Diphosphonate bone scans in patients with polyarthralgias.

Early detection of inflammatory arthropathy is notoriously difficult with standard radiographic techniques. We therefore assessed bone turnover with technetium Tc 99m medronate in 16 patients with persistent polyarthralgias who had no clinical synovitis, normal radiographs, and nondiagnostic results from laboratory evaluations. Abnormal scans were found in 11 of 16; five were unremarkable. Scan abnormality corresponded with symptomatic joints (11 of 11 patients). These 11 patients had normal test results for rheumatoid factor, antinuclear antibody, and HLA-B27. Patients with abnormal scans were treated with nonsteroidal antiinflammatory drugs or analgesics (11 of 11), hydroxychloroquine sulfate (four), or gold salts (one), with improvement (nine of 11); patients with normal scans (five of five) were treated successfully with nonsteroidal antiinflammatory drugs or analgesics and reassurance. One patient with a normal scan developed sarcoidosis; one, hypermobility syndrome; and one, a viral syndrome. Two patients had no diagnosis. Abnormal technetium Tc 99m medronate scans in patients with previously undiagnosed polyarthralgias suggested inflammatory arthropathy and influenced management decisions with favorable therapeutic outcomes.

Immunological assay of phytochrome in small sections of roots and other organs of maize (Zea mays L.) seedlings.

Phytochrome was determined in small sections of maize (Zea mays L.) seedlings by means of a highly specific double sandwich enzyme immunoassay which uses a monoclonal anti-phytochrome antibody for binding phytochrome and anti-phytochrome serum to detect the bound phytochrome. The distribution of phytochrome in maize seedlings was followed from germination to the 7th d after soaking the caryopses. Regions of high phytochrome accumulation were found in the coleoptile tip, the root cap and the shoot apex: the values for 5-d-old seedlings were 120, 80 and 70 µg phytochrome per g fresh weight (or 0.91, 0.61 and 0.53 nmol·g(-1)), respectively. The mesocotyl and the leaves contained relatively low amounts of phytochrome (less than 10 µg·g(-1)FW), which were almost uniformly distributed throughout these organs. As might be expected, regions of these organs adjacent to the shoot apex showed higher levels. The root, other than root tip, was almost devoid of phytochrome (0.2 to 0.5 µg·g(-1)). The general distribution of phytochrome in organs did not change during the development of seedlings. The amount of phytochrome, however, did fluctuate: up to the 5th or 6th d after soaking the caryopses, the levels increased in the regions of high phytochrome accumulation but thereafter decreased. After the 6th d the roots were 15 cm or longer and the coleoptiles became prone to penetration by primary leaves. The tips of adventitious roots, emerging after the 6th d, were also found to contain phytochrome. When the root cap was illuminated (4.3 W·m(-1)), phytochrome was degraded as in illuminated shoots. Degradation of phytochrome in coleoptile, mesocotyl and shoot apex started with a lag phase but phytochrome degradation in the root cap and the leaves started without a lag. In contrast to shoot phytochrome, which was almost completely degraded under continuous illumination, about 3% of initial phytochrome was measured in root caps after 24 h continuous illumination. Some of the data, obtained by immunological measurements, may indicate differences between phytochrome, or its synthesis or degradation, in the root cap and shoots. The results are discussed with a view to different red-light-mediated responses of grass seedlings.

Rheumatoid vasculitis: experience with 13 patients and review of the literature.

Rheumatoid vasculitis is an uncommon but potentially catastrophic complication of RA. There are few current extensive experiences and no consensus regarding the clinical, laboratory, histologic features, and management or prognosis of rheumatoid vasculitis. We therefore reviewed selected observations in 13 patients followed over the past decade and compared them with patients reported and with results of a survey of North American Rheumatologists. Our patients were seven men and six women (age, 33 to 70 years) who had had active RA for 4 to 36 years. They exhibited sensory neuropathy, mononeuritis multiplex, Felty syndrome, cutaneous lesions, leg ulcers, gangrene, anemia, leukocytosis, eosinophilia, high titers of RF, hypocomplementemia, and CICs or cryoglobulinemia approximately as frequently as other reported patients with rheumatoid vasculitis, but they displayed constitutional symptoms, subcutaneous nodules, ischemic changes, and proteinuria rather less consistently than in other series. These observations were not necessarily as expected by survey respondents. We, as in other series and suggested by survey respondents, tended to select penicillamine or cytotoxic drugs (or plasmapheresis) for patients with mononeuritis, gangrene, or leg ulcers, and nonsteroidal antiinflammatory drugs, antimalarials, gold, or penicillamine for sensory neuropathy or digital lesions. Four patients died, two deteriorated, and seven were stable or improved, a finding that was also similar to the experiences of others. Rheumatoid vasculitis is an uncommon, potentially catastrophic syndrome with varying clinico-pathologic features that have different prognostic implications and should be managed individually.

Properties of lectins from snails of the genus Helix probed by monoclonal antibodies.

Monoclonal antibodies were raised against the lectin of Helix pomatia (HPL). Besides antibodies bearing the more common gamma and kappa chains, antibodies with alpha, mu and lambda 2 chains were elicited. The anti-HPL antibodies are expected to be useful in studies on HPL biogenesis and HPL substructure and in studies concerned with the binding of HPL to cell surfaces. Binding of carbohydrates to HPL impaired the binding of anti-HPL antibodies. One to 3 mM GalNAc inhibited HPL-binding in two out of nine antibodies. None of the antibodies bound in the presence of micrograms per ml of the polyvalent blood group A-substance from hog stomach. Similarly, all anti-HPL antibodies were prevented from binding if non-inhibitory concentrations of A-substance were supplemented with GalNAc. Lectins from Helix aspersa (HAL) and Helix lucorum (HLL) differed from HPL in antigenic properties. Only one anti-HPL antibody each bound these lectins as well as HPL. Binding of lectins of Cepaea and Rapana was scarcely detectable. Most of the anti-HPL antibodies and the multivalent HPL-antigens formed precipitation lines in double diffusion tests. At least two antibodies (IgMs) did so with HLL but none with HAL. The possibility that antibodies were selected because of unknown interactions between HPL and the carbohydrate moieties of certain fractions of antibodies was excluded by raising the antibodies in the presence of tunicamycin to inhibit N-glycosylation.

Molecular properties of 5-aminolevulinic acid dehydratase from Spinacia oleracea.

5-Aminolevulinic acid dehydratase from spinach (Spinacia oleracea), highly purified by immunoprecipitation, was characterized by inhibitor studies, amino acid composition, the mode of substrate binding and electron photomicrography. The results show that the conversion of 5-aminolevulinate to porphobilinogen requires an active arginine residue and the formation of a Schiff base between the enzyme and 5-aminolevulinate. The formation of a Schiff base is well known for bacterial and animal dehydratases. Spinach dehydratase, however, is distinguished by its insensitivity to iodoacetamide, a low content of cysteine residues and a high proportion of acidic amino acids. In addition, electron photomicrographs of spinach dehydratase molecules do not resemble the corresponding images of beef liver dehydratase. The finding that an arginine residue is essential for substrate conversion corroborates the suggestion that the right orientation of the substrate in the active center is dependent on a positive charge.

Highly efficient purificaton of the labile plant enzyme 5-aminolevulinate dehydratase (EC 4.2.1.24) by means of monoclonal antibodies.

5-Aminolevulinate dehydratase (ALAD) from spinach (Spinatia oleracea) was isolated by affinity purification on an immunoabsorbens with a yield of 70 to 80% of the activity in the crude enzyme preparation. The enzyme eluted from the immunoabsorbens was pure as judged by polyacrylamide gel electrophoresis and is a hexamer with a subunit molecular weight of about 50 000. Enzyme bound to the immunoabsorbens was able to synthesize porphobilinogen in a continuous manner. Owing to the lability of the enzyme and its low abundance in plant tissue, we have been unable to obtain similar yields of purified enzyme using classical purification procedures. This highly efficient purification was made possible by using monoclonal antibodies as described by Köhler and Milstein (Nature 256, 495 (1975)). The availability of monoclonal antibodies meant that it was not necessary to purify the enzyme to homogeneity by classical means in order to raise an antiserum specific for ALAD. Sixteen clones of cells producing antibodies against ALAD were selected. They all expressed a chi light chain but differed in the heavy chain class which was eigher gamma 1 or gamma 2a. The availability of pure ALAD enzyme and of highly specific antibodies against the enzyme now enables us to answer questions concerning properties, localization, intercellular transport and evolution of ALAD. It is clear that the technique used and the questions asked are not restricted to ALAD.

Light-induced, Dark-reversible Absorbance Changes in Roots, Other Organs, and Cell-free Preparations.

Irradiation of maize (Zea mays) roots and coleoptiles with visible light causes dark-reversible absorbance changes in these organs. There is an increase in absorbance near 440 nanometers and smaller increases below 410 nanometers and about 595 nanometers. Decreases in absorbance are observed at about 420 nanometers and minor ones at 537 and 575 nanometers. These responses are also observed in cell-free preparations from roots and coleoptiles if dithionite, NADPH, or NADH is added prior to illumination. The dose curve for these effects has a distinct maximum at 420 nanometers and a minor one at 575 nanometers. Difference spectra and dose response curves indicate that heme compounds such as cytochromes or, more probably, peroxidase complexes are the photoreceptive and chemical reacting molecules. Siroheme-containing proteins may also be taken into consideration.The light-induced absorbance changes have half-lives of more than 200 seconds and 100 seconds in roots of maize and soybean, respectively. Two reactions, each with first order kinetics, appear to be superimposed. The respective rate constants for maize roots are about 0.004 and 0.04 seconds(-1). The generation of the effect has a much shorter half-life dependent on light intensity and wavelength. Little deviations from first order kinetics were detected. Rate constants for corn roots range between 0.05 and 0.01 seconds(-1).Apart from the problem in which hemoproteins are involved, there is the problem of correlating the reaction of the photoreactive and chemically reacting molecules to macroscopic responses such as phototropism.

Corneal thickness measurements following intraocular lens implantation.

Corneal thickness measurements were carried out on patients who had undergone combined phacoemulsification cataract extraction with simultaneous intraocular lens implantation in one eye at least six months prior to the date of measurement. The unoperated eye served as control. Another group of patients who had undergone simultaneous phacoemulsification cataract extraction combined with intraocular lens implantation were followed over a two-month period of time, serial corneal thickness measurements being performed. The results indicate that, in surgery performed by experienced surgeons on carefully selected patients, phacoemulsification cataract extraction using the Kelman technique combined with intraocular lens implantation had no significant effect on corneal endothelial function as reflected in corneal thickness changes.