Synthesis Of N-glycan mimetics and their evaluation as C-type lectin receptor (Clr) antagonists using microarray technology

209 p.Carbohydrate microarrays are well established tools for high-throughput analysis of carbohydrate-protein interactions and for screening the specificity of carbohydrate processing enzymes, In addition, a few groups have also explored the on-chip synthesis of ligands. The use of small amounts of substrates and the absence of long purification step make the microarray a time-saving approach for the parallel synthesis of hundred scaffolds.We developed a versatile platform to quickly synthesize significant number of glycomimetics via chemoenzymatic protocols and test them as potential ligands for C-type lectin receptors (CLRs). The transparent and the conductive hydrophobic-ITO surface allows to perform binding studies with labeled lectins by fluorescent spectroscopy as well as to estimate conversion rates of the on-chip reactions for every printed ligand by MALDI-TOF MS.CIC biomaGUN

Synthesis Of N-glycan mimetics and their evaluation as C-type lectin receptor (Clr) antagonists using microarray technology

209 p.Carbohydrate microarrays are well established tools for high-throughput analysis of carbohydrate-protein interactions and for screening the specificity of carbohydrate processing enzymes, In addition, a few groups have also explored the on-chip synthesis of ligands. The use of small amounts of substrates and the absence of long purification step make the microarray a time-saving approach for the parallel synthesis of hundred scaffolds.We developed a versatile platform to quickly synthesize significant number of glycomimetics via chemoenzymatic protocols and test them as potential ligands for C-type lectin receptors (CLRs). The transparent and the conductive hydrophobic-ITO surface allows to perform binding studies with labeled lectins by fluorescent spectroscopy as well as to estimate conversion rates of the on-chip reactions for every printed ligand by MALDI-TOF MS.CIC biomaGUN

Glomerular filtration rate and prevalence of chronic kidney disease in Wilms’ tumour survivors

Glomerular filtration rate (GFR) was evaluated in 32 Wilms’ tumour survivors (WTs) in a cross-sectional study using 99 Tc-diethylene triamine pentaacetic acid (99 Tc-DTPA) clearance, the Schwartz formula, the new Schwartz equation for chronic kidney disease (CKD), cystatin C serum concentration and the Filler formula. Kidney damage was established by beta-2-microglobulin (B-2-M) and albumin urine excretion, urine sediment and ultrasound examination. Blood pressure was measured. No differences were found between the mean GFR in 99 Tc-DTPA and the new Schwartz equation for CKD (91.8 ± 11.3 vs. 94.3 ± 10.2 ml/min/1.73 m2 [p = 0.55] respectively). No differences were observed between estimated glomerular filtration rate (eGFR) using the Schwartz formula and the Filler formula either (122.3 ± 19.9 vs. 129.8 ± 23.9 ml/min/1.73 m2 [p = 0.28] respectively). Increased urine albumin and B-2-M excretion, which are signs of kidney damage, were found in 7 (22%) and 3 (9.4%) WTs respectively. Ultrasound signs of kidney damage were found in 14 patients (43%). Five patients (15.6%) had more than one sign of kidney damage. Eighteen individuals (56.25%) had CKD stage I (10 with signs of kidney damage; 8 without). Fourteen individuals (43.75%) had CKD stage II (6 with signs of kidney damage; 8 without). The new Schwartz equation for CKD better estimated GFR in comparison to the Schwartz formula and the Filler formula. Furthermore, the WT survivors had signs of kidney damage despite the fact that GFR was not decreased below 90 ml/min/1.73 m2 with 99 Tc- DTPA

Cell-specific Bioorthogonal Tagging of Glycoproteins

Altered glycoprotein expression is an undisputed corollary of cancer development. Understanding these alterations is paramount but hampered by limitations underlying cellular model systems. For instance, the intricate interactions between tumour and host cannot be adequately recapitulated in monoculture of tumour-derived cell lines. More complex co-culture models usually rely on sorting procedures for proteome analyses and rarely capture the details of protein glycosylation. Here, we report a strategy termed Bio-Orthogonal Cell line-specific Tagging of Glycoproteins (BOCTAG). Cells are equipped by transfection with an artificial biosynthetic pathway that transforms bioorthogonally tagged sugars into the corresponding nucleotide-sugars. Only transfected cells incorporate bioorthogonal tags into glycoproteins in the presence of non-transfected cells. We employ BOCTAG as an imaging technique and to annotate cell-specific glycosylation sites in mass spectrometry-glycoproteomics. We demonstrate application in co-culture and mouse models, allowing for profiling of the glycoproteome as an important modulator of cellular function

Rapid On‐chip Synthesis of Complex Glycomimetics from N‐glycan Scaffolds for Improved Lectin Targeting

International audienceC-type lectin receptor (CLR) carbohydrate binding proteins found on immune cells with important functions in pathogen recognition as well as self and non-self-differentiation are increasingly moving into the focus of drug developers as targets for the immune therapy of cancer autoimmune diseases and inflammation and to improve the efficacy of vaccines. The development of molecules with increased affinity and selectivity over the natural glycan binders has largely focused on the synthesis of mono and disaccharide mimetics but glycan array binding experiments have shown increased binding selectivity and affinity for selected larger oligosaccharides that are able to engage in additional favorable interactions beyond the primary binding site. Here, a platform for the rapid preparation and screening of N-glycan mimetics on microarrays is presented that turns a panel of complex glycan core structures into structurally diverse glycomimetics by a combination of enzymatic glycosylation with a nonnatural donor and subsequent cycloaddition with a collection of alkynes. All surface-based reactions were monitored by MALDI-TOF MS to assess conversion and purity of spot compositions. Screening the collection of 374 N-glycomimetics against the plant lectin WFA and the 2 human immune lectins MGL ECD and Langerin ECD produced a number of high affinity binders as lead structures for more selective lectin targeting probes

Antitumor agents 7. Synthesis, antiproliferative activity and molecular modeling of new L-lysine-conjugated pyridophenoxazinones as potent DNA-binding ligands and topoisomerase IIa inhibitors

A series of L-lysine-conjugated pyridophenoxazinones 2e5 and 2′-5′ were designed and synthesized fordeveloping compounds with multimodal anticancer potentialities. All compounds inhibited the proliferation of a panel of human liquid and solid neoplastic cell lines. 2 and 5 were the most active compounds with IC50 values in the submicromolar range. UVevis, 1H NMR, unwinding, and docking experiments demonstrated that they intercalate between the middle 50-GC-30 base pairs with the carboxamide side chain lying into major groove. Charge-transfer contribution to the complex stability, evaluated by ab initio calculations, was found to correlate with cytotoxicity. Relaxation and cleavage assays showed that 2 and 5 selectively target Topo IIa over Topo IIb and stimulate the formation of covalent Topo IIeDNA complexes, functioning as poisons. Moreover, compound 5 induced DNA damage and arrested MCF-7 cells at the G2/M phase. Altogether, the work provides interesting structure-activity relationships in the pyridophenoxazinone-L-lysine conjugate series and identifies 5 as a promising candidate for further in vivo evaluation

Synthesis, Antiproliferative Activity, and Molecular Modelling of New DNA Intercalating Pyridophenoxazinone Carboxamides

Clinical studies have shown that multimodal therapy, a combination of anticancer treatments acting simultaneously on different biological targets is able to inhibit the proliferation of tumor cells present in different phases of the cell cycle. This new therapeutic approach improves the patient's survival and decreases the drug resistance.1 To obtain new antiproliferative compounds able to act through multimodal mechanisms on the cancerous cells, including DNA intercalation and topoisomerase inhibition,2 derivatives of pyridophenoxazinone carboxylic acids the carboxamides 1a-d and 2a-d, holding at C-9 and C-10 positions an aminoacidic chain, were designed by molecular modelling calculation, synthesized and evaluated for their inhibitory effect on a panel of solid and liquid tumor cell lines and found active at submicromolar concentration. Compound 2a was the most active, particularly on solid tumor

A INOVAÇÃO SOB A VISÃO DOS GESTORES DE DUAS INSTITUIÇÕES PÚBLICAS

O termo inovação liga-se a um contexto de mudança, de competição, de capacidade produtiva e de diferenciação. Nos estudos organizacionais, o tema permeia discussões e abre campos para atuações em diferentes níveis. Sendo assim, antes de praticar a inovação torna-se pertinente procurar auferir qual a percepção de inovação que determinados gestores têm. Este artigo busca demonstrar e comparar qual a percepção de inovação dos gestores de duas instituições tidas como públicas: uma Instituição de Ensino Superior da cidade de Blumenau/SC e uma Prefeitura Municipal localizada no Médio Vale do Itajaí/SC. Utilizou-se o questionário de Zhuang (1995) e foram pesquisados os 16 principais gestores da IES e os 12 principais gestores da Prefeitura Municipal. Os dados revelaram uma convergência na percepção de inovação de ambos os gestores e apontam o indivíduo como agente primordial para a inovação. Sugere-se expandir a pesquisa para novas organizações públicas. Palavras-Chave: inovação, percepção, gestão