Cloning, structure, and expression of a rat binding protein for polychlorinated biphenyls. Homology to the hormonally regulated progesterone-binding protein uteroglobin

Certain metabolites of polychlorinated biphenyls (PCBs) are retained in the Clara cells and in the airway lumen of rodent lung due to their interaction with a secretory 13-kDa protein. Here, we report the isolation of a cDNA encoding the rat lung PCB-binding protein. The identity of the PCB-binding protein is supported by expression of the cDNA in Cos-1 cells where the homogenates from transfected cells show specific binding of 4,4'-bis([ 3H]methylsulfonyl)-2,2',5,5'-tetrachlorobiphenyl, a high affinity ligand for the PCB-binding protein. Also a monospecific antiserum to the PCB-binding protein recognizes a 13-kDa protein in the homogenates of transfected cells but not in the corresponding fraction of mock-transfected cells. Northern blot analysis of total RNA from different rat tissues demonstrates that the cDNA detects a approximately 600-base pair mRNA which appears to be solely expressed in lung. Interestingly, DNA sequence analysis and prediction of the amino acid sequence reveals that the PCB-binding protein shares 53% positional amino acid identity with uteroglobin, a progesterone-binding protein found in rabbit uterus and lung. Furthermore, amino acids shown by x-ray crystallography to delineate the central cavity of uteroglobin, which fits progesterone, are highly conserved in the two proteins.     

The cell cycle of lymphocytes in Fanconi anemia

Summary BrdU-incorporation techniques were used to study the cell cycle in 18 cases of Fanconi's anemia (FA).By comparison with controls, a significant slowing of the cell cycle of lymphocytes in vitro was observed in all FA patients, and possibly in FA heterozygotes, although to a lesser degree. It is probable that the demonstration of the slowing is dependent on the culture conditions. No slowing was observed in other patients affected by at least one of the symptoms of FA. The slow cell cycle of FA cells is mostly due to a very long G₂-phase. A relationship between slow cell cycle and chromatid anomalies exists, the slower cells being significantly more frequently carriers of radial figures than the faster cells, in the same patient.     

Electron microscopy of lanthanum in plasma

Summary Aqueous solutions of lanthanum nitrate may be used as electron microscopic tracers in vivo to study vascular permeability in the experimental animal. However, with this technique the size of the tracer particles is not known. To gain information about the tracer size, we injected lanthanum nitrate into the blood circulation of living rabbits. The plasma obtained from such animals 30 min later, was studied with the electron microscope. The plasma contained an electron-dense material, readily visible in the electron microscope. A precipitate obtained after centrifugation of the whole blood to separate the cells, also contained the tracer. Lanthanum was found in large amounts in the fibrin clot obtained after treating the plasma with thrombin. The tracer was not detected in the serum (i.e. thrombin-treated plasma). The study indicates that ionic lanthanum injected into the blood circulation of living rabbits, is to a great extent bound to fibrinogen, and that the smallest possible size of the tracer is that of the fibrinogen,molecule (m. w. 330,000). Larger particles are present as well.     

Localization of Phloem Oxidases

Among oxidases, cytochrome oxidase has been localized in mitochondria of all phloem cells, catalase has been visualized in parenchyma peroxisomes and peroxidase has been localized in cell walls and in several cell organelles. In angiosperms, peroxidase is present in all phloem cell walls; it is sensitive to cyanide inhibition excepted in sieve areas and around plasmodesmata between sieve tubes and companion cells. In some species, this cyanide resistant oxidasic activity can be localized without exogenous H₂O₂. Peroxidase is localized on ribosomes, inside vacuoles, on the tonoplast and often on the plasmalemma in companion cells and differentiating sieve elements. In young sieve cells some dictyosomes can exhibit a strong peroxidasic activity. In mature parenchyma cells peroxidase can be associated with ER cisternae but not with vacuoles.     

Distribution of mRNAs of fibrinogen polypeptides and albumin in free and membrane-bound polyribosomes and induction of α-fetoprotein mRNA synthesis during liver regeneration after partial hepatectomy

To study the effect of regenerative response of the liver following partial hepatectomy on the synthesis of major plasma proteins (secretory proteins), we have determined the sequence contents and the distribution of albumin and fibrinogen polypeptide mRNAs in rat liver at intervals after partial hepatectomy and sham operation. Using a quantitative technique for the isolation of polyribosomes, we demonstrated that the distribution of RNA between free and membrane-bound polyribosomal fraction was unchanged in these experiments. There was no shift in the polyribosomal population to favor free polyribosomes after partial hepatectomy. However, there was a dramatic increase (5–6-fold) of the fibrinogen polypeptide mRNA concentration during the first 24 h after resection. In contrast, the albumin mRNA concentration decreased (2–3-fold). There were no α-fetoprotein mRNA sequences detectable in any liver RNA fraction in these experimental animals. In sham-operated rats with intact livers, similar changes of fibrinogen polypeptide and albumin mRNA concentrations as described in regenerating liver after partial hepatectomy, were observed. These results suggest that albumin and fibrinogen synthesis after partial hepatectomy is reciprocally regulated at the mRNA level and represents a nonspecific acute phase response to surgical trauma.     

Isolation and metabolic characteristics of rat and chicken enterocytes.

1. The recent recognition of the metabolic, as opposed to absorptive, functions of the small intestine prompted efforts the improve the preparation of metabolically competent columnar absorptive cells ('enterocytes') and to study their metabolic properties. 2. With this preparation, linear rates of O2 consumption are obtained for 40 min at 37 degrees C that are more than 50% higher than rates reported by other authors. 3. Among added substrates, glucose, glutamine and glutamate are the preferred fuels of respiration. The main nitrogenous products of glutamine metabolism are NH3, alanine and glutamate. Glutamine carbon was not detectable in citrulline or proline, in contrast with the findings of Windmueller & Spaeth [(1974) J. Biol. Chem. 249, 5070-5079] in the vascularly perfused small intestine. 4. The rates of O2 uptake in the presence of glutamine or glutamate are sufficient to account for the formation of the carbon skeleton of alanine from the amino acid substrate, i.e. the ratio of O2 used/alanine formed is greater than 1.5. 5. Added ADP and ATP are rapidly degraded to AMP and IMP to a large extent by release of hydrolytic enzymes from the enterocytes into the medium. 6. Chicken enterocytes isolated by the same method are more stable; linear rates of O2 uptake are maintained for 60-70 min.     

The structure of cyanobacterial phycobilisomes: a model

Phycobilisomes, supramolecular complexes of water-soluble accessory pigments, serve as the major light-harvesting antennae in cyanobacteria and red algae. Regular arrays of these organelles are found on the surface of the thylakoid membranes of these organisms. In the present study, the hemi-discoidal phycobilisomes of several species of cyanobacteria were examined in thin sections of cells and by negative staining after isolation and fixation. Their fundamental structures were found to be the same. Isolated phycobilisomes possessed a triangular core assembled from three stacks of disc-shaped subunits. Each stack contained two discs which were 12 nm in diameter and 6–7 nm thick. Each of these discs was probably subdivided into halves 3–3.5 nm thick. Radiating from each of two sides of the triangular core were three rods 12 nm in diameter. Each rod consisted of stacks of 2 to 6 disc-shaped subunits 6 nm thick. These discs were subdivided into halves 3 nm thick.The average number of discs of 6 nm thickness forming the peripheral rods varied among the strains studied. For certain chromatically adapting strains, the average rod length was dependent upon the wavelength of light to which cells were exposed during growth. Analyses of phycobilisomes by spectroscopic techniques, polyacrylamide gel electrophoresis, and electron microscopy were compared. These analyses suggested that the triangular core was composed of allophycocyanin and that the peripheral rods contained phycocyanin and phycoerythrin (when present). A detailed model of the hemi-discoidal phycobilisome is proposed. This model can account for many aspects of phycobiliprotein assembly and energy transfer.Abbreviations PBS phycobilisome(s) - PBP phycobiliprotein(s) - AP allophycocyanin - PC phycocyanin - PE phycoerythrin - PEC phycoerythrocyanin - AP-B allophycocyanin B - C- cyanobacterial - R- rhodophytan - B- Bangiophycean - SDS sodium dodecyl sulfate - LPP Lyngbya-Plectonema-Phormidium group - Na-KPO₄ buffers NaH₂PO₄ titrated with a solution of KH₂PO₄ of equivalent molarity to a given pH     

Enkephalin and substance P effects related to trigeminal pain

Iontophoretic applications of enkephalin (20-150 nA) reduced the spontaneous firing frequency of nociceptive neurons in the trigeminal nucleus caudalis of decerebrated cats. The response evoked by noxious stimulation (tooth pulp) was gradually inhibited during the 1st minute of application of the opioid and generally remained depressed for 5 min after the current was turned off. These effects of enkephalin were blocked by intravenously or iontophoretically administered naloxone. Nonnociceptive neurons or nociceptive neurons responding to nonnoxious inputs were less frequently inhibited by enkephalin. When tested on nonnociceptive cells, similar applications of substance P usually had little effect. Nociceptive neurons, however, were strongly excited by substance P. This action was not constant and was interrupted by periods of inactivation. Both types of peptide action were similar in temporal aspects. The results suggest a functional interrelationship between enkephalin and substance P in a trigeminal system mediating nociception.     

Automated adsorption isotherm device.

A device is described for the automated determination of the water adsorption isotherms of biological materials. The vapor pressure and weight of the adsorbate are measured directly with appropriate transducers, and equilibrium is defined on the basis of constant pressure. The accuracy of the device, determined on two samples with well-known water binding properties, is +/- 5%. Automation is achieved by electronic control.