Search for the rare decays K(L)→π0π0µ+µ- and K(L)→π0π0X0→π0π0µ+µ-.

The KTeV E799 experiment has conducted a search for the rare decays, K(L)→π(0)π(0)µ(+)µ(-) and K(L)→π(0)π(0)X(0)→π(0)π(0)µ(+)µ(-), where the X(0) is a possible new neutral boson that was reported by the HyperCP experiment with a mass of (214.3 ± 0.5) MeV/c(2). We find no evidence for either decay. We obtain upper limits of Br(K(L)→π(0)π(0)X(0)→π(0)π(0)µ(+)µ(-)) < 1.0 × 10(-10) and Br(K(L)→π(0)π(0)µ(+)µ(-)) < 9.2 × 10(-11) at the 90% confidence level. This result rules out the pseudoscalar X(0) as an explanation of the HyperCP result under the scenario that the dsX(0) coupling is completely real.

Messenger RNA structure: compatibility of hairpin loops with protein sequence.

Examination of the amino acid sequences of human cytochrome c and the alpha-chain variant of human hemoglobin Constant Spring has revealed the possiblity for base-paired hairpin loops in the messenger RNA's for these proteins. A similar analysis of the bacteriophage R17 coat protein suggests an additional unobserved loop in the R17 RNA. If such loops are present in messenger RNA's generally, it would suggest that DNA has more than one stable base-paired conformation.

Measurements of the decay KL-->e+ e- gamma.

The E799-II (KTeV) experiment at Fermilab has collected 83 262 K(L)-->e+ e- gamma(gamma) events above a background of 79 events. We measure a decay width, normalized to the K(L)-->pi0pi0pi(D)0 (pi0-->gammagamma, pi0-->gammagamma, pi(D0-->e+ e- gamma(gamma)) decay width, of Gamma(K(L)-->e+e-gamma(gamma))/Gamma(K(L)-->pi0pi0pi(D)0)=(1.3302+/-0.0046(stat)+/-0.0102(syst)) x 10(-3). We also measure parameters of two K(L)gamma*gamma form factor models. In the Bergström-Massó-Singer parametrization, we find Calpha(K*)= -0.517 +/- 0.030(stat) +/- 0.022(syst). We separately fit for the first parameter of the D'Ambrosio-Isidori-Portolés model and find alpha(DIP)= -1.729 +/- 0.043(stat) +/- 0.028(syst).

The effect of essential fatty acid deficiency upon fatty acid uptake by the brain.

Young adult rats, either control or essential fatty acid deficient, were administered either [3-H] oleic acid or [3-H] arachidonic acid by stomach tube. In addition, a group of control rats was given [3-H] palmitic acid. The rats were killed at various times therafter, and the radioactivity of the lipids of brain and plasma was examined. In confirmation of previous work, the blood lipid label was found to rise rapidly and then fall, wheras the activity of brain lipids increased slowly and did not show a decline through the 24-h period studied. Analysis of the brain uptake data according to first-order kinetics confirmed the impressions gained from visual inspection of the data. The initial rate of uptake of arachidonic acid was about 4.5 times that of oleic acid in control animals and in deficient animals. Essential fatty acid deficiency, however, did not induce an altered rate of uptake for either oleic acid or arachidonic acid. The rate of uptake of palmitic acid by control rats was not significantly different from that of oleic acid. Even though the initial rates of incorporation of oleic and arachidonic acids were not changed during essential fatty acid deficiency, the final levels of radioactivity obtained in brain lipids were higher in deficient rats with both fatty acids. The plateau value obtained with oleic acid was 1.5 times higher in deficient animals, while the plateau value for arachidonic acid was 1.7 times higher. An experiment in which deficient animals were allowed access to a control diet for 12 or 24 h prior to the labeling experiment suggested that the higher levels of radioactivity found in brain lipids of deficient animals was not due to an isotope dilution effect. Such animals still displayed the labeling pattern of deficient animals with arachidonic acid, while the results with oleic acid varied somewhat. Our results suggest that essential fatty acid deficiency does not alter the ability of the brain to take up the fatty acids studied. However, the fatty acids, especially arachidonic, are retained in the brain to a greater extent in the deficient animals.

Odd-and even-numbered fatty acids. Their contrasting behavior in normal and fowlpox virus-infected epithelium.

Upon infection with fowlpox virus, the amount of odd-numbered fatty acids in chick scalp epithelium shows a significant decrease compared with control values. This effect begins quite early and progresses throughout the period of infection. Individual members of the odd-numbered family (C15--C27 inclusive) were quantitatively related to the group as a whole during most of the infection. Experiments involving the administration of labeled acetate in vivo demonstrated an increase in the synthesis of even-numbered fatty acids and a decrease in the synthesis of odd-numbered fatty acids in infected epithelium. The reduced synthesis of odd-numbered fatty acids in infected epithelium could also be demonstrated with labeled propionate. The influence of the alpha-oxidation pathway was assayed in chick scalp epithelium in vivo by the administration of [1-14C,9,10-3H] stearic acid. The C17 acids formed had a 3H/14C ratio similar to that of the C16 acids, indicating that most label incorporation into C17 was due to beta-oxidation to acetate followed by resynthesis into fatty acids. C17 fatty acids from control and infected epithelium had similar 3H/14C ratios, indicating that the alpha-oxidation pathway probably does not contribute to the differences in odd-numbered fatty acid content observed. In assays for fatty acid synthetase activty, both [14C] acetyl-CoA and [14C]-propionyl-CoA were used as initial acceptors. The specific activities of preparations from infected scalp were similar to those of control preparations with both substrates. These results suggest that there is no decline in the ability to utilize propionate for fatty acid synthesis in infected epithelium.

Carbohydrate compositional effects on tissue distribution of chicken riboflavin-binding protein.

Riboflavin-binding proteins (RBP) purified from chicken egg white, yolk and the serum of laying hens differ in their carbohydrate compositions reflecting tissue-specific modifications of a single gene product. All three are complex glycoproteins having more than twice as many N-acetylglucosamine residues (greater than 12) as mannose residues (approx. 6). Egg white RBP is distinctive in having only one sialic acid and two galactose residues. Serum RBP contains approx. five sialic acid and seven galactose residues. In addition there is one residue of fucose. The carbohydrate composition of yolk RBP indicated the hydrolysis, respectively, of one, one, two and 3 residues of sialic acid, fucose, galactose, and N-acetylglucosamine from its precursor, serum RBP. The effect of these differing levels of glycosylation on plasma clearance, ovarian uptake and tissue distribution of 125I-labeled riboflavin-binding proteins in laying hens were compared. 2 h after intravenous injection, 19% of the egg white RBP, 29% of the yolk RBP, and 37% of the serum RBP remained in circulation. The kinetics of plasma clearance was distinctly biphasic for each of the radioiodinated proteins. The initial rapid-turnover component (t1/2 = 13 min) ranged from 27% of the serum RBP sample to 48% of the egg white RBP sample. The remaining slow-turnover components were cleared with half-lives of 81 min (egg white RBP), 101 min (yolk RBP), and 121 min (serum RBP). 16 h after injection, only 4% of the egg white RBP was deposited in the yolk of developing oocytes while about 12% of the serum RBP and yolk RBP was deposited. This highly significant difference is apparently due to preferential, carbohydrate-dependent clearance of egg white RBP by the liver rather than preferential uptake of serum and yolk RBP by the ovarian follicle. We find no evidence for carbohydrate-directed uptake of riboflavin-binding protein by the ovarian follicle.

Eye tracking in relation to age, sex, and illness.

Impaired eye-tracking performance in psychotic patients has been previously demonstrated. Nine groups of ten patients each were used to determine if, when equated for age and sex, some of the eye-tracking dysfunction observed by others could be alternately explained in terms of aging, sex, or the presence of Parkinson-like symptoms. Horizontal eye tracking was recorded while patients visually tracked a target moving at 5degrees/sec and 20degrees/sec. Both sinusoidal and triangular target movements were used. Results indicated that Parkinson patients were the poorest eye trackers and that young normal men were the best. Additionally, men were better eye trackers than women and normal controls were better eye trackers than were schizophrenic patients or their parents.

An oocytic membrane receptor for biotin-binding protein.

The chicken oocyte accumulates a biotin-binding protein (BBP) in the yolk that is distinct from the avidin in the 'egg white'. An identical BBP to that of the yolk is also present in the circulation of the laying hen. We report the first evidence for the existence of a BBP receptor in the oocyte vitelline membrane. Reduction of the 100 kDa receptor results in loss of BBP-binding activity; this suggests that a disulfide bonded region of the receptor is necessary for maintaining BBP-binding activity. We show further that the levels of serum BBP are strictly dependent on the presence of estrogen. As expected, BBP is not detected in significant quantities in rooster serum. Thus, these results suggest that circulatory BBP, like other estrogen-dependent components of serum, has a cognate binding activity on the oocyte membrane that may mediate its endocytosis.

Biotin-binding proteins and biotin transport to oocytes.

The eggs of chickens and other birds contain two proteins that bind biotin. Both are homotetrameric proteins of similar size. In contrast to the well-characterized egg white avidin, egg yolk biotin-binding protein has a very acidic isoelectric point, binds biotin with lower affinity, and is usually saturated with biotin. Like other egg yolk proteins, biotin-binding protein appears to be synthesized in the liver, transported by the blood stream to the ovary and deposited in the developing oocyte. Since the yolk of a chicken egg contains over 90% of the biotin in an egg and all of the biotin is bound to biotin-binding protein, the function of biotin-binding protein is undoubtedly to transport biotin to the egg for future use by the developing embryo. Avidin is produced by the oviduct and in the egg it is presumed to deter microbial growth around the oocyte by sequestering biotin. Among the eggs examined, those from turkeys have the lowest amount of biotin-binding protein and the highest amount of avidin. Furthermore, the majority of the biotin in turkey eggs can be bound to avidin in the egg white, suggesting a nutritional role for avidin in turkeys. An assay has been developed to conveniently measure apo- and holobiotin-binding proteins.

Riboflavin-deficient chicken embryos: hypoglycemia without dicarboxylic aciduria.

Chicken embryos in eggs laid by hens that are genetically unable to deposit riboflavin into their eggs die on or about the 13th day of incubation. We show that these riboflavin-deficient embryos grow normally until the day of death and that their heart rate is normal to within an hour of death. The embryos have symptoms of impaired fatty acid oxidation, including decreased activity of FAD-dependent medium-chain acyl CoA dehydrogenase in liver and heart along with a significant accumulation of intermediates of fatty acid oxidation (C10, C12, and C14 acids). Unlike riboflavin-deficient mammals, the embryos do not accumulate dicarboxylic acids derived from omega-oxidation of fatty acids. Blood glucose is near normal on day 10 but declines to undetectable levels by the time of death. Allantoic fluid from the riboflavin-deficient embryos of 11 days or older contains more lactate than 3-hydroxybutyrate, while in normal embryos the reverse is true. No appreciable amounts of glycine-conjugated acids were found. We conclude that the major and perhaps primary pathological effect of riboflavin deficiency in chicken embryos is the impairment of fatty acid beta-oxidation, and that the subsequent depletion of limited carbohydrate reserves leads to sudden death.

Relationship of biotin deposition in turkey eggs to dietary biotin and biotin-binding proteins.

The biotin and biotin-binding protein contents of egg yolk, egg albumen, and hen plasma were determined on eight groups of four turkey hens each that had been fed diets ranging from less than 10 to 3,475 micrograms available biotin per kilogram. Biotin deposition in the yolk was strongly dependent upon available dietary biotin below 100 micrograms/kg. Between 100 and 1,000 micrograms/kg the amount of biotin deposited in the yolk increased slightly and was directly related to and limited by a biotin-binding protein that transferred biotin from the plasma to the yolk. Over the entire dietary range, biotin deposition in yolk was proportional to the total biotin concentration in the plasma. In contrast, biotin deposition in the albumen, which was proportional to dietary biotin, increased several-fold over a very narrow range of plasma biotin concentration (56 to 62 micrograms/L). When dietary available biotin exceeded 160 micrograms/kg, there was more biotin deposited in the albumen than in the yolk. Although the concentration of unbound biotin in plasma is low, it appears to be the component of plasma biotin that is rapidly scavenged by avidin in the oviduct. It seems likely that avidin-bound biotin is available to the turkey embryo.