Suppressive effect of exogenous miR-16 and miR-34a on tumorigenesis of breast cancer cells

Recent investigations have shown tumor-suppressive roles for miR-16 and miR-34a. They also share some features in regard to targeting cancer cell signaling pathways which they control. Therefore, in this study, we aimed to further scrutinize whether exogenous induction of mature miR-34a and miR-16 can collaborate in breast tumor suppression. MDA-MB-231 and SK-BR-3 human breast cancer cell lines were cultured and transfected twice with hsa-miR-16-5p and hsa-miR-34a-5p mimics individually or in combination. The cells were analyzed for apoptosis rate and cell cycle indices by flow cytometry. Also, the expression of several invasion and the epithelial-mesenchymal transition markers was evaluated at gene and protein levels by quantitative real-time polymerase chain reaction and western blot analysis, respectively. Assessment of invasiveness and migratory potential of the transfected cells was performed using three-dimensional spheroid formation and wound-healing assay, respectively. In both cell lines, miR-16 and miR-34a induced apoptosis and cell-cycle arrest and also suppressed invasion and migration. Some of these effects, like cell-cycle arrest and induction of apoptosis, were significantly higher when using both microRNAs than when using them individually for transfection of the cells. Our results are indicating that miR-16 and miR-34a can collaborate in breast tumor suppression.     

The combined effect of water deficit stress and TiO2 nanoparticles on cell membrane and antioxidant enzymes in Helianthus annuus L.

Nanotechnology has become one of the several approaches attempting to ameliorate the severe effect of drought on plant''s production and to increase the plants tolerance against water deficit for the water economy. In this research, the effect of foliar application of TiO₂, nanoparticles or ordinary TiO₂, on Helianthus annuus subjected to different levels of water deficit was studied. Cell membrane injury increased by increasing the level of water deficit and TiO₂ concentration, and both types of TiO₂ affected the leaves in analogous manner. Ord-TiO₂ increased H₂O₂ generation by 67–240% and lipid peroxidation by 4–67% in leaves. These increases were more than that induced by Nano-TiO₂ and the effect was concentration dependent. Proline significantly increased in leaves by water deficit stress, reaching at 25% field capacity (FC) to more than fivefold compared to that in plants grown on full FC. Spraying plants with water significantly decreased the activities of enzymes in the water deficit stressed roots. The water deficit stress exerted the highest magnitude of effect on the changes of cell membrane injury, MDA, proline content, and activities of CAT and GPX. Nano-TiO₂ was having the highest effect on contents of H₂O₂ and GPX activity. In roots, the level of water deficit causes highest effect on enzyme activities, but TiO₂ influenced more on the changes of MDA and H₂O₂ contents. GPX activity increased by 283% in leaves of plants treated with 50 and 150 ppm Nano-TiO₂, while increased by 170% in those treated with Ord-TiO₂, but APX and CAT activities increased by 17–197%, in average, with Ord-TiO₂. This study concluded that Nano-TiO₂ didn’t ameliorate the effects of drought stress on H. annuus but additively increased the stress, so its use in nano-phytotechnology mustn’t be expanded without extensive studies.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-022-01153-z.     

Liquid chromatographic assay for a butenolide endothelin antagonist (PD 156707) in plasma

A sensitive and selective liquid chromatographic assay for determining the non-peptide endothelin A receptor antagonist PD 156707 (I) in rat plasma has been developed and validated. The analyte was isolated from matrix by solid-phase extraction. Liquid chromatographic separation was achieved isocratically ona 3.2 mm I.D., ODS column with a mobile hase of acetonitrile-ammonium phosphate (50 mM, pH 3.5) (44:56, v/v). Column effluent was monitored fluorometrically. Peak-height ratios (analyte/IS) were proportional to I concentrations in rat plasma from 25 to 1000 ng/ml. Assay precision and accuracy for I, based on quality controls, was 9.5% relative standard deviation, with relative error of ±6.5%. The quantitation limit was 25 ng/ml for a 200-μl sample aliquot.     

Comparative studies of chloroplastic and nuclear DNA repair abilities after ultraviolet irradiation of Euglena gracilis

Summary Studies of nuclear and chloroplastic-DNA repair after ultraviolet irradiation of Euglena gracilis show that photoreactivation is very efficient at both the nuclear and chloroplastic level. Liquid-holding or split-dose experiments and treatment with caffeine reveal, furthermore, that dark-repair is very efficient in nuclear DNA but not in chloroplastic DNA (ctDNA). The possibility of a chloroplastic dark-repair of restricted efficiency is discussed.Determination of chloroplastic DNA content by reassociation kinetics indicates that an important degradation follows UV irradiation during liquid holding in the dark.     

The T cell-specific CXC chemokines IP-10, Mig, and I-TAC are expressed by activated human bronchial epithelial cells

Recruitment of activated T cells to mucosal surfaces, such as the airway epithelium, is important in host defense and for the development of inflammatory diseases at these sites. We therefore asked whether the CXC chemokines IFN-induced protein of 10 kDa (IP-10), monokine induced by IFN-gamma (Mig), and IFN-inducible T-cell alpha-chemoattractant (I-TAC), which specifically chemoattract activated T cells by signaling through the chemokine receptor CXCR3, were inducible in respiratory epithelial cells. The effects of proinflammatory cytokines, including IFN-gamma (Th1-type cytokine), Th2-type cytokines (IL-4, IL-10, and IL-13), and dexamethasone were studied in normal human bronchial epithelial cells (NHBEC) and in two human respiratory epithelial cell lines, A549 and BEAS-2B. We found that IFN-gamma, but not TNF-alpha or IL-1 beta, strongly induced IP-10, Mig, and I-TAC mRNA accumulation mainly in NHBEC and that TNF-alpha and IL-1 beta synergized with IFN-gamma induction in all three cell types. High levels of IP-10 protein (> 800 ng/ml) were detected in supernatants of IFN-gamma/TNF-alpha-stimulated NHBEC. Neither dexamethasone nor Th2 cytokines modulated IP-10, Mig, or I-TAC expression. Since IFN-gamma is up-regulated in tuberculosis (TB), using in situ hybridization we studied the expression of IP-10 in the airways of TB patients and found that IP-10 mRNA was expressed in the bronchial epithelium. In addition, IP-10-positive cells obtained by bronchoalveolar lavage were significantly increased in TB patients compared with normal controls. These results show that activated bronchial epithelium is an important source of IP-10, Mig, and I-TAC, which may, in pulmonary diseases such as TB (in which IFN-gamma is highly expressed) play an important role in the recruitment of activated T cells.     

Saprolegniosis in salmonids and their eggs in Japan

An epizootic of the fungal infection saprolegniosis that occurred in freshwater-cultured salmons and their eggs at some hatcheries in Hokkaido (Japan) was investigated. In almost all cases, the initial clinical sign was characterized by the growth of cotton-like mycelia on the fishs' body surface, especially the head, adipose fin, and caudal fin, but the mycelia were not visible to the naked eye in the internal organs. Thirty-three strains isolated from lesions were classified in the genus Saprolegnia according to their morphological and biological characteristics on hemp seed cultures at various temperatures. Fifteen of the strains were identified as Saprolegnia parasitica, 16 were identified as S. salmonis, and two were identified as S. australis.     

Acylated flavonol diglucosides from Lotus polyphyllos

Three acylated flavonol diglucosides, kaempferol 3-O-β-(6″-O-E-p-coumaroylglucoside)-7-O-β-glucoside; quercetin 3-O-β-(6″-O-E-p-coumaroylglucoside)-7-O-β-glucoside; isorhamnetin 3-O-β-(6″-O-E-p-coumaroylglucoside)-7-O-β-glucoside were isolated from the whole plant aqueous alcohol extract of Lotus polyphyllos. The known 3,7-di-O-glucosides of the aglycones kaempferol, quercetin and isorhamnetin were also characterized. All structures were established on the basis of chemical and spectral evidence.     

Synthesis and biological evaluation of some acyclic alpha-(1H-pyrazolo-[3,4-d]pyrimidin-4-yl)thioalkylamide nucleosides

The chemical synthesis of some acyclic alpha-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)thioalkylamide nucleosides (10-12)a-c is described. The treatment of IH-pyrazolo[3,4-d]pyrimidin-4-thione 1 with compounds 2a-c gave, regioselectively, ethyl alpha-(1H-pyrazolo[3,4-d]pyrimidin-4-yl)thioalkylates 3a-c, respectively. These heterocycles were alkylated, separately, with alkylating agents 4, 5 and 6 to give, regioselectively, the N1-acyclic nucleosides (7-9)a-c which were deprotected to afford the desired products (10-12)a-c. All synthetic compounds were characterized on the basis of their physical and spectroscopic properties. The products (10-12)a-c were evaluated for their inhibitory effects against the replication of HIV-1 (III(B)), HIV-2 (ROD), various DNA viruses, a variety of tumor-cell lines and M. tuberculosis. No marked biological activity was found.     

Occurrence and distribution of Fusarium species in maize fields in New Zealand

Fusarium populations were investigated in maize grains and their husks about six weeks before harvest in three maize fields in the Manawatu region of New Zealand. The role of litter and soil as reservoirs for these fungi was also examined. Two techniques were used to examine populations, dilution plating and direct plating. Using the dilution plating technique the highest overall populations were found in husks (mean 2.2 x 10(5)/g) and litter (mean 1.4 x 10(5)/g), while similar lower numbers of viable propagules were obtained from grain (mean 2.1 x 10(3)/g) and soil (2.8 x 10(3)/g). With this technique five Fusarium spp. were commonly isolated; F. graminearum (Gibberella zeae), F. culmorum, F. subglutinans, F. oxysporum and F. acuminatum, of which F. graminearum was the most abundant. With the direct plating technique 87% of grains were infected with Fusarium spp., with some grains being infected with more than one species. Segments from husks and litter, 70% and 43% respectively, were colonised by Fusariumr spp. F. graminearum was the most frequent species isolated from maize grain and husk segments (48.3 and 37.7% colonisation respectively). Other species, particularly F. culmorum and F. acuminatum, were also found to be common contaminants. A total of 15 Fusarium spp. was recovered from all material examined by both techniques. Cultures with characteristics resembling those of F. moniliforme were rarely observed.