Simple micro-method for the isolation of gangliosides by reversed-phase chromatography

A simple and convenient technique has been developed for the isolation of gangliosides from small amounts of tissues or cells. A ganglioside fraction obtained by chromatography of the total lipid extract of DEAE-Sephadex was subjected to alkaline hydrolysis and salts and other non-lipid contaminants were removed by reversed-phase chromatography on a C₁₈ Sep-Pak cartridge. The purified gangliosides were then obtained by chromatography on a small Iatrobeads or Unisil column. This procedure yields a quantitative recovery of gangliosides that are free of contaminants which interfere with thin-layer chromatographic analysis. The procedure was used for the quantitative isolation of gangliosides from human brain white matter and human erythrocytes.     

Structural,photoluminescence, and optical characteristics of a Sm3+-doped lead borate–strontium–tungsten glass system: Evaluation of Judd–Ofelt intensity parameters and radiative properties

In this study, the melt quenching approach is used to synthesize a lead borate–strontium-based glass system doped with samarium ions. Modifications in the glass network structure arising from the addition of various concentrations of Sm³⁺ ions were investigated via Fourier transform infrared (FTIR) spectroscopy. FTIR analysis revealed B–O–B bridges, BO₃, and BO₄ units are present. UV–vis–NIR spectroscopic measurement was performed to study the optical absorption spectra. Optical constants such as optical bandgap energies, refractive indices, and other related parameters were evaluated. The lifetime fluorescence decay was measured and ranged between 1.04 and 1.88 ns. The photoluminescence spectra in the range 500–750 nm revealed four transitions from the ground state ⁶G_5/2 to the excited states ⁶H_5/2, ⁶H_7/2, ⁶H_9/2 and ⁶H_11/2 and J–O theory was utilized to study these optical transitions for Sm³⁺ ions. Calculations of the oscillator strengths and J–O intensity parameters were performed and the obtained J–O parameters followed the sequence Ω₄ > Ω₆ > Ω₂. The ratio O/R indicated a high lattice asymmetry around the samarium ions. The values of lifetimes and branching ratios for the fabricated samples emphasized their suitability to be used in laser applications. The current glass samples are good candidates for orange and red emission devices.     

Platform configuration, microfacies and cyclicities of the upper Albian to Turonian of west-central Jordan

We present a comprehensive facies scheme for west-central Jordan platform deposits of upper Albian to Turonian age, discuss Cenomanian and Turonian carbonate cycles, and reconstruct the paleogeographic evolution of the platform. Comparisons with adjacent shelf areas (Israel, Sinai) emphasize local characteristics as well as the regional platform development. Platform deposits are subdivided into fifteen microfacies types that define eight environments of deposition of three facies belts. Main facies differences between Cenomanian and Turonian platforms are: rudist-bearing packstones that characterise the higher-energy shallow subtidal (transition zone) during the Cenomanian, and fossiliferous (commonly with diverse foraminifer assemblages) wackestones and packstones of an open shallow subtidal environment. On Turonian platforms high-energy environments are predominantly characterised by oolithic or bioclastic grainstones and packstones, whereas peritidal facies are indicated by dolomitic wackestones with thin, wavy (cryptmicrobial) lamination. Rhythmic facies changes define peritidal or subtidal shallowing-up carbonate cycles in several Cenomanian and Turonian platform intervals. Cyclicities are also analysed on the base of accommodation plots (Fischer Plots). High-frequency accommodation changes within lower Cenomanian cyclic bedded limestones of the central and southern area exhibit two major cyclic sets (set I and II) each containing regionally comparable peaks. Accommodation patterns within cyclic set II coincide with the sequence boundary zone of CeJo1. The lateral and vertical facies distributions on the inner shelf allow the reconstruction of paleogeographic conditions during five time intervals (Interval A to E). An increased subsidence is assumed for the central study area, locally (area of Wadi Al Karak) persisting from middle Cenomanian to middle Turonian times. In contrast, inversion and the development of a paleo-high have been postulated for an adjacent area (Wadi Mujib) during late Cenomanian to early Turonian times, while small-scale sub-basins with an occasionally dysoxic facies developed northwards and further south during this time interval. A connection between these structural elements in Jordan with basins and uplift areas in Egypt and Israel during equivalent time intervals is assumed. This emphasises the mostly concordant development of that Levant Platform segment.     

Serum selenium predicts levels of F2-isoprostanes and prostaglandin F2alpha in a 27 year follow-up study of Swedish men

Low concentrations of selenium (Se) predict mortality and cardiovascular diseases in some populations. The effect of Se on in vivo indicators of oxidative stress and inflammation, two important features of atherosclerosis, in human populations is largely unexplored. This study investigated the longitudinal association between serum selenium (s-Se) and a golden standard indicator of oxidative stress in vivo (8-iso-prostaglandin F2alpha, a major F2-isoprostane), an indicator of cyclooxygenase (COX)-mediated inflammation (prostaglandin F2alpha), high sensitive C-reactive protein (hsCRP), interleukin-6 (IL-6) and serum amyloid A protein (SAA) in a follow-up study of 27 years. The s-Se was measured in 615 Swedish men at 50 years of age in a health investigation. The status of oxidative stress and inflammation was evaluated in a re-investigation 27 years later by quantification of urinary 8-iso-PGF2alpha and 15-keto-dihydro-PGF2alpha (a major metabolite of PGF2alpha) and serum hsCRP, SAA and IL-6. Men in the highest quartile of s-Se at age 50 had decreased levels of 8-iso-PGF2alpha compared to all lower quartiles and decreased levels of PGF2alpha compared to all lower quartiles at follow-up. These associations were independent of BMI, diabetes, hyperlipidemia, hypertension, smoking, alpha-tocopherol and beta-carotene at baseline. The s-Se was not associated with hsCRP, SAA or IL-6 at follow-up. In conclusion, high concentrations of s-Se predict reduced levels of oxidative stress and subclinical COX-mediated (but not cytokine-mediated) inflammation in a male population. The associations between Se, oxidative stress and inflammation, respectively, might be related to the proposed cardiovascular protective property of Se.     

Nestin-expressing neural stem cells identified in the scar following myocardial infarction

Nerve fiber innervation of the scar following myocardial damage may have occurred either via the growth of pre-existing fibers and/or the mobilization of neural stem cells. The present study examined whether neural stem cells were recruited to the infarct region of the rat heart following coronary artery ligation. The neural stem cell marker nestin was detected in the infarct region of 1-week post-myocardial infarct (MI) male rats and cultured scar-derived neural-like cells. By contrast, nestin staining was undetected in either scar myofibroblasts or cardiac myocytes residing in the non-infarcted left ventricle. Reactive astrocytes were isolated from the infarct region and characterized by the co-expression of nestin, glial fibrillary acidic protein, and vimentin. Specific staining of oligodendrocytes and neurons was also detected in the infarct region and cultured scar-derived neural-like cells. Furthermore, neurofilament-M positive fibers were identified in the scar and tyrosine hydroxylase immunoreactivity was observed in peripherin-positive neurons. Neurite formation was induced in PC12 cells treated with the conditioned-media of primary passage scar-derived cells, highlighting the synthesis and secretion of neurotrophic factors. Nerve growth factor (NGF) and brain-derived neurotrophic factor were detected in myofibroblasts and neural cells, and both cell types expressed the NGF receptors trkA and p75. These data highlight the novel observation that neural stem cells were recruited to the infarct region of the damaged rat heart and may contribute in part to nerve fiber growth and subsequent innervation of the scar.     

Synthesis of novel pyrroles and fused pyrroles as antifungal and antibacterial agents

Pyrroles and its fused forms possess antimicrobial activities, they can easily interact with biomolecules of living systems. A series of substituted pyrroles, and its fused pyrimidines and triazines forms have been synthesised, all newly synthesised compound structures were confirmed by spectroscopic analysis. Generally, the compounds inhibited growth of some important human pathogens, the best effect was given by: 2a, 3c, 4d on Gram-positive bacteria and was higher on yeast (C. albicans), by 5c on Gram-negative bacteria and by 5a then 3c on filamentous fungi (A. fumigatus and F. oxysporum). Such results present good antibacterial and antifungal potential candidates to help overcome the global problem of antibiotic resistance and opportunistic infections outbreak. Compound 3c gave the best anti-phytopathogenic effect at a 50-fold lower concentration than Kocide 2000, introducing a safe commercial candidate for agricultural use. The effect of the compounds on DNA was monitored to detect the mode of action.     

High Incidence of ACE/PAI-1 in Association to a Spectrum of Other Polymorphic Cardiovascular Genes Involving PBMCs Proinflammatory Cytokines in Hypertensive Hypercholesterolemic Patients: Reversibility with a Combination of ACE Inhibitor and Statin

Cardiovascular diseases (CVDs) are significantly high in the Lebanese population with the two most predominant forms being atherosclerosis and venous thrombosis. The purpose of our study was to assess the association of a spectrum of CVD related genes and combined state of hypertension hypercholesterolemia (HH) in unrelated Lebanese. Twelve polymorphisms were studied by multiplex PCR and reverse hybridization of DNA from 171 healthy individuals and 144 HH subjects. Two genes were significantly associated with HH: ACE (OR: 9.20, P<0.0001) and PAI-1 (OR: 2.29, P = 0.007), respectively with the occurrence of the risky alleles “Del” and “4G”. The frequencies of the Del and 4G alleles were found to be 0.98 and 0.90 in the HH group versus 0.84 and 0.79 in the healthy group, respectively. Serum ACE activity and PAI-I increased significantly with Del/Del and 4G/5G genotypes. The co-expression of Del/4G(+/+) was detected in 113 out of 171 (66.0%) controls and 125 out of 144 (86.8%) HH subjects. Del/4G(-/-) was detected in only 6 (3.5%) controls and undetected in the HH group. Three venous thrombosis related genes [FV(Leiden), MTHFR(A1298C) and FXIII(V34L)] were significantly related to the prominence of the co-expression of Del/4G(+/+). A range of 2 to 8 combined polymorphisms co-expressed per subject where 5 mutations were the most detected. In Del/4G(+/+) subjects, peripheral blood mononuclear cells (PBMCs) produced significant elevated levels of IFN-γ and TNF-α contrary to IL-10, and no variations occurred for IL-4. ACE inhibitor (ramipril) in combination with statin (atorvastatin) and not alone reversed significantly the situation. This first report from Lebanon sheds light on an additional genetic predisposition of a complex spectrum of genes involved in CVD and suggests that the most requested gene FVL by physicians may not be sufficient to diagnose eventual future problems that can occur in the cardiovascular system. Subjects expressing the double mutations (Del/4G) are at high risk for the onset of CVDs.     

Acid phosphatase activity in the chick ovary during embryonic development

Acid phosphatase activity was studied in the left and right ovaries of the chick during embryonic development. The cytochemical study indicated that local enzymatic activity is localized mainly in germ and somatic cells. From the results obtained in the study on the specific activity of this acid hydrolase, it can be inferred that the higher concentration of this enzyme in the right ovary would be determined by a decrease of total proteins in the total homogenate and in the cellular fractions of this organ. Besides, a decrease in the enzymatic activity of this ovary is not observed, but it occurs in the left ovary. This finding would indicate a lack of enzymatic segregation that could be related to the phenomenon of ovarian atrophy. Finally, the electrophoretic study indicated that it would apparently exist only one molecular form of the enzyme. Our results support the hypothesis that acid phosphatase would be involved in the atrophic processes of the right ovary during embryonary differentiation.     

Cellular Expression of Cyclooxygenase,Aromatase, Adipokines,Inflammation and Cell Proliferation Markers in Breast Cancer Specimen

Current evidences suggest that expression of Ki67, cyclooxygenase (COX), aromatase, adipokines, prostaglandins, free radicals, β-catenin and α-SMA might be involved in breast cancer pathogenesis. The main objective of this study was to compare expression/localization of these potential compounds in breast cancer tissues with tissues collected adjacent to the tumor using immunohistochemistry and correlated with clinical pathology. The breast cancer specimens were collected from 30 women aged between 49 and 89 years who underwent breast surgery following cancer diagnosis. Expression levels of molecules by different stainings were graded as a score on a scale based upon staining intensity and proportion of positive cells/area or individually. AdipoR1, adiponectin, Ob-R, leptin, COX-1, COX-2, aromatase, PGF_2α, F₂-isoprostanes and α-SMA were localised on higher levels in the breast tissues adjacent to the tumor compared to tumor specimens when considering either score or staining area whereas COX-2 and AdipoR2 were found to be higher considering staining intensity and Ki67 on score level in the tumor tissue. There was no significant difference observed on β-catenin either on score nor on staining area and intensity between tissues adjacent to the tumor and tumor tissues. A positive correlation was found between COX-1 and COX-2 in the tumor tissues. In conclusion, these suggest that Ki67, COXs, aromatase, prostaglandin, free radicals, adipokines, β-catenin and α-SMA are involved in breast cancer. These further focus the need of examination of tissues adjacent to tumor, tumor itself and compare them with normal or benign breast tissues for a better understanding of breast cancer pathology and future evaluation of therapeutic benefit.     

EGFR kinase activity is required for TLR4 signaling and the septic shock response

Mammalian Toll-like receptors (TLR) recognize microbial products and elicit transient immune responses that protect the infected host from disease. TLR4—which signals from both plasma and endosomal membranes—is activated by bacterial lipopolysaccharides (LPS) and induces many cytokine genes, the prolonged expression of which causes septic shock in mice. We report here that the expression of some TLR4-induced genes in myeloid cells requires the protein kinase activity of the epidermal growth factor receptor (EGFR). EGFR inhibition affects TLR4-induced responses differently depending on the target gene. The induction of interferon-β (IFN-β) and IFN-inducible genes is strongly inhibited, whereas TNF-α induction is enhanced. Inhibition is specific to the IFN-regulatory factor (IRF)-driven genes because EGFR is required for IRF activation downstream of TLR—as is IRF co-activator β-catenin—through the PI3 kinase/AKT pathway. Administration of an EGFR inhibitor to mice protects them from LPS-induced septic shock and death by selectively blocking the IFN branch of TLR4 signaling. These results demonstrate a selective regulation of TLR4 signaling by EGFR and highlight the potential use of EGFR inhibitors to treat septic shock syndrome.