipoposomes and TEAC. According to the results obtained in this study, the fermentation of cowpeas (naturally or with L. plantarum) and fermentation and subsequent heat treatment in an autoclave are good processes to obtain functional cowpea flours having higher antioxidant capacity than the raw legume.     

Vitamin C,provitamin A carotenoids,and other carotenoids in high-pressurized orange juice during refrigerated storage

Vitamin C, provitamin A carotenoids, and other carotenoids were measured in freshly squeezed juices from oranges (Citrus sinensis L. var. Valencia late) that were subjected to high-pressure (HP) treatment. Also, the stability of these compounds was studied during refrigerated storage at 4 degrees C. HP treatment is an alternative to heat preservation methods for foods; therefore, it is essential to assess the impact of HP on bioactive compounds. Several processes that combine HP treatment with heat treatment for various time periods were assayed: T0, fresh juice (without treatment); T1, 100 MPa/60 degrees C/5 min; T2, 350 MPa/30 degrees C/2.5 min; T3, 400 MPa/40 degrees C/1 min. Fresh and treated samples were kept refrigerated (4 degrees C) over 10 days. After application of HP and during the refrigeration period, the qualitative and quantitative determination of vitamin C, provitamin A carotenoids (beta- and alpha-carotene; beta- and alpha-cryptoxanthin), and the xanthophylls zeaxanthin and lutein was achieved by high-performance liquid chromatography. T1 and T3 juices showed a decrease in ascorbic acid and total vitamin C just after HP treatment (D0) compared with T0 juices. On the contrary, T2 juices, just after HP treatment (D0), had the same levels of both compounds compared to untreated juices. T1, T2, and T3 treatments led to an increase in the extraction of carotenoids and provitamin A carotenoids. Total carotenoid content after the 10-day refrigerated storage period resulted in no significant quantitative changes in T1 juices, whereas in T2 and T3 juices small losses were found at the end of the storage period (20.56 and 9.16%, respectively). These losses could be influenced by the depleted protection of vitamin C toward carotenoid oxidation during the same period. A similar trend was found in provitamin A carotenoids for the different treated juices.     

Recycling of Two-Phase Olive-Mill Cake “Alperujo” by Co-composting with Animal Manures

The use of poultry manure or goat/sheep manure in the co-composting of the two-phase olive-mill cake “alperujo” (ALP) with olive leaf (OL) is compared by studying organic-matter mineralization and humification processes during composting and the characteristics of the end products. For this, two different piles (P1 and P2) were prepared using ALP with OL mixed with poultry manure (PM) and goat/sheep manure (GSM), respectively, and composted by the turned windrow composting system. Throughout the composting process, a number of parameters were monitored, such as temperature, pH, electrical conductivity (EC), organic matter (OM), OM losses, total organic carbon (C_org), total nitrogen (N_t), C_org/N_t ratio, and the germination index (GI). In both piles, the temperature exceeded 55 °C for more than 2 weeks, which ensured maximum pathogen reduction. Organic-matter losses followed a first-order kinetic equation in both piles. The final composts presented a stabilized OM and absence of phytotoxins, as observed in the evolution and final values of the C_org/N_t ratio (C_org/N_t < 20) and the germination index (GI > 50 percent). Therefore, composting can be considered as an efficient treatment to recycle this type of waste, obtaining composts with suitable properties that can be safely used in agriculture.