Energy-dependent formation of free ATP in yeast submitochondrial particles, and its stimulation by oligomycin

Yeast submitochondrial particles, in a P_i- and NADH-dependent reaction, produced low concentrations of free ATP in the absence of added ADP. This formation of free ATP, as measured by the luciferin-luciferase method, was strongly stimulated by oligomycin. For maximal stimulation, oligomycin was to be added not earlier than 5–10 min after the addition of NADH. Upon addition of antimycin or FCCP the system was completely inhibited. The amount of free ATP formed corresponded to one-third of the amount of bound ATP in submitochondrial particles. The stimulatory effect of oligomycin disappeared if the submitochondrial particles were spun down after oligomycin stimulation and then resuspended in the reaction medium, whereas submitochondrial particles with no oligomycin added initially were stimulated by oligomycin after the same procedure. A different picture emerged with addition of ADP. If the submitochondrial particles were preenergized with NADH in the presence of oligomycin before the addition of ADP the formation of free ATP upon subsequent addition of ADP was inhibited by oligomycin. In the presence of oligomycin, but lacking preenergization with NADH, a stimulation of free ATP formation was achieved with added ADP. A possible explanation for the stimulating effect of oligomycin on ATP formation in the absence of added ADP is that it enhances the release of bound ATP in an energy-requiring process. The release of only about one-third of the bound ATP could indicate that one of three nucleotide-binding subunits involved in the mechanism of ATP formation by ATP synthase is in a state suitable for such an energy-dependent release of ATP.     

Tumor promoter chloroform is a potent protein kinase C activator

The major interaction site for tumor-promoting phorbol esters is the calcium-activated, phospholipid-dependent protein kinase (protein kinase C), a key-element in signal transduction. Binding of phorbol esters results in enzyme activation which mediates, at least in part, the action of these agents. We have investigated the effects of tumor promoter chloroform on protein kinase C activity. Like thrombin and 12-O-tetradecanoylphorbol-13-acetate (TPA), chloroform was able to activate protein kinase C in intact rabbit platelets. In addition, chloroform stimulated enzyme activity as well as TPA binding capacity in cell-free system. Scatchard analysis of the data has shown that chloroform increased the number of phorbol ester binding sites. Structurally related compounds, carbon tetrachloride and methylene chloride, activated the enzyme similarly.     

5'-modified agonist and antagonist (2'-5')(A)n analogues: synthesis and biological activity

Two 5'-modified (2'-5')(A)4 oligomers with an increased resistance to phosphatase degradation were synthesized and evaluated for their ability to develop an antiviral response when introduced into intact cells by microinjection or by chemical conjugation to poly(L-lysine). The enzymatic synthesis of 5'-gamma-phosphorothioate and beta,gamma-difluoromethylene (2'-5')(A)4 from adenosine 5'-O-(3-thiotriphosphate) and adenosine beta,gamma-difluoromethylenetriphosphate by (2'-5')-oligoadenylate synthetase is described. The isolation and characterization of these (2'-5')(A)4 analogues were achieved by high-performance liquid chromatography. The structures of 5'-modified tetramers were corroborated by enzyme digestion. These two 5'-modified tetramers compete as efficiently as natural (2'-5')(A)4 for the binding of a radiolabeled (2'-5')(A)4 probe to ribonuclease (RNase) L. Nevertheless, at the opposite to 5'-gamma-phosphorothioate (2'-5')(A)4, beta,gamma-difluoromethylene (2'-5')(A)4 failed to induce an antiviral response after microinjection in HeLa cells. In addition, it behaves as an antagonist of RNase L as demonstrated by its ability to inhibit the antiviral properties of 5'-gamma-phosphorothioate (2'-5')(A)4 when both are microinjected in HeLa cells. The increased metabolic stability of 5'-gamma-phosphorothioate (2'-5')(A)4 as compared to that of (2'-5')(A)4 was first demonstrated in cell-free extracts and then confirmed in intact cells after introduction in the form of a conjugate to poly(L-lysine). Indeed, 5'-gamma-phosphorothioate (2'-5')(A)4-poly(L-lysine) conjugate induces protein synthesis inhibition and characteristic ribosomal RNA cleavages for longer times than unmodified (2'-5')(A)4-poly(L-lysine) in the same cell system.(ABSTRACT TRUNCATED AT 250 WORDS)     

A complex balanced chromosomal rearrangement in repeated abortions

Summary A double balanced reciprocal translocation involving four chromosomes, t(1;19;6;14) (1p11; 19p11; 6q25; 14q21), was found in the phenotypically normal husband in a couple referred because of repeated abortions. Reciprocal translocations, t(6;14), had been transmitted by his mother, his father being apparently homozygous for a translocation comprising pairs 1 and 19-t(1;19)(1;19). The genetic consequences of this complex chromosomal rearrangement are analyzed.     

Hydrolysis of rice straw to fermentable sugars byTrichoderma enzymes

Summary ATrichoderma sp. (IMB-Tr) isolated from rice straw possessed cellulolytic and xylanolytic activity, comparable to those produced byTrichoderma reesei QM 9414 (a proven cellulolytic fungus). IMB-Tr produced 2.9 and 1.9 times, respectively, greater -glucosidase activity compared toT. reesei when grown on microcrystalline cellulose and rice straw. Percentage enzymic hydrolysis increased with increase in the sodium hydroxide concentration used in the pretreatment of rice straw and with the increase of enzyme concentration used in the hydrolysis. The extracellular enzyme fraction ofT. reesei possessed greater hydrolytic power than that of IMB-Tr. However, when a combined enzyme preparation from the two organisms was used, an appreciable degree of synergism was observed; an increase in reducing sugars up to 39% was seen. The reducing sugar produced by enzymic hydrolysis was mainly glucose, xylose and cellobiose. Fermentation of a 4.8% (w/v) sugar hydrolysate (produced by the enzymic hydrolysis of rice straw) bySaccharomyces cerevisiae produced 10.7 g/l of ethanol compared to 18.8 g/l produced by the fermentation of 4.8% (w/v) pure glucose.

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Resumen Se ha aíslado a partir de paja de arroz una cepa deTrichoderma sp. (IMB-Tr) que posee actividades celulolíticas y xilanolíticas comparables a las deTrichoderma reesei QM 9414 (un hongo probadamnete celulolítico). IMB-Tr produjo 2.9 y 1.9 veces más actividad -glucosidásica queT. reesei cuando ambos se hicieron crecer en celulosa microcristalina y en paja de arroz respectivamente. El porcentaje de hidrolisis enzimática se incrementó con el aumento en la concentración del hidróxido sódico empleado en el pretratamiento de la paja de arroz y con el aumento de la concentración enzimática utilizada en la hidrolisis. La fracción extracelular enzimática deT. reesei poseía un mayor poder hidrolítico que la de IMB-Tr, sin embargo cuando se usó un preparado enzimático combinado de ambos microorganismos se obtuvo un apreciable efecto sinérgico, observándose un incremento de hasta un 39% de los azucares reductores producidos. Estos azucares fueron principalmente glucosa, xilosa y celobiosa. La fermentación de un 4.8% (p/v) del hidrolisado azucarado (producido por la hidrolisis enzimática de la paja de arroz) porSaccharomyces cerevisiae produjo 10.7 g/l de etanol comparado a 18.8 g/l obtenidos de la fermentación de 4.8% (p/v) de glucosa pura.

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Résumé Une souche deTrichoderma sp. (IMB-Tr), isolée à partir de paille de riz, a une activité cellulolytique et xylanolytique comparable à celle deTrichoderma reesei QM 9414 (champignon cellulolytique reconnu). L'activité -glucosidase d'IMB-Tr cultivé sur cellulose micro-cristalline ou sur paille de riz est, respectivement, 2.9 et 1.9 fois plus élevée que celle deT. reesei. Le pourcentage d'hydrolyse enzymatique croit avec la concentration de la soude employée pour le pré-traitement de la paille et avec la concentration d'enzyme utilisée pour l'hydrolyse. La fraction exocellulaire de l'enzyme a une activité hydrolysante plus élevée dans le cas deT. reesei que dans celui de IMB-Tr. Cependant, si on emploie un mélange des activités enzymatiques des deux organismes, on constate une nette synergie et un accroissement des sucres réducteurs allant jusqu'à 39%. Les sucres réducteurs obtenus par hydrolyse enzymatique comprennent principalement du glucose, du xylose et du cellobiose. La fermentation parSaccharomyces cerevisiae d'un hydrolysat enzymatique de paille de riz contenant 4.8% (poids/vol.) de sucres fournit 10.7 g/l d'éthanol, au lieu de 18.8 g/l obtenus par fermentation de glucose pur à la même concentration.

     

Sialic acid is a cell surface component of Entamoeba invadens trophozoites

The surface anionic groups of Entamoeba invadens were analysed by cell electrophoresis, by ultrastructural cytochemistry, and by identification of sialic acids using paper and gas-liquid chromatography. Binding of colloidal iron hydroxide (CIH) and of cationized ferritin (CF) particles at pH 1.8 and 7.2, respectively, was observed on the cell surface. E. invadens has a highly negative surface charge (-0.96 microns s-1 V-1 cm). Treatment of the cells with trypsin and neuraminidase significantly reduced the electrophoretic mobility by 24% and 40%, respectively. Treatment of the amoebae with neuraminidase also markedly decreased the binding of CIH to the cell surface. This finding suggests that sialic acid residues are the major anionogenic groups exposed on the surface of E. invadens. Paper and gas-liquid chromatography showed that N-acetylneuraminic acid was the only derivative characterized in E. invadens.     

Inheritance of resistance to bacterial blight in common bean

Summary Inheritance of resistance to common bacterial blight in the trifoliate leaf, plant canopy, and pods was controlled by a single major gene. Additive followed by dominance effects were more important than epistatic interactions. Narrow-sense heritability values ranged from 0.18 to 0.87 for trifoliate leaf, from 0.26 to 0.76 for canopy, and from 0.11 to 0.36 for pods. Observed gains from selection for resistance were higher than expected gains. Implications of these results in breeding for resistance are discussed.     

Isolation of Corynebacterium diphtheriae from sperm fluid

The isolation of toxigenic Corynebacterium diphtheriae from sperm is reported. The organism was identified through the investigation of fluorescence under the UV light, the presence of pirazinecarboxilamidase enzyme (Pyz), in vitro and in vivo and virulence methods (single radial immunodiffusion, cell culture, guinea pig intradermic test). The strain was initially considered nontoxinogenic by single radial immunodiffusion, but its virulence was observed afterwards, when we applied the tests already mentioned. The strain could be considered a "Diphtheroid" without adequate specification.