Effect of particle size on the anthelmintic efficacy of mebendazole against Nippostrongylus brasiliensis in the rat.

The effect of reduction in particle size on the anthelmintic efficacy of mebendazole (methyl 5 (6)- benzoyle 1–2 benzimidazole carbamate) was studied in rats undergoing a primary infection with N. brasiliensis. A single oral treatment with fine ground mebendazole (particle size spectrum—54·95 per cent of particles less than 10·62 μ dia.; 86·06 per cent less than 21·27 μ) removed more than 98 per cent of adult worms from the intestine at a dose rate of 12·5 mg/kg body wt. On the other hand the best result achieved with coarse ground mebendazole (18·47 per cent of particles less than 10·62 μ dia; 42·26 per cent less than 21·27 μ dia) was 58 per cent of adult worms removed at a dose rate of 100 mg/kg body wt. It was also shown that fine ground mebendazole adversely affected migrating third stage larvae of N. brasiliensis.     

Photophosphorylation and related properties of reaggregated vesicles from spinach Photosystem I particles

The small Photosystem I particles prepared from spinach chloroplasts by the action of Triton X-100 (TSF 1 particles) reaggregate into membrane structures when they are incubated with soybean phospholipids and cholate and then subjected to a slow dialysis. The membranes so formed are vesicular in nature and show the capability of catalyzing phenazine methosulfate-mediated cyclic photophosphorylalation at rates which are usually about 20% of those observed with chloroplasts, but higher rates have been obtained. When coupling factor is removed from the chloroplasts by treatment with EDTA, a requirement for coupling factor can be shown for the subsequent ATP formation. The uncouplers carbonylcyanide 3-chlorophenyl-hydrazone, valinomycin, Triton X-100 and NH⁺₄ are effective with the reformed vesicles, which do not show the typical light-induced pH gradient observed with chloroplasts. Incubation of the TSF 1 particles with phospholipids alone allows for the formation of membrane vesicles, but such vesicles are only slightly active in ATP formation. In most properties investigated, the reformed membrane vesicles resemble the original chloroplast membrane so far as phenazine methosulfate-mediated cyclic photophosphorylation is concerned, which indicates a high degree of selectivity in the reaggregation process. The major difference between chloroplasts and the reformed vesicles is the failure of the latter to show a light-induced pH gradient.     

Designing cross-linked lipase aggregates for optimum performance as biocatalysts

Cross-linked enzyme aggregates (CLEAs) are prepared by precipitation of an enzyme and then chemical cross-linking the precipitate. Three CLEAs of lipase with glutaraldehyde concentrations of 10 mM (CLEA A), 40 mM (CLEA B) and 60 mM (CLEA C) were prepared. Studies show that there is a trade-off between thermal stability vs transesterification/hydrolysis rate vs enantioselectivity. The initial rates for transesterification of β-citronellol for the uncross-linked enzyme and CLEAs A, B and C were 243, 167, 102 and 40 µmol mg^?1 h^?1, respectively. Their thermal stabilities in aqueous media, as reflected by their half-life values at 55°C, were 6, 9, 13 and 16 h, respectively. The enantioselectivity, E values (for kinetic resolution of β-citronellol by transesterification) were 19, 74, 11 and 6, respectively. These results show that CLEA C was the most thermostable; the uncross-linked enzyme was best at obtaining the highest transesterification rate; and CLEA A was best suited for the enantioselective synthesis. Scanning electron microscopy (SEM) showed that the morphology of CLEA was dependent upon the extent of cross-linking.     

Effects of rosuvastatin on electronegative LDL as characterized by capillary isotachophoresis: the ROSARY Study

Electronegative LDL, a charge-modified LDL (cm-LDL) subfraction that is more negatively charged than normal LDL, has been shown to be inflammatory. We previously showed that pravastatin and simvastatin reduced the electronegative LDL subfraction, fast-migrating LDL (fLDL), as analyzed by capillary isotachophoresis (cITP). The present study examined the effects of rosuvastatin on the more electronegative LDL subfraction, very-fast-migrating LDL (vfLDL), and small, dense charge-modified LDL (sd-cm-LDL) subfractions. Patients with hypercholesterolemia or those who were being treated with statins (n = 81) were treated with or switched to 2.5 mg/d rosuvastatin for 3 months. Rosuvastatin treatment effectively reduced cITP cm-LDL subfractions of LDL (vfLDL and fLDL) or sdLDL (sd-vfLDL and sd-fLDL), which were closely related to each other but were different from the normal subfraction of LDL [slow-migrating LDL (sLDL)] or sdLDL (sd-sLDL) in their relation to the levels of remnant-like particle cholesterol (RLP-C), apolipoprotein (apo) C-II, and apoE. The percent changes in cm-LDL or sd-cm-LDL caused by rosuvastatin were correlated with those in the particle concentrations of LDL or sdLDL measured as LDL-apoB or sdLDL-apoB and the levels of HDL-C, RLP-C, apoC-II, and apoE. In conclusion, rosuvastatin effectively reduced both the vfLDL subfraction and sd-cm-LDL subfractions as analyzed by cITP.     

Cryo-electron microscopy reveals the membrane insertion mechanism of V. cholerae hemolysin

Vibrio cholerae hemolysin (HlyA) is a 65?kDa pore-forming toxin which causes lysis of target eukaryotic cells by forming heptameric channels in the plasma membrane. Deletion of the 15?kDa C-terminus β-prism carbohydrate-binding domain generates a 50?kDa truncated variant (HlyA50) with 1000-fold-reduced pore-forming activity. Previously, we showed by cryo-electron microscopy that the two toxin oligomers have central channels, but the 65?kDa toxin oligomer is a seven-fold symmetric structure with bowl-, ring-, and arm-like domains, whereas the 50?kDa oligomer is an asymmetric jar-like heptamer. In the present study, we determined three-dimensional(3D) structures of HlyA and HlyA50 in presence of erythrocyte stroma and observed that interaction of the 65?kDa toxin with the stroma induced a significant decrease in the height of the β-barrel oligomer with a change in conformation of the ring- and arm-like domains of HlyA. These features were absent in interaction of HlyA50 with stroma. We propose that this conformational transition is critical for membrane-insertion of the toxin.     

Performance,rumen fermentation,blood minerals,leukocyte and antioxidant capacity of young Holstein calves receiving high-surface ZnO instead of common ZnO

ABSTRACT

This study was conducted to assess the effect of feeding high-surface ZnO instead of common ZnO on the performance, rumen fermentation, blood minerals, leukocytes and antioxidant capacity of pre- and post-weaning calves. Thirty male suckling Holstein calves were allotted to one of three experimental groups (10 replicates) in a completely randomised design. Calves received: (1) a low Zn diet without Zn supplementation (control diet), (2) a high Zn diet containing 50 mg supplementary Zn/kg dry matter (DM) as common ZnO or (3) a high Zn diet containing 50 mg supplementary Zn/kg DM as high-surface ZnO (nano-ZnO). The control diet contained a native Zn content of 35.5, 34.7 or 33.7 mg/kg DM for the age periods of 7 to 30, 31 to 70 and 71 to 100 d, respectively. Supplementation of the diet with Zn did not change the dry matter intake (DMI) of calves during d 7 to 30 but increased the ADG in this period (p < 0.05). During age periods of 31 to 70 and 71 to 100 d, DMI and ADG of the Zn supplemented calves were higher (p < 0.05) than the control animals. The nutrient digestibility and the concentration of rumen volatile fatty acids were positively affected (p < 0.05) and the rumen ammonia-N concentration decreased (p < 0.05) by dietary Zn supplementation. Furthermore, the incidence of diarrhoea and pneumonia was lower in calves receiving the Zn supplemented diets. Irrespective of ZnO source, the blood total antioxidant capacity, leukocyte and haematocrit levels significantly increased (p < 0.05) with the ZnO supplemented diets. The post-weaning DMI, nutrient digestibility and blood haematocrit levels were higher in calves receiving high-surface ZnO, compared to those supplemented with common ZnO. With inclusion of the Zn sources in pre- and post-weaning diets, the blood Zn concentration increased (p < 0.05), but the blood Cu, Fe, Ca, P and Mg levels remained unchanged. Regardless of source, dietary supplementation of young calves with ZnO improved the performance and decreased rumen ammonia-N and the incidence of diseases. Moreover, high-surface ZnO had advantages over common ZnO in increasing the post-weaning feed intake, digestibility and blood Zn concentration.     

“Protein aggregates” contain RNA and DNA,entrapped by misfolded proteins but largely rescued by slowing translational elongation

All neurodegenerative diseases feature aggregates, which usually contain disease‐specific diagnostic proteins; non‐protein constituents, however, have rarely been explored. Aggregates from SY5Y‐APP_Sw neuroblastoma, a cell model of familial Alzheimer''s disease, were crosslinked and sequences of linked peptides identified. We constructed a normalized “contactome” comprising 11 subnetworks, centered on 24 high‐connectivity hubs. Remarkably, all 24 are nucleic acid‐binding proteins. This led us to isolate and sequence RNA and DNA from Alzheimer''s and control aggregates. RNA fragments were mapped to the human genome by RNA‐seq and DNA by ChIP‐seq. Nearly all aggregate RNA sequences mapped to specific genes, whereas DNA fragments were predominantly intergenic. These nucleic acid mappings are all significantly nonrandom, making an artifactual origin extremely unlikely. RNA (mostly cytoplasmic) exceeded DNA (chiefly nuclear) by twofold to fivefold. RNA fragments recovered from AD tissue were ~1.5‐to 2.5‐fold more abundant than those recovered from control tissue, similar to the increase in protein. Aggregate abundances of specific RNA sequences were strikingly differential between cultured SY5Y‐APP_Sw glioblastoma cells expressing APOE3 vs. APOE4, consistent with APOE4 competition for E‐box/CLEAR motifs. We identified many G‐quadruplex and viral sequences within RNA and DNA of aggregates, suggesting that sequestration of viral genomes may have driven the evolution of disordered nucleic acid‐binding proteins. After RNA‐interference knockdown of the translational‐procession factor EEF2 to suppress translation in SY5Y‐APP_Sw cells, the RNA content of aggregates declined by >90%, while reducing protein content by only 30% and altering DNA content by ≤10%. This implies that cotranslational misfolding of nascent proteins may ensnare polysomes into aggregates, accounting for most of their RNA content.     

A kinematic method for computing the motion of the body centre-of-mass (CoM) during walking: a Bayesian approach

The gait pattern of a particular patient can be altered in a large set of pathologies. Tracking the body centre-of-mass (CoM) during the gait allows a quantitative evaluation of these diseases at comparing the gait with normal patterns. A correct estimation of this variable is still an open question because of its non-linearity and inaccurate location. This paper presents a novel strategy for tracking the CoM, using a biomechanical gait model whose parameters are determined by a Bayesian strategy. A particle filter is herein implemented for predicting the model parameters from a set of markers located at the sacral zone. The present approach is compared with other conventional tracking methods and decreases the calculated root mean squared error in about a 56% in the x-axis and 59% in the y-axis.     

All roads lead to Rome (but some may be harder to travel): SRP-independent translocation into the endoplasmic reticulum

Abstract

Translocation into the endoplasmic reticulum (ER) is the first biogenesis step for hundreds of eukaryotic secretome proteins. Over the past 30 years, groundbreaking biochemical, structural and genetic studies have delineated one conserved pathway that enables ER translocation- the signal recognition particle (SRP) pathway. However, it is clear that this is not the only pathway which can mediate ER targeting and insertion. In fact, over the past decade, several SRP-independent pathways have been uncovered, which recognize proteins that cannot engage the SRP and ensure their subsequent translocation into the ER. These SRP-independent pathways face the same challenges that the SRP pathway overcomes: chaperoning the preinserted protein while in the cytosol, targeting it rapidly to the ER surface and generating vectorial movement that inserts the protein into the ER. This review strives to summarize the various mechanisms and machineries which mediate these stages of SRP-independent translocation, as well as examine why SRP-independent translocation is utilized by the cell. This emerging understanding of the various pathways utilized by secretory proteins to insert into the ER draws light to the complexity of the translocational task, and underlines that insertion into the ER might be more varied and tailored than previously appreciated.