Tissue distribution and subcellular localization of the family of Kidney Ankyrin Repeat Domain (KANK) proteins

Kidney Ankyrin Repeat-containing Proteins (KANKs) comprise a family of four evolutionary conserved proteins (KANK1 to 4) that localize to the belt of mature focal adhesions (FAs) where they regulate integrin-mediated adhesion, actomyosin contractility, and link FAs to the cortical microtubule stabilization complex (CMSC). The human KANK proteins were first identified in kidney and have been associated with kidney cancer and nephrotic syndrome. Here, we report the distributions and subcellular localizations of the four Kank mRNAs and proteins in mouse tissues. We found that the KANK family members display distinct and rarely overlapping expression patterns. Whereas KANK1 is expressed at the basal side of epithelial cells of all tissues tested, KANK2 expression is mainly observed at the plasma membrane and/or cytoplasm of mesenchymal cells and KANK3 exclusively in vascular and lymphatic endothelial cells. KANK4 shows the least widespread expression pattern and when present, overlaps with KANK2 in contractile cells, such as smooth muscle cells and pericytes. Our findings show that KANKs are widely expressed in a cell type-specific manner, which suggests that they have cell- and tissue-specific functions.     

Better alone? A demographic case study of the hemiparasite Castilleja tenuiflora (Orobanchaceae): A first approximation

Castilleja tenuiflora is a facultative root hemiparasitic plant that has colonized a disturbed lava field in central Mexico. To determine the effects of hemiparasitism on the population dynamics of the parasite, we identified a set of potential hosts and quantified their effects on the vital rates of C. tenuiflora during 2016–2018. Connections between the roots of the hemiparasite and the hosts were confirmed with a scanning electron microscope. Annual matrices considering two conditions (with and without potential hosts) were built based on vital rates for each year, and annual stochastic finite rate growth rates (λ_s) were calculated. Plants produced more reproductive structures with hosts than without hosts. A Life Table Response Experiment (LTRE) was performed to compare the contributions of vital rates between conditions. We identified 19 species of potential hosts for this generalist hemiparasite. Stochastic lambda with hosts λ_s = 1.02 (CI = 0.9999, 1.1) tended to be higher than without them λ_s = 0.9503 (CI = 0.9055, 0.9981). The highest elasticity values correspond to survival. LTRE indicated that the most important parameters are survival and fecundity; the total contribution of fecundity (0.0192) to the difference in growth was three times lower than that of survival (0.0603). Piqueria trinervia was the most abundant host, and C. tenuiflora had a higher lambda with it than with other species. Individuals can grow alone, but hosts can have a positive effect on the vital parameters of C. tenuiflora and on λ.     

Separate roles of IQGAP Rng2p in forming and constricting the Schizosaccharomyces pombe cytokinetic contractile ring

Eukaryotic cells require IQGAP family multidomain adapter proteins for cytokinesis, but many questions remain about how IQGAPs contribute to the process. Here we show that fission yeast IQGAP Rng2p is required for both the normal process of contractile ring formation from precursor nodes and an alternative mechanism by which rings form from strands of actin filaments. Our work adds to previous studies suggesting a role for Rng2p in node and ring formation. We demonstrate that Rng2p is also required for normal ring constriction and septum formation. Systematic analysis of domain-deletion mutants established how the four domains of Rng2p contribute to cytokinesis. Contrary to a previous report, the actin-binding calponin homology domain of Rng2p is not required for viability, ring formation, or ring constriction. The IQ motifs are not required for ring formation but are important for ring constriction and septum formation. The GTPase-activating protein (GAP)–related domain is required for node-based ring formation. The Rng2p C-terminal domain is the only domain essential for viability. Our studies identified several distinct functions of Rng2 at multiple stages of cytokinesis.     

Frammenti cotiledonari di Aesculus hippocastanum L. coltivati in vitro. Primi dati sul comportamento dell'amido e dell'escina

Abstract

Fragments of Aesculus hippocastanum L. cotyledons grown in vitro. First results about starch and aescin characteristic features.—Cotyledon fragments of Aesculus hippocastanum grown in vitro in different media have been able to form callus and roots. The starch granules in the new cells are compound in structure and morphologically different from the simple cotyledon granules, whereas they are similar to the granules of the other parts of the plant in toto. Moreover, the callus has no aescin even though it originates from the cotyledor tissues.     

The Inhibition of Nucleoside Transport in Human Erythrocytes by Mioflazine Analogues

Abstract

Kinetic analysis of the transport protein (both influx and efflux), usually performed with radiolabelled nucleosides such as adenosine and uridine, has provided a wealth of information regarding the various kinetic and equilibrium parameters (1).     

Lipophilic Prodrugs and Formulations of Conventional (Deoxy)Nucleoside and Fluoropyrimidine Analogs in Cancer

Many drugs that are currently used for the treatment of cancer have limitations, such as induction of resistance and/or poor biological half-life, which reduce their clinical efficacy. To overcome these limitations, several strategies have been explored. Chemical modification by the attachment of lipophilic moieties to (deoxy)nucleoside analogs should enhance the plasma half-life, change the biodistribution, and improve cellular uptake of the drug. Attachment of a lipophilic moiety to a phosphorylated (deoxy)nucleoside analog will improve the activity of the drugs by circumventing the rate-limiting activation step of (deoxy)nucleoside analogs. Encapsulating drugs in nanoparticles or liposomes protects the drug against enzymatic breakdown in the plasma and makes it possible to get lipophilic compounds to the tumor site. In this review, we discuss the considerable progress that has been made in increasing the efficacy of classic (deoxy)nucleoside and fluoropyrimidine compounds by chemical modifications and alternative delivery systems.     

In Vivo and in Vitro Activity And Mechanism Of Action of the Multidrug Cytarabine-L-Glycerylyl-Fluorodeoxyuridine

Multidrugs have the potential to bypass resistance. We investigated the in vitro activity and resistance circumvention of the multidrug cytarabine-L-fluorodeoxyuridine (AraC-L-5FdU), linked via a glycerophospholipid linkage. Cytotoxicity was determined using sensitive (A2780, FM3A/0) and resistant (AG6000, AraC resistant, deoxycytidine kinase deficient; FM3A/TK-, 5FdU resistant, thymidine kinase deficient) cell lines. Circumvention of nucleoside transporter and activating enzymes was determined using specific inhibitors, HPLC analysis and standard radioactivity assays. AraC-L-5FdU was active (IC50: 0.03 μM in both A2780 and FM3A/0), had some activity in AG6000 (IC50: 0.28 μ M), but no activity in FM3A/TK^? (IC50: 18.3 μM). AraC-nucleotides were not detected in AG6000. 5FdU-nucleotides were detected in all cell lines. AraC-L-5FdU did not inhibit TS in FM3A/TK^? (5%). Since phosphatase/nucleotidase-inhibition reduced cytotoxicity 7–70-fold, cleavage seems to be outside the cell, presumably to nucleotides, and then to nucleosides. The multidrug was orally active in the HT-29 colon carcinoma xenografts which are resistant toward the single drugs.