Rational identification of a Cdc42 inhibitor presents a new regimen for long- term hematopoietic stem cell mobilization

Mobilization of hematopoietic stem cells (HSCs) from bone marrow (BM) to peripheral blood (PB) by cytokine granulocyte colony-stimulating factor (G-CSF) or the chemical antagonist of CXCR4, AMD3100, is important in the treatment of blood diseases. Due to clinical conditions of each application, there is a need for continued improvement of HSC mobilization regimens. Previous studies have shown that genetic ablation of the Rho GTPase Cdc42 in HSCs results in their mobilization without affecting survival. Here we rationally identified a Cdc42 activity-specific inhibitor (CASIN) that can bind to Cdc42 with submicromolar affinity and competitively interfere with guanine nucleotide exchange activity. CASIN inhibits intracellular Cdc42 activity specifically and transiently to induce murine hematopoietic stem/progenitor cell egress from the BM by suppressing actin polymerization, adhesion, and directional migration of stem/progenitor cells, conferring Cdc42 knockout phenotypes. We further show that, although, CASIN administration to mice mobilizes similar number of phenotypic HSCs as AMD3100, it produces HSCs with better long-term reconstitution potential than that by AMD3100. Our work validates a specific small molecule inhibitor for Cdc42, and demonstrates that signaling molecules downstream of cytokines and chemokines, such as Cdc42, constitute a useful target for long-term stem cell mobilization

Characterization and phylogenetic analysis of the mitochondrial genome of Sarcocheilichthys sinensis (Bleeker) from Baima Hu Lake

Sarcocheilichthys sinensis (Bleeker), is a small-sized benthopelagic fish with ornamental value. In the present study, the complete mitochondrial genome of S. sinensis was sequenced and determined. The complete mitogenome of S. sinensis was 16,683 bp in length, consisting of 22 tRNA genes, 13 protein-coding genes, 2 rRNA genes, and 2 non-coding regions. The overall base composition of the S. sinensis mitogenome is 30.50% A, 26.28% T, 26.60% C, and 16.61% G, exhibiting obvious AT bias (56.79%). The phylogenetic analysis showed that S. sinensis clustered in genus Sarcocheilichthys. Present study provides useful data to population genetics and conservation biology of Sarcocheilichthys fishes

Quantifying within-plant spatial heterogeneity in carbohydrate availability in cotton using a local-pool model

Background and Aims Within-plant spatial heterogeneity in the production of and demand for assimilates may have major implications for the formation of fruits. Spatial heterogeneity is related to organ age, but also to position on the plant. This study quantifies the variation in local carbohydrate availability for the phytomers in the same cohort using a cotton growth model that captures carbohydrate production in phytomers and carbohydrate movement between phytomers. Methods Based on field observations, we developed a functional-structural plant model of cotton that simulates production and storage of carbohydrates in individual phytomers and transport of surplus to other phytomers. Simulated total leaf area, total above-ground dry mass, dry mass distribution along the stem, and dry mass allocation fractions to each organ at the plant level were compared with field observations for plants grown at different densities. The distribution of local carbohydrate availability throughout the plant was characterized and a sensitivity analysis was conducted regarding the value of the carbohydrate transport coefficient. Key Results The model reproduced cotton leaf expansion and dry mass allocation across plant densities adequately. Individual leaf area was underestimated at very high plant densities. Best correspondence with measured plant traits was obtained for a value of the transport coefficient of 0.1 d -1. The simulated translocation of carbohydrates agreed well with results from C-labelling studies. Moreover, simulation results revealed the heterogeneous pattern of local carbohydrate availability over the plant as an emergent model property. Conclusions This modelling study shows how heterogeneity in local carbohydrate production within the plant structure in combination with limitations in transport result in heterogeneous satisfaction of demand over the plant. This model provides a tool to explore phenomena in cotton that are thought to be determined by local carbohydrate availability, such as branching pattern and fruit abortion in relation to climate and crop management

Rhamnose-Containing Compounds: Biosynthesis and Applications

Rhamnose-associated molecules are attracting attention because they are present in bacteria but not mammals, making them potentially useful as antibacterial agents. Additionally, they are also valuable for tumor immunotherapy. Thus, studies on the functions and biosynthetic pathways of rhamnose-containing compounds are in progress. In this paper, studies on the biosynthetic pathways of three rhamnose donors, i.e., deoxythymidinediphosphate-L-rhamnose (dTDP-Rha), uridine diphosphate-rhamnose (UDP-Rha), and guanosine diphosphate rhamnose (GDP-Rha), are firstly reviewed, together with the functions and crystal structures of those associated enzymes. Among them, dTDP-Rha is the most common rhamnose donor, and four enzymes, including glucose-1-phosphate thymidylyltransferase RmlA, dTDP-Glc-4,6-dehydratase RmlB, dTDP-4-keto-6-deoxy-Glc-3,5-epimerase RmlC, and dTDP-4-keto-Rha reductase RmlD, are involved in its biosynthesis. Secondly, several known rhamnosyltransferases from Geobacillus stearothermophilus, Saccharopolyspora spinosa, Mycobacterium tuberculosis, Pseudomonas aeruginosa, and Streptococcus pneumoniae are discussed. In these studies, however, the functions of rhamnosyltransferases were verified by employing gene knockout and radiolabeled substrates, which were almost impossible to obtain and characterize the products of enzymatic reactions. Finally, the application of rhamnose-containing compounds in disease treatments is briefly described

Synthetic and Immunological Studies of Mycobacterial Lipoarabinomannan Oligosaccharides and Their Protein Conjugates

Lipoarabinomannan (LAM) is one of the major constituents of the Mycobacterium tuberculosis cell wall and an attractive molecular scaffold for antituberculosis drug and vaccine development. In this paper, a convergent strategy was developed for the synthesis of LAM oligosaccharides with an α-1,2-linked dimannopyranose cap at the nonreducing end. The strategy was highlighted by efficient coupling of separately prepared nonreducing end and reducing end oligosaccharides. Glycosylations were mainly achieved with thioglycoside donors, which gave excellent yields and stereoselectivity even for reactions between complex oligosaccharides. The strategy was utilized to successfully synthesize tetra-, hepta-, and undecasaccharides of LAM from d-arabinose in 10, 15, and 14 longest linear steps and 7.84, 7.50, and 2.59% overall yields, respectively. The resultant oligosaccharides with a free amino group at their reducing end were effectively conjugated with carrier proteins, including bovine serum albumin and keyhole limpet hemocyanin (KLH), via a bifunctional linker. Preliminary immunological studies on the KLH conjugates revealed that they could elicit robust antibody responses in mice and that the antigen structure had some influence on their immunological property, thus verifying the potential of the oligosaccharides for vaccine development and other immunological studies