Biological activities of the LXRα and β agonist, 4β-hydroxycholesterol,and of its isomer, 4α-hydroxycholesterol,on oligodendrocytes: Effects on cell growth and viability,oxidative and inflammatory status

The biochemical and biological properties of 4β-hydroxycholesterol and of its isomer, 4α-hydroxycholesterol, are not well known. So, we determined the ability of 4α- and 4β-hydroxycholesterol to react with LXRα and LXRβ, and we characterized the activities of these oxysterols on oligodendrocytes which are myelin synthesizing cells. The effects of 4α- and 4β-hydroxycholesterol were studied on 158N murine oligodendrocytes to assess their activities on cell growth and viability, oxidative and inflammatory status. To this end different parameters were used: cell counting with trypan blue; identification of dead cells and cell cycle analysis with propidium iodide; evaluation of mitochondrial depolarization, lysosomal membrane integrity, actin depolimerization, nuclear morphology, and superoxide anion production after staining with JC-1, acridine orange, rhodamine-phalloidin, Hoechst 33342, and dihydroethidium, respectively; evaluation of ultrastructural changes by transmission electron microscopy, and cytokine quantification with a cytometric bead array. Only 4β-hydroxycholesterol is a LXRα and β agonist. No cytotoxic effects were found with 4α-hydroxycholesterol except a slight inhibition of cell growth at elevated concentrations. At high concentrations, 4β-hydroxycholesterol was not only able to inhibit cell growth, but also to induce cell death associated with a loss of mitochondrial transmembrane potential, dysfunctions of lysosomal membrane integrity, and superoxide anion overproduction. These side effects were lower than those observed with 7-ketocholesterol and 25-hydroxycholesterol used as positive controls. On oligodendrocyte murine primary cultures, only lysosomal membrane integrity was slightly affected under treatment with 4α- and 4β-hydroxycholesterol. So, 4α- and 4β-hydroxycholesterol have different biological activities. Their ability to induce cytotoxic effects on oligodendrocytes can be considered as weak comparatively to 7-ketocholesterol and 25-hydroxycholesterol.     

Computational Analysis of AMPK-Mediated Neuroprotection Suggests Acute Excitotoxic Bioenergetics and Glucose Dynamics Are Regulated by a Minimal Set of Critical Reactions

Loss of ionic homeostasis during excitotoxic stress depletes ATP levels and activates the AMP-activated protein kinase (AMPK), re-establishing energy production by increased expression of glucose transporters on the plasma membrane. Here, we develop a computational model to test whether this AMPK-mediated glucose import can rapidly restore ATP levels following a transient excitotoxic insult. We demonstrate that a highly compact model, comprising a minimal set of critical reactions, can closely resemble the rapid dynamics and cell-to-cell heterogeneity of ATP levels and AMPK activity, as confirmed by single-cell fluorescence microscopy in rat primary cerebellar neurons exposed to glutamate excitotoxicity. The model further correctly predicted an excitotoxicity-induced elevation of intracellular glucose, and well resembled the delayed recovery and cell-to-cell heterogeneity of experimentally measured glucose dynamics. The model also predicted necrotic bioenergetic collapse and altered calcium dynamics following more severe excitotoxic insults. In conclusion, our data suggest that a minimal set of critical reactions may determine the acute bioenergetic response to transient excitotoxicity and that an AMPK-mediated increase in intracellular glucose may be sufficient to rapidly recover ATP levels following an excitotoxic insult.     

ANTIMICROBIAL AND ANTIHYPERLIPIDEMIC ACTIVITIES OF ISOLATED QUERCETIN FROM ANABAENA AEQUALIS1

This study aimed to isolate quercetin (for the first time) from Anabaena aequalis Borge, which inhabits soil surface of Wadi El‐Alaqui Protectorate located in Aswan city, Egypt. The isolated compound showed significant antibacterial activity against the gram‐positive bacteria Sarcina maxima and Micrococcus kristinae, the gram‐negative bacterium Klebsiella pneumoniae, as well as against the filamentous fungus Aspergillus flavus. The isolated compound was identified as quercetin using the structure elucidation based on UV, electrospray ionization mass spectrometry (ESIMS), ¹H and ¹³C NMR, proton–proton correlation spectroscopy (¹H‐¹H COSY), distortionless enhancement by polarization transfer (DEPT), heteronuclear single quantum coherence (HSQC), and heteronuclear multiple bond correlations spectrum (HMBC). Medium lethal dose (LD₅₀) of the isolated compound and its side effects against hyperlipidemia induced by ethanol intake in albino rats were carried out. No deaths were reported in rats within 72 h, which suggests that the isolated compound plays a beneficial role as an antihyperlipidemic agent in the treatment of alcohol‐induced hepatic tissue damage, which can be described as one of the therapeutic values.     

Localizing confined epileptic foci in patients with an unclear focus or presumed multifocality using a component-based EEG-fMRI method

Precise localization of epileptic foci is an unavoidable prerequisite in epilepsy surgery. Simultaneous EEG-fMRI recording has recently created new horizons to locate foci in patients with epilepsy and, in comparison with single-modality methods, has yielded more promising results although it is still subject to limitations such as lack of access to information between interictal events. This study assesses its potential added value in the presurgical evaluation of patients with complex source localization. Adult candidates considered ineligible for surgery on account of an unclear focus and/or presumed multifocality on the basis of EEG underwent EEG-fMRI. Adopting a component-based approach, this study attempts to identify the neural behavior of the epileptic generators and detect the components-of-interest which will later be used as input in the GLM model, substituting the classical linear regressor. Twenty-eight sets interictal epileptiform discharges (IED) from nine patients were analyzed. In eight patients, at least one BOLD response was significant, positive and topographically related to the IEDs. These patients were rejected for surgery because of an unclear focus in four, presumed multifocality in three, and a combination of the two conditions in two. Component-based EEG-fMRI improved localization in five out of six patients with unclear foci. In patients with presumed multifocality, component-based EEG-fMRI advocated one of the foci in five patients and confirmed multifocality in one of the patients. In seven patients, component-based EEG-fMRI opened new prospects for surgery and in two of these patients, intracranial EEG supported the EEG-fMRI results. In these complex cases, component-based EEG-fMRI either improved source localization or corroborated a negative decision regarding surgical candidacy. As supported by the statistical findings, the developed EEG-fMRI method leads to a more realistic estimation of localization compared to the conventional EEG-fMRI approach, making it a tool of high value in pre-surgical evaluation of patients with refractory epilepsy. To ensure proper implementation, we have included guidelines for the application of component-based EEG-fMRI in clinical practice.     

Genetic structure and functioning of alien ship rat populations from a Corsican micro-insular complex

The detrimental effects of the introduced ship rat on bird species on the Lavezzi Mediterranean archipelago has led to the decision to eradicate the rodent from the main island, Lavezzu (73 ha), as well as from several neighbouring islets. A genetic study using eight microsatellite markers has revealed some of the dynamics of this rat population. First, it has been shown that the rat population was genetically stable (no departure from Hardy-Weinberg equilibrium and no linkage disequilibrium) and as suggested by paleontological data, established for a long time. This information is encouraging in term of viability of the eradication campaign in the long term, even if rare immigration events cannot be excluded. Second, this study shows that rats are likely to swim between the main island and the islets quite regularly since no clear genetic differentiation has been detected between them. This result is quite surprising since the ship rat is not known for its swimming abilities. Third, a cryptic genetic structure has been detected on the main island, with the north peninsula differentiated from the rest. This result correlates the observation of particular predation behaviours only observed in this part of the island. Finally, evolutionary hypotheses (e.g., dispersal limitation, emergence of family groups, local adaptation) are discussed to explain the genetic patterns observed and the population functioning inferred. These results should be of particular interest to wildlife managers concerned with rat eradications, and also provide clear insights into the study of other biological invasions.     

Tomato (Solanum lycopersicum) growth and fruit quality affected by organic fertilization and ozonated water

Protoplasma - The use of modern and safe techniques to increase plant growth and yield is of significance. There is little data, to our knowledge, on the use of organic fertilization and ozonated...     

Cloning and Characterization of a Plasmid Encoded ACC Deaminase from an Indigenous Pseudomonas fluorescens FY32

In addition to the characterized mechanisms responsible for many direct effects of plant growth promoting bacteria (PGPB) on plants, it has been suggested that a number of PGPB contain the enzyme ACC deaminase that catalyzes degradation of 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, into α-ketobutyrate and ammonia. As part of an effort to obtain an ACC deaminase encoding gene from a collection of soil samples, only one bacterial isolate, Pseudomonas fluorescens FY32 was capable of growing on ACC as a sole source of nitrogen. The ACC deaminase gene was amplified from the above isolate by polymerase chain reaction (PCR) giving an expected DNA fragment, 1017 bp. Sequence analysis of the fragment showed that it was highly homologous (94% and 98% identities at nucleotide and amino acid levels, respectively) to the previously characterized acdS gene from Pseudomonas sp. 6G5. Furthermore, fusion of the ACC deaminase ORF with lacZ gene resulted in the expression of active enzyme in Escherichia coli. In addition, further analyses revealed that the acdS gene was plasmid-encoded so that a large plasmid (pFY32) with almost 50 kb in size was identified from this bacterium. Furthermore, transfer of pFY32 into E. coli DH5α proved its ACC deaminase activity. This result was in accordance with previous reports suggesting horizontal transfer of the acdS gene. However, it needs more investigation to identify whether this pFY32 plasmid has undergone lateral gene transfer during the evolutionary process.     

Quantifying the HIV-1 integrase inhibitor raltegravir in female genital tract secretions using high-performance liquid chromatography with ultraviolet detection

Understanding the pharmacokinetics of drugs in peripheral body compartments, such as the genital tract, is particularly important in the infectious diseases arena. However, extracting drugs from small volumes of viscous, proteinacious substances like cervicovaginal fluid is particularly challenging. The goal of this study was to develop a method to quantify raltegravir, an HIV-1 integrase inhibitor, in the female genital tract. The method included sample preparation with perchloric acid followed by solid-phase extraction, separation with reverse-phase high-performance liquid chromatography, and detection with an ultraviolet wavelength of 218 nm. The method was linear from 0.05 to 10.0 mg/L, with minimal endogenous interference. The method was accurate (1.2–11.0% deviation) and precise (1.1–12.6% CV) for both within and between-day analyses. The ability to detect raltegravir in the female genital tract is essential for future investigations of raltegravir as an agent for prevention of HIV acquisition, and this method will be used for clinical studies further evaluating pharmacokinetic–pharmacodynamic relationships in this body compartment.     

Effects of Graded Inclusion of Bioactive Peptides Derived from Sesame Meal on the Growth Performance,Internal Organs,Gut Microbiota and Intestinal Morphology of Broiler Chickens

International Journal of Peptide Research and Therapeutics - The aim of this experiment was to evaluate the effects of bioactive peptides derived from sesame meal (BPSM) compared with...