The gene for the alpha polypeptide of pyruvate dehydrogenase is X-linked in humans.

The chromosomal location of the gene for the alpha polypeptide of the pyruvate dehydrogenase (alpha E1), a major component of the pyruvate dehydrogenase complex, was determined by using a cloned cDNA for alpha E1. This 1-kb cDNA was isolated from a human liver lambda gt11 expression library with specific antibodies and included the coding (from amino acid 144 to the carboxy terminus) and the 3' untranslated regions. Southern blot analysis of the DNA from a panel of rodent-human hybrid cells showed that the absence or the presence of the major EcoRI fragment that hybridized with this cDNA probe was concordant with the presence of the Xq24-p22 region of the human X chromosome. The result of in situ hybridization with human metaphase chromosomes further mapped the alpha E1 gene to the Xp arm.     

Increased sensitivity of the rapid hydrophobic grid membrane filter enzyme-labeled antibody procedure for Escherichia coli O157 detection in foods and bovine feces.

Several strains of Escherichia coli O157:H7 artificially inoculated into vegetables and dairy products were recovered on hydrophobic grid membrane filters and enumerated by an enzyme-labeled antibody assay. The mean of the recoveries from 12 fresh vegetables was 108.8%, whereas that from 10 dairy products was 93.2%. Modified tryptic soy broth at 43 degrees C with shaking at 100 rpm provided optimum recovery of the organism from meat, with a sensitivity of less than or equal to 1 CFU/g, which is 10 times more sensitive than direct plating. The method performed equally well with vegetable and dairy products. Tryptic soy broth, however, under the same conditions gave the best results for fecal samples. Of 22 asymptomatic dairy cattle, reported as having positive Brucella titers when assayed with polyclonal antibodies, eight were found to contain E. coli O157 in their feces as demonstrated by the enzyme-labeled antibody assay by using monoclonal antibodies. This finding may explain some of the false-positive Brucella tests.     

The time resolved emission spectra of peptide conformers measured by pulsed laser excitation

The fluorescence decays of a number of peptides were measured using pulsed laser excitation and time correlated single photon counting techniques. In all cases double exponential kinetics were observed with the short lifetime component (0.5 – 0.85 ns) predominating over the long lifetime componet (1.62 – 2.21 ns). The lifetimes varied with the peptides measured. The time resolved emission spectra of LysTrpLys shows both components have similar spectral maxima at 345 nm. It is suggested that the dual exponential kinetics originate from different conformers of the peptides.     

Planar bilayer membranes from pure lipids.

Planar lipid bilayer membranes are formed from mixtures of pure lipids in the absence of non-biological solvents. The solventless bilayers are characterized by a large specific capacitance (586-957 nF/cm2) comparable to that of cell membranes but considerably greater than that of conventional lipid/decane bilayers. Hydrocarbon solvents, such as n-alkanes or squalene, thicken the bilayer. Membrane dielectric thickness is used as an indicator of bilayer lipid composition. For membranes made from pure monoglyceride/triglyceride mixtures the thickness of the solventless lipid bilayer is independent of both the chain length (11-22 carbons) and mol fraction (0.1-0.9) of triglyceride in the bulk mixture. In contrast, the thickness of the bilayer (2.0-3.3 nm) depends strongly upon the length (16-24 carbons) of the monoglyceride component. Molecular volume considerations lead to the conclusion that the bulk lipid mixture disproportionates to yield bilayer membranes composed of nearly pure monoglyceride. The dielectric thickness of the monoglyceride bilayer is consistent with the notion that the lipid fatty acyl chains are fluid.     

Development of the gigantocerebellum of the weakly electric fish Pollimyrus

The morphogenesis of the "hypertrophied" mormyrid cerebellum was investigated in Pollimyrus (Pisces). Two adults and 36 larvae and young fish raised in captivity were used. Two Gnathonemus petersii adults were taken for comparison. The ontogenetic development of the various cerebellar structures was analysed in inverse chronological order with the aid of serial sagittal and frontal brain sections. Special attention was given to the trilobed corpus cerebelli (C1, C2, C3), the lobi transitorii et caudales, the valvula, the crista cerebelli, the eminentia granularis and the lobus lineae lateralis. 1. The cerebellar structures are of bilateral origin; they develop from the cerebellar and acoustico-lateral "anlage" of the rhombencephalon behind the rhombomesencephalic fissure, either through budding or individualisation and appear between the 4th and 11th day after spawning. The midline fusion of the symmetrical structures is accomplished somewhat later, between the 8th and 23rd days. 2. The cerebellar structures acquire their definitive spatial organisation within 38 days, except for the valvula whose development takes much longer. Recognisable from the 11th day, the valvula upon which ridges are visible from the beginning continues to grow after the 38th day beyond the mesencephalic ventricle, finally overlying the telencephalon frontally and the different rhombencephalic structures caudally. This development, which includes a large antero-lateral folding of the valvula, takes 240 days. 3. Cytological differentiation is just as complex as the general development of the cerebellar structures. Cortical stratification first begins on the 8th to the 11th day in the corpus cerebelli and in the valvula from day 21 to 23 onwards. This differentiation is characterised throughout almost the entire cerebellum by a downward migration of the superficial undifferentiated cells which then constitute the granular layer. In the valvula, the majority of the undifferentiated cells leave the ridges to form a continuous granular layer at the base of the ridges. 4. A differentiation gradient was observed on the antero-posterior axis. 5. In spite of its complexity, the mormyrid cerebellum develops much more rapidly than the cerebellum of the trout.     

Localization of 5S RNA and rRNA genes in the Norway rat

The genes coding for the two classes of ribosomal RNA molecules, 5S RNA and 18+28S RNA, have been localized in the Norway rat (Rattus norvegicus). The 18+28S RNA cistrons are found on three chromosomes, at secondary constrictions on the short arms of chromosomes 3 and 12 and at the telomere of the short arm of chromosome 11. These sites were confirmed using the silver staining technique for nucleolar organizer regions. Two sites were found for the 5S RNA genes; one is closely linked to the 18+28S gene site on chromosome 12. The second site is at or near the telomere of the long arm of chromosome 19.     

Analysis of the interaction of organic phosphates with hemoglobin.

The interaction of organic phosphates with hemoglobin is studied by use of a simple thermodynamic approach. A model-independent analysis is employed to evaluate the accuracy of Adair constants determined in the presence of 2,3-diphosphoglycerate (DPG). The change of oxygen affinity in the presence of phosphates is related to the macroscopic phosphate binding constants of oxy- and deoxyhemoglobin and used to extract such binding constants from oxygen equilibrium measurements. The change of the Bohr effect in the presence of phosphates and the competitive binding of carbon dioxide and DPG are treated quantitatively. The binding of organic phosphates is incorporated into an allosteric model, in which the effect of phosphate on both tertiary and quaternary structure changes is included. By use of this model, the factors which can be responsible for the increased functional heterogeneity of alpha and beta chains in the presence of phosphates are clarified.     

Selective enrichment of Shigella in the presence of Escherichia coli by use of 4-chloro-2-cyclopentylphenyl beta-D-galactopyranoside.

A procedure involving the use of citrate-buffered lactose broth (pH 6.5) containing an analogue of a beta-galactoside (4-chloro-2-cyclopentylphenyl beta-D-galactopyranoside) has been developed for the enrichment of Shigella in competition with a 100-fold higher population of Escherichia coli. The system makes use of the beta-galactosidase activity of E. coli which hydrolyzes the phenolic derivative of beta-galactoside to galactose and an aglycone moiety (4-chloro-2-cyclopentylphenol) which is toxic to E. coli but is tolerated by Shigella. The procedure is particularly effective in the enrichment of S. sonnei and S. flexneri; S. dynsenteriae and S. boydii are enriched to a lesser extent.     

Analysis of cooperativity in hemoglobin. Valency hybrids, oxidation, and methemoglobin replacement reactions.

An allosteric model proposed previously for structure-function relations in hemoglobin is applied to the analysis of low- and high-spin valency hybrids. By assuming that the low-spin oxidized chains have the tertiary structure of oxygenated chains while the high-spin oxidized chains have a tertiary structure intermediate between that of deoxygenated and oxygenated chains, the model parameters associated with the different valency hybrids can be obtained, and their equilibrium properties can be estimated. The hybrid results are used also to provide an interpretation of methemoglobin and its ligand replacement reactions and of the oxidation-reduction equilibrium of normal hemoglobin. For the various systems studied it is found that the effects of pH and 2,3-diphosphoglycerate are in agreement with the model.