IDENTIFICATION OF MYCOPLASMATACEAE BY THE FLUORESCENT ANTIBODY METHOD

Clark, Harold W. (The George Washington University, Washington, D.C.), Jack S. Bailey, Richard C. Fowler, and Thomas McP. Brown. Identification of Mycoplasmataceae by the fluorescent antibody method. J. Bacteriol. 85:111-118. 1963.-The conditions of the fluorescent antibody reactions were studied in relation to their application to Mycoplasmataceae or pleuropneumonia-like organisms (PPLO). Mycoplasma hominis type 1 and 2 antigens and their homologous antisera were used to determine the activity and specificity of these and other strains. Fluorescein isothiocyanate conjugated antiserum globulin preparations were used in both the direct and indirect fluorescent antibody methods. A direct tube technique was used for the detection and measurement of growth in broth cultures by the addition of conjugated antiserum. The specific fluorescent staining and recognition of hot water fixed M. hominis colonies was presented as a suitable identification standard. The antigenic activity was found to remain in the insoluble residue after exposure of M. hominis strains to sonic vibration (9 kc) for 30 min and centrifugation. Brief 2-min exposures of tissue cells to vibration (9 kc) caused the disruption of tissues, with the release of viable and "bound" nonwashable strains that reacted specifically with fluorescent antibody. It is proposed to apply both the sonic vibration and the fluorescent antibody techniques for the identification of Mycoplasmataceae in human tissues.     

SEROLOGICAL GROUPING OF ACTINOMYCES BY MEANS OF FLUORESCENT ANTIBODIES.

Slack, John M. (West Virginia University, Morgantown), Ann Winger, and Dane W. Moore, Jr. Serological grouping of actinomyces by means of fluorescent antibodies. J. Bacteriol. 82:54-65. 1961.-Serological groups A, B, C and D of actinomyces were established using fluorescent antibody techniques. One hundred and thirty-eight cultures were included in the study. Eighty-nine were classed in group A, 15 in B, 13 in C, and 21 in D.The isolates were from patients and animals with actinomycosis and from healthy human beings. There was no correlation between source of the isolate and serological group. Furthermore, no one species could be placed exclusively in one group although the majority of those designated as Actinomyces bovis were in group A.Seventeen anaerobic diphtheroids and seven Corynebacterium acnes isolates were placed in group A. One diphtheroid was in each of groups B and D. On this basis it is suggested that these organisms be included in the genus Actinomyces.Additional species of Corynebacterium as well as Lactobacillus Propionibacterium, Streptomyces, and Nocardia did not fluoresce with any of the group antisera.     

THE CONFORMATION OF CLATHRIN LIGHT-CHAINS ON TRISKELIONS PROBED BY ANTIBODY INHIBITION

The conformation of clathrin light-chains along the proximal arm of the clathrin triskelion was studied by using rabbit anti-(light-chain peptides) to inhibit the binding of a mouse monoclonal antibody against an epitope in the amino-terminal region. Prior incubation of triskelions with rabbit antisera raised against the extreme carboxyl-terminal of the light-chains partially inhibited binding. The inhibition was largely removed when tested on light-chains that had been freed from triskelions. This suggests that when the light-chains bind the heavy-chain, they adopt a conformation in which the amino and carboxyl-terminal domains are not fully extended, but fold such that these two domains face each other.     

DNA ORGANIZATION IN THE MATURE SPERM OF SEVERAL ORTHOPTERA BY THE METHOD OF POLARIZED FLUORESCENCE MICROSCOPY

Mature sperm of Acheta domesticus, Acheta assimilis, Nemobius fasciatus, Nemobius confusus, Melanoplus femur-rubrum, Romalea microptera, Scudderia curvicauda, and Ceuthophilus nigricans were examined for DNA configuration by means of polarized fluorescence microscopy. In all cases, the results suggest that the DNA lies in unsupercoiled array, predominantly parallel to the elongate sperm head axes. Detailed calculations of the factors influencing the degree of polarization of the fluorescent emission from supercoiled DNA are contained in an appendix.     

INHIBITION BY APOMORPHINE OF DOPAMINE DEAMINATION IN THE RAT BRAIN

Abstract— Apomorphine (A) inhibited dopamine deamination by rat brain mitochondria, but did not influence catechol- O -methyltransferase (COMT) activity by brain homogenates. The administration of apomorphine (10mg/kg i.p.) to normal rats increased brain dopamine (DA) by 34 per cent and decreased homovanillic acid (HVA) and dihydroxyphenylacetic acid (DOPAC) by 60 per cent. In rats treated with reserpine 15 min prior to A, the latter prevented the rise of cerebral HVA and DOPAC and the depletion of DA produced by the former. Finally, A decreased the L-DOPA-induced accumulation of HVA and DOPAC in the rat basal ganglia. These results indicate that A inhibits DA deamination by monoamine oxidase.
This inhibition seems to be specific since apomorphine did not influence 5-HIAA levels in normal rats and prevented neither central 5-HT depletion nor 5-HIAA rise induced by reserpine.     

INHIBITION OF PHOTOSYNTHESIS IN CHLORELLA PYRENOIDOSA BY THE IODO-ACETYL RADICAL

Photosynthesis in Chlorella pyrenoidosa is inhibited by iodo-acetic acid and iodo-acetamide, both of which attack the Blackman reaction. Since acetamide is without effect, the iodo-acetyl radical must be responsible. The study of the action of the acid is complicated by the fact that its ions penetrate slowly, if at all, so that negative results with this agent are without significance unless penetration can be established. The absorption spectrum of the cells is not affected by concentrations of iodo-acetamide which completely inhibit photosynthesis. This establishes that the chromophore groups of chlorophyll are not involved, and renders it unlikely that any other part of the molecule is. Inasmuch as cyanide likewise inhibits by way of the Blackman reaction, it would seem necessary to postulate that this complex can be attacked at two different loci, which may or may not be on the same molecule. The presence of the iodo-acetyl radical also gives rise to three other effects. (1) Concentrations (10^–5 M or less) too small to inhibit photosynthesis may increase the rate by interacting with the photochemical complex. (2) Concentrations (ca. 10^–4 M) which inhibit photosynthesis increase the rate of respiration. (3) Concentrations (10^–3 M or more) higher than those required to inhibit photosynthesis inhibit respiration.     

酶抑制动力学模型判断中的统计分析

文章用数理统计的方法导出了酶抑制动力学研究中双倒数作图法的统计模型,克服了作图法中主观因素的干扰,建立了判断竞争抑制,非竞争抑制,反竞争抑制,线性混合抑制的统计分析方法。     

铁离子抑止金微素产量的原因和去除抑止作用的研究

在不同酦酵培养基上,铁离子抑止金霉素产量的作用浓度并不相同。生长加有铁离子的酦酵培养基上的菌体,一般对葡萄糖的利用较慢,生长延迟。如将生长在加有铁离子的培养基中的菌体,接种到不加铁离子的新鲜培养基中,或用置换培养的方法,菌体能正常酦酵,不受前阶段铁离子的影响。在酦酵过程中,行分期加铁的试验结果,发现铁离子能普遍地使金霉素效价降低至同一水平,而铁离子抑止金霉素效价的作用,在10分钟行已经完成。用嵌合剂三甘氨酸处理这种酦酵溶液,能使金霉素效价恢复至80-90%,同样亦相应的将铁离子摄取下来。这些工作的结果,说明铁离子和金霉素分子的结合,是促使金霉素效价落降的主要原因。作者在进行嵌合剂处理时,特别指出当铁离子和附着在菌丝细胞上的金霉素分子结合后,必须先使嵌合剂将这个结合物离解后,然后酸化培养基,使金霉素分子释放至培养溶液中,因为金霉素分子与铁离子的结合物是不能因培养溶液的酸化,而从菌体细胞表面释放出来的。作者认为在金霉素酦酵工业上可以利用铁罐进行发酵,只要注意在提炼时利用嵌合剂的问题。