A new approach to genetic alteration of soybean protein composition and quality

Although soybeans produce high-quality meal, modern animal and fish production systems often require synthetic essential amino acid supplements to fortify feed rations. However, biotechnology may enable development of soybeans with naturally adequate levels of certain essential amino acids for advanced feed formulations. One approach involves genetic manipulation of glycinin (11S) and β-conglycinin (7S) contents, the principal components of soybean storage proteins. Because 11S contains more cysteine and methionine than 7S protein, a higher 11S:7S ratio could lead to beneficial changes in the nutritional quality of soybean meal. Although genotypic variation for 11S:7S may be low among soybean [Glycine max (L.) Merr.] germplasm, ratios ranging from 1.7–4.9 were observed among accessions of the wild ancestor of cultivated soybean (Glycine soja Sieb, and Zucc.). Thus, wild soybean germplasm was evaluated as a potential source of genes that govern protein synthesis that may have been lost during the domestication of G. max. Change in the amount of 11S protein accounts for a significant portion of the genotypic variation in protein concentration and composition among wild soybeans. Strong positive correlation exists between the 11S:7S ratio and methionine or cysteine concentration of total protein. Moderate positive associations were found for threonine or tyrosine. A moderate negative correlation was found between lysine and 11S:7S. No association was found for leucine and phenylalanine or for total essential amino acid concentration. Based on these data, G. soja may contain a different complement of genes that influence expression of 11S and 7S proteins than G. max germplasm. Thus, through interspecific hybridization, wild soybeans may be a useful genetic resource for the further improvement of protein quality in cultivated soybeans.     

Genetic regulation of linolenic acid concentration in wild soybean Glycine soja accessions

Soybean [Glycine max (L.) Merr.] oil from current commercial cultivars typically contains ca. 8% linolenic acid. Inheritance studies have shown that linolenic acid concentration in soybean seed is determined by at least two genes which govern activity of the predominant ω-6 and ω-3 desaturases. Selection of germplasm exhibiting homozygous recessive alleles that encode these desaturases has enabled development of soybeans having less than 3.0% linolenic acid. However, accessions of the wild ancestor of modern soybean cultivars, Glycine soja (Sieb. and Zucc.), have oils containing twice the highest linolenic acid concentration found in normal G. max cultivars. Although little is known about inheritance of linolenic acid in wild soybean, it would appear that additional or alternative forms of genes may govern its synthesis. To test this hypothesis, cultivated soybean germplasm was hybridized with wild soybean genotypes having significant differences in linolenic acid concentration. Seed of F₃ progeny from these G. max x G. soja populations exhibited distinct segregation patterns for relative estimates of ω-6 and ω-3 desaturase activity. Frequency class distribution analyses of the segregation patterns, and linear relations between median ω-6 or ω-3 desaturation estimates and corresponding linolenic acid concentration among allelic classes from these populations suggested the high-linolenic acid trait in wild soybean genotypes was determined by a set of desaturase alleles that were different from corresponding alleles in G. max. Introgression of these alternative alleles in G. max germplasm opens a new avenue of research on the genitic regulation of linolenic acid, and may lead to the production of highly polyunsaturated soybean oils for various industrial applications.     

Use of NMR for predicting protein concentration in soybean seeds based on oil measurements

Increasing the level of protein in soybean seeds has been a major target for soybean [Glycine max (L.) Merr.] breeders. The objective of this study was to examine the potential of predicting soybean seed protein based on oil values as determined by NMR. Seed protein and oil concentrations were determined in an F₂ population generated from the cross between a G. max (NK, S08-80), and a G. soja (PI 458536) cultivar. The protein concentration in the population ranged from 40.4 to 52.6%. Protein-oil regression analysis was used to generate an equation for predicting seed protein concentration based on oil readings. The regression equation Protein=62.3–1.3 [Oil] (R ²=0.46) was developed, with a corresponding correlation of −0.69 between the traits. With this equation, the mean protein concentration of the selected 25% of the population (a simulated breeding pressure) was greater than the mean of the unselected population (46.1%, SE=0.13) by about 1.9%. Individual F₂ plants that exceeded the mean protein value of the population constituted 86.4% of the selected samples. Selection based on oil concentration, however, failed to include 27.1% of the plants that were among the top 25% for protein concentration. Selection of high-protein plants based on NMR oil measurement was reasonably effective in the test population and might offer a new and rapid method of selecting high-protein individuals in soybean populations derived from the wild soybean progenitor, G. soja. If further tested on other populations and samples, it might be used as an analytical alternative for an indirect measurement of protein concentration based on NMR measurements of the oil.     

Comparison of PPAR Ligands as Modulators of Resolution of Inflammation,via Their Influence on Cytokines and Oxylipins Release in Astrocytes

Neuroinflammation is a key process of many neurodegenerative diseases and other brain disturbances, and astrocytes play an essential role in neuroinflammation. Therefore, the regulation of astrocyte responses for inflammatory stimuli, using small molecules, is a potential therapeutic strategy. We investigated the potency of peroxisome proliferator-activated receptor (PPAR) ligands to modulate the stimulating effect of lipopolysaccharide (LPS) in the primary rat astrocytes on (1) polyunsaturated fatty acid (PUFAs) derivative (oxylipins) synthesis; (2) cytokines TNFα and interleukin-10 (IL-10) release; (3) p38, JNK, ERK mitogen-activated protein kinase (MAPKs) phosphorylation. Astrocytes were exposed to LPS alone or in combination with the PPAR ligands: PPARα (fenofibrate, GW6471); PPARβ ({"type":"entrez-nucleotide","attrs":{"text":"GW501516","term_id":"289075981","term_text":"GW501516"}}GW501516, GSK0660); PPARγ (rosiglitazone, GW9662). We detected 28 oxylipins with mass spectrometry (UPLC-MS/MS), classified according to their metabolic pathways: cyclooxygenase (COX), cytochrome P450 monooxygenases (CYP), lipoxygenase (LOX) and PUFAs: arachidonic (AA), docosahexaenoic (DHA), eicosapentaenoic (EPA). All tested PPAR ligands decrease COX-derived oxylipins; both PPARβ ligands possessed the strongest effect. The PPARβ agonist, {"type":"entrez-nucleotide","attrs":{"text":"GW501516","term_id":"289075981","term_text":"GW501516"}}GW501516 is a strong inducer of pro-resolution substances, derivatives of DHA: 4-HDoHE, 11-HDoHE, 17-HDoHE. All tested PPAR ligands decreased the release of the proinflammatory cytokine, TNFα. The PPARβ agonist {"type":"entrez-nucleotide","attrs":{"text":"GW501516","term_id":"289075981","term_text":"GW501516"}}GW501516 and the PPARγ agonist, rosiglitazone induced the IL-10 release of the anti-inflammatory cytokine, IL-10; the cytokine index, (IL-10/TNFα) was more for {"type":"entrez-nucleotide","attrs":{"text":"GW501516","term_id":"289075981","term_text":"GW501516"}}GW501516. The PPARβ ligands, {"type":"entrez-nucleotide","attrs":{"text":"GW501516","term_id":"289075981","term_text":"GW501516"}}GW501516 and GSK0660, are also the strongest inhibitors of LPS-induced phosphorylation of p38, JNK, ERK MAPKs. Overall, our data revealed that the PPARβ ligands are a potential pro-resolution and anti-inflammatory drug for targeting glia-mediated neuroinflammation.     

Exploring seed and meal composition and protein solubility in soybean genotypes from different domestication periods

Traditionally, commercial soybean breeding has focused on increasing seed yield by crossing elite cultivars and limiting the genetic diversity within commercial germplasm. Wild and ancestral soybean genotypes have higher seed protein concentrations than commercial ones. Different seed protein concentrations and compositions result in diverse functional properties of soybean meal, in particular solubility is important for beverages and protein isolates production. The objectives of our study were (i) to characterize seed protein concentration and composition in genotypes from different soybean domestication periods (types) and (ii) to evaluate the protein concentration and solubility profiles of the defatted meals obtained from these genotypes. Variation within seed and meal protein concentration, composition, and solubility was evident along the domestication process. Wild relative (G. soja) and Elite genotypes had the maximum and minimum seed protein concentrations, respectively (42.9 and 36.3 g 100 g⁻¹). Soybean meal protein concentrations were 55.1, 47.7, 48.4 and 44.1 g 100 g⁻¹ for Wild relative (G. soja), Asian landraces, North American (Nam) ancestors and Elite, respectively. Ample genotypic variation was observed for β-conglycinin components, such as for β, α, and α′ subunits and for total glycinin and its components. Asian landraces had the highest protein solubility. Wild and ancestral germplasm are a reservoir of useful traits to improve soybean seed quality. This study opens the gates to the introduction of ancestral germplasm to breeding programs focused on protein quality and functionality.     

Wild Soybean Oxalyl-CoA Synthetase Degrades Oxalate and Affects the Tolerance to Cadmium and Aluminum Stresses

Acyl activating enzyme 3 (AAE3) was identified as being involved in the acetylation pathway of oxalate degradation, which regulates the responses to biotic and abiotic stresses in various higher plants. Here, we investigated the role of Glycine soja AAE3 (GsAAE3) in Cadmium (Cd) and Aluminum (Al) tolerances. The recombinant GsAAE3 protein showed high activity toward oxalate, with a K_m of 105.10 ± 12.30 μM and V_max of 12.64 ± 0.34 μmol min⁻¹ mg⁻¹ protein, suggesting that it functions as an oxalyl–CoA synthetase. The expression of a GsAAE3–green fluorescent protein (GFP) fusion protein in tobacco leaves did not reveal a specific subcellular localization pattern of GsAAE3. An analysis of the GsAAE3 expression pattern revealed an increase in GsAAE3 expression in response to Cd and Al stresses, and it is mainly expressed in root tips. Furthermore, oxalate accumulation induced by Cd and Al contributes to the inhibition of root growth in wild soybean. Importantly, GsAAE3 overexpression increases Cd and Al tolerances in A. thaliana and soybean hairy roots, which is associated with a decrease in oxalate accumulation. Taken together, our data provide evidence that the GsAAE3-encoded protein plays an important role in coping with Cd and Al stresses.     

Relationship between seed mass and linolenic acid in progeny of crosses between cultivated and wild soybean

Soybean [Glycine max (L.) Merr.] oil from current commercial cultivars typically contains ca. 8% linolenic acid (18:3). Applications of plant biotechnology have enabled plant breeders to develop germplasm having as low as 2.0% 18:3. Oils that are naturally low in 18:3 exhibited improved flavor characteristics and greater oxidative stability in high-temperature frying applications compared to hydrogenated soybean oil. As an extension of that research, efforts are underway to characterize genes in soybean that govern expression of higher than normal 18:3 concentration. Such oils may be of interest to the oleochemicals industry for various nonfood applications. Relatively high 18:3 in seed oil is a characteristic trait of the ancestor of modern soybean cultivars, Glycine soja (Sieb. and Zucc.). Accessions of this species have rarely been utilized in soybean improvement, and thus represent a virtually untapped genetic resource for genes governing 18:3 synthesis. We have hybridized cultivated soybean with wild soybean plant introductions. F_3:4 seed from the resultant G. max × G. soja populations exhibited a wide segregation pattern for 18:3 and seed mass. A strong negative association was found between 18:3 concentration and seed mass. Oil concentration was positively correlated with seed mass. Evaluation of glycerolipid composition revealed that high 18:3 was not associated with an altered proportion of phospholipid and triacylglycerol among lines segregating for seed mass. Thus, smaller seed mass may be a convenient trait to distinguish future soybean cultivars with highly polyunsaturated oils from other cultivars in production.     

Biallelic Variants in KIF17 Associated with Microphthalmia and Coloboma Spectrum

Microphthalmia, anophthalmia, and coloboma (MAC) are a group of congenital eye anomalies that can affect one or both eyes. Patients can present one or a combination of these ocular abnormalities in the so called “MAC spectrum”. The KIF17 gene encodes the kinesin-like protein Kif17, a microtubule-based, ATP-dependent, motor protein that is pivotal for outer segment development and disc morphogenesis in different animal models, including mice and zebrafish. In this report, we describe a Sicilian family with two siblings affected with congenital coloboma, microphthalmia, and a mild delay of motor developmental milestones. Genomic DNA from the siblings and their unaffected parents was sequenced with a clinical exome that revealed compound heterozygous variants in the KIF17 gene ({"type":"entrez-nucleotide","attrs":{"text":"NM_020816.4","term_id":"1677537661","term_text":"NM_020816.4"}}NM_020816.4: c.1255C > T (p.Arg419Trp); c.2554C > T (p.Arg852Cys)) segregating with the MAC spectrum phenotype of the two affected siblings. Variants were inherited from the healthy mother and father, are present at a very low-frequency in genomic population databases, and are predicted to be deleterious in silico. Our report indicates the potential co-segregation of these biallelic KIF17 variants with microphthalmia and coloboma, highlighting a potential conserved role of this gene in eye development across different species.     

Effects of TGF-β1 Receptor Inhibitor GW788388 on the Epithelial to Mesenchymal Transition of Peritoneal Mesothelial Cells

We investigated the effectiveness of the transforming growth factor beta-1 (TGF-β) receptor inhibitor {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388 on the epithelial to mesenchymal transition (EMT) using human peritoneal mesothelial cells (HPMCs) and examined the effectiveness of {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388 on the peritoneal membrane using a peritoneal fibrosis mouse model. HPMCs were treated with TGF-β with or without {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388. Animal experiments were conducted on male C57/BL6 mice. Peritoneal fibrosis was induced by intraperitoneal injection of chlorhexidine gluconate. {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388 was administered by once-daily oral gavage. The morphological change, cell migration, and invasion resulted from TGF-β treatment, but these changes were attenuated by cotreatment with {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388. TGF-β-treated HPMCs decreased the level of the epithelial cell marker and increased the levels of the mesenchymal cell markers. Cotreatment with {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388 reversed these changes. Phosphorylated Smad2 and Smad3 protein levels were stimulated with TGF-β and the change was attenuated by cotreatment with {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388. For the peritoneal fibrosis mice, thickness and collagen deposition of parietal peritoneum was increased, but this change was attenuated by cotreatment with {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388. {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388, an orally available potent TGF-β receptor type 1 inhibitor, effectively attenuated TGF-β-induced EMT in HPMCs. Cotreatment with {"type":"entrez-nucleotide","attrs":{"text":"GW788388","term_id":"293585730","term_text":"GW788388"}}GW788388 improved peritoneal thickness and fibrosis, and recovered peritoneal membrane function in a peritoneal fibrosis mouse model.     

Effect of phosphate rock sources on biological nitrogen fixation by soybean

Very little information is available concerning the effect of phosphate rock (PR) sources on biological nitrogen fixation (BNF) in legume crops. In a greenhouse study, the¹⁵N isotopic dilution technique was used to compare the effectiveness of three sources of PR (Hahotoe rock, Togo; Tilemsi rock, Mali; and Sechura rock, Peru) with that of triple superphosphate (TSP) in increasing soybean seed yield and the amounts of N fixed by the soybean crop. The acid Hartsells slit loam was limed to pH 5.2 and incubated with 8.5 mg N kg^–1 as K¹⁵NO₃ and sucrose for 2 months prior to planting. Then fertilizer P was incorporated into the soil at 12.5, 25, 50, and 100 mg P kg^–1 rates.The relative agronomic effectiveness (RAE) of the three PRs with respect to TSP (RAE = 100%) in terms of increasing seed yield was Hahotoe rock = 6.0%, Tilemsi rock = 45.9%, and Sechura rock = 75.2%; this trend followed the same trend as PR reactivity, i.e., Sechura rock > Tilemsi rock > Hahotoe rock. BNF was affected significantly by all the P treatments. Of the total N derived from the three N sources (atmosphere, N_dfa; fertilizer K¹⁵NO₃, N_dff; and soil, N_dfs), N_dfa was highest with TSP and lowest with Hahotoe rock, whereas the reverse was found with N_dfs. Among various plant parts, more N_dfa was translocated and stored in seeds than in stems + leaves and roots. The RAE values of the three PRs with respect to TSP (RAE = 100%) in terms of influencing the amount of BNF were Hahotoe rock = 3.0%, Tilemsi rock = 43.4%, and Sechura rock = 71.2%. A linear relationship was found between the amount of BNF by the whole soybean plant and the soybean seed yield.     

Translational Read-Through Therapy of RPGR Nonsense Mutations

X-chromosomal retinitis pigmentosa (RP) frequently is caused by mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene. We evaluated the potential of PTC124 (Ataluren, Translama^TM) treatment to promote ribosomal read-through of premature termination codons (PTC) in RPGR. Expression constructs in HEK293T cells showed that the efficacy of read-through reagents is higher for UGA than UAA PTCs. We identified the novel hemizygous nonsense mutation c.1154T > A, p.Leu385* ({"type":"entrez-nucleotide","attrs":{"text":"NM_000328.3","term_id":"1677500221"}}NM_000328.3) causing a UAA PTC in RPGR and generated patient-derived fibroblasts. Immunocytochemistry of serum-starved control fibroblasts showed the RPGR protein in a dot-like expression pattern along the primary cilium. In contrast, RPGR was no longer detectable at the primary cilium in patient-derived cells. Applying PTC124 restored RPGR at the cilium in approximately 8% of patient-derived cells. RT-PCR and Western blot assays verified the pathogenic mechanisms underlying the nonsense variant. Immunofluorescence stainings confirmed the successful PTC124 treatment. Our results showed for the first time that PTC124 induces read-through of PTCs in RPGR and restores the localization of the RPGR protein at the primary cilium in patient-derived cells. These results may provide a promising new treatment option for patients suffering from nonsense mutations in RPGR or other genetic diseases.     

Genome-Wide Identification of Soybean ABC Transporters Relate to Aluminum Toxicity

ATP-binding cassette (ABC) transporter proteins are a gene super-family in plants and play vital roles in growth, development, and response to abiotic and biotic stresses. The ABC transporters have been identified in crop plants such as rice and buckwheat, but little is known about them in soybean. Soybean is an important oil crop and is one of the five major crops in the world. In this study, 255 ABC genes that putatively encode ABC transporters were identified from soybean through bioinformatics and then categorized into eight subfamilies, including 7 ABCAs, 52 ABCBs, 48 ABCCs, 5 ABCDs, 1 ABCEs, 10 ABCFs, 111 ABCGs, and 21 ABCIs. Their phylogenetic relationships, gene structure, and gene expression profiles were characterized. Segmental duplication was the main reason for the expansion of the GmABC genes. Ka/Ks analysis suggested that intense purifying selection was accompanied by the evolution of GmABC genes. The genome-wide collinearity of soybean with other species showed that GmABCs were relatively conserved and that collinear ABCs between species may have originated from the same ancestor. Gene expression analysis of GmABCs revealed the distinct expression pattern in different tissues and diverse developmental stages. The candidate genes GmABCB23, GmABCB25, GmABCB48, GmABCB52, GmABCI1, GmABCI5, and GmABCI13 were responsive to Al toxicity. This work on the GmABC gene family provides useful information for future studies on ABC transporters in soybean and potential targets for the cultivation of new germplasm resources of aluminum-tolerant soybean.     

Restriction fragment length polymorphism analysis of soybean fatty acid content

The quality of soybean [Glycine max (L.) Merr.] oil depends greatly upon its fatty acid composition. The objective of this study was to map quantitative trait loci affecting the relative content of the fatty acids composing soybean seed oil. The mapping was done in a population formed from a cross between aG. max experimental line and aG. soja plant introduction. Sixty F2-derived lines from this population were genotyped with 243 restriction fragment length polymorphism, five isozyme, one storage protein and three morphological markers. The genotyped lines were grown in a replicated trial, and the seed harvested was analyzed for palmitic, stearic, oleic, linoleic and linolenic fatty acids. Markers significantly (P>0.005) associated with each fatty acid were found. Individual markers were associated with up to 38% of the total variance in specific fatty acids among the F2 lines. The greatest associations between markers and fatty acid content were observed with markers on two major linkage groups. Joint contribution of North Central Region, USDA-ARS and Journal Paper no. J-14849 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011; Project 2974.     

Clinical Considerations for a Family with Dilated Cardiomyopathy,Sudden Cardiac Death,and a Novel TTN Frameshift Mutation

Dilated cardiomyopathy (DCM) is the leading indication for heart transplantation. TTN gene truncating mutations account for about 25% of familial DCM cases and for 18% of sporadic DCM cases. The clinical relevance of specific variants in TTN has been difficult to determine because of the sheer size of the protein for which TTN encodes, as well as existing extensive genetic variation. Clinicians should communicate novel clinically-relevant variants and genotype–phenotype associations, so that animal studies evaluating the molecular mechanisms are always conducted with a focus on clinical significance. In the present study, we report for the first time the novel truncating heterozygous variant {"type":"entrez-nucleotide","attrs":{"text":"NM_001256850.1","term_id":"378925624","term_text":"NM_001256850.1"}}NM_001256850.1:c.72777_72783del (p.Phe24259Leufs*51) in the TTN gene and its association with DCM in a family with sudden death. This variant occurs in the A-band region of the sarcomere, in a known mutational hotspot of the gene. Truncating titin variants that occur in this region are the most common cause of DCM and have been rarely reported in asymptomatic individuals, differently from other pathogenic TTN gene variants. Further studies are warranted to better understand this particular clinically-relevant variant.     

Gene-Diet Interaction between SIRT6 and Soybean Intake for Different Levels of Pulse Wave Velocity

Soybean is a common food for the Chinese people. We aimed to investigate the risk for brachial ankle pulse wave velocity (baPWV) with inflammatory-related SNPs and soybean. baPWV was measured, and 16 inflammatory-related SNPs located on ADIPOQ, CDH13, SIRT3, SIRT6, CXCL12, CXCR4, NOS1, PON1 and CDKN2B were genotyped in 1749 Chinese participants recruited from various communities. ADIPOQ rs12495941 (GT/TT vs. GG: crude OR = 1.27, p = 0.044) and SIRT6 rs107251 (CT/TT vs. CC: crude OR = 0.74, p = 0.009) were associated with abnormal baPWV (baPWV ≥ 1700 cm/s). After adjustment for conventional environmental risk factors, rs12495941 was associated with abnormal baPWV (GT/TT vs. GG: adjusted OR = 1.43, p = 0.011), but the association between rs107251 and abnormal baPWV was not significant (CT/TT vs. CC: adjusted OR = 0.83, p = 0.173). The interaction between rs107251 and soybean intake for different levels of baPWV was statistically significant (p = 0.017). Compared with a high level of soybean intake, a low level of soybean intake can significantly decrease the risk of abnormal baPWV in individuals of rs107251 CT/TT genotypes (≤100 vs. >100 g/week: adjusted OR = 0.542, p = 0.003). In this study, associations between ADIPOQ rs12495941, SIRT6 rs107251 and baPWV, as well as an interaction between SIRT6 rs107251 and soybean intake for different levels of baPWV were found.     

Long-Lasting Activity of ERK Kinase Depends on NFATc1 Induction and Is Involved in Cell Migration-Fusion in Murine Macrophages RAW264.7

Macrophages are mononuclear cells that become osteoclasts (OCs) in the presence of two cytokines, macrophage colony-stimulating factor (M-CSF), and receptor activator of NF-κB ligand (RANKL). RANKL binding to its specific receptor RANK leads to OCs differentiation mainly by nuclear factor of activated T-cells cytoplasmic 1 (NFATc1). In our previous study, the analysis of the protein network in NFATc1-knockdown cells, using the Ingenuity Pathway Analysis (IPA), showed a link between NFATc1 and Mitogen-activated protein kinase kinase (MEK)-extracellular receptor kinase (ERK) signaling pathway. Therefore, this study aimed to extend our knowledge of the relationship between NFATc1 and the ERK. Here, we demonstrate that delayed ERK1/2 phosphorylation in pre-OC RANKL-induced depends on NFATc1. Indeed, the knockdown of NFATc1 reduced the phosphorylation of ERK1/2 (60%) and the pharmacological inhibition of the ERK1/2 kinase activity impairs the expression of NFATc1 without preventing its translocation into the nucleus. Furthermore, silencing of NFATc1 significantly reduced RANKL-induced migration (p < 0.01), and most pre-OCs are still mononuclear after 48 h (80 ± 5%), despite the presence of actin rings. On the other hand, the inhibitors {"type":"entrez-nucleotide","attrs":{"text":"FR180204","term_id":"258307209","term_text":"FR180204"}}FR180204 and PD98059 significantly reduced RANKL-induced cell migration (p < 0.01), leading to a reduction in the number of multinucleated cells. Finally, we suggest that long-lasting ERK activity depends on NFATc1 induction and is likely linked to cell migration, fusion, and OC differentiation.     

Fat content and fatty acid composition of oils extracted from selected wild-gathered tropical plant seeds from Nigeria

As the search for alternative sources of food to alleviate hunger continues, this study was undertaken to determine the fat content and the fatty acid composition of 15 lesser-known wild tropical seeds gathered in Nigeria. Results were contrasted with five tropical soybean varieties (Glycine max). The fat content varies from less than 1% (Pterocarpus santalinoides, Daniellia ogea) to 59% (Entandrophragma angolense). The fatty acid composition of most of the wild and mostly leguminous seeds differed considerably, compared to the composition of tropical soybeans. The oil of Adansonia digitata, Prosopis africana, Afzelia lebbeck, Enterolobium cyclocarpium, and Sesbania pachycarpa contained high proportions of linoleic and oleic acid as well as palmitic and linolenic acid. Seeds of Milletia thonningii, Lonchocarpus sericeus, and S. pachycarpa were much higher in linolenic acid and relatively poor in linoleic acid, compared to soybeans. The content of saturated fatty acids was higher than that of soybeans, resulting in lower polyunsaturated/saturated (P/S) ratios (0.83–2.12) than observed in soybeans (P/S=3.4), with the exception of the composition of S. pachycarpa (P/S=3.15). Some of these less familiar wild seeds could be used as sources for industrial or edible oils, provided that possible toxic constituents could be removed.