The Trail, 1988-09-08

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Critical Points in the Management of Pseudohypoaldosteronism Type 1

Pseudohypoaldosteronism type 1 (PHA-1, MIM #264350) is caused by defective transepithelial sodium transport. Affected patients develop life-threatening neonatal-onset salt loss, hyperkalemia, acidosis, and elevated aldosterone levels due to end-organ resistance to aldosterone. In this report, we present a patient diagnosed as PHA-1 who had clinical and laboratory findings compatible with the diagnosis and had genetically proven autosomal recessive PHA-1. The patient received high doses of sodium supplementation and potassium-lowering therapies; however, several difficulties were encountered in the management of this case. The aim of this presentation was to point out the potential pitfalls in the treatment of such patients in the clinical practice and to recommend solutions

Dual LSD1 and HDAC6 Inhibition Induces Doxorubicin Sensitivity in Acute Myeloid Leukemia Cells

Defects in epigenetic pathways are key drivers of oncogenic cell proliferation. We developed a LSD1/HDAC6 multitargeting inhibitor (iDual), a hydroxamic acid analogue of the clinical candidate LSD1 inhibitor GSK2879552. iDual inhibits both targets with IC50 values of 540, 110, and 290 nM, respectively, against LSD1, HDAC6, and HDAC8. We compared its activity to structurally similar control probes that act by HDAC or LSD1 inhibition alone, as well as an inactive null compound. iDual inhibited the growth of leukemia cell lines at a higher level than GSK2879552 with micromolar IC50 values. Dual engagement with LSD1 and HDAC6 was supported by dose dependent increases in substrate levels, biomarkers, and cellular thermal shift assay. Both histone methylation and acetylation of tubulin were increased, while acetylated histone levels were only mildly affected, indicating selectivity for HDAC6. Downstream gene expression (CD11b, CD86, p21) was also elevated in response to iDual treatment. Remarkably, iDual synergized with doxorubicin, triggering significant levels of apoptosis with a sublethal concentration of the drug. While mechanistic studies did not reveal changes in DNA repair or drug efflux pathways, the expression of AGPAT9, ALOX5, BTG1, HIPK2, IFI44L, and LRP1, previously implicated in doxorubicin sensitivity, was significantly elevated

Promising anticancer agents based on 8-hydroxyquinoline hydrazone copper(II) complexes

We report the synthesis and characterization of a group of benzoylhydrazones (L n ) derived from 2-carbaldehyde-8-hydroxyquinoline and benzylhydrazides containing distinct para substituents (R = H, Cl, F, CH 3 , OCH 3 , OH and NH 2 , for L 1-7 , respectively; in L 8 isonicotinohydrazide was used instead of benzylhydrazide). Cu(II) complexes were prepared by reaction of each benzoylhydrazone with Cu(II) acetate. All compounds were characterized by elemental analysis and mass spectrometry as well as by FTIR, UV-visible absorption, NMR or electron paramagnetic resonance spectroscopies. Complexes isolated in the solid state ( 1–8 ) are either formulated as [Cu(HL)acetate] (with L 1 and L 4 ) or as [Cu(L n )] 3 ( n = 2, 3, 5, 6, 7 and 8). Single crystal X-ray diffraction studies were done for L 5 and [Cu(L 5 )] 3 , confirming the trinuclear formulation of several complexes. Proton dissociation constants, lipophilicity and solubility were determined for all free ligands by UV-Vis spectrophotometry in 30% (v/v) DMSO/H 2 O. Formation constants were determined for [Cu(LH)], [Cu(L)] and [Cu(LH −1 )] for L = L 1 , L 5 and L 6 , and also [Cu(LH −2 )] for L = L 6 , and binding modes are proposed, [Cu(L)] predominating at physiological pH. The redox properties of complexes formed with L 1 , L 5 and L 6 are investigated by cyclic voltammetry; the formal redox potentials fall in the range of +377 to +395 mV vs. NHE. The binding of the Cu(II)-complexes to bovine serum albumin was evaluated by fluorescence spectroscopy, showing moderate-to-strong interaction and suggesting formation of a ground state complex. The interaction of L 1 , L 3 , L 5 and L 7 , and of the corresponding complexes with calf thymus DNA was evaluated by thermal denaturation. The antiproliferative activity of all compounds was evaluated in malignant melanoma (A-375) and lung (A-549) cancer cells. The complexes show higher activity than the corresponding free ligand, and most complexes are more active than cisplatin. Compounds 1, 3, 5 , and 8 were selected for additional studies: while these complexes induce reactive oxygen species and double-strand breaks in both cancer cells, their ability to induce cell-death by apoptosis varies. Within the set of compounds tested, 8 emerges as the most promising one, presenting low IC 50 values, and high induction of oxidative stress and DNA damage, which eventually lead to high rates of apoptosis

Hypoxia induces an early primitive streak signature, enhancing spontaneous elongation and lineage representation in gastruloids

The cellular microenvironment, together with intrinsic regulators, shapes stem cell identity and differentiation capacity. Mammalian early embryos are exposed to hypoxia in vivo and appear to benefit from hypoxic culture in vitro. Yet, how hypoxia influences stem cell transcriptional networks and lineage choices remain poorly understood. Here, we investigated the molecular effects of acute and prolonged hypoxia on embryonic and extra-embryonic stem cells as well as the functional impact on differentiation potential. We find a temporal and cell type-specific transcriptional response including an early primitive streak signature in hypoxic embryonic stem cells mediated by HIF1α. Using a 3D gastruloid differentiation model, we show that hypoxia-induced T expression enables symmetry breaking and axial elongation in the absence of exogenous WNT activation. When combined with exogenous WNT activation, hypoxia enhances lineage representation in gastruloids, as demonstrated by highly enriched signatures of gut endoderm, notochord, neuromesodermal progenitors and somites. Our findings directly link the microenvironment to stem cell function and provide a rationale supportive of applying physiological conditions in models of embryo development

Multitargeting epi-epi drugs for multidrug reistance

Epigenetic therapy is now a clinical reality with eight approved drugs that target DNA methyltransferases, histone deacetylases (HDACs) and lysine methyltransferases. A further recent development is the concept of epigenetic multitargeting through the rational design of novel agents that combine the inhibition of an epigenetic pathway with a second non-epigenetic target and five such compounds have advanced to clinical development. We are investigating the even newer concept of „epi-epi‟ drugs that inhibit two separate epigenetic pathways. Such dual targeting agents have the potential to achieve higher efficacy against proliferating cancer cells while reducing tunor resistance. In this presentation, we report a selective dual histone deacetylase and demethylase inhibitor with an IC50 LSD1 respectively. The compound was biologically profiled together with control compounds that were either single inhibitors or inactive against either enzyme. The dua inhibitor was active against a panel of leukemia cell lines at a micromolar level and induced apoptosis. Target engagement asays such as CETSA were employed to confirm the inhibition of HDAC6 and LSD1 in cells. Further experiments were carried out to identify synergistic effects with clinically approved agents and promising results were observed with doxorubicin

Solution chemical properties and anticancer potential of 8-hydroxyquinoline hydrazones and their oxidovanadium(IV) complexes

Funding Information: This work was supported by Centro de Química Estrutural, which is financed by national funds from Fundação para a Ciência e Tecnologia (FCT), projects UIDB/00100/2020, UIDP/00100/2020 and LA/P/0056/2020, and Programa Operacional Regional de Lisboa 2020. We also thank project PTDC/QUI-QIN/0586/2020 and N. Ribeiro acknowledges FCT for SFRH/BD/135797/2018 grant. The Portuguese NMR and Mass spectrometry IST-UL are acknowledged for the access to the equipment. This work was supported by the Portuguese-Hungarian Scientific & Technological CooperationTÉT-PT-2018-00002, ÚNKP-21-3-SZTE-455 (to V. Pósa) New National Excellence Program Ministry of Human Capacities. The support of the ‘Lendület’ Programme (ELKH, LP2019-6/2019) and the COST ActionCA18202, NECTAR-Network for Equilibria and Chemical Thermodynamics Advanced Research is also acknowledged. This work was also supported by Koç University School of Medicine (KUSOM) and the authors gratefully acknowledge use of the services and facilities of the Koç University Research Center for Translational Medicine (KUTTAM), funded by the Presidency of Turkey, Presidency of Strategy and Budget. The content is solely the responsibility of the authors and does not necessarily represent the official views of the Presidency of Strategy and Budget. G. Sciortino, V. Ugone, E. Garribba thank Fondazione di Sardegna (grant FdSGarribba2017) and Regione Autonoma della Sardegna (grant RASSR79857); G. Sciortino also thanks MICINN’ Juan de la Cierva program, FJC2019-039135-I for the financial support. L. Maia thanks the Associate Laboratory for Green Chemistry - LAQV, which is financed by national funds from Fundação para a Ciência e a Tecnologia, MCTES (FCT/MCTES; UIDB/50006/2020 and UIDP/50006/2020). Funding Information: This work was supported by Centro de Química Estrutural , which is financed by national funds from Fundação para a Ciência e Tecnologia (FCT), projects UIDB/00100/2020 , UIDP/00100/2020 and LA/P/0056/2020 , and Programa Operacional Regional de Lisboa 2020 . We also thank project PTDC/QUI-QIN/0586/2020 and N. Ribeiro acknowledges FCT for SFRH/BD/135797/2018 grant. The Portuguese NMR and Mass spectrometry IST-UL are acknowledged for the access to the equipment. This work was supported by the Portuguese-Hungarian Scientific & Technological Cooperation TÉT-PT-2018-00002 , ÚNKP-21-3-SZTE-455 (to V. Pósa) New National Excellence Program Ministry of Human Capacities . The support of the ‘Lendület’ Programme (ELKH, LP2019-6/2019 ) and the COST Action CA18202 , NECTAR-Network for Equilibria and Chemical Thermodynamics Advanced Research is also acknowledged. This work was also supported by Koç University School of Medicine (KUSOM) and the authors gratefully acknowledge use of the services and facilities of the Koç University Research Center for Translational Medicine (KUTTAM), funded by the Presidency of Turkey, Presidency of Strategy and Budget. The content is solely the responsibility of the authors and does not necessarily represent the official views of the Presidency of Strategy and Budget. G. Sciortino, V. Ugone, E. Garribba thank Fondazione di Sardegna (grant FdSGarribba2017 ) and Regione Autonoma della Sardegna (grant RASSR79857 ); G. Sciortino also thanks MICINN’ Juan de la Cierva program, FJC2019-039135-I for the financial support. L. Maia thanks the Associate Laboratory for Green Chemistry - LAQV , which is financed by national funds from Fundação para a Ciência e a Tecnologia, MCTES (FCT/MCTES; UIDB/50006/2020 and UIDP/50006/2020 ). Publisher Copyright: © 2022 Elsevier Inc.We report the synthesis and characterization of a family of benzohydrazones (Ln, n = 1–6) derived from 2-carbaldehyde-8-hydroxyquinoline and benzylhydrazides containing different substituents in the para position. Their oxidovanadium(IV) complexes were prepared and compounds with 1:1 and 1:2 metal-to-ligand stoichiometry were obtained. All compounds were characterized by elemental analyses and mass spectrometry as well as FTIR, UV–visible absorption, NMR (ligand precursors) and EPR (complexes) spectroscopies, and by DFT computational methods. Proton dissociation constants, lipophilicity and solubility in aqueous media were determined for all ligand precursors. Complex formation with V(IV)O was evaluated by spectrophotometry for L4 (Me-substituted) and L6 (OH-substituted) and formation constants for mono [VO(HL)]+, [VO(L)] and bis [VO(HL)2], [VO(HL)(L)]−, [VO(L)2]2− complexes were determined. EPR spectroscopy indicates the formation of [VO(HL)]+ and [VO(HL)2], with this latter being the major species at the physiological pH. Noteworthy, the EPR data suggest a different behaviour for L4 and L6, which confirm the results obtained in the solid state. The antiproliferative activity of all compounds was evaluated in malignant melanoma (A-375) and lung (A-549) cancer cells. All complexes show much higher activity on A-375 (IC50 20 μM). Complex 3 (F-substituted) shows the lowest IC50 on both cell lines and lower than cisplatin (in A-375). Studies identified this compound as the one showing the highest increase in Annexin-V staining, caspase activity and induction of double stranded breaks, corroborating the cytotoxicity results. The mechanism of action of the complexes involves reactive oxygen species (ROS) induced DNA damage, and cell death by apoptosis.publishersversionpublishe

In vitro Antimicrobial Susceptibility of Urinary Tract Infection Pathogens in Children

Aim:Urinary tract infection (UTI) is one of the most common bacterial infections in children. Empirical treatment is commenced according to the patient’s characteristics and the antimicrobial susceptibility patterns in the region. Therefore, a determination of antimicrobial resistance patterns has a great importance in effective treatment. The aim of this study was to determine the pathogens which cause UTIs in patients admitted to a university hospital in Izmir and to determine their antimicrobial susceptibility pattern.Materials and Methods:The files of patients aged between 0-18 years, followed up with a diagnosis of UTI, vesicoureteral reflux and neurogenic bladder in Ege University Faculty of Medicine Paediatric Nephrology Unit between February, 2013 and November, 2018 were retrospectively reviewed.Results:A total of 1,126 positive urine cultures from 729 patients (65% female) were included in this study. Gram-negative pathogens constituted 88.2% of the cultures. Escherichia coli (E. coli) was the most commonly isolated bacteria with a prevalence of 59.1%, followed by Klebsiella pneumonia with 17.9%, and Enterococcus faecalis with 8.3% (n=93). Ampicillin, cefuroxime and trimethoprim-sulfamethoxazole with susceptibility rates of 18.6%, 39.6%, 49.0% respectively, constituted the highest resistant antimicrobials to Enterobacteriaceae. Enterococcus spp. showed the highest resistance to gentamycin with 50% resistance in tested cases. Pseudomonas spp. with 64.3% susceptibility showed the highest resistance to piperacillin-tazobactam.Conclusion:This study revealed that bacterial resistance to commonly used antimicrobials in UTI is an important and challenging problem which requires planning

Promising anticancer agents based on 8-hydroxyquinoline hydrazone copper(II) complexes

We report the synthesis and characterization of a group of benzoylhydrazones (L n ) derived from 2-carbaldehyde-8-hydroxyquinoline and benzylhydrazides containing distinct para substituents (R = H, Cl, F, CH 3 , OCH 3 , OH and NH 2 , for L 1-7 , respectively; in L 8 isonicotinohydrazide was used instead of benzylhydrazide). Cu(II) complexes were prepared by reaction of each benzoylhydrazone with Cu(II) acetate. All compounds were characterized by elemental analysis and mass spectrometry as well as by FTIR, UV-visible absorption, NMR or electron paramagnetic resonance spectroscopies. Complexes isolated in the solid state ( 1–8 ) are either formulated as [Cu(HL)acetate] (with L 1 and L 4 ) or as [Cu(L n )] 3 ( n = 2, 3, 5, 6, 7 and 8). Single crystal X-ray diffraction studies were done for L 5 and [Cu(L 5 )] 3 , confirming the trinuclear formulation of several complexes. Proton dissociation constants, lipophilicity and solubility were determined for all free ligands by UV-Vis spectrophotometry in 30% (v/v) DMSO/H 2 O. Formation constants were determined for [Cu(LH)], [Cu(L)] and [Cu(LH −1 )] for L = L 1 , L 5 and L 6 , and also [Cu(LH −2 )] for L = L 6 , and binding modes are proposed, [Cu(L)] predominating at physiological pH. The redox properties of complexes formed with L 1 , L 5 and L 6 are investigated by cyclic voltammetry; the formal redox potentials fall in the range of +377 to +395 mV vs. NHE. The binding of the Cu(II)-complexes to bovine serum albumin was evaluated by fluorescence spectroscopy, showing moderate-to-strong interaction and suggesting formation of a ground state complex. The interaction of L 1 , L 3 , L 5 and L 7 , and of the corresponding complexes with calf thymus DNA was evaluated by thermal denaturation. The antiproliferative activity of all compounds was evaluated in malignant melanoma (A-375) and lung (A-549) cancer cells. The complexes show higher activity than the corresponding free ligand, and most complexes are more active than cisplatin. Compounds 1, 3, 5 , and 8 were selected for additional studies: while these complexes induce reactive oxygen species and double-strand breaks in both cancer cells, their ability to induce cell-death by apoptosis varies. Within the set of compounds tested, 8 emerges as the most promising one, presenting low IC 50 values, and high induction of oxidative stress and DNA damage, which eventually lead to high rates of apoptosis