Antifungal Effect of Penicillamine Due to the Selective Targeting of L-Homoserine O-Acetyltransferase

Due to the apparent similarity of fungal and mammalian metabolic pathways, the number of established antifungal targets is low, and the identification of novel ones is highly desirable. The results of our studies, presented in this work, indicate that the fungal biosynthetic pathway of L-methionine, an amino acid essential for humans, seems to be an attractive perspective. The MET2 gene from Candida albicans encoding L-homoserine O-acetyltransferase (CaMet2p), an enzyme catalyzing the first step in that pathway, was cloned and expressed as the native or the oligo-His-tagged fusion protein in Escherichia coli. The recombinant enzymes were purified and characterized for their basic molecular properties and substrate specificities. The purified MET2 gene product revealed the appropriate activity, catalyzed the conversion of L-homoserine (L-Hom) to O-acetyl-L-homoserine (OALH), and exhibited differential sensitivity to several L-Hom or OALH analogues, including penicillamine. Surprisingly, both penicillamine enantiomers (L- and D-Pen) displayed comparable inhibitory effects. The results of the docking of L- and D-Pen to the model of CaMet2p confirmed that both enantiomeric forms of the inhibitor are able to bind to the catalytic site of the enzyme with similar affinities and a similar binding mode. The sensitivity of some fungal cells to L-Pen, depending on the presence or absence of L-Met in the medium, clearly indicate Met2p targeting. Moreover, C. glabrata clinical strains that are resistant to fluconazole displayed a similar susceptibility to L-Pen as the wild-type strains. Our results prove the potential usefulness of Met2p as a molecular target for antifungal chemotherapy.     

The Influence of Angiotensin Peptides on Survival and Motility of Human High-Grade Serous Ovarian Cancer Cells in Serum Starvation Conditions

High-grade serous ovarian carcinoma (HGSOC) is the most frequent and malignant form of ovarian cancer. A local renin–angiotensin system (RAS) has been found in the ovary, and changes in selected components of this system were observed in pathological states and also in ovarian cancer. In the present study, we examined the effect of three peptides, Ang-(1-7), Ang-(1-9) and Ang-(3-7), on proliferation and motility of the OVPA8 cell line, a new well-defined and preclinical model of HGSOC. We confirmed the presence of mRNA for all angiotensin receptors in the tested cells. Furthermore, our findings indicate that all tested angiotensin peptides increased the metabolic serum in the medium by activation of cell defense mechanisms such as nuclear factor kappaB signaling pathway andapoptosis. Moreover, tested angiotensin peptides intensified serum starvation-induced cell cycle arrest at the G0/G1 phase. In the case of Ang-(3-7), a significant decrease in the number of Ki67 positive cells (Ki67+) and reduced percentage of activated ERK1/2 levels in ovarian cancer cells were additionally reported. The angiotensin-induced effect of the accumulation of cells in the G0/G1 phase was not observed in OVPA8 cells growing on the medium with 10% FBS. Moreover, in the case of Ang-(3-7), the tendency was quite the opposite. Ang-(1-7) but not Ang-(1-9) or Ang-(3-7) increased the mobility of reluctant-to-migrate OVAP8 cells cultured in the serum-free medium. In any cases, the changes in the expression of VIM and HIF1A gene, associated with epithelial–mesenchymal transition (EMT), were not observed. In conclusion, we speculate that the adaptation to starvation in nutrient-deprived tumors can be modulated by peptides from the renin–angiotensin system. The influence of angiotensin peptides on cancer cells is highly dependent on the availability of growth factors and nutrients.     

The Involvement of RAGE and Its Ligands during Progression of ALS in SOD1 G93A Transgenic Mice

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by a progressive degeneration of upper and lower motor neurons that causes paralysis and muscle atrophy. The pathogenesis of the disease is still not elucidated. Receptor for Advanced Glycation End Product (RAGE) is a major component of the innate immune system and has implications in ALS pathogenesis. Multiple studies suggest the role of RAGE and its ligands in ALS. RAGE and its ligands are overexpressed in human and murine ALS motor neurons, astrocytes, and microglia. Here, we demonstrated the expression of RAGE and its ligands during the progression of the disease in the transgenic SOD1 G93A mouse lumbar spinal cord. We observed the highest expression of HMGB1 and S100b proteins at ALS onset. Our results highlight the potential role of RAGE and its ligands in ALS pathogenesis and suggest that some of the RAGE ligands might be used as biomarkers in early ALS diagnosis and potentially be useful in targeted therapeutic interventions at the early stage of this devastating disease.     

Effect of processing treatments (frozen,frying) on contents of minerals in tissues of ‘frutti di mare’

The objective of this study was to determine the effect of thermal treatment (frying) of frozen seafood on the content of minerals and heavy metals. An analysis was conducted to compare concentration of sodium, magnesium, potassium, calcium, copper, iron, manganese, zinc, as well as lead, cadmium and mercury in the analysed tissues of invertebrates (shellfish – blue mussel, Japanese squid, white shrimp and octopus). Thermal processing (in this case, frying) applied to the frozen invertebrates was found to affect significantly the content of macro‐ and microelements as well as the content of heavy metals, except for potassium and manganese content in mussels and shrimps, respectively. Frozen and thermally treated (frying) mussels are characterised by significantly higher concentrations of the analysed minerals (except Na⁺, Ca⁺², Cu⁺²), as well as higher concentrations of cadmium and mercury. In turn, shrimps and squids are invertebrates with lower content of both macro‐ and microelements.     

Metabolic profiles of cow's blood; a review

The term ‘metabolic profile’ refers to the analysis of blood biochemical parameters that are useful to assess and prevent metabolic and nutritional disorders in dairy herds. In the higher standards of milk production, the priority in modern breeding is keeping dairy cows in high lactation and healthy. The proper analysis, as well as control. of their feeding and metabolic status is immensely important for the health condition of the herd. The disproportion between the genetically determined ability for milk production and the limitations in improving the energy value of the ration may be the cause of metabolic disorders. Negative energy balance has a major impact on the body's hormonal balance and organ functions and mostly appears during transition periods: from 3 to 2 weeks prepartum until 2–3 weeks postpartum. The term ‘transition’ is used to underscore the important physiological, metabolic and nutritional changes occurring in this time. The manner in which these changes occur and how they are diagnosed and detected are extremely important, as they are closely related to clinical and subclinical postpartum diseases, lactation and reproductive performance – factors that significantly shape the profitability of production. Therefore the priority for intensive milk production is prevention of metabolic diseases and other disorders. It is the intent of this review to synthesize and summarize the information currently available on metabolic status and physiological changes in the cow's body that occur during lactation, as well as to discuss the interpretation of the results, which will be a useful diagnostic tool in nutritional evaluations of the dairy herd. © 2016 Society of Chemical Industry     

Screening of lactic acid bacteria for antimicrobial properties from mayonnaise-based products and raw materials

A collection of lactic acid bacteria isolated from both mayonnaise-based products and raw materials used to manufacture them was tested for antimicrobial activity. Out of 144 strains (97 lactobacilli, 23 lactococci and 24 enterococci) only three supernatants of Enterococcus spp. strains (EN3, EN14 and EN15) exhibited activity against lactobacilli and lactococci. The supernatant of the strain EN3 exclusively inhibited the growth of Bacillus cereus and Listeria monocytogenes. Enterococcus strains EN3, EN14 and EN15 produced thermostable bacteriocins, which had antibacterial activity.     

Sustainable nickel catalyst for the conversion of lignocellulosic biomass to H2-rich gas

The objective of this paper was to design sustainable nickel catalysts supported on selected fly ash based zeolites to thermal processing of lignocellulosic feedstock towards hydrogen-rich gas. Moreover, in order to increase its catalytic performance in the studied process the catalyst supported on the most promising fly ash based zeolite was modified by selected rare-earth and transition metals (La, Pr, Ce, Y, Gd, Zr). The performed measurements exhibited that incorporation of nickel into the structure of zeolite A modified by lanthanum resulted in the most effective production of H₂. The characterization of its physicochemical properties (XRD, TPR, SEM-EDS, TPD-NH₃, BET and TGA-DTA) suggested that large pore size, moderate acidity, increased reducibility of an active phase and higher resistance to coke formation are the main factors responsible for increased activity of this catalyst.     

The effect of La2O3 and CeO2 modifiers on properties of Ni–Al catalysts for LNG prereforming

Ni/La–Al₂O₃ and Ni/Ce–Al₂O₃ catalysts with a small amount of promoters intended for prereforming of LNG were characterized by XRF, N₂ adsorption-desorption, XRD, H₂ chemisorption, HRTEM and XPS. The catalytic activity was evaluated in methane steam reforming both in the kinetic and diffusion regime, at temperatures characteristic of pre-reforming. Carbonaceous deposit was analysed by TPO-MS method. The nature and location of the coke were studied by HRTEM.La or Ce addition into Ni–Al system causes the increase of the active surface area of Ni by enhancing its dispersion. Studies at kinetic regime have shown that the promoted catalysts have almost twice the activity than reference Ni–Al catalyst. This effect was not confirmed by measurements in the diffusion regime on whole catalyst tablets. Almost identical textural properties of catalysts and diffusive limitations related to them but not the catalytic properties of the material itself appeared to be crucial factors. The presence of La (but not Ce) causes a significant increase in resistance to coking.     

Fiducial markers for combined 3-dimensional mass spectrometric and optical tissue imaging

Mass spectrometric imaging (MSI) has become widely used in the analysis of a variety of biological surfaces. Biological samples are spatially, morphologically, and metabolically complex. Multimodal molecular imaging is an emerging approach that is capable of dealing with this complexity. In a multimodal approach, different imaging modalities can provide precise information about the local molecular composition of the surfaces. Images obtained by MSI can be coregistered with images obtained by other molecular imaging techniques such as microscopic images of fluorescent protein expression or histologically stained sections. In order to properly coregister images from different modalities, each tissue section must contain points of reference, which are visible in all data sets. Here, we report a newly developed coregistration technique using fiducial markers such as cresyl violet, Ponceau S, and bromophenol blue that possess a combination of optical and molecular properties that result in a clear mass spectrometric signature. We describe these fiducial markers and demonstrate an application that allows accurate coregistration and 3-dimensional reconstruction of serial histological and fluorescent microscopic images with MSI images of thin tissue sections from a breast tumor model.