Purification of bovine conglutinin using pepsin digestion

This paper describes a new method for the purification of bovine conglutinin based on the relative resistance of this protein to pepsin digestion. First, conglutinin is purified by absorption on yeast, then the preparation is treated with 2% pepsin (w/w) at 4°C for 18 hr, and finally gel filtrated on agarose A5m. The yield is 60–75% and conglutinin thus prepared appears physically, immunochemically and functionally intact. This procedure allows for a rapid production of sufficient amounts of conglutinin for immune complex detection or purification methods.     

Differentiation of Peptococcus and Peptostreptococcus by gas-liquid chromatography of cellular fatty acids and metabolic products

Gas-liquid chromatographic (GLC) profiles of cellular fatty acids and metabolic products were useful in identifying strains of Peptococcus saccharolyticus, Peptococcus asaccharolyticus, Peptostreptococcus anaerobius, Peptostreptococcus micros, and Streptococcus intermedius. The GLC results supported the recent taxonomic decision to transfer aerotolerant Peptostreptococcus species to the genus Streptococcus. Because inconsistencies in the results prevented our differentiating Peptococcus prevotii. Peptococcus magnus, and Peptococcus variabilis by GLC, additional strains will have to been examined. These GLC techniques are amenable to routine use; however, for interlaboratory results to be meaningful, the classification and nomenclature of the anaerobic gram-positive cocci should be standardized.     

Differences in assembly or stability of complex I and other mitochondrial OXPHOS complexes in inherited complex I deficiency

NADH-ubiquinone oxidoreductase (complex I) deficiency is amongst the most encountered defects of the mitochondrial oxidative phosphorylation (OXPHOS) system and is associated with a wide variety of clinical signs and symptoms. Mutations in complex I nuclear structural genes are the most common cause of isolated complex I enzyme deficiencies. The cell biological consequences of such mutations are poorly understood. In this paper we have used blue native electrophoresis in order to study how different nuclear mutations affect the integrity of mitochondrial OXPHOS complexes in fibroblasts from 15 complex I-deficient patients. Our results show an important decrease in the levels of intact complex I in patients harboring mutations in nuclear-encoded complex I subunits, indicating that complex I assembly and/or stability is compromised. Different patterns of low molecular weight subcomplexes are present in these patients, suggesting that the formation of the peripheral arm is affected at an early assembly stage. Mutations in complex I genes can also affect the stability of other mitochondrial complexes, with a specific decrease of fully-assembled complex III in patients with mutations in NDUFS2 and NDUFS4. We have extended this analysis to patients with an isolated complex I deficiency in which no mutations in structural subunits have been found. In this group, we can discriminate between complex I assembly and catalytic defects attending to the fact whether there is a correlation between assembly/activity levels or not. This will help us to point more selectively to candidate genes for pathogenic mutations that could lead to an isolated complex I defect.     

Gill repartition of two Monogenea: Diplectanum aequans (Wagener 1857), Diesing 1958 and D. laubieri Lambert and Maillard 1974 (Monogenea, Monopisthocotylea) simultaneous parasites of the sea bass Dicentrarchus labrax (author's transl)]

The close observation of the gill habitat of two Monogeneans Diplectanum aequans (Wagener 1857) Diesing 1858 and Diplectanum laubieri Lambert and Maillard 1974 shows the microbiotopes of these two simultaneous parasites of the Sea-bass: Dicentrarchus labrax. The authors propose to study the distribution of the Monogeneans with references to three theoretic axes in the gill chambers. For the Diplectanum, the populations spread on each of these axes according to preferential gradients.     

Interference of oral immunization with the intestinal absorption of heterologous albumin

Rats were immunized with human serum albumin (HSA) by a single intragastric administration of 200 mg of HSA. Two weeks later their capacity to absorb a similar intragastric test dose of HSA was found to be greatly impaired, the concentrations of HSA in mesenteric venous serum having been reduced to 33 %, 25 % and 60 % of those in similarly tested but unprimed animals, respectively 1, 2 and 3 hours after the intragastric test dose. Rats given antigen (HSA) together with intestinal secretions from intragastric immunized rats also showed a striking decrease in intestinal absorption of a test dose, compared to controls given the antigen together with nonimmune intestinal secretions. From these data it is concluded that local immunization of the gut impairs its capacity to absorb the corresponding antigen, and that this effect is largely due to the failure to absorb antigen bound to secretory antibodies.     

The mutational specificity of l-nitroso-6-nitropyrene in the lacI gene of Escherichia coli strains deficient in nucleotide excision repair

We have examined the mutational specificity of 1-nitroso-6-nitropyrene(1,6-NONP), an activated metabolite of the carcinogen 1,6-dinitropyrene,in the lacI gene of Escherichia coli strains which are deficientin nucleotide excision repair (strain NR6113,     

Analysis of mycolic acid cleavage products and cellular fatty acids of Mycobacterium species by capillary gas chromatography.

After growth and experimental conditions were established, the mycolic acid cleavage products, constituent fatty acids, and alcohols of representative strains of Mycobacterium tuberculosis, M. smegmatis, M. fortuitum complex, M. kansasii, M. gordonae, and M. avium complex were determined by capillary gas chromatography. Reproducible cleavage of mycolic acid methyl esters to tetracosanoic (24:0) or hexacosanoic (26:0) acid methyl esters was achieved by heating the sample in a high-temperature muffle furnace. The major constituent fatty acids in all species were hexadecanoic (16:0) and octadecenoic (18:1 omega 9-c, oleic) acids. With the exception of M. gordonae, 10-methyloctadecanoic acid was found in all species; moreover, M. gordonae was the only species tested which contained 2-methyltetradecanoic acid. M. kansasii was characterized by the presence of 2,4-dimethyltetradecanoic acid, M. avium complex by 2-eicosanol, and M. tuberculosis by 26:0 mycolic acid cleavage product. The mycolic acid cleavage product in the other five species tested was 24:0. Although a limited number of strains and species were tested, preliminary results indicate that this gas chromatographic method can be used to characterize mycobacterial cultures by their mycolic acid cleavage products and constituent fatty acid and alcohol content.     

Comparison of the direct radiolabeled antiglobulin assay and the direct immunobead binding test for detection of sperm-associated antibodies

Thirty-seven semen samples were assayed for sperm-associated IgG and IgA using the immunobead test. Portions of these sperm samples were sent for testing with a direct radiolabeled antiglobulin assay and the testing results were compared. If the results of the immunobead test when only tail-tip bead binding was noted are regarded as negative, there was close correlation between the two assay methodologies.     

Intrafractional motion during proton beam scanning

Patient and internal organ motion during treatment with a scanned proton beam can introduce unplanned heterogeneities in the dose distribution throughout the irradiated volume. With static beam techniques, a margin around the target volume is added to compensate for patient and organ motion. This margin may not provide the solution with dynamic beam scanning. Intrafractional motion parallel and perpendicular to the beam axis is studied using two different scanning methods on a cubic water phantom. The direction of motion relative to the beam scanning direction as well as the method of scanning the proton beam across the target has a significant effect on the resulting dose distribution within the target volume. In the extreme cases studied here up to 100% of the target receives a dose outside the recommended limits, with a minimum dose as low as 34% of the prescribed dose.