Self-association of isolated large cytoplasmic domain of plasma membrane H+-ATPase from Saccharomyces cerevisiae: Role of the phosphorylation domain in a general dimeric model for P-ATPases

AbstractLarge cytoplasmic domain (LCD) plasma membrane H+-ATPase from S. cerevisiae was expressed as two fusion polypeptides in E. coli: a DNA sequence coding for Leu353–Ileu674 (LCDh), comprising both nucleotide (N) and phosphorylation (P) domains, and a DNA sequence coding for Leu353–Thr543 (LCDΔh, lacking the C-terminus of P domain), were inserted in expression vectors pDEST-17, yielding the respective recombinant plasmids. Overexpressed fusion polypeptides were solubilized with 6 M urea and purified on affinity columns, and urea was removed by dialysis. Their predicted secondary structure contents were confirmed by CD spectra. In addition, both recombinant polypeptides exhibited high-affinity 2′,3′-O-(2,4,6-trinitrophenyl)adenosine-5′-triphosphate (TNP-ATP) binding (Kd=1.9 μM and 2.9 μM for LCDh and LCDΔh, respectively), suggesting that they have native-like folding. The gel filtration profile (HPLC) of purified LCDh showed two main peaks, with molecular weights of 95 kDa and 39 kDa, compatible with dimeric and monomeric forms, respectively. However, a single elution peak was observed for purified LCDΔh, with an estimated molecular weight of 29 kDa, as expected for a monomer. Together, these data suggest that LCDh exist in monomer–dimer equilibrium, and that the C-terminus of P domain is necessary for self-association. We propose that such association is due to interaction between vicinal P domains, which may be of functional relevance for H+-ATPase in native membranes. We discuss a general dimeric model for P-ATPases with interacting P domains, based on published crystallography and cryo-electron microscopy evidence

Effects of feed particle size on energy values for broiler chickens at various ages

The objective of this study was to evaluate the effects of various geometric mean diameters (GMDs) of particles of corn, pelleted soybean meal and a corn-soy mixture in the proportion of 70% and 30%, respectively, on the nutritional value of the feeds. The study evaluated energy consumption, the contents of apparent metabolizable energy (AME) and AME corrected for nitrogen balance (AMEn) and the metabolizability coefficients for broiler chickens at various ages. A total of 540 Cobb 500 male broilers were housed in metabolic cages (experimental units). Trials were performed separately with each feed. A completely randomized design was used with four treatments, namely corn with 573, 636, 851, and 1012 μm GMDs; pelleted soybean meal with 538, 550, 665, and 741 μm GMDs; and the corn-soy mixture with 627, 658, 893, and 1040 μm GMDs. Birds were evaluated on days 1 - 10, 11 - 20, 21 - 30, and 31 - 40. Larger GMDs resulted in lower energy consumption. From 1 to 10 days, birds consumed less metabolizable energy than older birds. Birds fed corn from days 1 to 10 had higher metabolizable energy (P <0.05) with increasing GMD up to 1042 μm. However, the results varied, depending on the feed and its combinations. The use of coarse particles could reduce the costs of grinding, and would have few effects on the metabolizable energy of broiler chickens

Effect on Broiler Production Performance and Meat Quality of Feeding Ulva lactuca Supplemented with Carbohydrases

Macroalgae have been increasingly exploited worldwide for feed, food and biofuel applications, due to their nutritive and bioactive compounds. Green seaweeds belonging to the genus Ulva have high growth rates, which makes them suitable for being cultured in sustainable algae production, such as an integrated multi-trophic aquaculture system. This is expected to increase the use of Ulva sp. as an alternative source to conventional feedstuffs (e.g., cereals and soybean meal) for poultry. The objective of the current study was to assess if the incorporation of 15% Ulva lactuca in broiler chickens’ diet, combined or not with carbohydrate-active enzymes, would enhance meat nutritional quality without compromising animal growth performance. Overall, U. lactuca led to an accumulation of antioxidant carotenoids, n-3 PUFA and macrominerals, including magnesium, potassium and phosphorus, in the breast muscle, with likely health benefits, without significantly impairing growth performance. The supplementation of macroalgae with a recombinant ulvan lyase reduced ileal viscosity with possible beneficial effects on broiler digestibility. Although dietary U. lactuca showed potential to increase meat quality, it reduced meat overall acceptability, which suggests the use of a lower algae inclusion level to prevent a negative meat sensory perception for consumersinfo:eu-repo/semantics/publishedVersio

Dynamic Critical Behavior of an Extended Reptation Dynamics for Self-Avoiding Walks

We consider lattice self-avoiding walks and discuss the dynamic critical behavior of two dynamics that use local and bilocal moves and generalize the usual reptation dynamics. We determine the integrated and exponential autocorrelation times for several observables, perform a dynamic finite-size scaling study of the autocorrelation functions, and compute the associated dynamic critical exponents $z$. For the variables that describe the size of the walks, in the absence of interactions we find $z \approx 2.2$ in two dimensions and $z\approx 2.1$ in three dimensions. At the $\theta$-point in two dimensions we have $z\approx 2.3$.Comment: laTeX2e, 32 pages, 11 eps figure

Fungal Infections in Neonatal Intensive Care

Neonates represent a unique and highly vulnerable patient population. Advances in medical technology have improved the survival and quality of life of newborns, particularly those with extreme prematurity or with congenital defects. Furthermore, immunologic immaturity and altered cutaneous barriers play some role in the vulnerability of neonates to nosocomial infections. In this context, the incidence of invasive fungal infections has increased significantly worldwide, representing an important infective complication in patients hospitalized in intensive care units. Invasive fungal infections in Neonatal Intensive Care Unit (NICUs) show high mortality; being species of Candida, the most isolates etiologic agents. The better prognosis of the patient is associated with the early diagnosis and fast treatment. However, guidelines to facilitate the optimal therapy choice for the treatment of neonatal fungal disease do not exist. The current antifungal agents that are available to treat fungemia among newborns and children are based on clinical trials in adults, since there are few comparative studies of antifungal agents in infants. The most commonly used drugs for the treatment of invasive fungal infections in neonates are classified in four different classes: polyene, azoles, analogs of pyrimidines and echinocandins

Influence of Feeding Weaned Piglets with Laminaria digitata on the Quality and Nutritional Value of Meat

Laminaria digitata is a novel feedstuff that can be used in pig diets to replace conventional feedstuffs. However, its resilient cell wall can prevent the monogastric digestive system from accessing intracellular nutrients. Carbohydrate-active enzyme (CAZyme) supplementation is a putative solution for this problem, degrading the cell wall during digestion. The objective of this work was to evaluate the effect of 10% L. digitata feed inclusion and CAZyme supplementation on the meat quality and nutritional value of weaned piglets. Forty weaned piglets were randomly allocated to four experimental groups (n = 10): control, LA (10% L. digitata, replacing the control diet), LAR (LA + CAZyme (0.005% Rovabio® Excel AP)) and LAL (LA + CAZyme (0.01% alginate lyase)) and the trial lasted for two weeks. The diets had no effect on any zootechnical parameters measured (p > 0.05) and meat quality traits, except for the pH measured 24 h post-mortem, which was higher in LAL compared to LA (p = 0.016). Piglets fed with seaweed had a significantly lower n-6/n-3 PUFA ratio compared to control, to which the higher accumulation of C20:5n-3 (p = 0.001) and C18:4n-3 (p < 0.0001) contributed. In addition, meat of seaweed-fed piglets was enriched with bromine (Br, p < 0.001) and iodine (I, p < 0.001) and depicted a higher oxidative stability. This study demonstrates that the nutritional value of piglets’ meat could be improved by the dietary incorporation of L. digitata, regardless of CAZyme supplementation, without negatively affecting growth performance in the post-weaning stageinfo:eu-repo/semantics/publishedVersio

Amerindian genetic ancestry and INDEL polymorphisms associated with susceptibility of childhood B-cell Leukemia in an admixed population from the Brazilian Amazon

AbstractAcute lymphoblastic leukemia (ALL) is a malignant tumor common in children. Studies of genetic susceptibility to cancer using biallelic insertion/deletion (INDEL) type polymorphisms associated with cancer development pathways may help to clarify etymology of ALL. In this study, we investigate the role of eight functional INDEL polymorphisms and influence of genetic ancestry to B-cell ALL susceptibility in children of Brazilian Amazon population, which has a high degree of inter-ethnic admixture. Ancestry analysis was estimated using a panel of 48 autosomal ancestry informative markers. 130 B-cell ALL patients and 125 healthy controls were included in this study. The odds ratios and 95% confidence intervals were adjusted for confounders. The results indicated an association between the investigated INDEL polymorphisms in CASP8 (rs3834129), CYP19A1 (rs11575899) e XRCC1 (rs3213239) genes in the development of B-cell ALL. The carriers of Insertion/Insertion (Ins/Ins) genotype of the polymorphism in CASP8 gene presented reduced chances of developing B-cell ALL (P=0.001; OR=0.353; 95% CI=0.192–0.651). The Deletion/Deletion (Del/Del) genotype of the polymorphism in CYP19A1 gene was associated to a lower chance of developing B-cell ALL (P=3.35×10−6; OR=0.121; 95% CI=0.050–0.295), while Del/Del genotype of the polymorphism in XRCC1 gene was associated to a higher chance of developing B-cell ALL (P=2.01×10−4; OR=6.559; 95% CI=2.433–17.681). We also found that Amerindian ancestry correlates with the risk of B-cell ALL. For each increase of 10% in the Amerindian ancestry results in 1.4-fold chances of developing B-cell ALL (OR=1.406; 95% IC=1.123–1.761), while each increase of 10% in the European ancestry presents a protection effect in the development of B-cell ALL (OR=0.666; 95% IC=0.536–0.827). The results suggest that genetic factors influence leukemogenesis and might be explored in the stratification of B-cell ALL risk in admixed populations

Enhancing meat quality of weaned piglets with the dietary incorporation of Ulva lactuca and carbohydrases supplementation

Pubmed Free articleThe impact of the dietary incorporation of 7% Ulva lactuca, a green seaweed, on the quality and nutritional value of piglet’s meat was assessed. U. lactuca is rich in nutrients and bioactive compounds but its cell wall is composed of complex polysaccharides that reduce their bioavailability. Therefore, the effect of supplementing piglet diets with exogenous carbohydrases was also assessed here. A total of 40 male weaned piglets were divided into four dietary groups, each with 10 piglets: control (wheat, maize and soybean meal-based diet), UL (7% U. lactuca replacing the control diet), UL +R (UL and 0.005% Rovabio®), and UL +E (UL and 0.01% ulvan lyase). The piglets were fed the diets for 2 weeks. The results showed that incorporating U. lactuca in piglet diets did not influence most of the meat quality traits (P >0.05). However, the incorporation of U. lactuca with the commercial carbohydrase (UL +R) increased the amount of the docosahexaenoic acid (DHA; 22:6n-3) in their meat (P =0.011) compared with the control, by 54%. In addition, meat from piglets fed seaweed diets showed a nearly two-fold increase in iodine contents (P <0.001). Meat tenderness, juiciness and overall acceptability of piglets fed the control diet and the UL diet were lower than those fed the diets containing seaweed and carbohydrases (P <0.001). Overall, the findings indicate that 7% U. lactuca in the diets of weaned piglets had no major detrimental effects on meat quality and their carbohydrase supplementation has the potential to improve meat sensory traits.Funding: This research was funded by Fundação para a Ciência e a Tecnologia (FCT), Portugal, through PTDC/CAL-ZOO/30238/2017 grant, associated with a post-doc contract to M.C., and PhD fellowships to J.M. P. (SFRH/BPD/116816/2016) and D.M.R. (SFRH/BD/143992/2019). CIISA (UIDB/00276/2020), AL4AnimalS (LA/P/0059/2020) and LEAF (UIDB/04129/2020) grants, also from FCT, are also acknowledged. The authors acknowledge Teresa Costa from Indukern, Lda. (Sintra, Portugal), for the Rovabio® Excel AP kind donation. The graphical abstract was created using BioRender.com (https://biorender.com/).info:eu-repo/semantics/publishedVersio