MicroRNA and mRNA Expression Changes in Glioblastoma Cells Cultivated under Conditions of Neurosphere Formation.

Glioblastoma multiforme (GBM) is one of the most highly metastatic cancers. The study of the pathogenesis of GBM, as well as the development of targeted oncolytic drugs, require the use of actual cell models, in particular, the use of 3D cultures or neurospheres (NS). During the formation of NS, the adaptive molecular landscape of the transcriptome, which includes various regulatory RNAs, changes.

Transcriptome Changes in Glioma Cells Cultivated under Conditions of Neurosphere Formation.

Glioma is the most common and heterogeneous primary brain tumor. The development of a new relevant preclinical models is necessary. As research moves from cultures of adherent gliomas to a more relevant model, neurospheres, it is necessary to understand the changes that cells undergo at the transcriptome level.

Are Small Nucleolar RNAs "CRISPRable"? A Report on Box C/D Small Nucleolar RNA Editing in Human Cells.

CRISPR technologies are nowadays widely used for targeted knockout of numerous protein-coding genes and for the study of various processes and metabolic pathways in human cells. Most attention in the genome editing field is now focused on the cleavage of protein-coding genes or genes encoding long non-coding RNAs (lncRNAs), while the studies on targeted knockout of intron-encoded regulatory RNAs are sparse. Small nucleolar RNAs (snoRNAs) present a class of non-coding RNAs encoded within the introns of various host genes and involved in post-transcriptional maturation of ribosomal RNAs (rRNAs) in eukaryotic cells.

Characterization of primary normal and malignant breast cancer cell and their response to chemotherapy and immunostimulatory agents.

Background: The phenomenon of chemotherapy-resistant cancers has necessitated the development of new therapeutics as well as the identification of specific prognostic markers to predict the response to novel drugs. Primary cancer cells provide a model to study the multiplicity of tumourigenic transformation, to investigate alterations of the cellular response to various molecular stimuli, and to test therapeutics for cancer treatment.

Methods: Here, we developed primary cultures of human breast tissue - normal cells (BN1), cancer cells (BC5), and cells from a chemotherapy-treated tumour (BrCCh1) to compare their response to conventional chemotherapeutics and to innate immunity stimulators with that of the immortalized breast cells MCF7, MDA-MB-231, and MCF10A.

Variety of RNAs in Peripheral Blood Cells, Plasma, and Plasma Fractions.

Human peripheral blood contains RNA in cells and in extracellular membrane vesicles, microvesicles and exosomes, as well as in cell-free ribonucleoproteins. Circulating mRNAs and noncoding RNAs, being internalized, possess the ability to modulate vital processes in recipient cells. In this study, with SOLiD sequencing technology, we performed identification, classification, and quantification of RNAs from blood fractions: cells, plasma, plasma vesicles pelleted at 16,000 and 160,000, and vesicle-depleted plasma supernatant of healthy donors and non-small cell lung cancer (NSCLC) patients.

Sensitivity of endometrial cancer cells from primary human tumor samples to new potential anticancer peptide lactaptin.

Purpose: Endometrial carcinoma is the most common gynecologic malignancy which is associated with a poor prognosis when diagnosed at an advanced stage; therefore, the discovery of efficacious new drugs is required to reinforce conventional chemotherapy. Short-term cultures of primary cells from endometrial tumors could be used for testing new anticancer therapeutics as well as for the development of personalized cancer therapy strategy. Here, the antitumor effect of a recombinant analogue of lactaptin (RL2), a new potential anticancer molecule, was examined against primary human endometrial cancer cells.

Regulatory role of small nucleolar RNAs in human diseases.

Small nucleolar RNAs (snoRNAs) are appreciable players in gene expression regulation in human cells. The canonical function of box C/D and box H/ACA snoRNAs is posttranscriptional modification of ribosomal RNAs (rRNAs), namely, 2'-O-methylation and pseudouridylation, respectively. A series of independent studies demonstrated that snoRNAs, as well as other noncoding RNAs, serve as the source of various short regulatory RNAs.

Lactaptin induces p53-independent cell death associated with features of apoptosis and autophagy and delays growth of breast cancer cells in mouse xenografts.

Lactaptin, the proteolytic fragment of human milk kappa-casein, induces the death of various cultured cancer cells. The mechanisms leading to cell death after lactaptin treatment have not been well characterized. In this study the in vivo and in vitro effects of a recombinant analogue of lactaptin (RL2) were examined.

Artificial box C/D RNAs affect pre-mRNA maturation in human cells.

Box C/D small nucleolar RNAs (snoRNAs) are known to guide the 2'-O-ribose methylation of nucleotides in eukaryotic ribosomal RNAs and small nuclear RNAs. Recently snoRNAs are predicted to regulate posttranscriptional modifications of pre-mRNA. To expand understanding of the role of snoRNAs in control of gene expression, in this study we tested the ability of artificial box C/D RNAs to affect the maturation of target pre-mRNA.

A novel pro-apoptotic effector lactaptin inhibits tumor growth in mice models.

Lactaptin, a human milk-derived protein, induces apoptosis in cultured tumor cells. We designed a recombinant analog of lactaptin (RL2) and tested its antitumor activity. The sensitivity of hepatocarcinoma A-1 (HA-1), Lewis lung carcinoma, and Ehrlich carcinoma to RL2 were tested to determine the most reliable in vitro animal model.

Unbiased approach to profile the variety of small non-coding RNA of human blood plasma with massively parallel sequencing technology.

Objective: Understanding structures of circulating RNA expands fundamental knowledge of cell communications and signaling pathways as well as allows developing new molecular diagnostic approaches. The aim of this study was to deploy a new approach to sequencing cDNA library construction which expands the capabilities of high-throughput sequencing analysis of small non-coding RNAs. With the approach, we performed massively parallel sequencing of human blood plasma RNA to document profile of common and peculiar RNA species normally circulating in blood of healthy individuals.

Radionuclide transfer to reptiles.

Reptiles are an important, and often protected, component of many ecosystems but have rarely been fully considered within ecological risk assessments (ERA) due to a paucity of data on contaminant uptake and effects. This paper presents a meta-analysis of literature-derived environmental media (soil and water) to whole-body concentration ratios (CRs) for predicting the transfer of 35 elements (Am, As, B, Ba, Ca, Cd, Ce, Cm, Co, Cr, Cs, Cu, Fe, Hg, K, La, Mg, Mn, Mo, Na, Ni, Pb, Po, Pu, Ra, Rb, Sb, Se, Sr, Th, U, V, Y, Zn, Zr) to reptiles in freshwater ecosystems and 15 elements (Am, C, Cs, Cu, K, Mn, Ni, Pb, Po, Pu, Sr, Tc, Th, U, Zn) to reptiles in terrestrial ecosystems. These reptile CRs are compared with CRs for other vertebrate groups.

Characterisation of circulating DNA by parallel tagged sequencing on the 454 platform.

Background: Fragments of genomic DNA that can be isolated from the blood and body fluids of vertebrates are also known as circulating DNA. This DNA has widely been investigated as a biomarker for cancer and other diseases but the origin and significance of circulating DNA have not been elucidated to date.

Methods: We used a parallel tagged sequencing method to sequence circulating DNA obtained from control individuals as well as cancer patients on the GSFLX sequencer (454 life sciences).

Fragments of noncoding RNA in plasma of human blood.

Mammalian extracellular fluids, such as plasma of blood or plasma of milk, contain cell-free RNAs. Fragments of ubiquitously expressed mRNAs as well as tumor-specific and viral RNAs have been previously revealed in human plasmas by template-defined RT-PCR. In the present work we aimed to detect major forms of human blood plasma RNAs or to reveal new forms by using two approaches which were independent of context of RNA templates.

Splicing by exon exclusion impaired by artificial box c/d RNA targeted to branch-point adenosine.

Box C/D small nucleolar RNAs guide the site-specific 2'-O-ribose methylation of nucleotides in target rRNAs and snRNAs. In this study we used the ability of C/D box snoRNAs to guide modification of target RNAs with the aim of affecting the expression of predetermined human genes. We constructed an analogue of the human U24 box C/D RNA, the first antisense element of which was directed to induce 2'-O-ribose methylation of branch-point adenosine in the intron of the human heat-shock cognate protein (HSC8) pre-mRNA.

Deamination of adenosines in extracellular RNA spontaneously internalized by human cells.

Ribonucleic acids circulating in mammalian extracellular fluids as well as RNAs accumulating in culture medium condensed by mammalian cells are internalized by acceptor cells and distributed among cellular compartments. The internalized RNA can be involved in the induction and regulation of cellular processes as guide or signaling molecules. The internalization of RNA may be accompanied by covalent modifications influencing the stability and functionality of this RNA.

Accuracy and resolution of a dynamic-speckle profilometer.

We propose and demonstrate a new technique for fast noncontact and continuous profile measurement of a rough surface. The technique is based on frequency tracking of the power modulation of spatially filtered scattered light. A dynamic speckle pattern is created when the laser beam scans the surface under study.

Dynamic speckle effect induced by an acousto-optic deflector for fast range sensing.

A technique for fast distance measurements based on continuous frequency measurements of the power modulation of spatially filtered scattered light is proposed. For what is to our knowledge the first time, it is shown that the technique works when laser beam scanning is performed with an acousto-optic deflector. The most impressive feature of the proposed technique is that it works at very high scanning speed, providing an extremely fast response time.

Fast distance measurements by use of dynamic speckles.

A technique for fast measurements of the distance to an object's surface based on spatial filtering of dynamic speckle patterns is proposed. Exploitation of two spatial filters (Ronchi rulings) enables measurements to be independent of surface speed. Experimental verification of the technique is demonstrated at speeds of the surface as high as 50 m/s.

Ribonucleic acids of human milk.

The milk feeding is the most essential process laying the foundation of human health at the postnatal development. However little is known about nucleic acids secreted into mother's milk during lactation. In order to investigate the composition and abundance of human milk NA we adapted the conventional isolation method to achieve high yield of total nucleic acids from milk samples.

Extracellular ribonucleic acids of human milk.

Human milk has been shown to contain heterogeneous oligoribonucleotides varying in size from dimers to 100 mers. The sets of long oligonucleotides in milk samples from different donors and from different stages of lactation have some conservative elements. Sequences of some RNA oligonucleotides correspond to the 3'-part of 5.

Catalytic nucleotide-hydrolyzing antibodies in milk and serum of clinically healthy human mothers.

Background: In humans, pregnancy and lactation are associated with the production of catalytically active antibodies (abzymes) in serum and breast milk. However, the substrate specificities of the abzymes in these biological fluids, particularly breast milk, have not been studied

Material/methods: IgG fractions were isolated from human milk by subsequent steps of chromatographic purification on Protein-A Sepharose, DEAE-cellulose, and anti-IgG Sepharose. The nucleotide-hydrolyzing activity of electrophoretically homogeneous IgG antibodies was measured using 32P-labeled nucleotides and TLC.

Multiple enzymic activities of human milk lactoferrin.

Lactoferrin (LF) is a Fe3+-binding glycoprotein, first recognized in milk and then in other human epithelial secretions and barrier fluids. Many different functions have been attributed to LF, including protection from iron-induced lipid peroxidation, immunomodulation and cell growth regulation, DNA binding, and transcriptional activation. Its physiological role is still unclear, but it has been suggested to be responsible for primary defense against microbial and viral infection.

Phosphorylation of Minor Lipids of Human Milk Tightly Bound to Secretory Immunoglobulin A.

Catalytic antibodies or abzymes possessing the different catalytic activities were detected in the sera of patients with various autoimmune pathologies, where their presence is most probably associated with autoimmunization. Normal humans are generally considered to have no abzymes, since no obvious immunizing factors are present. The ability of small fraction of sIgA from human milk to phosphorylate selectively casein in the presence of gamma(32)P-ATP was previously shown to be a property of the Abs.