Structural Basis for the Function of the C-Terminal Proton Release Pathway in the Calcium Pump

The calcium pump (sarco/endoplasmic reticulum Ca²⁺-ATPase, SERCA) plays a major role in calcium homeostasis in muscle cells by clearing cytosolic Ca²⁺ during muscle relaxation. Active Ca²⁺ transport by SERCA involves the structural transition from a low-Ca²⁺ affinity E2 state toward a high-Ca²⁺ affinity E1 state of the pump. This structural transition is accompanied by the countertransport of protons to stabilize the negative charge and maintain the structural integrity of the transport sites and partially compensate for the positive charges of the two Ca²⁺ ions passing through the membrane. X-ray crystallography studies have suggested that a hydrated pore located at the C-terminal domain of SERCA serves as a conduit for proton countertransport, but the existence and function of this pathway have not yet been fully characterized. We used atomistic simulations to demonstrate that in the protonated E2 state and the absence of initially bound water molecules, the C-terminal pore becomes hydrated in the nanosecond timescale. Hydration of the C-terminal pore is accompanied by the formation of water wires that connect the transport sites with the cytosol. Water wires are known as ubiquitous proton-transport devices in biological systems, thus supporting the notion that the C-terminal domain serves as a conduit for proton release. Additional simulations showed that the release of a single proton from the transport sites induces bending of transmembrane helix M5 and the interaction between residues Arg762 and Ser915. These structural changes create a physical barrier against full hydration of the pore and prevent the formation of hydrogen-bonded water wires once proton transport has occurred through this pore. Together, these findings support the notion that the C-terminal proton release pathway is a functional element of SERCA and also provide a mechanistic model for its operation in the catalytic cycle of the pump.     

Corrosion behavior of T24, T92, VM12, and AISI 304 steels exposed to KCl–NaCl–K2SO4–Na2SO4 salt mixtures

The corrosion mechanisms of T24, T92, VM12, and AISI 304 steels are studied under the influence of NaCl–KCl, NaCl–Na₂SO₄, and KCl–K₂SO₄ salt mixtures in a dry air atmosphere at 650°C for 15 days. NaCl–KCl was the most aggressive deposit and AISI 304 stainless steel exhibited the highest corrosion resistance. There was no relation between the Cr content of the ferritic steels and their corrosion resistance in NaCl–KCl. In contrast, the resistance of high-Cr steels was better when exposed to NaCl–Na₂SO₄ and KCl–K₂SO₄. The high-Cr and the low-Cr steels were more susceptible to NaCl–Na₂SO₄ and to KCl–K₂SO₄, respectively.     

Fat Distribution and Lipid Profile of Young Adults with Congenital Adrenal Hyperplasia Due to 21-Hydroxylase Enzyme Deficiency

We aimed to compare detailed fat distribution and lipid profile between young adults with congenital adrenal hyperplasia due to 21-hydroxylase enzyme deficiency and a control group. We also verified independent associations of treatment duration and daily hydrocortisone dose equivalent (HDE) with lipid profile within patients. This case–control study included 23 patients (7 male and 16 female) matched by an age range of young adults (18–31 years) with 20 control subjects (8 male and 12 female). Dual energy X-ray absorptiometry was used to measure the fat distribution. Male patients demonstrated elevated indices of fat mass for total (7.7 ± 2.1 vs. 4.5 ± 1.3 kg/m², p = 0.003), trunk (4.0 ± 1.2 vs. 2.2 ± 0.8 kg/m², p = 0.005), android (0.63 ± 0.24 vs. 0.32 ± 0.15 kg/m², p = 0.008), gynoid (1.34 ± 0.43 vs. 0.74 ± 0.24 kg/m², p = 0.005), arm (0.65 ± 0.16 vs. 0.39 ± 0.10 kg/m², p = 0.009), and leg regions (2.7 ± 0.8 vs. 1.6 ± 0.4 kg/m², p = 0.005) than the control group, but not in females. However, female patients demonstrated elevated ratio of low-density lipoprotein cholesterol to high-density lipoprotein cholesterol (1.90 ± 0.46 vs. 1.39 ± 0.47, p = 0.009) than the control group, but not in males. Total fat mass was inversely correlated with total testosterone (r = −0.64, p = 0.014) and positively correlated with leptin in males (r = 0.75, p = 0.002). An elevated daily HDE (β = 0.43, p = 0.038 and β = 0.47, p = 0.033) and trunk to total fat mass ratio (β = 0.46, p = 0.025, and β = 0.45, p = 0.037) were independently correlated with impaired lipid profile markers. Although there is no altered lipid profile, male patients demonstrated an increased fat distribution. However, female patients presented with an impaired lipid profile marker but demonstrated close values of normal fat distribution. Interestingly, the dose of glucocorticoid therapy can have some role in the lipid mechanisms.     

Development of low-alcohol isotonic beer by interrupted fermentation

Alcohol-free beer with isotonic properties is getting more popular and its production can be carried out by different production strategies; however, interrupted fermentation is still a challenge. Therefore, the objective of this study was to develop a low-alcohol isotonic beer (<0.5% v/v) by interrupted fermentation. Moreover, the major objective is to compare the developed product to commercial beverages (sports drinks, ‘Pilsen' regular beer, alcohol-free beers and low-alcohol isotonic beer). The beverages were evaluated based on pH, alcohol content (% v/v), total titratable acidity (mEq L⁻¹), osmolality (mOsmol kg⁻¹), bitterness International Bitterness Units, colour European Brewery Convention, total phenolic compounds (mg L⁻¹ gallic acid), reducing and total sugars (%) and Na and K contents (mg L⁻¹). The developed low-alcohol isotonic beer presented characteristics similar to sports drinks, with the advantage of being richer in phenolic compounds and suitable osmolality. Despite salts were added in its formulation, the grades attributed to all beers employed in the sensory evaluation, as well as the purchase intention did not present significant differences.     

An index structure supporting rule activation in pervasive applications

World Wide Web - Rule mechanism has been widely used in many areas, such as databases, artificial intelligent and pervasive computing. In a rule mechanism, rule activation decides which rules are...     

Effect of temperature on raw whole milk density and its potential impact on milk payment in the dairy industry

The objective of this study was to determine the effect of temperature on whole milk density measured at four different temperatures: 5, 10, 15, and 20 °C. A total of ninety-three individual milk samples were collected from morning milking of thirty-two Holstein Friesian dairy cows, of national average genetic merit, once every two weeks over a period of 4 weeks and were assessed by Fourier transform infrared spectroscopy for milk composition analysis. Density of the milk was evaluated using two different analytical methods: a portable density meter DMA35 and a standard desktop model DMA4500M (Anton Paar GmbH, UK). Milk density was analysed with a linear mixed model with the fixed effects of sampling period, temperature and analysis method; triple interaction of sampling period x analysis method x temperature; and the random effect of cow to account for repeated measures. The effect of temperature on milk density (ρ) was also evaluated including temperature (t) as covariate with linear and quadratic effects within each analytic method. The regression equation describing the curvature and density–temperature relationship for the DMA35 instrument was ρ = 1.0338−0.00017T−0.0000122T² (R² = 0.64), while it was ρ = 1.0334 + 0.000057T−0.00001T² (R² = 0.61) for DMA4500 instrument. The mean density determined with DMA4500 at 5 °C was 1.0334 g cm⁻³, with corresponding figures of 1.0330, 1.0320 and 1.0305 g cm⁻³ at 10, 15 and 20 °C, respectively. The milk density values obtained in this study at specific temperatures will help to address any bias in weight–volume calculations and thus may also improve the financial and operational control for the dairy processors in Ireland and internationally.     

Development of a new ion-imprinted polymer (IIP) with Cd2+ ions based on divinylbenzene copolymers containing amidoxime groups

Polymer Bulletin - In this study, we synthesized a new ion-imprinted polymer (IIP) based on introduction of amidoxime groups in acrylonitrile, complexation with Cd2+ ions and polymerization with...