Immunogenic and antigenic epitopes of Ig. XXV. Monoclonal antibodies that differentiate the Mcg+/Mcg- and Oz+/Oz- C region isotypes of human lambda L chains

The C region of human lambda L chains is specified by multiple C lambda genes of which three--C lambda 1, C lambda 2, and C lambda 3--encode for the isotypes designated Mcg+, Kern- Oz-, and Kern- Oz+, respectively. The Mcg, Kern, and Oz factors have been characterized by sequence differences involving specific C lambda amino acid residues. They have also been recognized serologically by polyclonal antisera but, with rare exception, these reagents are no longer available. We have obtained two murine anti-human lambda-chain mAb, 14G1 and 14D1, that recognize antigenic determinants specific for the C lambda isotypes Mcg and Oz, respectively. These antisera have been used to classify as Mcg+/Mcg- or Oz+/Oz- monoclonal lambda-chains (Bence Jones proteins) and intact Ig lambda proteins. There was complete concordance between the chemical and serologic assignment of lambda-chains as Mcg+/Mcg- or as Oz+/Oz-; no single protein expressed both isotypes. There was no evident association between the C region isotype Mcg or Oz and the V region subgroup of the protein tested. However, our finding that four of seven amyloid-associated lambda VI Bence Jones proteins were Oz+ suggests a predominant expression of the C lambda 3 gene product among proteins of this uncommon V lambda subgroup.     

The cytochrome oxidase II gene in mitochondria of the sugar-beetBeta vulgaris L.

We have cloned and analyzed the sugar-beet mitochondrial gene for cytochrome oxidase subunit II (coxII). The sugar-beet and its deduced amino acid sequence were compared to its homologouscoxII gene sequences from both monocot and dicot plants. It was found to be highly conserved (89–95%) compared to homologue in other plant species. The 780 bp coding sequence of the sugar beetcoxII gene is interrupted at position 383 by a 1463 bp intron. This intron contains an additional 107 bp sequence that is not found in any of the plantcoxII genes studied thus far. The structure of the intron suggests that a large intron existed in an ancestralcoxII gene before monocots and dicots diverged in evolution. Three CGG codons in the sugar-beetcoxII coding sequence align with conserved tryptophan residues in the homologous gene of other species, suggesting that RNA editing takes place also in sugar-beet mitochondria. In 13 out of 24 codons ofcoxII mRNA that were found to be edited in four other plants, the sugar-beet gene already utilizes the edited codons. This phenomenon may indicate that the mitochondrial genome in sugar-beet is phylogenetically more archaic relative to these plants. An additional sequence of 279 bp that is identical to the first exon ofcoxII was identified in the mtDNA of the sugar-beet. This pseudo-gene is transcribed and its existence in the mitochondrial genome is unexplained.     

Calcium-Dependent Transglutaminase of Rat Sympathetic Ganglion in Development and After Nerve Injury

The activity of transglutaminase (TG) was examined in the rat superior cervical ganglion (SCG) during development and after postganglionic nerve crush. During postnatal development the enzyme activity is increased by sevenfold in parallel to protein content of the ganglion and reaches adult levels by day 35 after birth. The endogenous activity (enzyme activity assayed in the absence of the exogenous substrate) during development is transiently elevated with a peak at day 21 postnatal. In the adult ganglion the enzyme specific activity is evenly distributed in all subcellular compartments, but most of it is contained in the cytosol. Within the first hour after axotomy TG activity is rapidly and transiently elevated. The peak value, 80% above control levels, is attained by 30 min postoperative. At this time the activity is increased in all subcellular fractions, but the endogenous activity is selectively increased in the fraction containing nuclei. The enhanced TG activity after axotomy can be prevented by topical treatments with verapamil, an inhibitor of voltage-dependent calcium fluxes across excitable membranes, or with the calcium chelator EGTA. The results show that intracellular TG activity is present in the SCG and that it increases with postnatal growth of the ganglion. After axotomy the enzyme activity is rapidly and transiently increased in the ganglion and this elevation critically depends on calcium fluxes.     

Electron microscopic observations on isolated mitochondrial membranes,prepared by various histological methods

Summary The pictures of isolated mitochondrial membranes, as seen on the electron-microscope, depend very much on the method of specimen preparation. Subunits of linear dimensions of about 25 m, (electron transport particles) are observed in carbon-replicas of the membranes and in specimens treated with trypsin or pepsin (0.02% for 30 mins) and shadowed with platinum. A three-layered structure of the unit membrane is seen in sections of specimens fixed with osmium tetroxide or formalin followed by post-fixation with osmium tetroxide. But fixation with potassium permanganate or with formalin, followed by post-fixation with potassium permanganate reveals an electron-dense globular structural element in the unit membrane. An electron-transparent ultrastructural element of the unit membrane is observed after treatment with trypsin (0.2% for 5 mins) and fixation with osmium tetroxide. Unsectioned specimens treated with 0.02% trypsin for 30 mins show a honeycomb-like structure of the membrane. Thus, part of the results appear to support the concept of a mosaic-like structure of the unit membrane, whereas other results are in agreement with the classical concept of a three-layered structure.The authors wish to express their gratitude to Dr. Sina Rosenthal, Department of Physiological Chemistry, Humboldt University, Berlin, who prepared the isolated membranes, to Mr. E. Fischer, Head Technician of the Department of Electron Microscopy, Greifswald University, who took most of the electron micrographs, to Mr. G. Bartsch, Department of Electron Microscopy, Greifswald University, and especially to Prof. W. Bargmann and to Doz. E. Lindner, Department of Anatomy, Kiel University, for many valuable suggestions.     

The Molecular Weight and Aggregation of DNA

The effects of enzymatic attack and of shear during the isolation and deproteinization of DNA have been investigated. Different methods of disaggregating DNA have been studied, and conditions under which reaggregation can occur are discussed. It was found that shaking with chloroform-octanol does not degrade DNA from the seven sources studied; that light scattering yields valid weight-average molecular weights for these samples; and that, when disaggregated, the molecular weights of these samples are in the range 1.2-2.4 million and the length-to-mass ratios are high.     

Localization of a New Type of X-Linked Liver Glycogenosis to the Chromosomal Region Xp22 Containing the Liver α-Subunit of Phosphorylase Kinase (PHKA2)

We describe here a new type of X-linked liver glycogen storage disease. The main symptoms include liver enlargement and growth retardation. The clinical and biochemical abnormalities of this glycogenosis are similar to those of classical X-linked liver glycogenosis due to phosphorylase kinase deficiency (XLG). However, in contrast to patients with XLG, the patients described here have no reduced phosphorylase kinase activity in erythrocytes and leukocytes, and no enzyme deficiency could be found. Linkage analysis of four families with this X-linked type of liver glycogenosis assigned the disease gene to Xp22. Lod scores obtained with the markers DXS987, DXS207, and DXS999 were 3.97, 2.71, and 2.40, respectively, all at 0% recombination. Multipoint linkage analysis localized the disease gene between DXS143 and DXS989 with a maximum lod score of 4.70 at θ = 0, relative to DXS987. As both the classical XLG gene and the liver α-subunit of PHK (PHKA2) are also located in Xp22, this variant type of XLG may be allelic to classical XLG, and both diseases may be caused by mutations in PHKA2. Therefore, we propose to classify XLG as XLG type I (the classical type of XLG) and XLG type II (the variant type of XLG).     

Morphological effects of tributyltin (TBT) in vitro on the genital system of the mesogastropodaLittorina littorea (Prosobranchia)

This ultrastructural study investigates the pathological changes in the penial, the sperm groove and the glandular cushion epithelium in maleLittorina littorea (Mesogastropoda) related to TBT (tributyltin) contamination. The results are compared with those onOcinebrina aciculata (Neogastropoda), which shows a wide range of cell changes in the penis epithelia of male and imposex affected females. The investigation of the different penis epithelia ofL. littorea revealed that the cells analysed show a low sensibility towards TBT. The cells display normal metabolism. Certain atypical structures like swelling microvilli and cristae which tend to lie parallel to the long axis in the mitochondria, were detected as pathological effects.