Response to treatment in hereditary metabolic disease: 1993 survey and 10-year comparison.

Knowledge about cause, pathogenesis, and manifestations of hereditary metabolic diseases puts them among the best known of all human diseases. On the other hand, outcomes of treatment are cause for uncertainty and concern. In 1985, Hayes et al. analyzed efficacy of treatment up to 1983 in 65 of these diseases selected randomly from the McKusick catalogs. Disease scores were calculated for seven parameters: longevity; reproductive capability; somatic and cognitive development; and handicaps affecting schooling, work, and cosmetic appearance. Scores of the untreated and treated phenotypes were then compared. We have now measured progress over the past decade by calculating scores on the same 65 diseases from data in several hundred new reports published since 1983. All seven parameters in the 1993 survey reflect improved efficacy of treatment in the 10-year interval. However, the percent of diseases for which all manifestations of the disease were removed by treatment has not changed (12% in 1983; 12% in 1993). The group in which manifestations were untouched by treatment has become smaller (48% in 1983; 31% in 1993), and the group partially ameliorated by treatment had increased reciprocally (40% in 1983; 57% in 1993). Progress in the treatment of hereditary metabolic disease is thus better than it was, but it is still only a partial success. The advances are attributable to greater success with organ and tissue transplantation, better pharmacotherapy, and better support systems. Restoration of normal homeostasis, the key to successful treatment, remains an elusive challenge and is a logical, major focus for research in human genetics.     

Hydroponic Seedling Bioassay for the Bioherbicides Colletotrichum truncatum and Alternaria cassiae

A rapid bioassay was developed to measure the bioherbicidal efficacy of spore preparations of the pathogens Colletotrichum truncatum (Schwein.) Andrus and W. D. Moore and Alternaria cassiae Jurair and Khan on hemp sesbania (Sesbania exaltata) and sicklepod (Cassia obtusifolia), respectively. The system uses 4-day-old dark-grown seedlings (grown hydroponically in paper towel cylinders) which were sprayed with spore suspensions. Shoot lengths were monitored non-destructively, and recorded over time under conditions of dark growth, 90-100% relative humidity and 25 C. Shoot growth inhibition and stem collapse (mortality) were directly related to the spore concentration applied. Generally, at 10 3 - 10 4 spores ml-1, these pathogens caused significant shoot growth inhibition within 25-30 h and seedling death within 40-50 h. This bioassay has been used to study herbicide-pathogen interactions, and may be extended to determine the bioherbicidal efficacy of different pathogen isolates, pathovars or spore formulations. This technique is more rapid, uses a lower inoculum volume, requires less space and is performed under more controlled conditions than conventional greenhouse bioassay methods. The data obtained are more quantitative than those obtained from bioassays relying on visual rating systems.     

Association of the type II cAMP-dependent protein kinase with a human thyroid RII-anchoring protein. Cloning and characterization of the RII-binding domain.

The type II cAMP-dependent protein kinase (PKA) is localized to specific subcellular environments through binding of the dimeric regulatory subunit (RII) to anchoring proteins. Subcellular localization is likely to influence which substrates are most accessible to the catalytic subunit upon activation. We have previously shown that the RII-binding domains of four anchoring proteins contain sequences which exhibit a high probability of amphipathic helix formation (Carr, D. W., Stofko-Hahn, R. E., Fraser, I. D. C., Bishop, S. M., Acott, T. E., Brennan, R. G., and Scott J. D. (1991) J. Biol. Chem. 266, 14188-14192). In the present study we describe the cloning of a cDNA which encodes a 1015-amino acid segment of Ht 31. A synthetic peptide (Asp-Leu-Ile-Glu-Glu-Ala-Ala-Ser-Arg-Ile-Val-Asp-Ala-Val-Ile-Glu-Gln-Val -Lys-Ala-Ala-Tyr) representing residues 493-515 encompasses the minimum region of Ht 31 required for RII binding and blocks anchoring protein interaction with RII as detected by band-shift analysis. Structural analysis by circular dichroism suggests that this peptide can adopt an alpha-helical conformation. Both Ht 31 (493-515) peptide and its parent protein bind RII alpha or the type II PKA holoenzyme with high affinity. Equilibrium dialysis was used to calculate dissociation constants of 4.0 and 3.8 nM for Ht 31 peptide interaction with RII alpha and the type II PKA, respectively. A survey of nine different bovine tissues was conducted to identify RII binding proteins. Several bands were detected in each tissues using a 32P-RII overlay method. Addition of 0.4 microM Ht 31 (493-515) peptide to the reaction mixture blocked all RII binding. These data suggest that all anchoring proteins bind RII alpha at the same site as the Ht 31 peptide. The nanomolar affinity constant and the different patterns of RII-anchoring proteins in each tissue suggest that the type II alpha PKA holoenzyme may be specifically targeted to different locations in each type of cell.     

Electromyographic changes in work-related myalgia of the trapezius muscle.

In 11 patients, all women, 21-55 years of age, with unilateral work-related myalgia of the trapezius muscle, the right and left trapezius muscles were examined simultaneously for electromyogram (EMG) signs of localized muscle fatigue. All patients were tested with 0-kg hand load for 5 min, holding the arms straight at 90 degrees of elevation in the scapular plane. Only 4 of the patients tolerated exposure to higher load levels. They were tested with 1 kg hand load for 3 min and 2 kg hand load for 2 min, with a period of rest of 30 min between the trials. The EMG mean power frequency (MPF) and root mean square (rms) were calculated. Data were normalized with the initial value as a reference and regression analyses were performed. On both sides a decrease of MPF and an increase of rms were found with increasing time and load, i.e. classical EMG signs of localized muscle fatigue. Compared with the nonaffected side smaller changes were found on the affected side, possibly due to pain inhibition, impaired microcirculation and biochemical changes along the muscle fibres. At 0-kg hand load we found no change of MPF on either side despite subjective feelings of fatigue and pain. We interpreted these findings as an indication of reduced capacity of the affected trapezius muscle to sustain static load with early development of pain-associated local fatigue.     

The expression of plasmalemma transcobalamin-II receptors on human blood lymphocytes stimulated by mitogens]

The influence of mitogens on the expression of surface membrane TC-II receptors of human blood lymphocytes and internalization of (TS-II+57Co-CNCbl) complex into cytoplasm were investigated. Mature lymphocytes have a very small number of surface receptors to plasma TC-II but their expression is increased significantly by PHA or Con-A stimulation. CBl transport to cytoplasma is activated in definite sequence by two different mechanisms. Stimulated cells take free CBl without participation of TC-II in early hours of mitogen action (12-42 hrs) before maximal 3H-thymidine incorporation into DNA. On day 3 of cultivation, specific mechanism of CBl transport triggers and the number of lymphoblast receptors is increased manifold. Radioactive CBl enters cytoplasma due to interaction of TC-II-CN [57Co] CBl of the medium with surface membrane receptor of the cells. Thus, the definition of TC-II receptors as an important functional parameter may serve a marker of proliferating cells.