A missense mutation (G197→A) in theα-l-fucosidase gene of fucosidosis patients leads to loss ofα-l-fucosidase

Fucosidosis is an autosomal recessive lysosomal storage disease resulting from the absence of -l-fucosidase activity. Two natural missense mutations (G¹⁹⁷A) and (A⁸⁶⁰G) within the -l-fucosidase gene have been reported to be homozygous in four patients with fucosidosis. Expression of wild-type and mutated -l-fucosidase cDNAs in COS-1 cells revealed complete deficiency of -l-fucosidase for the G¹⁹⁷A transition and a normal level of enzyme for A⁸⁶⁰G. We therefore conclude that the change of G¹⁹⁷A is responsible for fucosidosis in the patients while A⁸⁶⁰G is a normal polymorphic variant of -l-fucosidase.     

Regulation of the synthesis of human chorionic gonadotropin by strains of HeLa cells in culture

Summary Thirty-seven strains of HeLa cells were examined for their ability to synthesize human chorionic gonadotropin (hCG) and its alpha subunit (hCG-α) in culture. Synthesis of hCG-α and hCG also was investigated in the presence of sodium butyrate and 5-bromo-2′-deoxyuridine (BrdUrd). All HeLa strains synthesized hCG-α in culture. Sodium butyrate increased the synthesis of hCG-α in all HeLa cells; BrdUrd increased synthesis in 32 of the 37 strains examined. Although few HeLa strains synthesized hCG in the absence of inducers, hCG was detected in most strains in the presence of sodium butyrate. The synthesis of hCG and its alpha subunit is, therefore, a stable genetic characteristics of HeLa cells. Certain preparations of hCG and its subunits were generously provided through the Center for Population Research of the National Institute of Child Health and Human Development, NIH.     

The peritrophic membrane in adult simuliids (Diptera) before and after feeding on blood and blood-sucrose mixtures

A peritrophic membrane formed in the posterior midgut of female simuliids of seven species within 30 min of feeding on blood of various avian or mammalian hosts, and 6–12 hr thereafter it displayed up to seven concentric laminae. Within the first 48 hr of blood digestion, the posterior part of the peritrophic membrane (where digestion was most advanced) began to decompose and PM disappearance was complete at the end of blood digestion. In different species blood digestion at 18°–20° required 120–180 hr which was increased to over 200 hr by microsporidan infection. If a blood-sucrose mixture went directly to the midgut, a thin membrane formed around it, but not if it went indirectly by way of the crop. A prefeeding secretion in the anterior midgut is considered to be the peritrophic membrane of the pharate adult surrounding a small, but variable, amount of material, the meconium.

---

Résumé Sept espèces de simulies ont été étudiées: 5 ornithophiles, 2 mammalophiles. Trente minutes après un repas de sang sur divers oiseaux (poulet, canard) ou mammifères (homme, cerf, élan d'Amérique) on constate qu'une membrane péritrophique est sécrétée par toute le surface épithéliale dans la région moyenne de l'intestin moyen postérieur dilaté des femelles. Six à douze heures après le repas cette membrane comporte jusqu'à sept lamelles concentriques. Une sécrétion moins importante se forme autour de la petite quantité de sang que l'on trouve dans l'estomac antérieur, non dilaté et étroit; cette sécrétion glisse vers l'arrière et forme une capsule recouvrant le pôle antérieur du bol alimentaire sanguin dans la partie postérieure de l'intestin moyen, ceci pendant le premier jour. La digestion commence à la périphérie et à l'extrémité postérieure de la masse sanguine et se poursuit enfin à l'extrémité antérieure, où la membrane péritrophique conserve le plus longtemps sa structure en contact avec le sang non digéré. Dans les premières 48 heures de la digestion du sang, la région postértieure de la membrane péritrophique (là où la digestion est la plus avancée) commence à se décomposer, et sa disparition est complète quand la digestion du sang est achevée. Chez les différentes espèces étudiées, la digestion du sang à 18–20° exige 120–180 heures, pouvant demander plus de 200 heures chez des insectes atteints d'une infection par des microsporidies. Si un mélange de sang et sucrose parvient directement dans la région moyenne de l'intestin moyen, une membrane mince se forme autour, mais non si cet aliment passe par le jabot. Une sécrétion précédant l'alimentation s'observe dans l'intestin moyen antérieur des mouches non nourries et doit représenter, chez l'adulte venant d'éclore, la membrane péritrophique entourant le méconium: petite quantité de matière d'importance variable. Cette matière se retrouve souvent au centre de la masse sanguine après alimentation et est lente à digérer.

     

A four-iron, four-sulfide ferredoxin with high thermostability from Clostridium thermoaceticum.

A ferredoxin containing only one [Fe4S4] cluster was purified from Clostridium thermoaceticum. It has a molecular weight of about 7,300, a partial specific volume of 0.67, and an isoelectric point of 3.25. Its absorption spectrum has two maxima at 390 nm (epsilon = 16.8 X 10(3)M-1cm-1) and at 280 nm (epsilon = 24.2 X 10(3)M-1cm-1). The absorption at 390 nm is almost half that of other clostridial ferredoxins, which have two [Fe4S4] clusters, and is similar to other ferredoxins with only one [Fe4S4] cluster. The ferredoxin had high thermal stability and retained over 50% of its activity after treatment at 80 degrees C. It functions in the transfer of electrons from pyruvate to nicotinamide adenine dinucleotide phosphate (NADP), which indicates the presence of pyruvate:ferredoxin oxidoreductase and reduced ferredoxin-NADP reductase in C, thermoaceticum. NADPH is used in the synthesis of acetate from CO2 in this organism.     

Resolution of optical isomers by high-pressure liquid chromatography: The separation of benzo[a]pyrene trans-diol derivatives

The optical isomers of (±)r-7,t-8-dihydroxy-7,8-dihydrobenzo[a]pyrene and its synthetic precursor (±)r-7,t-8-dihydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene were resolved as their di-(−)menthoxyacetates using high-pressure liquid chromatography. Saponification of the resolved diesters yielded the corresponding enantiomers. The specific rotation, CD spectra, and ORD curves are reported. The resolution of these optical isomers permits detailed studies on the enzymatic intermediates and the mechanism of benzo[a]pyrene activation to its carcinogenic form. The method is of general usefulness for the resolution of optical isomers.     

The shape of myosin subfragment-1. A equivalent oblate ellipsoid model based on hydrodynamic properties

The molecular weights of the two heads of myosin subfragment-1, S-1(A1) and S-1(A2), based on sedimentation equilibrium are 120 000 and 110 000. Hydrodynamically, the two heads are indistinguishable, with intrinsic viscosity, [eta], of 0.064-0.065 dL/g and sedimentation coefficient, s(0)20,w, of 5.8 S.Together with the rotational correlation time taken from the literature (235 ns), all three hydrodynamic properties can be better fitted with an equivalent oblate ellipsoid of revolution than a prolate model. The width of the equatorial axis of the ellipsoid is about 135 A (the axial ratio is about 6). Probably, the S-1(A1) and S-1(A2) molecules have a half-doughnutlike or a flattened pearlike shape rather than an elongated one.     

Sulfite-induced lipid peroxidation in chloroplasts as determined by ethane production

Ethane formation, as a measure of lipid peroxidation, was studied in spinach (Spinacia oleracea L.) chloroplasts exposed to sulfite. Ethane formation required sulfite and light, and occurred with concomitant oxidation of sulfite to sulfate. In the dark, both ethane formation and sulfite oxidation were inhibited. Ethane formation was stimulated by ferric or ferrous ions and inhibited by ethylenediamine tetraacetate. The photosynthetic electron transport modulators, 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and phenazine methosulfate, inhibited both sulfite oxidation and ethane formation. Methyl viologen greatly stimulated ethane formation, but had little effect on sulfite oxidation. Methyl viologen, in the absence of sulfite, caused only a small amount of ethane formation in comparison to that produced with sulfite alone. Sulfite oxidation and ethane formation were effectively inhibited by the radical scavengers, 1,2-dihydroxybenzene-3,5-disulfonic acid and ascorbate. Ethanol, a hydroxyl radical scavenger, inhibited ethane formation only to a small degree; formate, which converts hydroxyl radical to superoxide radical, caused a small stimulation in both sulfite oxidation and ethane formation. Superoxide dismutase inhibited ethane formation by 50% when added at a concentration equivalent to that of the endogenous activity. Singlet oxygen did not appear to play a role in ethane formation, inasmuch as the singlet oxygen scavengers, sodium azide and 1,4-diazobicyclo-[2,2,2]-octane, were not inhibitory. These data are consistent with the view that O₂ is reduced by the photosynthetic electron transport system to superoxide anion, which in turn initiates the free radical oxidation of sulfite, and the free radicals produced during sulfite oxidation were responsible for the peroxidation of membrane lipids, resulting in the formation of ethane.