Living cells are constantly exchanging momentum with their surroundings. So far, there is no consensus regarding how cells respond to such external stimuli, although it reveals much about their ...internal structures, motility as well as the emergence of disorders. Here, we report that twelve cell lines, ranging from healthy fibroblasts to cancer cells, hold a ubiquitous double power-law viscoelastic relaxation compatible with the fractional Kelvin-Voigt viscoelastic model. Atomic Force Microscopy measurements in time domain were employed to determine the mechanical parameters, namely, the fast and slow relaxation exponents, the crossover timescale between power law regimes, and the cell stiffness. These cell-dependent quantities show strong correlation with their collective migration and invasiveness properties. Beyond that, the crossover timescale sets the fastest timescale for cells to perform their biological functions.
Mitochondria are central key players in cell metabolism, and mitochondrial DNA (mtDNA) instability has been linked to metabolic changes that contribute to tumorigenesis and to increased expression of ...pro-tumorigenic genes. Here, we use melanoma cell lines and metastatic melanoma tumors to evaluate the effect of mtDNA alterations and the expression of the mtDNA packaging factor, TFAM, on energetic metabolism and pro-tumorigenic nuclear gene expression changes. We report a positive correlation between mtDNA copy number, glucose consumption, and ATP production in melanoma cell lines. Gene expression analysis reveals a down-regulation of glycolytic enzymes in cell lines and an up-regulation of amino acid metabolism enzymes in melanoma tumors, suggesting that TFAM may shift melanoma fuel utilization from glycolysis towards amino acid metabolism, especially glutamine. Indeed, proliferation assays reveal that TFAM-down melanoma cell lines display a growth arrest in glutamine-free media, emphasizing that these cells rely more on glutamine metabolism than glycolysis. Finally, our data indicate that TFAM correlates to VEGF expression and may contribute to tumorigenesis by triggering a more invasive gene expression signature. Our findings contribute to the understanding of how TFAM affects melanoma cell metabolism, and they provide new insight into the mechanisms by which TFAM and mtDNA copy number influence melanoma tumorigenesis.
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•A GH18 chitinase from Chromobacterium violaceum was produced in Escherichia coli.•The recombinant protein was purified and partially characterized.•The purified chitinase inhibited ...conidia germination of two Fusarium species.•A mechanism of action for the antifungal activity is suggested.
The biocontrol activity of some soil strains of Chromobacterium sp. against pathogenic fungi has been attributed to secreted chitinases. The aim of this work was to characterize biochemically a recombinant chitinase (CvChi47) from C. violaceum ATCC 12472 and to investigate its effects on phytopathogenic fungi. CvChi47 is a modular enzyme with 450 amino acid residues, containing a type I signal peptide at the N-terminal region, followed by one catalytic domain belonging to family 18 of the glycoside hydrolases, and two type-3 chitin-binding domains at the C-terminal end. The recombinant enzyme was expressed in Escherichia coli as a His-tagged protein and purified to homogeneity. The native signal peptide of CvChi47 was used to direct its secretion into the culture medium, from where the recombinant product was purified by affinity chromatography on chitin and immobilized metal. The purified protein showed an apparent molecular mass of 46 kDa, as estimated by denaturing polyacrylamide gel electrophoresis, indicating the removal of the signal peptide. CvChi47 was a thermostable protein, retaining approximately 53.7% of its activity when heated at 100 °C for 1 h. The optimum hydrolytic activity was observed at 60 °C and pH 5. The recombinant chitinase inhibited the conidia germination of the phytopathogenic fungi Fusarium oxysporum and F. guttiforme, hence preventing mycelial growth. Furthermore, atomic force microscopy experiments revealed a pronounced morphological alteration of the cell surface of conidia incubated with CvChi47 in comparison to untreated cells. Taken together, these results show the potential of CvChi47 as a molecular tool to control plant diseases caused by these Fusarium species.
Scoliosis is a condition that affects the spine and causes chest rotation and trunk distortion. Individuals with severe deformities may experience dyspnea on exertion and develop respiratory failure. ...Respiratory oscillometry is a simple and non- invasive method that provides detailed information on lung mechanics. This work aims to investigate the potential of oscillometry in the evaluation of respiratory mechanics in patients with scoliosis and its association with physical performance. We analyzed 32 volunteers in the control group and 32 in the scoliosis group. The volunteers underwent traditional pulmonary function tests, oscillometry, and the 6-minute walk test (6MWT). Oscillometric analysis showed increased values of resistance at 4 Hz (R4, P<0.01), 12 Hz (R12, P<0.0001), and 20 Hz (R20, P<0.01). Similar analysis showed reductions in dynamic compliance (Cdyn, P<0.001) and ventilation homogeneity, as evaluated by resonance frequency (fr, P<0.001) and reactance area (Ax, P<0.001). Respiratory work, described by the impedance modulus, also showed increased values (Z4, P<0.01). Functional capacity was reduced in the group with scoliosis (P<0.001). A significant direct correlation was found between Cobb angle and R12, AX, and Z4 (P=0.0237, P=0.0338, and P=0.0147, respectively), and an inverse correlation was found between Cdyn and Cobb angle (P=0.0190). These results provided new information on respiratory mechanics in scoliosis and are consistent with the involved pathophysiology, suggesting that oscillometry may improve lung function tests for patients with scoliosis. Key words: Respiratory oscillometry; Forced oscillation technique; Scoliosis; Functional capacity
Hoodia gordonii is a plant species used traditionally in southern Africa to suppress appetite. Recently, it has been associated with a significant increase in blood pressure and pulse rate in women, ...suggesting sympathomimetic activity. The present study investigated the possible antidepressant-like effects of acute and repeated (15 days) administration of H. gordonii extract (25 and 50 mg/kg, po) to mice exposed to a forced swimming test (FST). Neurochemical analysis of brain monoamines was also carried out to determine the involvement of the monoaminergic system on these effects. Acute administration of H. gordonii decreased the immobility of mice in the FST without accompanying changes in general activity in the open-field test during acute treatment, suggesting an antidepressant-like effect. The anti-immobility effect of H. gordonii was prevented by pretreatment of mice with PCPA an inhibitor of serotonin (5-HT) synthesis, NAN-190 (a 5-HT1A antagonist), ritanserin (a 5-HT2A/2C antagonist), ondansetron (a 5-HT3A antagonist), prazosin (an α1-adrenoceptor antagonist), SCH23390 (a D1 receptor antagonist), yohimbine (an α2-adrenoceptor antagonist), and sulpiride (a D2 receptor antagonist). A significant increase in 5-HT levels in the striatum was detected after acute administration, while 5-HT, norepinephrine and dopamine were significantly elevated after chronic treatment. Results indicated that H. gordonii possesses antidepressant-like activity in the FST by altering the dopaminergic, serotonergic, and noradrenergic systems.
Mo-CBP3 is a chitin-binding protein from M. oleifera seeds that inhibits the germination and mycelial growth of phytopathogenic fungi. This protein is highly thermostable and resistant to pH changes, ...and therefore may be useful in the development of new antifungal drugs. However, the relationship of MoCBP3 with the known families of carbohydrate-binding domains has not been established. In the present study, full-length cDNAs encoding 4 isoforms of Mo-CBP3 (Mo-CBP3-1, Mo-CBP3-2, Mo-CBP3-3 and Mo-CBP3-4) were cloned from developing seeds. The polypeptides encoded by the Mo-CBP3 cDNAs were predicted to contain 160 (Mo-CBP3-3) and 163 amino acid residues (Mo-CBP3-1, Mo-CBP3-2 and Mo-CBP3-4) with a signal peptide of 20-residues at the N-terminal region. A comparative analysis of the deduced amino acid sequences revealed that Mo-CBP3 is a typical member of the 2S albumin family, as shown by the presence of an eight-cysteine motif, which is a characteristic feature of the prolamin superfamily. Furthermore, mass spectrometry analysis demonstrated that Mo-CBP3 is a mixture of isoforms that correspond to different mRNA products. The identification of Mo-CBP3 as a genuine member of the 2S albumin family reinforces the hypothesis that these seed storage proteins are involved in plant defense. Moreover, the chitin-binding ability of Mo-CBP3 reveals a novel functionality for a typical 2S albumin.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
This work has developed and evaluated a microemulsion-based drilling fluid formulation with characteristics to be applied in oil wells. The microemulsion was formulated with a solution of ...water/glycerol, pine oil, and Tween 80, a nonionic and biodegradable surfactant. The physical and chemical properties of the drilling fluid obtained in this work were investigated through rheology and filtration analysis, solids content, aging, lubricity, toxicity, and thermal degradation. A non-toxic microemulsion-based drilling fluid oil-in-water (O/W) with high lubricity (0.07638) and thermal stability was obtained with suitable viscosity, gel strength and low fluid loss (4.0 mL), low solids content (6%), stability in a wide range of salinity conditions, and the possibility of high water content (above 85% in mass fraction). The fluid presented a pseudoplastic behavior, and statistically significant Herschel–Bulkley parameters were obtained.
Aims
This study aimed to evaluate the in vitro cytotoxicity and efficacy of synthetic host defence peptides (HDPs), alone or in combination with florfenicol (FFC), oxytetracycline (OTC) or ...thiamphenicol (TAP), against different pathogenic bacteria isolated from diseased fish.
Methods and Results
Solid‐phase synthesis, purification and characterization of several HDPs were performed manually, using the fluorenylmethyloxycarbonyl protecting group in different resins and via high‐performance liquid chromatography‐mass spectrometry, respectively. The in vitro cytotoxicity and antimicrobial activity of HDPs, FFC, OTC and TAP against Nile tilapia red blood cells (RBCs) and relevant fish pathogenic bacteria (Aeromonas, Citrobacter, Edwardsiella, Streptococcus, Lactococcus and Vibrio) was determined using the haemolysis assay and broth microdilution method, respectively. The checkerboard assay was used to evaluate the synergy between the most active HDPs and other antimicrobials against the tested strains. MUC 7 12‐mer, FFC, OTC and TAP were not cytotoxic to Nile tilapia RBCs, in all tested concentrations. LL‐37, (p‐BthTX‐I)2 and Hylin‐a1 were not cytotoxic at concentrations up to 78·13, 19·53 and 9·77 μg ml−1, respectively. HDPs demonstrated potent antimicrobial activity (minimum inhibitory concentration ≤31·25 µg ml−1) against Aeromonas jandaei (KR‐12‐a5), Citrobacter freundii (Kr‐12‐a5; (p‐BthTX‐I)2; LL‐37; and Hylin a1), Streptococcus agalactiae (Hylin a1; (p‐BthTX‐I)2 and LL‐37), Lactococcus garviae (Hylin a1), and Vibrio fluvialis (KR‐12‐a5). The combinations of (p‐BthTX‐I)2 with TAP and LL‐37 with FFC showed synergistic activity against C. freundii (fractional inhibitory concentration index of 0·25 and 0·50, respectively).
Conclusions
Synthetic HDPs have the potential as a good treatment option for bacterial diseases in aquaculture.
Significance and Impact of the Study
The in vivo effectiveness of synthetic HDPs such as KR‐12‐a5; LL‐37; (p‐BthTX‐I)2 and Hylin a1 can be tested alone or in combination with conventional antimicrobials as a treatment option to reduce the use of antimicrobials in aquaculture.
The chalcone-like series 1a–1g was efficiently synthesized from Morita–Baylis–Hillman reaction (52–74% yields). Compounds 1a–1g were designed by molecular hybridization based on the anti-inflammatory ...drug methyl salicylate (3) and the antileishmanial moiety of the Morita–Baylis–Hillman adducts 2a–2g. The 1a–1g compounds were much more actives than precursor series 2a–2g, for example, IC50=7.65μM on Leishmania amazonensis and 10.14μM on Leishmania chagasi (compound 1c) when compared to IC50=50.08μM on L. amazonensis and 82.29μM on L. chagasi (compound 2c). The IC50 values of compound 3 (228.49μM on L. amazonensis and 261.45μM on L. chagasi) and acryloyl salicylate 4 (108.50μM on L. amazonensis and 118.83μM on L. chagasi) were determined here, by the first time, on Leishmania.
In cull ewes, energy supply by time short periods can affect growth, carcass traits, and meat quality. The objective of this study was to evaluate the effect of biscuit bran (BB) and cashew nut bran ...(CNB) in diets with different total digestible nutrient (TDN) content on animal performance and carcass and meat traits of cull ewes. Twenty Morada Nova cull ewes, with the initial BW of 30.1 ± 3.56 kg, and three years of age were distributed in a completely randomized design into factorial scheme 2 × 2, with two energy sources (BB and CNB) × two levels of TDN. All diets were isoproteic with variations in TDN content increased by 10% (TDN10) and 25% (TDN25) above the recommended requirement for the study category, which was 66.0% TDN. The experimental period lasted 56 days, in which daily intake in relation to metabolic weight and weight performance of the animals was determined. The hot carcass, the cold carcass, commercial cuts, loin eye area, and subcutaneous fat thickness were measured, obtained, or calculated. A sample of the Longissimus lumborum (LL) muscle was used by determination of physical–chemical composition, sensory, and fatty acid (FA) profile analysis. The ANOVA was performed to access the effect of diet and energy level on the characteristics studied, and means were compared by the Turkey and Kruskal Wallis tests at 5% of probability. A higher daily gain weight was observed from TDN25 (P = 0.006). Among by-products, higher nutrient intakes were observed in animals fed with BB diets (P < 0.01), except for crude protein. The same patterns have been observed for live weight, reconstituted half-carcass, higher yield in fat thickness, and internal carcass length. Cashew nut bran produced darker colored meat (P = 0.017) and higher cooking weight loss (P = 0.008). Higher lipid content was observed in animals submitted to diets with CNB and TDN25 (P = 0.001). The internal length of the carcass was higher in animals fed with BB (P = 0.041). All diets did not effect on sensory characteristics (P > 0.05). Total digestible nutrient in 25% above recommended promotes higher weight gain and meat with higher lipid content and less weight loss by cooling. The inclusion of BB, compared to CNB, provides a higher weight of carcasses and better conformation, in addition to lower total lipid content with higher proportion for monounsaturated FAs in meat, providing more attractive features for the consumer market.
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