Vaccination of lambs against Haemonchus contortus infection with a somatic protein (Hc23) from adult helminths

A somatic protein from adult Haemonchus contortus (Hc23), the most abundant component in a low molecular weight fraction with known immunizing effect against experimental haemonchosis, has been purified by immunochromatography. The immunoprophylactic value of Hc23 was tested in groups of 5–6 months old Assaf lambs using Al(OH)₃ or Escherichia coli lipopolysaccharide + inactivated Propionibacterium acnes as adjuvant and the results compared with uninfected control, uninfected and challenged or infected and challenged lambs. Immunization with Hc23 in either adjuvant elicited significant reductions in fecal egg counts after challenge with 15,000 L3s (70.67%–85.64%, respectively) and reduced (67.1% and 86%) abomasal worm counts (45 days post-challenge). Immunized lambs displayed higher peripheral eosinophil counts, were less anaemic and had weight gains than challenged controls. The results suggest that the Hc23 antigen can induce a partially protective response against haemonchosis in lambs.     

Calcium and EGTA Alleviate Cadmium Toxicity in Germinating Chickpea Seeds

Impact of exogenous calcium and ethylene glycol tetraacetic acid (EGTA) supplement on chickpea (Cicer arietinum L.) germinating seeds exposed to cadmium stress for 6 days was studied. Ca and EGTA late treatment (3 days) alleviated growth inhibition and decreased Cd accumulation as well as lipid peroxidation and protein carbonylation in both root and shoot cells. Exogenous effector application relieved Cd-induced cell death which was associated with a constant level of ATP, which was considered as an apoptotic-like process. Redox balance was examined through the study of the redox state of pyridine nucleotide couples NAD⁺/NADH and NADP⁺/NADPH as well as their related oxidative [NAD(P)H-oxidase] and dehydrogenase (glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and malate dehydrogenase) enzyme activities. The present research illustrated an ameliorative effect of Ca and EGTA on growth of Cd-exposed chickpea seedlings that occurs through the protection of sensitive cell sites from Cd-induced oxidation, namely membrane lipids and proteins, rather than the improvement of recycling capabilities of the cellular reducing power.     

Effect of some progestational steroids on lactation in Egyptian women. I. Milk yield during the first year of lactation.

The effect of monthly injectios of 300 mg Depo-Provera or 150 mg Deladroxone, and of daily oral administration of .5 mg chlormadinone acetate or .3 mg quingestanol acetate on lactation was studied in Egyptian women during the 1st year of lactation. Women receiving Depo-Provera had the highest milk yield, followed by those taking Deladroxone and quingestanol acetate. The milk yields while taking these hormonal preparations were higher than in untreated controls. However, those women taking chlormadinone acetate had lower milk yields than untreated controls. The increased milk yield is probably due to the progestagenic activity, and minimal estrogenicity, of these drugs.     

N‐terminal acetylation modestly enhances phase separation and reduces aggregation of the low‐complexity domain of RNA‐binding protein fused in sarcoma

The RNA‐binding protein fused in sarcoma (FUS) assembles via liquid–liquid phase separation (LLPS) into functional RNA granules and aggregates in amyotrophic lateral sclerosis associated neuronal inclusions. Several studies have demonstrated that posttranslational modification (PTM) can significantly alter FUS phase separation and aggregation, particularly charge‐altering phosphorylation of the nearly uncharged N‐terminal low complexity domain of FUS (FUS LC). However, the occurrence and impact of N‐terminal acetylation on FUS phase separation remains unexplored, even though N‐terminal acetylation is the most common PTM in mammals and changes the charge at the N‐terminus. First, we find that FUS is predominantly acetylated in two human cell types and stress conditions. Next, we show that recombinant FUS LC can be acetylated when co‐expressed with the NatA complex in Escherichia coli. Using NMR spectroscopy, we find that N‐terminal acetylated FUS LC (FUS LC Nt‐Ac) does not notably alter monomeric FUS LC structure or motions. Despite no difference in structure, Nt‐Ac‐FUS LC phase separates more avidly than unmodified FUS LC. More importantly, N‐terminal acetylation of FUS LC reduces aggregation. Our findings highlight the importance of N‐terminal acetylation of proteins that undergo physiological LLPS and pathological aggregation.     

Skeletal muscle morphology in power-lifters with and without anabolic steroids

The morphological appearance of the vastus lateralis (VL) muscle from high-level power-lifters on long-term anabolic steroid supplementation (PAS) and power-lifters never taking anabolic steroids (P) was compared. The effects of long- and short-term supplementation were compared. Enzyme-immunohistochemical investigations were performed to assess muscle fiber type composition, fiber area, number of myonuclei per fiber, internal myonuclei, myonuclear domains and proportion of satellite cells. The PAS group had larger type I, IIA, IIAB and IIC fiber areas (p<0.05). The number of myonuclei/fiber and the proportion of central nuclei were significantly higher in the PAS group (p<0.05). Similar results were seen in the trapezius muscle (T) but additionally, in T the proportion of fibers expressing developmental myosin isoforms was higher in the PAS group compared to the P group. Further, in VL, the PAS group had significantly larger nuclear domains in fibers containing ≥5 myonuclei. The results of AS on VL morphology in this study were similar to previously reported short-term effects of AS on VL. The initial effects from AS appear to be maintained for several years.     

Optimization of DsRed production in Escherichia coli: effect of ribosome binding site sequestration on translation efficiency

DsRed-Express, a popular reporter protein, cannot be expressed in Escherichia coli using a consensus ribosome binding site (RBS) potentially due to basepairing in the RBS that inhibits translation initiation. Saturation mutagenesis was used to probe for a gene sequence that minimized basepairing in the RBS while maintaining the same spectral properties and maturation characteristics as DsRed-Express. The new DsRed, designated here as RFP(EC) for E. coli optimized red fluorescent protein, fluoresces 2.5 times greater than DsRed-Express when expressed from the same vector.     

2-(2-Furanyl)-7-phenyl[1,2,4]triazolo[1,5-c]pyrimidin-5-amine analogs: highly potent, orally active, adenosine A2A antagonists. Part 1

The structure-activity relationship of this novel class of compounds based on 2-(2-furanyl)-7-phenyl[1,2,4]-triazolo[1,5-c]pyrimidin-5-amine, 1, and its analogs was evaluated for their in vitro and in vivo adenosine A(2A) receptor antagonism. Several compounds displayed oral activity at 3 mg/kg in a rat catalepsy model. Specifically, compound 8g displayed an excellent in vitro profile, as well as a highly promising in vivo profile.     

The solvent extraction of metal ions from aqueous solutions containing NNN′N′-Ethylenediaminetetraacetic and related ccids. Part 1. Uranium(IV)

The uranium(IV) complexes [U(EDTA)(H₂O)₂], [U(HOEDTA)]⁺, and [U(DTPA)]^? are well-formed in the pH fange 2–3 ([DTPA]^5- = diethylenetriaminepentaacetate; [HOEDTA]^3-  N-(2-hydroxyethyl)ethylenediaminetriacetate). Of these, only [U(DTPA)]- is extracted from an aqueous phase at pH 2 by the perchlorate salt of the primary amine, Primene JM-T. As the aqueous phase pH was raised, extraction occurred in all three cases and hydrolysed species may be extracted from EDTA and HOEDTA solutions but [U(DTPA)]^? resists hydrolysis. The addition of sulphate had a marked effect on the extraction of U(IV) from EDTA and HOEDTA through the formation of [U(EDTA)(SO₄)(H₂O)]^2- and [U(HOEDTA)(SO₄)(H₂O)_n]^?. The equilibrium constant, log β₁, for: [(U(EDTA)(H₂O)₂] 2 [SO₄]^2? ? [U(EDTA)(SO₄)(H₂O)]^2- 2 H₂O was found to be 2.43 ± 0.04 (I = 1 mol dm^?3, NaClO₄; pH 2.0; 20 °C) from spectrophotometric data.With tri-n-octylphosphine oxide (TOPO) electronic spectroscopy showed that the same U(IV) complex was extracted at pH 2 for Cs₂UCl₆, U(IV)/ HOEDTA, and U(IV)/DTPA and the aminepoly- carboxylates were aqueous phase masking agents but with [U(EDTA)(H₂O)₂] oxidation gave a uranyl(VI) organic phase species.Uranium(IV) is strongly extracted from aqueous solutions of HOEDTA at pH 2 or 3 by bis(2-ethyl- hexyl)phosphoric acid (HBEHP) but less so from EDTA and DTPA. Since U(IV) is completely extracted from Cs₂UCl₆ it could be that the amine- polycarboxylates were aqueous phase masking agents although spectral evidence did not support this.     

Phosphorylation of the FUS low‐complexity domain disrupts phase separation,aggregation, and toxicity

Neuronal inclusions of aggregated RNA‐binding protein fused in sarcoma (FUS) are hallmarks of ALS and frontotemporal dementia subtypes. Intriguingly, FUS's nearly uncharged, aggregation‐prone, yeast prion‐like, low sequence‐complexity domain (LC) is known to be targeted for phosphorylation. Here we map in vitro and in‐cell phosphorylation sites across FUS LC. We show that both phosphorylation and phosphomimetic variants reduce its aggregation‐prone/prion‐like character, disrupting FUS phase separation in the presence of RNA or salt and reducing FUS propensity to aggregate. Nuclear magnetic resonance spectroscopy demonstrates the intrinsically disordered structure of FUS LC is preserved after phosphorylation; however, transient domain collapse and self‐interaction are reduced by phosphomimetics. Moreover, we show that phosphomimetic FUS reduces aggregation in human and yeast cell models, and can ameliorate FUS‐associated cytotoxicity. Hence, post‐translational modification may be a mechanism by which cells control physiological assembly and prevent pathological protein aggregation, suggesting a potential treatment pathway amenable to pharmacologic modulation.