Status of yam (Dioscorea spp.) in the Democratic Republic of Congo

Abstract

Yam is an important tuber crop with enormous potential to enhance rural sustenance and livelihood in DRC. However, studies to enhance its genetic improvement are very far from sufficient with only a handful of information available on the crop. Yam has been treated as an orphan crop compared to contemporary crops such as cassava and sweet potato which have adapted to different cropping systems and become widespread in production. The lack of research attention to address the major production challenges has further decreased the value and potential of the crop compared to its contemporaries. These production constraints include lack of adequate quality planting materials, low yield potential, poor resistance/tolerance to yam mosaic and anthracnose diseases and ultimately poor tuber quality attributes focusing on tuber taste, flesh oxidation and dry matter contents of the majority of the farmers preferred varieties. In this review, we evaluated the status of yam in DRC and presented the needful activities to be incorporated for its improvement. Diversity has however been maintained mainly through ennoblement efforts in house gardens and small farmlands using traditional farming methods. Studies from other nations where yam has been successful with prominence in characterization and genetic improvement brought to light the need for DRC to consider yam as a staple carbohydrate food source, even to the extent of modifications in food public policy. Reversal of the yam's current stigma is a challenge to the scientific community and the population in general.

In‐Cell Arrestin‐Receptor Interaction Assays

Abstract

G protein-coupled receptors (GPCRs) represent ∼30% of current drug targets. Ligand binding to these receptors activates G proteins and arrestins, which function in different signaling pathways. Given that functionally selective or biased ligands preferentially activate one of these two groups of pathways, they may be superior medications for certain disease states. The identification of such ligands requires robust drug screening assays for both G protein and arrestin activity. This unit describes protocols for assays that monitor reversible arrestin recruitment to GPCRs in living cells using either bioluminescence resonance energy transfer (BRET) or nanoluciferase complementation (NanoLuc). Two types of assays can be used: one configuration directly measures arrestin recruitment to a GPCR fused to a protein tag at its intracellular C-terminus, whereas the other configuration detects arrestin translocation to the plasma membrane in response to activation of an unmodified GPCR. Together, these assays are powerful tools for studying dynamic interactions between GPCRs and arrestins. © 2023 Wiley Periodicals LLC.

Basic Protocol 1: Receptor-arrestin BRET assay to measure ligand-induced recruitment of arrestin to receptors

Basic Protocol 2: Receptor-arrestin NANOBIT assay to measure ligand-induced recruitment of arrestin to receptors

Alternative Protocol 1: BRET assay to measure ligand-induced recruitment of arrestin to the plasma membrane

Alternative Protocol 2: NANOBIT assay to measure ligand-induced recruitment of arrestin to the plasma membrane

Support Protocol 1: Optimization of polyethylenimine (PEI) concentration for transfection

Current Strategies of Modeling Human Trophoblast Using Human Pluripotent Stem Cells in vitro

Abstract

We previously established a trophoblast differentiation protocol from primed human pluripotent stem cells (PSC). To induce this lineage, we use a combination of Bone Morphogenetic Protein-4 (BMP4) and the WNT inhibitor IWP2. This protocol has enabled us to obtain a pure population of trophectoderm (TE)-like cells that could subsequently be terminally differentiated into syncytiotrophoblasts (STB) and extravillous trophoblasts (EVT). However, the resulting TE-like cells could only be terminally differentiated to a variable mixture of STB and EVT, with a bias toward the STB lineage. Recently, methods have been developed for derivation and culture of self-renewing human trophoblast stem cells (TSC) from human embryos and early gestation placental tissues. These primary TSCs were further able to differentiate into either STB or EVT with high efficiency using the lineage specific differentiation protocols. Based partly on these protocols, we have developed methods for establishing self-renewing TSC-like cells from PSC, and for efficient lineage-specific terminal differentiation. Here, we describe in detail the protocols to derive and maintain PSC-TSC, from both embryonic stem cells (ESC) and patient-derived induced pluripotent stem cells (iPSC), and their subsequent terminal differentiation to STB and EVT. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.

Basic Protocol 1: Trophoblast Differentiation into TE-like Cells

Basic Protocol 2: Conversion of PSC-Derived TE-like Cells to TSC

Basic Protocol 3: Passaging PSC-Derived TSC in iCTB Complete Medium

Basic Protocol 4: STB Differentiation from PSC-derived TSC

Basic Protocol 5: EVT Differentiation from PSC-derived TSC

Support Protocol 1: Geltrex-coated tissue culture plate preparation

Support Protocol 2: Collagen IV-coated tissue culture plate preparation

Support Protocol 3: Fibronectin-coated tissue culture plate preparation

miR‐34a/SIRT1/HIF‐1α axis is involved in cardiac angiogenesis of type 2 diabetic rats: The protective effect of sodium butyrate combined with treadmill exercise

miR-34a/SIRT1/HIF-1α axis is involved in cardiac angiogenesis of type 2 diabetic rats: The protective effect of sodium butyrate combined with treadmill exercise

Sodium butyrate and treadmill exercise, alone or together, significantly reduced the blood glucose levels in type 2 diabetic rats. Combination of sodium butyrate and treadmill exercise, improved oral glucose tolerance test in type 2 diabetic rats. Diabetes increased miR-34a expression and FOXO1 levels, and decreased SIRT1 and HIF-1α levels that were measured by real-time PCR and ELISA method. A combination of sodium butyrate and treadmill exercise significantly decreased miR-34a expression and FOXO1 levels, and improved SIRT1 and HIF-1α levels in the heart tissue of diabetic rats. Sodium butyrate and treadmill exercises, alone or together, increased angiogenesis in the heart tissue of diabetic rats. So, those may be useful in the treatment of diabetes through the mir-34a/SIRT1/FOXO1-HIF-1α pathway.


Abstract

Type 2 diabetes mellitus (T2DM) is one of the most common metabolic disorders worldwide. Recent research has indicated that sodium butyrate (NaB) affects glucose metabolism and exercise has an anti-hyperglycemic effect in diabetes. This study aimed to evaluate the effects of NaB and treadmill exercise on heart angiogenesis through the miR-34a/SIRT1/FOXO1-HIF-1α pathway. Diabetic animals received NaB (400 mg/kg daily, orally) and treadmill exercise for 6 weeks. The effect of NaB and treadmill exercise, alone or combined, on miR-34a expression, SIRT1, FOXO1, HIF-1α levels, and angiogenesis in diabetic heart tissue was measured. Diabetes caused increased miR-34a (p < 0.01) and FOXO1 (p < 0.001) expression levels. Also, SIRT1 (p < 0.001) and HIF-1α (not significant) expression levels were reduced in diabetic rats. NaB and treadmill exercise decreased miR-34a (respectively p < 0.05 and not significant) and FOXO1 (both p < 0.001) expression levels and improved SIRT1 (both not significant) and HIF-1α (respectively p < 0.01 and p < 0.001) levels. Also, NaB combined with treadmill exercise decreased miR-34a (p < 0.001) and FOXO1 (p < 0.001) expression levels, and elevated SIRT1 (p < 0.05) and HIF-1α (p < 0.001) levels in comparison with the diabetic group. NaB and treadmill exercises modulate the expression of miR-34a and the levels of SIRT1, FOXO1, and HIF-1α proteins, thus increasing angiogenesis in the heart tissue of diabetic rats. It can be concluded that NaB and treadmill exercise, alone or combined, may be useful in the treatment of diabetes through the miR-34a/SIRT1/FOXO1-HIF-1α pathway.

Torpor‐responsive microRNAs in the heart of the Monito del monte, Dromiciops gliroides

Torpor-responsive microRNAs in the heart of the Monito del monte, Dromiciops gliroides

Monito del monte (Dromiciops gliroides) undergoes metabolic rate depression to conserve energy during torpor. Differential microRNA expression plays a key role in metabolic reorganization by regulating key signaling and metabolic pathways.


Abstract

The marsupial Monito del monte (Dromiciops gliroides) utilizes both daily and seasonal bouts of torpor to preserve energy and prolong survival during periods of cold and unpredictable food availability. Torpor involves changes in cellular metabolism, including specific changes to gene expression that is coordinated in part, by the posttranscriptional gene silencing activity of microRNAs (miRNA). Previously, differential miRNA expression has been identified in D. gliroides liver and skeletal muscle; however, miRNAs in the heart of Monito del monte remained unstudied. In this study, the expression of 82 miRNAs was assessed in the hearts of active and torpid D. gliroides, finding that 14 were significantly differentially expressed during torpor. These 14 miRNAs were then used in bioinformatic analyses to identify Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways that were predicted to be most affected by these differentially expressed miRNAs. Overexpressed miRNAs were predicted to primarily regulate glycosaminoglycan biosynthesis, along with various signaling pathways such as Phosphoinositide-3-kinase/protein kinase B and transforming growth factor-β. Similarly, signaling pathways including phosphatidylinositol and Hippo were predicted to be regulated by the underexpression of miRNAs during torpor. Together, these results suggest potential molecular adaptations that protect against irreversible tissue damage and enable continued cardiac and vascular function despite hypothermia and limited organ perfusion during torpor.

Platelet‐activating factor as an endogenous cofactor of food anaphylaxis

Platelet-activating factor as an endogenous cofactor of food anaphylaxis

PAF is an endogenous factor of food anaphylaxis.


Abstract

Anaphylaxis is a severe, acute, life-threatening generalized or systemic hypersensitivity reaction. The incidence of anaphylaxis is increasing worldwide, with medications and food contributing to most cases. Physical exercise, acute infections, drugs, alcohol, and menstruation are the external cofactors associated with more severe systemic reaction. The aim of this review is to show that platelet-activating factor contributes to the development of severe anaphylactic reaction, and even to anaphylactic shock.