Small non-coding RNA transcriptomic profiling in adult and fetal human brain.

Small non-coding RNAs (sncRNAs) make up ~1% of the transcriptome; nevertheless, they play significant roles in regulating cellular processes. Given the complexity of the central nervous system, sncRNAs likely hold particular importance in the human brain. In this study, we provide sncRNA transcriptomic profiles in a range of adult and prenatal brain regions, with a focus on piRNAs, due to their underexplored expression in somatic cells and tissue-specific nature.

Transcriptome analysis of macrophages during Brucella abortus infection clarifies the survival mechanisms of the bacteria.

Brucellosis is a critical zoonotic disease impacting humans and animals globally, causing symptoms like fever and arthritis in humans and reproductive issues in animals. The disease stems from the Brucella genus, adept at evading the immune system and proliferating within host cells. This study explores how Brucella abortus manipulates host cellular mechanisms to sustain infection, focusing on the interaction with murine macrophages over 24 h.

Inhibition of platelet-activating factor (PAF)-induced platelet aggregation by fatty acids from human saliva.

Experiments were undertaken to identify the nature of a previously identified inhibitor of PAF-induced platelet aggregation (PA) in human saliva. Human saliva fractionated by preparative thin layer chromatography (TLC) yielded a fraction that co-migrated with fatty acids (FAs) and inhibited PAF-induced aggregation of platelets. Synthetic FAs tested for their capacities to inhibit 0.

DNA Polymerase and dRP-lyase activities of polymorphic variants of human Pol ι.

Y-family DNA polymerase iota (Pol ι) is involved in DNA damage response and tolerance. Mutations and altered expression level of POLI gene are linked to a higher incidence of cancer. We biochemically characterized five active site polymorphic variants of human Pol ι: R71G (rs3218778), P118L (rs554252419), I236M (rs3218784), E251K (rs3218783) and P365R (rs200852409).

Translesion DNA Synthesis and Reinitiation of DNA Synthesis in Chemotherapy Resistance.

Many chemotherapy drugs block tumor cell division by damaging DNA. DNA polymerases eta (Pol η), iota (Pol ι), kappa (Pol κ), REV1 of the Y-family and zeta (Pol ζ) of the B-family efficiently incorporate nucleotides opposite a number of DNA lesions during translesion DNA synthesis. Primase-polymerase PrimPol and the Pol α-primase complex reinitiate DNA synthesis downstream of the damaged sites using their DNA primase activity.

Translesion DNA Synthesis and Carcinogenesis.

Tens of thousands of DNA lesions are formed in mammalian cells each day. DNA translesion synthesis is the main mechanism of cell defense against unrepaired DNA lesions. DNA polymerases iota (Pol ι), eta (Pol η), kappa (Pol κ), and zeta (Pol ζ) have active sites that are less stringent toward the DNA template structure and efficiently incorporate nucleotides opposite DNA lesions.

BER gene polymorphisms associated with key molecular events in bladder cancer.

Aim: Base excision repair (BER) gene polymorphisms are known to play an independent role in predisposition to developing different cancers as well as to be associated with clinicopathological traits of the disease modifying its clinical outcomes. One of the underlying mechanisms is presumed to include interplay between BER gene polymorphisms and key mutational, epigenetic and chromosomal events in tumor tissues. The present study was aimed at elucidating potential gene-gene interaction and assessing their mutual effects in bladder cancer (BC).

FGFR3 and TP53 mutations in a prospective cohort of Belarusian bladder cancer patients.

Aim: The aim of this study was to determine the frequencies of FGFR3 and TP53 mutations in a prospective cohort of 150 bladder cancer patients and to assess the relationship between their mutational status and clinicopathological variables.

Materials And Methods: The FGFR3 and TP53 mutations were detected by the SNaPshot method and PCR-single-strand conformational polymorphism analysis followed by DNA sequencing.

Results: The activating FGFR3 mutations were found in 71 (47.

DNA-damage response associated with occupational exposure, age and chronic inflammation in workers in the automotive industry.

The evaluation of genome integrity in populations occupationally exposed to combine industrial factors is of medical importance. In the present study, the DNA-damage response was estimated by means of the alkaline comet assay in a sizeable cohort of volunteers recruited among workers in the automotive industry. For this purpose, freshly collected lymphocytes were treated with hydrogen peroxide (100μM, 1min, 4°C) in vitro, and the levels of basal and H(2)O(2)-induced DNA damage, and the kinetics and efficiency of DNA repair were measured during a 180-min interval after exposure.

Biomarkers for genome instability in some genetic disorders: a pilot study.

Context: The study of genome integrity in some genetic disorders has diagnostic and prognostic importance because of the evident relationship between genome instability and both DNA repair deficiencies and cancer predisposition.

Objective: The objective was to compare the chromosomal and DNA damage responses in lymphocytes from patients with Nijmegen breakage syndrome (NBS), Fanconi anemia (FA) and Williams-Beuren syndrome (WBS) to find additional biomarkers of genome instability.

Methods: The cytogenetic approaches were combined with the alkaline Comet assay to estimate genome integrity in cultured or freshly isolated and H(2)O(2)-treated lymphocytes.

Chromosomal instability at the 7q11.23 region impacts on DNA-damage response in lymphocytes from Williams-Beuren syndrome patients.

Williams-Beuren syndrome (WBS) is the chromosomal disorder arising from a hemizygous microdeletion at 7q11.23. The present study was focused on a comparative investigation of genomic integrity in WBS patients by use of cytogenetic methods and the alkaline comet assay.

Pharmacokinetics of monepantel and its sulfone metabolite, monepantel sulfone, after intravenous and oral administration in sheep.

The pharmacokinetic properties of the developmental Amino-Acetonitrile Derivative (AAD), monepantel and its sulfone metabolite, monepantel sulfone were investigated in sheep following intravenous (i.v.) and oral administrations.

Activation and cytotoxicity of 2-alpha-aminoacyl prodrugs of methotrexate.

In an effort to improve the selectivity of the anticancer drug methotrexate (MTX), a series of potential prodrugs in which the 2-amino group was acylated with various alpha-amino acids (as well as L-pyroglutamic acid) was synthesized. Such derivatives are anticipated to be hydrolysed to MTX by appropriate aminopeptidases localized (over-expressed naturally or targeted as anti-tumor antibody conjugates) in the vicinity of the tumor. The L-leucyl, L-valyl, L-isoleucyl, D-alanyl and L-pyroglutamyl derivatives were assessed as to their suitability as prodrugs.

Inhibition of platelet aggregation by diterpene acids from Pinus massoniana resin.

The acidic fraction of the resin of Pinus massoniana Lamb. from China was converted to the p-nitrophenyl esters, and the esters separated by chromatography. The separated p-nitrophenyl esters were individually hydrolysed by potassium hydroxide in acetone-water at room temperature to 8 diterpene acids of the pimarane and abietane groups: pimaric acid (8(14),15-pimaradien-18-oic acid) (1), levopimaric acid (8(14),12-abietadien-18-oic acid) (2), palustric acid (8,13-abietadien-18-oic acid) (3), neobietic acid (8(14),13(15)-abietadien-18-oic acid) (4), abietic acid (7,13-abietadien-18-oic acid) (5), dehydroabietic acid (8,11,13-abietatrien-18-oic acid) (6), 7-oxodehydroabietic acid (7-oxo-8,11,13-abietatrien-18-oic acid) (7) and 7 alpha-hydroxydehydroabietic acid (7 alpha-hydroxy-8,11,13-abietatrien-18-oic acid) (8).

Measurement of PAF in blood by radioimmunoassay. Examination of interfering factors and problems involved in accurate quantification.

A recently developed radioimmunoassay for PAF was applied to blood extracts with the aims of defining and overcoming problems that lead to erroneous results and establishing optimum conditions for the accurate determination of PAF levels. The high lipid content of blood was found to interfere with the assay, and it appeared that phosphatidylcholine and/or sphingomyelin might be among the lipids responsible. Interference was eliminated by either dilution or preparative TLC of blood extract prior to RIA although dilution is unlikely to be generally useful due to the low amounts of PAF normally present in human blood.

Inhibitor(s) of platelet-activating factor (PAF) in human saliva.

Quantitation of platelet-activating factor (PAF) in human saliva samples by radioimmunoassay indicated there was, at times, sufficient PAF present to aggregate platelets. However, in certain samples, we observed little or no aggregation, and furthermore, these samples were found to inhibit aggregation induced by PAF (200 pg). Chromatographic fractionation of pooled saliva increased the PAF activity 4-fold, and the observed inhibitory activity was found to co-migrate with the fatty acids.

Quantitation by radioimmunoassay of PAF in human saliva.

Approximately 75% of the PAF present in saliva is recovered on extraction of whole saliva (0.8 vol) with chloroform/methanol/water (2:2:1, v/v/v). PAF levels, determined by our recently developed radioimmunoassay, in saliva extracts ranged from 0.

A specific, sensitive and high-capacity immunoassay for PAF.

A specific radioimmunoassay for platelet-activating factor (PAF) sensitive in the range 10-1000 pg (0.02-2 pmoles) has been developed. Detailed quantitative hapten inhibition studies showed specificity for the acetyl group at C-2 of PAF, a requirement for the ether linkage at C-1 and some tolerance for substituents on the choline nitrogen.

Synthesis of a PAF immunogen and production of PAF-specific antibodies.

Platelet activating factor (PAF), a naturally occurring phospholipid with many potent physiological and pharmacological activities, is implicated as a mediator of many diseases. An immunoassay for PAF would greatly improve quantitation, and hence PAF-specific antibodies were required. Chemically-reactive analogs of PAF, containing an aldehyde group at the end of the 1-O-alkyl chain (hexyl or dodecyl), were synthesized from readily available materials.

Antibodies to platelet-activating factor (PAF) inhibit PAF-induced platelet aggregation.

Platelet-activating factor (PAF) is the most potent platelet agonist known. PAF-induced platelet aggregation was blocked by pre-incubation of PAF with the immunoglobulin fraction from sheep or rabbit anti-PAF anti-serum. Inhibition was specific for PAF and was dependent upon immunoglobulin and PAF concentrations.

Stability of platelet activating factor (PAF) in human saliva. Quantitation by radioimmunoassay.

Platelet activating factor (PAF) is thought to mediate many inflammatory processes and its involvement in health and disease may be clarified by examining PAF levels in human secretions. The known presence of PAF, the ease of obtaining samples and the relative stability of PAF in saliva, makes this fluid a preferred source for examination of PAF in health and disease. The activity of PAF-acetylhydrolase (the PAF degrading enzyme) in saliva was 1,000-fold lower than that found in human plasma.

Examination for platelet-activating factor production by preimplantation mouse embryos using a specific radioimmunoassay.

A specific and highly sensitive radioimmunoassay was used to measure platelet-activating factor (PAF) production by preimplantation mouse embryos in vitro. Levels of PAF greater than 1 pg per embryo were not observed in 24-h culture medium from 2-cell embryos, compacted morulae or blastocysts, or in extracts from these embryos. Synthetic PAF added to embryos at the start of culture could be almost totally recovered after the incubation period, indicating negligible degradation of PAF during culture.

The specificity of the binding of platelet activating factor (PAF) to anti-PAF antibodies.

Quantitative hapten inhibition experiments employing sheep anti-PAF antibodies and selected PAF analogues were undertaken with the aim of defining the antigenic determinant structures complementary to the antibody combining sites. The most important fine structural features for inhibition of antibody to PAF were shown to be an acetyl group at position 2 of the phospholipid glycerol backbone and an ether group at position 1. Of the naturally occurring compounds, C16- and C18:1-PAF proved to be the most potent inhibitors and more active than C18-PAF while phospholipids with a propionyl, butyryl or hexanoyl group at position 2 showed either weak or no inhibitory activity.

Combining site specificities of rabbit antibodies to platelet-activating factor (PAF).

Anti-PAF sera from six different rabbits, immunized with C12- or C6-PAF as immunogen, were examined in hapten inhibition experiments in an attempt to define the fine structural recognition specificities of the antibody combining sites. Using a selection of naturally occurring lipids and PAF analogues, no significant cross-reactivity was observed with the lipids or with the inactive metabolite, lyso-PAF. Comparison of the structural specificity requirements of the antibodies from each rabbit showed some heterogeneity, with one antiserum demonstrating a different recognition specificity at position 1 on the glycerol backbone of the PAF molecule.