Peptide structure stabilization by membrane anchoring and its general applicability to the development of potent cell-permeable inhibitors

Isolated protein motifs that are involved in interactions with their binding partners can be used to inhibit these interactions. However, peptides corresponding to protein fragments tend to have no defined secondary or tertiary structure in the absence of scaffolding by the rest of protein molecule. This results in low inhibitor potency. NMR and CD spectroscopy studies of lipopeptide inhibitors of the Hedgehog pathway revealed that membrane anchoring allows the cell membrane to function as a scaffold and facilitate the folding of short peptides. In addition, lipidation enhances cell permeability and increases the concentration of the compounds near the membrane, thus facilitating potent inhibition. The general applicability of this rational approach was further confirmed by the generation of selective antagonists of the insulin-like growth factor 1 receptor with GI(50) values in the nanomolar range. Lipopeptides corresponding to protein fragments were found to serve as potent and selective inhibitors of a number of nondruggable molecular targets.     

Milking machine function and analysis

Milking system evaluation is facilitated by use of a pulsator recorder, airflow meter, and a vacuum stability gauge. The most important points of a milking system are vacuum, inflations, and pulsation. An evaluation sheet for milking systems as proposed by the National Mastitis Council is presented. Paramount to evaluation of any milking system is that the parameters must relate to milking performance or teat and udder health.     

Automatic inductive voltage divider bridge for operation from 10 Hzto 100 kHz

A bridge to calibrate programmable and manual inductive voltage dividers is described. The bridge is based on a programmable 30-b binary inductive voltage divider with terminal linearity of ±0.1 ppm in phase and ±2 ppm quadrature at 400 Hz. Measurements of programmable test dividers can be automated using software developed to align the bridge components and perform an automatic balance     

Synthesis and properties of some polyamides containing the sulphonyl diphenylene unit

Homopolyamides of two isomeric di(aminophenyl) sulphones with α,ω-polymethylene dicarboxylic acids have been prepared by low-temperature solution polycondensation and by a high-temperature melt procedure. The glass transition temperatures (T_g) of these polymers are strongly dependent on the positions of the amine residues in the aromatic ring, and they fall progressively as the length of the methylene sequence in the dicarboxylic acid residue increases. The polymers are not crystallised by thermal treatment although in certain cases some crystallinity can be induced by treatment with an organic liquid. The presence of the bulky, polar sulphone group plays an important part in the success of the high-temperature polymerisation and makes a significant contribution to the high level of mechanical and thermal properties of the resulting polymers.     

A new assay for endocytosis in erythrocyte ghosts based on loss of acetylcholinesterase activity

A new method for assaying endocytosis in erythrocyte ghosts is presented. The method involves measuring the percentage loss of acetylcholinesterase activity which occurs when vacuoles form, making the acetylcholinesterase on the vacuole surface inaccessible. This method is compared to other methods of measuring endocytosis in this system, including phase contrast microscope estimation of vesiculation, stereological analysis of electron micrographs to determine vesiculation and loss of sialic acid accessible to neuraminidase due to endocytosis. Comparison of the percentage loss of acetylcholinesterase activity with the electron micrographic and sialic acid methods showed that all three methods gave a quantitative measure of the percentage of total membrane area taken in as vesicles. Since the acetylcholinesterase method was fast, easy, inexpensive, and quantitative, it was the preferred method for assay of endocytosis. The inhibition of endocytosis by Ca2+ was observed with this method; the success of this experiment demonstrated the applicability of the method to the study of inhibitors of endocytosis.     

Short communication: Effect of abomasal inorganic phosphorus infusion on phosphorus absorption in large intestine,milk production,and phosphorus excretion of dairy cattle

The objective of the study was to evaluate the effect of inorganic phosphorus (Pi) infusion on P absorption in large intestine, milk production, and phosphorus excretion. Four ruminally and ileally cannulated crossbred cows were used in a 4 × 4 Latin square with 21-d periods. Cows were fed a total mixed ration containing 0.21% P, providing 50% of the cows' P requirement. Cobalt-EDTA was used as marker to measure large intestine digesta flow. On d 13 to 21 of each period, each cow was infused daily with 0, 20.1, 40.2, or 60.3 g of Pi into the abomasum and total collection was conducted on d 18 to 21. Ileal samples were collected every 9 h on d 18 to 21. Feed, digesta, and fecal samples were analyzed for total P and Pi using the molybdovanadate yellow method and blue method, respectively. All data were analyzed using PROC GLIMMIX in SAS 9.3 (SAS Institute Inc., Cary, NC) using contrasts to evaluate linear, quadratic, and cubic effects of Pi infusion dose. Dry matter intake, apparent dry matter digestibility, milk yield, and milk total P were unaffected by Pi infusion. Ileal flow and fecal excretion of total P and Pi increased linearly with increasing infused Pi. In the large intestine, net absorption of TP and Pi was increased linearly with increasing infused Pi. The magnitude of absorption from the large intestine was greater than reflected in current models, raising questions that could be evaluated with longer infusion periods or dietary alteration.     

Determination of Heat Conductivity and Thermal Diffusivity of Waste Glass Melter Feed: Extension to High Temperatures

The heat conductivity (λ) and the thermal diffusivity (a) of reacting glass batch, or melter feed, control the heat flux into and within the cold cap, a layer of reacting material floating on the pool of molten glass in an all‐electric continuous waste glass melter. After previously estimating λ of melter feed at temperatures up to 680°C, we focus in this work on the λ(T) function at T > 680°C, at which the feed material becomes foamy. We used a customized experimental setup consisting of a large cylindrical crucible with an assembly of thermocouples, which monitored the evolution of the temperature field while the crucible with feed was heated at a constant rate from room temperature up to 1100°C. Approximating measured temperature profiles by polynomial functions, we used the energy equation to estimate the λ(T) approximation function, which we subsequently optimized using the finite‐volume method combined with least‐squares analysis. The heat conductivity increased as the temperature increased until the feed began to expand into foam, at which point the conductivity dropped. It began to increase again as the foam turned into a bubble‐free glassmelt. We discuss the implications of this behavior for the mathematical modeling of the cold cap.