Waste generated by the processing of natural stone poses environmental and economic problems. One promising use for stone waste is soil stabilization. In this paper, a number of tests including ...unconfined compressive strength (UCS) and California bearing ratio (CBR) of crushed stone were performed to study the effect of curing time, soaked and unsoaked conditions and freeze-thaw cycles on the geotechnical characteristics of cement-stabilized specimens. The results indicate that crushed stone waste can be combined with cement as a stabilizer to improve soil properties. Also the optimum moisture content (OMC) and maximum dry density (MDD) are dependent on the cement content and almost independent of the crushed stone waste content. The CBR increased significantly as the crushed stone waste and cement contents increased under both soaked and unsoaked conditions, but these had little effect on the UCS at curing times of less than 14 days. Cement-stabilized specimens with 10% crushed stone waste showed a significant increase in UCS. The CBR and UCS of unsoaked specimens were higher than for soaked specimens at different crushed stone waste and cement contents. The CBR and UCS values increased with an increase in curing time. However, the curing time had a more obvious effect on UCS than on CBR.
Research on patient-derived induced pluripotent stem cells (iPSCs) could immensely benefit from the implementation of CRISPR/Cas9 genome editing of iPSCs, creating unique opportunities such as the ...establishment of isogenic iPSC lines for disease modeling or personalized patient-specific drug screenings. Here we describe a stepwise protocol of safe, efficient, and selection-free CRISPR/Cas9-mediated gene correction or knockout in human iPSCs followed by 3D spin-embryoid body (EB)-based hematopoietic/neutrophilic iPSC-differentiation.
Understanding the list of plant species in watersheds is important for executive and managerial actions and phyto-geographical studies. The floristic richness gives the policy and operation of the ...natural communities and enhances a whole accepting of the design and procedure of their structure. The studies of vegetation were conducted throughout 16,000 km
2
of Qom Rood Watershed of Central Iran. Plant phenotypes were defined based on the Raunkiaer’s life form characteristics. The Asteraceae showed the most family with 22 species (18.6%). Poaceae (20 species, 16.9%), Brassicaceae (10 species, 8.4%), Lamiaceae and Papilionaceae (8 species, 6.7%), Caryophyllaceae (6 species, 5%), Apiaceae and Liliaceae (5 species, 4.2%), Chenopodiaceae and Rosaceae (4 species, 3.3%), Euphorbiaceae, Rubiaceae, Plantaginaceae, and Polygonaceae (3 species, 2.5%), Geraniaceae, Zygophyllaceae, and Ranunculaceae (2 species, 1.6), and Convolvulaceae, Dipsacaceae, Fumariaceae, Iridaceae, Malvaceae, Plumbaginaceae, Resedaceae, and Solanaceae (1 species, 0.84%) respectively showed the percentage of species presence in the case study. The results also showed that hemicryptophytes with 40.6% (49 species) showed the highest life form followed by therophyte 30.5% (36 species), chamaephytes 10.1% (12 species), geophyte 10.1% (12 species), and phanerophyte 7.6% (9 species). Iran-Turanian region had the prevailing chorotypes (70 species, 70.8%). According to the IUCN Red List Categories, 19 threatened plant taxa were found in the study area.
Mutations in the ELANE gene, encoding the neutrophil elastase (NE) protein, are responsible for most cyclic neutropenia (CyN) cases and approximately 25% of congenital neutropenia (CN) cases. In CN ...and in CyN, a median of 2.8% of CD34+ cells were early CD49f+ hematopoietic stem cells (eHSC) that did not express ELANE and thus escape from the unfolded protein response (UPR) caused by mutated NE. In CyN, the CD49f+ cells respond to granulocyte colony-stimulating factor (G-CSF) with a significant upregulation of the hematopoietic stem cell-specific transcription factors, C/EBPα, MLL1, HOXA9, MEIS1, and HLF during the ascending arm of the cycle, resulting in the differentiation of myeloid cells to mature neutrophils at the cycle peak. However, NE protein released by neutrophils at the cycle's peak caused a negative feedback loop on granulopoiesis through the proteolytic digestion of G-CSF. In contrast, in CN patients, CD49f+ cells failed to express mRNA levels of HSC-specific transcription factors mentioned above. Rescue of C/EBPα expression in CN restored granulopoiesis.
A Autosomal-dominant
mutations are the most common cause of severe congenital neutropenia. Although the majority of congenital neutropenia patients respond to daily granulocyte colony stimulating ...factor, approximately 15 % do not respond to this cytokine at doses up to 50 μg/kg/day and approximately 15 % of patients will develop myelodysplasia or acute myeloid leukemia. "Maturation arrest," the failure of the marrow myeloid progenitors to form mature neutrophils, is a consistent feature of
associated congenital neutropenia. As mutant neutrophil elastase is the cause of this abnormality, we hypothesized that
associated neutropenia could be treated and "maturation arrest" corrected by a CRISPR/Cas9-sgRNA ribonucleoprotein mediated
knockout. To examine this hypothesis, we used induced pluripotent stem cells from two congenital neutropenia patients and primary hematopoietic stem and progenitor cells from four congenital neutropenia patients harboring
mutations as well as HL60 cells expressing mutant
We observed that granulocytic differentiation of
knockout induced pluripotent stem cells and primary hematopoietic stem and progenitor cells were comparable to healthy individuals. Phagocytic functions, ROS production, and chemotaxis of the
KO (knockout) neutrophils were also normal. Knockdown of
in the mutant
expressing HL60 cells also allowed full maturation and formation of abundant neutrophils. These observations suggest that
CRISPR/Cas9 RNP based
knockout of patients' primary hematopoietic stem and progenitor cells followed by autologous transplantation may be an alternative therapy for congenital neutropenia.
Protein therapeutics frequently face major challenges, including complicated production, instability, poor solubility, and aggregation. De novo protein design can readily address these challenges. ...Here, we demonstrate the utility of a topological refactoring strategy to design novel granulopoietic proteins starting from the granulocyte-colony stimulating factor (G-CSF) structure. We change a protein fold by rearranging the sequence and optimising it towards the new fold. Testing four designs, we obtain two that possess nanomolar activity, the most active of which is highly thermostable and protease-resistant, and matches its designed structure to atomic accuracy. While the designs possess starkly different sequence and structure from the native G-CSF, they show specific activity in differentiating primary human haematopoietic stem cells into mature neutrophils. The designs also show significant and specific activity in vivo. Our topological refactoring approach is largely independent of sequence or structural context, and is therefore applicable to a wide range of protein targets.
Computational protein design is rapidly becoming more powerful, and improving the accuracy of computational methods would greatly streamline protein engineering by eliminating the need for empirical ...optimization in the laboratory. In this work, we set out to design novel granulopoietic agents using a rescaffolding strategy with the goal of achieving simpler and more stable proteins. All of the 4 experimentally tested designs were folded, monomeric, and stable, while the 2 determined structures agreed with the design models within less than 2.5 Å. Despite the lack of significant topological or sequence similarity to their natural granulopoietic counterpart, 2 designs bound to the granulocyte colony-stimulating factor (G-CSF) receptor and exhibited potent, but delayed, in vitro proliferative activity in a G-CSF-dependent cell line. Interestingly, the designs also induced proliferation and differentiation of primary human hematopoietic stem cells into mature granulocytes, highlighting the utility of our approach to develop highly active therapeutic leads purely based on computational design.
Nicotinamide phosphoribosyltransferase (NAMPT) regulates cellular functions through the protein deacetylation activity of nicotinamide adenine dinucleotide (NAD
)-dependent sirtuins (SIRTs). SIRTs ...regulate functions of histones and none-histone proteins. The role of NAMPT/SIRT pathway in the regulation of maintenance and differentiation of human-induced pluripotent stem (iPS) cells is not fully elucidated.
We evaluated the effects of specific inhibitors of NAMPT or SIRT2 on the pluripotency, proliferation, survival, and hematopoietic differentiation of human iPS cells. We also studied the molecular mechanism downstream of NAMPT/SIRTs in iPS cells.
We demonstrated that NAMPT is indispensable for the maintenance, survival, and hematopoietic differentiation of iPS cells. We found that inhibition of NAMPT or SIRT2 in iPS cells induces p53 protein by promoting its lysine acetylation. This leads to activation of the p53 target, p21, with subsequent cell cycle arrest and induction of apoptosis in iPS cells. NAMPT and SIRT2 inhibition also affect hematopoietic differentiation of iPS cells in an embryoid body (EB)-based cell culture system.
Our data demonstrate the essential role of the NAMPT/SIRT2/p53/p21 signaling axis in the maintenance and hematopoietic differentiation of iPS cells.
Quantitative estimation of canopy biophysical variables are very important in different studies such as meteorology, agriculture and ecology, so knowledge of the spatial and temporal distribution of ...these variables would be highly beneficial. Meanwhile, remote sensing is known as an important source of information to estimate fractional vegetation cover in large areas. Today spectral indices have been very popular in the remote sensing of vegetation features. But often reflections of soil and rocks are much more than reflections of sparse vegetation in these areas, that makes separation of plant signals difficult. So in this study measured fractional vegetation cover of a desert area were evaluated with 20 vegetation indices in five different categories as the most appropriate category, or indicator for desert vegetation to be identified. The five categories were including: (1) conventional ratio and differential indices such as NDVI; (2) indices corrected and derived from the traditional indicators such as NDVIc and GNDVI; (3) soil reflectance adjusted indices such as SAVI; (4) triangle indices based on three discreet bands in their equation (Green, Red and NIR) like TVI; and (5) non-conventional ratio and differential indices such as CI. According to the results of this research, DVI index with 0.668 the coefficient of determination (
R
2) showed the best fractional vegetation cover estimation. But according to the sparse vegetation in desert areas and the results of this research it seems none of these indicators alone can accurately estimate the percentage of vegetation cover, however, to do a proper estimation it is possible to enter data of these indices in a multivariate regression model. Using this method enabled us to increase the coefficient of determination of fractional vegetation cover estimation model up to 0.797.
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