KrioBlastTM as a new technology of hyper-fast cryopreservation of cells and tissues. Part 2. Kinetic vitrification of human pluripotent stem cells and spermatozoa

Pilot experiments on kinetic vitrification of human pluripotent stem cells and spermatozoa using a KrioBlastTM-2 without penetrating cryoprotectants have shown high survival of cells (75-85% in both cases

KrioBlastTM as a new technology of hyper-fast cryopreservation of cells and tissues. Part 1. Thermodynamic aspects and potential applications in reproductive and regenerative medicine

Kinetic (dynamic) vitrification is a promising trend in cryopreservation of biological materials because it allows avoiding the formation of lethal intracellular ice and minimizes harmful effects of highly toxic penetrating cryoprotectants. A uniform cooling protocol and the same instruments can be used for practically all types of cell

Tripodal oxazolidine-N-oxyl diradical complexes of Dy3+ and Eu3+

Two diradical complexes of the formula [LnRad2(CF3SO3)3] c (Ln(III) = Dy, Eu, Rad = 4,4-dimethyl-2,2-bis(pyridin-2-yl)-1,3-oxazolidine-3-oxyl) were obtained in air conditions. These are the first examples of diradical compounds of lanthanides and oxazolidine nitroxide. The complexes were characterized crystallographically and magnetically. Single crystal XRD analysis revealed that their coordination sphere is composed of three monodentate triflates and two tripodal Rad, which coordinate the central atom in a tridentate manner via two N atoms of the pyridine groups and the O atom of a nitroxide group. The LnO5N4 polyhedron represents a spherical capped square antiprism with point symmetry close to C4v. The data of static magnetic measurements are compatible with the presence of two paramagnetic ligands in the coordination sphere of the metal

Reparative Spheroids in HPV-Associated Chronic Cervicitis

Background: Spheroid cell structures (SCS) described in cell culture are used to study cell-cell and cell-matrix interactions. However, the role of the SCS in the repair process in vivo remains unexplored. The aim of the study was to examine the cellular composition of the spherical structures and their functional significance in the repair of the squamous epithelium in human papilloma virus-associated chronic cervicitis (HPV-CC). Methods and Results: The cytology and biopsy materials from 223 patients with HPV-CC were subjected to molecular testing for HPV DNA by Real-Time Polymerase Chain Reaction (Real-Time PCR) with genotyping and chromogenic in situ hybridization (CISH), as well as immunocytological and immunohistochemical analyses of p16INK4A, Ki67, SMA, Vimentin, CD34, E-cadherin, Oct4, CD44, CKW markers. In the stem cell niche zone, these spheroid structures were discovered having proliferative activity and showing signs of producing stem cells involved in the repair of the cervical mucosa in HPV-CC. Conclusion: The persistence of the HPV in the stem cell niche zone cells in the cervix determines the chronization of inflammation in this area, with the ability to perform pathological repair. The immunophenotype of the spheroid cell structures in the HPV-CC includes cells with signs of stem cells (‘stemness’) and the mesenchymal-epithelial transition

Endothelial Dysfunction as a Consequence of Endothelial Glycocalyx Damage: A Role in the Pathogenesis of Preeclampsia

The endothelial glycocalyx is an intravascular compartment which consists of carbohydrate part of membrane glycoconjugates, free proteoglycans and associated proteins. It is thought to play an important role in the vascular tone regulation, vascular permeability and thromboresistance. It was suggested that the leading cause of endothelial dysfunction in various cardiovascular, inflammatory, and kidney diseases is the damage of the endothelial glycocalyx. This review presents the changes in the composition and structure of the endothelial glycocalyx in the settings of damage and under systemic inflammatory response, and the impact of these changes on the functions of endothelial cells and intercellular contacts, mediating the interaction of endothelium and the immune cells. The second issue, discussed in this article is a possible role of endothelial glycocalyx in the pathogenesis of preeclampsia—a complication of pregnancy associated with hypertension, proteinuria and edema. The reviewed data contribute a new insight in the endothelial dysfunction pathogenesis

Angiogenesis-Related Factors in Early Pregnancy Loss

The habitual loss of early pregnancy is one of the major problems of obstetrics nowadays, provided that the cause of more than 50% of all early pregnancy losses is unknown. Adequate angiogenesis is one of the main indicators of proper formation of placental system, making the basis of fetal life support. The objective description of angiogenesis in physiological development of pregnancy and in pathological conditions is complicated by the difficulties in obtaining and characterizing placental tissue in early pregnancy. Thus, angiogenesis‐related factors are promising indicators to characterize angiogenesis in pregnancy. This chapter draws attention to alteration in angiogenesis‐related factors in peripheral blood of patients with habitual early pregnancy losses. Investigation of factors (vascular endothelial growth factor (VEGF), sFlt‐1, sKDR, metalloproteinase (MMP)‐2, MMP‐9, tissue inhibitor (TIMP)‐1, TIMP‐2 and placental growth factor (PLGF)), which specifically and nonspecifically regulate angiogenesis in pregnancy, was performed in the most significant terms for placentogenesis: 6 weeks, 7–8 weeks and 11–14 weeks of pregnancy. It was found that in a missed abortion there was a significant imbalance of angiogenesis‐related factors compared with normal pregnancy. These results reflect a disturbance of angiogenesis in a missed abortion and point to the importance of the studied factors in the pathogenesis of early pregnancy losses

Two novel mutations associated with ataxia-telangiectasia identified using an ion ampliSeq inherited disease panel

© 2017 Kuznetsova, Trofimov, Shubina, Kochetkova, Karetnikova, Barkov, Bakharev, Gusev and Sukhikh. Ataxia-telangiectasia (A-T), or Louis-Bar syndrome, is a rare neurodegenerative disorder associated with immunodeficiency. For families with at least one affected child, timely A-T genotyping during any subsequent pregnancy allows the parents to make an informed decision about whether to continue to term when the fetus is affected. Mutations in the ATM gene, which is 150 kb long, give rise to A-T; more than 600 pathogenic variants in ATM have been characterized since 1990 and new mutations continue to be discovered annually. Therefore, limiting genetic screening to previously known SNPs by PCR or hybridization with microarrays may not identify the specific pathog enic genotype in ATM for a given A-T family. However, recent developments in next-generation sequencing technology offer prompt high-throughput full-length sequencing of genomic fragments of interest. This allows the identification of the whole spectrum of mutations in a gene, including any novel ones. We report two A-T families with affected children and current pregnancies. Both families are consanguineous and originate from Caucasian regions of Russia and Azerbaijan. Before our study, no ATM mutations had been identified in the older children of these families. We used ion semiconductor sequencing and an Ion AmpliSeq ™ Inherited Disease Panel to perform complete ATM gene sequencing in a single member of each family. Then we compared the experimentally determined genotype with the affected/normal phenotype distribution in the whole family to provide unambiguous evidence of pathogenic mutations responsible for A-T. A single novel SNP was allocated to each family. In the first case, we found a mononucleotide deletion, and in the second, a mononucleotide insertion. Both mutations lead to truncation of the ATM protein product. Identification of the pathogenic mutation in each family was performed in a timely fashion, allowing the fetuses to be tested and diagnosed. The parents chose to continue with both pregnancies as both fetuses had a healthy genotype and thus were not at risk of A-T

New NIR-emissive tetranuclear Er(III) complexes with 4-hydroxo-2,1,3-benzothiadiazolate and dibenzoylmethanide ligands: Synthesis and characterization

© The Royal Society of Chemistry 2015. New tetranuclear heteroleptic complexes [Er4(dbm)6(O-btd)4(OH)2] (1) and [Er4(dbm)4(O-btd)6(OH)2] (2) (O-btd = 4-hydroxo-2,1,3-benzothiadiazolate and dbm = dibenzoylmethanide) and their solvates with toluene, THF and CH2Cl2 were prepared using two synthetic approaches. The structures of the products were confirmed by single-crystal X-ray diffraction. Magnetic properties of 1 and 2 are in good agreement with X-ray data. The effective magnetic moment (μeff) values at 300 K for 1 and 2 corresponds to a system of 4 non-interacting Er(iii) ions in the ground state 4J15/2 with g = 6/5. At ambient temperature and upon excitation with λexc = 450 nm, complexes 1 and 2 exhibit luminescence at ∼1530 nm, i.e. in the near infra-red (NIR) region. The luminescence intensity grows with increasing the number of the (O-btd)- ligands in the complexes. This observation suggests (O-btd)- as a new efficient antenna ligand for the lanthanide-based NIR luminescence

Synthesis, luminescent and magnetic properties of new tetranuclear lanthanide complexes with 4-hydroxy-2,1,3-benzothiadiazolate and dibenzoylmethanide ligands

© 2017 Elsevier LtdFour new tetranuclear complexes, [Ln4(dbm)4(O-btd)6(OH)2] 1a (Ln = Yb), 1b (Ln = Dy) and [Ln4(dbm)6(O-btd)4(OH)2] 2a (Ln = Yb), 2b (Ln = Dy) (O-btd = 4-hydroxo-2,1,3-benzothiadiazolate and dbm = dibenzoylmethanide), and their solvates with toluene and tetrahydrofuran were synthesized. The magnetic properties of the complexes 1a, 1b, 2a and 2b have been studied, revealing different magnetic susceptibility patterns for the Yb(III) and Dy(III) complexes in the temperature range 80–300 K. Upon lowering the temperature, a weak antiferromagnetic ordering is observed for both Yb compounds, while the Dy derivatives are assumed to show weak ferromagnetic interactions. The Yb complexes 1a and 2a demonstrate strong NIR luminescence at ∼1000 nm in CH2Cl2 solution upon excitation of the (O-btd)− ligand. This observation suggests the (O-btd)− ligand to be an efficient antenna ligand for lanthanide-based NIR luminescence