Characterization of the Particles of Purified κ-Casein: Trypsin as a Probe of Surface-Accessible Residues

κ-Casein as purified from bovine milk exhibits a rather unique disulfide bonding pattern as revealed by SDS–PAGE. The disulfide-bonded caseins present range from dimer to octamer and above and preparations contain about 10% monomer. All of these heterogeneous polymers, however, self-associate into nearly spherical particles with an average diameter of 13 nm at pH 8.0, as revealed by negatively stained transmission electron micrographs and dynamic light scattering. The weight-average molecular weight of the aggregates at pH 8.0, as judged by analytical ultracentrifugation, is 648,000. Trypsin digestion at pH 8.0 was used to probe the surface groups of the κ-casein A polymers. The reaction with trypsin was rapid and the peptides liberated were identified by separation with reverse-phase HPLC, amino acid analysis, and protein sequencing. The most rapidly released peptides (t _1/2 < 30 sec) were from cleavage at Arg 97 and Lys residues 111 and 112. These results suggest a surface orientation for these residues, and the data are in accord with earlier proposed 3D predictive models for κ-casein. It is speculated that Arg 97, together with adjacent His residues (98 and 100) and Lys residues 111 and 112, form two positively charged clusters on the surface of the otherwise negatively charged casein. These clusters bracket the neutral chymosin cleavage site (whose hydrolysis triggers a well-known digestive process) and so these clusters may facilitate docking of the substrate caseins with chymosin.     

Management of kudzu by the bioherbicide,Myrothecium verrucaria,herbicides and integrated control programmes

Replicated field plots were established and monitored for two years to evaluate management practices for kudzu. The bioherbicidal plant pathogen, Myrothecium verrucaria, several herbicides and a variety of integrated control programmes achieved a high level of kudzu suppression, although no system tested reliably achieved eradication in this time frame.     

Spatial distribution of the active surveillance of sheep scrapie in Great Britain: an exploratory analysis

Background  

This paper explores the spatial distribution of sampling within the active surveillance of sheep scrapie in Great Britain. We investigated the geographic distribution of the birth holdings of sheep sampled for scrapie during 2002 – 2005, including samples taken in abattoir surveys (c. 83,100) and from sheep that died in the field ("fallen stock", c. 14,600). We mapped the birth holdings by county and calculated the sampling rate, defined as the proportion of the holdings in each county sampled by the surveys. The Moran index was used to estimate the global spatial autocorrelation across Great Britain. The contributions of each county to the global Moran index were analysed by a local indicator of spatial autocorrelation (LISA).     

Effects of converting enzyme inhibitors on renal P-450 metabolism of arachidonic acid

The effects of blockade of the renin-angiotensin system on the renal metabolism of arachidonic acid (AA) were examined. Male Sprague-Dawley rats were treated with vehicle, captopril (25 mg x kg(-1) x day(-1)), enalapril (10 mg x kg(-1) x day(-1)), or candesartan (1 mg x kg(-1) x day(-1)) for 1 wk. The production of 20-hydroxyeicosatetraenoic acid (20-HETE) and epoxyeicosatrienoic acids (EETs) by renal cortical microsomes increased in rats treated with captopril by 59 and 24% and by 90 and 58% in rats treated with enalapril. Captopril and enalapril increased 20-HETE production in the outer medulla by 100 and 143%, respectively. In contrast, blockade of ANG II type 1 receptors with candesartan had no effect on the renal metabolism of AA. Captopril and enalapril increased cytochrome P-450 (CYP450) reductase protein levels in the renal cortex and outer medulla and the expression of CYP450 4A protein in the outer medulla. The effects of captopril on the renal metabolism of AA were prevented by the bradykinin-receptor antagonist, HOE-140, or the nitric oxide (NO) synthase inhibitor, N(G)-nitro-L-arginine methyl ester. These results suggest that angiotensin-converting enzyme inhibitors may increase the formation of 20-HETE and EETs secondary to increases in the intrarenal levels of kinins and NO.     

REVIVAL OF MAMMALIAN SPERM AFTER IMMERSION IN LIQUID NITROGEN

1. A wide variety of procedures was used to test the motility of mammalian sperm after plunging them into liquid nitrogen at –195°C. and later rapidly warming them to 35°C. by plunging them into a suitable balanced and isotonic medium. 2. Using seminal fluid sperm from the same human donor, maximal numbers of motile sperm survived vitrification when the samples were (a) very fresh, (b) untreated with plasmolysing solutions, (c) plunged into the refrigerant in the form of a foam. The maximum yield of motile human sperm recoverable from the liquid nitrogen was 50 per cent. Since in this sample only 75 per cent of the sperm were alive before immersion, 67 per cent of the living sperm survived vitrification. 3. Experiments with sperm from 31 rabbits were made with a variety of conditions of pretreatment to obtain maximal yields of recoverable, motile sperm after vitrification by liquid nitrogen. (a) A consistent recoverable yield of about 0.5 per cent was obtained when the untreated suspension of sperm was smeared on cellophane and partially dried in air before immersing in liquid nitrogen. (b) On a few out of many occasions plasmolysis for several minutes with hypertonic Ringer solution gave a recoverable yield of 0.1 per cent as did (c) pretreatment with hypertonic Ringer and butyric acid.     

ACCUMULATION OF SALT AND PERMEABILITY IN PLANT CELLS

1. Comparisons are made of concentrations of K and Br in exudates of barley roots and in expressed sap from roots, under conditions favorable for aerobic metabolism. Both methods lead to the same general viewpoint concerning metabolically governed transport of solutes by living plant cells. 2. Cyanide in low concentration prevented salt accumulation by barley roots. Methylene blue, without decrease of CO₂ production by roots, destroyed power of salt accumulation. 3. K and Br ions entered roots to only a slight extent under an anaerobic condition, even with an inward gradient of ionic concentration. 4. Lactate or alcohol, under aerobic conditions, did not prevent rapid accumulation of salt by root cells. 5. Experiments on fluids obtained by suction from tomato roots gave evidence of loss of salt-accumulating power under the influence of N₂ gas or CO₂ gas, together with probable effects on cell permeability. 6. Several experiments on Nitella cells in which radioactive isotopes were used are reported. Bromide gradually moved into vacuolar sap until the concentration appeared to exceed that of the protoplasm, on the basis of the results of the several types of experiments. Accumulation of salt in the vacuole did not occur anaerobically. 7. Some views of interrelations of permeability, salt accumulation, and metabolism are suggested for further discussion.     

Phytoplankton community response to reservoir aging, 1968–92

The effects of reservoir aging on the phytoplankton community of amidwestern U.S. reservoir constructed in 1965 (Pawnee Reservoir) werestudied by comparing algal biovolume and species composition from April 1992through November 1992 to surveys conducted in 1968–73 and 1990. Meansummer total phosphorus, nitrate-nitrogen, Secchi disk depth, totalsuspended solids, chlorophyll a, and phytoplankton species composition datacharacterized Pawnee Reservoir during 1968–69 as a high nutrient,relatively clear water environment. Phytoplankton biomass was relativelylow, consisting mainly of cyanophytes and non-flagellated chlorophytes.During 1970–73, water clarity was poor, total suspended solids werehigh, and total phosphorus was lower, but was still greater than 100 µgl^–1. The 1970–73 phytoplankton biomass was high and wasdominated by cyanophytes. Mean summer total phosphorus remained >100µg l^–1, water clarity remained poor, but phytoplanktonbiomass decreased significantly during 1990–92. The dramatic drop inchlorophyll a and low mean volatile suspended solids indicated thatinorganic suspended sediments, rather than phytoplankton, accounted for themajority of the turbidity in 1990-92. In addition to lower phytoplanktonbiomass, community composition shifted away from buoyancy-regulatingcyanophytes toward flagellated chlorophytes. These data suggest that asreservoirs located in agricultural watersheds age, (1) inorganic suspendedsediments have a significant effect on the light environment as well asphytoplankton biomass and species composition, (2) the control ofphytoplankton biomass and species composition shifts away from nutrients tolight and suspended sediments, and (3) there is a 1–2 year lag in theresponse of phytoplankton biomass to maximum nutrient loading during thetrophic upsurge period. Thus, sedimentation has been shown to be a primarydeterminant of plankton and benthic macroinvertebrate community compositionas Pawnee Reservoir aged.     

Die Genetik der bipolaren Störung

With a life-time prevalence of about 0.5–1.5%, bipolar (manic depressive) disorder represents a common psychiatric disease. Family, twin, and adoption studies have consistently shown that genetic factors contribute to disease development. Genome-wide linkage studies have detected chromosomal regions that are very likely to harbor predisposing genes. Meta-analyses suggest, however, that the genetic contribution of the individual loci must be relatively small which could be one reason for the difficulties in identifying the genes responsible. Very recently, genome-wide association analyses, investigating hundreds of thousands of single nucleotide polymorphisms (SNPs) in phenotypically well-characterized patient and control cohorts, promise a major breakthrough in search of disease-associated genes.