Ammonia transport by the urinary bladder of Bufo marinus

Among cyanobacteria, the heterocystous, N2-fixing Anabaena variabilis and the unicellular Anacystis nidulans have recently been shown to possess an NAD+-dependent, bidirectional hydrogenase. A 5.0 kb DNA segment of the A. nidulans genome is now identified to harbor the structural genes hoxUYH coding for three subunits of the bidirectional hydrogenase. The gene arrangement in A. nidulans and in A. variabilis is remarkably dissimilar. In A. nidulans, but not in A. variabilis, the four accessory genes hoxW, hypA, hypB and hypF could be identified downstream of hoxH. An insertional homozygous mutant in hoxH from A. nidulans was completely inactive in performing Na2S204-dependent H2 evolution but could utilize the gas with almost 50% of the activity of the wild type. These findings with the first defined hydrogenase mutant in any photosynthetic, 02-evolving microorganism indicate that the unicellular cyanobacterium A. nidulans possesses both an uptake and a bidirectional hydrogenase. The physiological role(s) of the two hydrogenases in unicellular non-N2-fixing cyanobacteria is not yet understood.     

Prediction of major depression and dysthymia from CES-D scores among ethnic minority adolescents

Two tumour cell clones, 6D1 and 4C2 cells, which are defective both in the major histocompatibility gene complex (MHC) class I expression and in the endogenous antigen presentation, are recovered with interferon (IFN)-gamma treatment. The present study describes the ultrastructure of these cells by using scanning and transmission electron microscopy in relation to the effect of IFN-gamma treatment. The general morphology of these cells was found to be similar to each other and comparable to that of a tumour cell clone, 4A1 cells, of the same origin, normal in MHC class I expression; they exhibited a fibroblast-like appearance and had many blebs on all the cell surfaces, with desmosome-like junctions between cells. On IFN-gamma treatment, surface fine blebs appeared less, and mitochondria became more densely stained. Expression of MHC class I molecules on the cell surface was much higher in the IFN-gamma treated 6D1 and 4C2 cells than in untreated cells, when estimated by immunoelectron microscopy. The addition of an epitope peptide to these cells did not enhance the class I expression, which differed from other antigen presentation-defective cells such as RMA-S cells, nor change the cell surface morphology.     

Expression of inducible nitric oxide synthase in human burn wounds

A water-soluble synthetic peptide with only nine amino acid residues, comprising the 131-139 sequence region of the cytotoxic protein alpha-sarcin (secreted by the mold Aspergillus giganteus), interacts with large unilamellar vesicles composed of acid phospholipids. It promotes lipid mixing between bilayers and leakage of vesicle aqueous contents, and it also abolishes the phospholipid phase transition. Other larger peptides containing such an amino acid sequence also produce these effects. These peptides acquire alpha-helical conformation in the presence of trifluoroethanol, but display beta-strand conformation in the presence of sodium dodecyl sulfate. The interaction of these peptides with the lipid vesicles also results in beta-structure. The obtained data are discussed in terms of the involvement of the 131-139 stretch of alpha-sarcin in its interaction with lipid membranes.     

The effect of combined external beam radiotherapy and brachytherapy on local control and wound complications in patients with high-grade soft tissue sarcomas of the extremity with positive microscopic margin

PURPOSE: A previously reported randomized trial from out institution demonstrated a local control advantage to adjuvant brachytherapy (BRT) for completely resected high grade soft tissue sarcoma (STS). In recent years, BRT boost has been combined with wide field external beam radiotherapy (EBRT) for selected patients in whom the margin(s) of resection was positive. This study evaluates the impact of BRT boost plus EBRT on local control in this subset of patients and on wound complication rates. METHODS AND MATERIALS: Between January, 1987 and December, 1992, 105 adult patients with primary or locally recurrent high grade STS of the extremity were treated with wide local excision and BRT alone (87 patients; dose: 45 Gy) or BRT plus EBRT (18 patients; dose: 15-20 Gy BRT + 45-50 Gy EBRT). The margin(s) of resection was positive in 10 out of 18 patients in the BRT + EBRT group vs. 17 out of 87 patients in the BRT alone group. Wound complications were classified as major if they required further operative intervention; moderate if there was purulent discharge, hematoma > 25 ml, wound separation > 2 cm, and persistent seroma requiring drainage; or minor if less than moderate. Median follow-up was 22 months. RESULTS: The overall 2-year actuarial local control rate was 86%. There was no difference in the 2-year actuarial local control rate between the BRT + EBRT group (90%) and the BRT alone group (82%) (9 = 0.32). However, for patients with positive resection margins the use of BRT + EBRT produced better local control than BRT alone [9 out of 10 (90%) vs. 10 out of 17 (59%)]. This difference approached but did not reach statistical significance (p = 0.08). No difference was seen in patients with negative margins. There was no significant difference in the overall wound complication rate (26% BRT vs. 38% BRT + EBRT, p = 0.31) nor in the combined major and moderate wound complication rate (16% BRT vs. 27% BRT + EBRT, p = 0.39). CONCLUSION: Our preliminary data suggest a trend in favor of BRT boost + EBRT as the optimal adjuvant local strategy for STS with positive resection margins. There is no significant difference in the wound complication rate with either technique.     

On-the-fly fluorescence lifetime detection of dye-labeled DNA primers for multiplex analysis

Mixtures of dye-labeled, M13-forward DNA primers were separated by capillary gel electrophoresis and detected on-the-fly, using fluorescence lifetime measurements, to evaluate four-decay detection for multiplex DNA sequencing. Three different four-dye systems were used, two that were excited at 488 nm and one that was excited at 514 nm. Each dye-labeled primer was identified on the basis of the lifetime of the conjugated dye using nonlinear least squares or the maximum entropy method to analyze the lifetime data. Overlapping electrophoretic peaks were generated by making multiple injections of mixtures of the dye-labeled primers. The overlapping peaks were resolved by fitting the data to two-, three- or four-component lifetime models used in nonlinear least-squares analysis in which each lifetime component was fixed to the predetermined lifetime of the corresponding dye-labeled primer. In two of the dye systems, the lifetimes of the four dye-labeled primers were sufficiently different to allow peak resolution. In the other dye system, addition of 10% DMSO to the run buffer changed the lifetime of one dye-labeled primer, allowing it to be resolved from another dye-labeled primer with similar lifetime.     

A method for rapid filling of fine-tipped micropipettes with electrolyte solutions including those containing horseradish peroxidase

Cooling the tips of micropipettes pulled from filament-containing capillary glass dramatically increases the rate and completeness of filling with a variety of electrolyte solutions. In tests with solutions containing horseradish peroxidase this method increased the completeness of filling by more than two orders of magnitude.     

Tissue and plasma peptidase activity is altered during hypothermic hibernation in the 13 lined ground squirrel (Spermophilus tridecemlineatus)

Adult 13 lined ground squirrels were monitored for entry into a state of hypothermic hibernation or arousal in a cold room on a photoperiod LD 2:22. Once animals developed predictable hibernation patterns, animals were killed at the mid point of hypothermic hibernation or arousal for determination of plasma and tissue angiotensin-1-converting enzyme (Kininase II) activity. Enzyme was extracted from plasma, lung, kidney, liver, forebrain and brainstem and assayed in vitro. During hypothermic hibernation enzyme activity is significantly decreased in all tissues examined. These data suggest that the activity of tissue and plasma peptidases are altered during the cyclic torporous periods characteristics of hibernation in this species.