Characterization of ovarian nuclei with the parameter of power ratio

The nucleoli and chromatin clumps of ovarian cells contain important features in discriminating malignant cells from normal ones. In geometric properties, the ovarian nucleoli and chromatin clumps appear as irregularly shaped dark spots in the nuclear images from specimens immunohistochemically stained with antibody to Mib-1. Malignant cells often have more active and larger nucleoli and chromatin clumps. However, estimating the size of the nucleoli or chromatin clumps is a difficult task since it is not easy to recognize and accurately separate the regions of nucleoli and chromatin clumps from the rest of the nuclei that are highly irregular and variant in contents and intensities. In this paper, we develop a method to derive a parameter called power ratio that is proportionally related to the size of nucleoli and chromatin clumps based on an ideal nuclear model without the region segmentation of nucleoli or chromatin clumps. Results of characterization of the parameter and comparison between malignant and normal cells are provided.     

Hybrid composites of ABS with carbonaceous fillers for electromagnetic shielding applications

This work evaluates the influence of two types of carbonaceous fillers, carbon black (CB) and carbon nanotubes (CNTs), on the electrical, electromagnetic, and rheological properties of composites based on poly(acrylonitrile‐co‐butadiene‐co‐styrene) (ABS) prepared by the melt mixing. Electrical conductivity, electromagnetic shielding efficiency (EMI SE) in the X‐band frequency range (8–12.4 GHz), and melt flow index (MFI) results showed that ABS/CNT composites exhibit higher electrical conductivity and EMI SE, but lower MFI when compared to ABS/CB composites. The electrical conductivity of the binary composites showed an increase of around 16 orders of magnitude, when compared to neat ABS, for both fillers. Binary composites with 5 and 15 wt % of filler showed an EMI SE of, respectively, ?44 and ?83 dB for ABS/CNT, and ?9 and ?34 dB for ABS/CB. MFI for binary composites with 5 wt % were 15.45 and 0.55 g/10 min for CB and CNT, respectively. Hybrid composites ABS/CNT.CB with 3 wt % total filler and fraction 50:50 and 75:25 showed good correlation between EMI SE and MFI. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 46546.     

Binding of pyrene to hydrophobic fractions of dissolved organic matter: effect of polyvalent metal complexation

The effects of polyvalent metal cations on pyrene binding to hydrophobic acid and neutral fractions (HoA and HoN, respectively) of dissolved organic matter (DOM) were elucidated. The DOM was isolated from sewage sludge; pyrene binding was estimated from fluorescence measurements. Isotherms of pyrene binding to both fractions were nonlinear. Pyrene binding was higher for HoN due to the combined effect of greater hydrophobicity, aromaticity and the large molecular size of this fraction relative to HoA. The complexation of HoA with Cu2+, Al3+, and Fe3+ increased the binding of pyrene only when the HoA was equilibrated with polyvalent cations before pyrene was added. The maximal increase in pyrene binding to HoA was 56%, 64%, and 118% when pre-equilibrated with Cu2+, Fe3+, and Al3+, respectively. Pyrene binding to HoN was not affected by the presence of metal cations. HoA complexation with metal cations increased the apparent molecular size of this fraction. We suggest that the presence of metal cations induces the formation of pseudomicelles, which are more efficient in binding pyrene than the low-molecular-weight components. Our results demonstrate that HoA and HoN components can significantly affect the transport of organic contaminants in soils irrigated with treated wastewater or amended with sewage sludge.     

Periwinkle (Littorina littorea) as a sentinel species: a field study integrating chemical and biological analyses

The gastropod Littorina littorea (common periwinkle) is an abundant and widespread North Atlantic species. The characteristic development of Intersex in L. littorea has been widely applied as a biomarker for tributyltin (TBT) contamination. Here, we assess the potential of L. littorea as a novel sentinel species for evaluating the sublethal effects in wild populations of widely distributed contaminants. We collected animals from six sites across the South coast of England. Tissue concentrations of polycyclic aromatic hydrocarbons (PAHs), organotin compounds (OTCs), polychlorinated biphenyls (PCBs), and organochlorine pesticides (OCPs) were measured and compared with biomarkers of damage to DNA (Comet assay), lysosomal stability (NRR assay), and endocrine disruption (Intersex development). There was a strong correlation between DNA damage and PAH bioaccumulation (n=6, r=0.84, p<0.05), as well as that between Intersex development and OTC pollution (n=6, r=0.91, p<0.05). The relationship between PAH bioaccumulation and DNA strand breaks was nonlinear, highlighting the need to consider the role of adaptive mechanisms in the interpretation of field results. These results illustrate the potential use of periwinkles for monitoring a wide range of priority pollutants.     

Micro-kinetic model of fructo-oligosaccharide synthesis for prebiotic products

A new micro-kinetic model of the enzyme-catalyzed synthesis of fructo-oligosaccharides (FOS) was developed. A commercial enzyme mixture Pectinex® Ultra SP-L derived from Aspergillus aculeatus was used. A variety of initial enzyme concentrations (1–5 vol%) and sucrose concentrations (400–600 g/L) were experimentally investigated and included in kinetic modeling. Several variations of kinetic mechanisms and corresponding models have been examined. A hybrid genetic algorithm was used to predict the kinetic parameters simultaneously for all experimental data. The best fitting model has been adopted, and with an average error of 13.34%, it describes the experimental data very well. The influence of initial concentrations on the conversion of sucrose and production of FOS is being carefully investigated. It was shown that the initial sucrose concentration significantly affects the highest level of FOS concentration, but the enzyme concentration controls the time at which maximum is reached as well as the rate of FOS decomposition.     

Antioxidant effectiveness of coffee extracts and selected constituents in cell‐free systems and human colon cell lines

Scope: Epidemiological studies suggest that coffee can reduce the risk of degenerative diseases such as diabetes type 2, cardiovascular disease and cancer. These beneficial effects have partly been attributed to the antioxidant activity of coffee. We determined composition and antioxidant potential of differentially roasted coffee extracts and investigated the impact of selected original constituents and roast products. Methods and results: Parameters studied were direct antioxidant activity (trolox equivalent antioxidant capacity/oxygen radical absorbing capacity), cellular reactive oxygen species (ROS) level, DNA damage and protein expression of NAD(P)H: quinone oxidoreductase, γ‐glutamylcysteine ligase and glutathione reductase in HT‐29/Caco‐2 cells at 24‐h incubation. All extracts showed distinct direct antioxidant activity: medium roasts>light roast AB1 (caffeoylquinic acid (CQA)‐rich Arabica Brazil extract); dark roast AB2 (N‐methylpyridinium (NMP)‐rich Arabica Brazil extract), and diminished t‐butylhydroperoxide‐induced ROS level in HT‐29 cells (AB2>medium roasts>AB1). NAD(P)H:quinone oxidoreductase 1 expression and γ‐glutamylcysteine ligase expression were distinctly induced by AB1 and 5‐CQA, but not by AB2 and NMP. 5‐CQA and caffeic acid exhibited highest trolox equivalent antioxidant capacity/oxygen radical absorbing capacity values (5‐CQA: 1.3/3.5 mM and caffeic acid: 1.3/3.9 mM trolox); ROS level was distinctly diminished by 5‐CQA (≥3 μM), catechol (30 μM) and trigonelline (≥30 μM), whereas menadione‐induced DNA damage in Caco‐2 cells was reduced by NMP compounds (1–30 μM). Conclusion: The results emphasize that both original constituents and roast products contribute to the cellular antioxidant effectiveness of coffee.     

Mitochondrial Side Effects of Surgical Prophylactic Antibiotics Ceftriaxone and Rifaximin Lead to Bowel Mucosal Damage

Despite their clinical effectiveness, a growing body of evidence has shown that many classes of antibiotics lead to mitochondrial dysfunction. Ceftriaxone and Rifaximin are first choice perioperative antibiotics in gastrointestinal surgery targeting fundamental processes of intestinal bacteria; however, may also have negative consequences for the host cells. In this study, we investigated their direct effect on mitochondrial functions in vitro, together with their impact on ileum, colon and liver tissue. Additionally, their impact on the gastrointestinal microbiome was studied in vivo, in a rat model. Rifaximin significantly impaired the oxidative phosphorylation capacity (OxPhos) and leak respiration in the ileal mucosa, in line with increased oxidative tissue damage and histological changes following treatment. Ceftriaxone prophylaxis led to similar changes in the colon mucosa. The composition and diversity of bacterial communities differed extensively in response to antibiotic pre-treatment. However, the relative abundances of the toxin producing species were not increased. We have confirmed the harmful effects of prophylactic doses of Rifaximin and Ceftriaxone on the intestinal mucosa and that these effects were related to the mitochondrial dysfunction. These experiments raise awareness of mitochondrial side effects of these antibiotics that may be of clinical importance when evaluating their adverse effects on bowel mucosa.     

Microglial CD74 Expression Is Regulated by TGFβ Signaling

Simple SummaryIn the present study, we provided evidence that TGFβ signaling regulated the expression of the microglia activation marker CD74. Our data demonstrated that TGFβ1 inhibited LPS-induced upregulation of CD74. Moreover, inhibition of microglial TGFβ signaling in vitro and silencing of TGFβ signaling by deletion of Tgfbr2 in vivo resulted in marked upregulation of microglial CD74.AbstractMicroglia play important roles during physiological and pathological situations in the CNS. Several reports have described the expression of Cd74 in disease-associated and aged microglia. Here, we demonstrated that TGFβ1 controled the expression of Cd74 in microglia in vitro and in vivo. Using BV2 cells, primary microglia cultures as well as Cx3cr1^CreERT2:R26-YFP:Tgfbr2^fl/fl in combination with qPCR, flow cytometry, and immunohistochemistry, we were able to provide evidence that TGFβ1 inhibited LPS-induced upregulation of Cd74 in microglia. Interestingly, TGFβ1 alone was able to mediate downregulation of CD74 in vitro. Moreover, silencing of TGFβ signaling in vivo resulted in marked upregulation of CD74, further underlining the importance of microglial TGFβ signaling during regulation of microglia activation. Taken together, our data indicated that CD74 is a marker for activated microglia and further demonstrated that microglial TGFβ signaling is important for regulation of Cd74 expression during microglia activation.     

In Silico Genome-Scale Analysis of Molecular Mechanisms Contributing to the Development of a Persistent Infection with Methicillin-Resistant Staphylococcus aureus (MRSA) ST239

The increasing frequency of isolation of methicillin-resistant Staphylococcus aureus (MRSA) limits the chances for the effective antibacterial therapy of staphylococcal diseases and results in the development of persistent infection such as bacteremia and osteomyelitis. The aim of this study was to identify features of the MRSA_ST239 0943-1505-2016 (SA943) genome that contribute to the formation of both acute and chronic musculoskeletal infections. The analysis was performed using comparative genomics data of the dominant epidemic S. aureus lineages, namely ST1, ST8, ST30, ST36, and ST239. The SA943 genome encodes proteins that provide resistance to the host’s immune system, suppress immunological memory, and form biofilms. The molecular mechanisms of adaptation responsible for the development of persistent infection were as follows: amino acid substitution in PBP2 and PBP2a, providing resistance to ceftaroline; loss of a large part of prophage DNA and restoration of the nucleotide sequence of beta-hemolysin, that greatly facilitates the escape of phagocytosed bacteria from the phagosome and formation of biofilms; dysfunction of the AgrA system due to the presence of psm-mec and several amino acid substitutions in the AgrC; partial deletion of the nucleotide sequence in genomic island vSAβ resulting in the loss of two proteases of Spl—operon; and deletion of SD repeats in the SdrE amino acid sequence.     

Stereodivergent Biocatalytic Formal Reduction of α-Angelica Lactone to (R)- and (S)-γ-Valerolactone in a One-Pot Cascade

The formal asymmetric and stereodivergent enzymatic reduction of α-angelica lactone to both enantiomers of γ-valerolactone was achieved in a one-pot cascade by uniting the promiscuous stereoselective isomerization activity of Old Yellow Enzymes with their native reductase activity. In addition to running the cascade with one enzyme for each catalytic step, a bifunctional isomerase-reductase biocatalyst was designed by fusing two Old Yellow Enzymes, thereby generating an unprecedented case of an artificial enzyme catalyzing the reduction of nonactivated C=C bonds to access (R)-valerolactone in overall 41 % conversion and up to 91 % ee. The enzyme BfOYE4 could be used as single biocatalyst for both steps and delivered (S)-valerolactone in up to 84 % ee and 41 % overall conversion. The reducing equivalents were provided by a nicotinamide recycling system based on formate and formate dehydrogenase, added in a second step. This enzymatic system provides an asymmetric route to valuable chiral building blocks from an abundant bio-based chemical.