Fibronectin-degrading proteases from the membranes of transformed cells

The local degradation of fibronectin substrata by Rous sarcoma virus-transformed chick embryonic fibroblasts requires cell-contact-related metalloendoprotease and serine-protease activities. Using fibronectin-containing SDS gels, two large proteases with apparent molecular weights of 120K and 150K were found only in the membrane fraction of transformed cells and were absent in normal cells. Both 120K and 150K proteases were active at neutral pH, but showed preferential inhibitor sensitivities of serine and metal proteases, respectively. The 150K protease appeared to account for most of the proteolytic activity since metalloendoprotease inhibitors completely blocked proteolytic activity of the 150K in fibronectin gels, more than 80% of the fibronectin-degrading activity of solubilized membranes, and largely suppressed the appearance of fibronectin degradation spots in cultures of transformed cells.     

Transpososomes: stable protein-DNA complexes involved in the in vitro transposition of bacteriophage Mu DNA

We report that two types of stable protein-DNA complexes, or transpososomes, are generated in vitro during the Mu DNA strand transfer reaction. The Type 1 complex is an intermediate in the reaction. Its formation requires a supercoiled mini-Mu donor plasmid, Mu A and HU protein, and Mg2+. In the Type 1 complex the two ends of Mu are held together, creating a figure eight-shaped molecule with two independent topological domains; the Mu sequences remain supercoiled while the vector DNA is relaxed because of nicking. In the presence of Mu B protein, ATP, target DNA, and Mg2+, the Type 1 complex is converted into the protein-associated product of the strand transfer reaction. In this Type 2 complex, the target DNA has been joined to the Mu DNA ends held in the synaptic complex at the center of the figure eight. Supercoils are not required for the latter reaction.     

Critical considerations in the development of an assay for sulfidopeptide leukotrienes in plasma

A sensitive and specific assay has been developed for measurement of total sulfidopeptide leukotrienes (LT) in plasma. LTC4 and LTD4 in plasma are converted to LTE4 which is then extracted by C18 Sep-Pak binding and elution. Total LTE4 is resolved by reverse phase high performance liquid chromatography (RP-HPLC) and quantitated by radioimmunoassay (RIA). A [3H]LTE4 internal standard is added to the starting plasma sample to allow overall recovery to be calculated and to define the fractions from RP-HPLC to be assayed for LTE4-like immunoreactivity. The correlation between the measured increase in LTE4 concentration after addition of incremental amounts of LTC4 and LTE4 to plasma was 0.989 and 0.978, respectively, with slopes of 1.05 and 1.11. Addition of 51 pg/ml LTE4 to 5 ml plasma was detectable; the measured increase was 48 +/- 12 pg/ml (mean +/- SE, n = 7). The intra-assay coefficient of variation for 341 pg/ml of added LTC4 was 3.2% (n = 6). Sulfidopeptide leukotrienes could not be detected in blood samples taken from 12 normal volunteers in whom the theoretical detection limit, calculated from the sensitivity of the RIA, the overall recovery of LTE4, and the volume of plasma extracted, was 83 +/- 4 pg LTE4/ml plasma (0.19 +/- 0.01 pmol sulfidopeptide leukotriene/ml plasma; mean +/- SE).     

Age-specific Chaoborus predation on rotifer prey

SUMMARY. 1. This is the first study to examine predator-prey interactions between Chaoborm instars and rotifer prey. The predatory behaviour of instars I–III of Chaoborus pimctipennis and the diet selectivity of instars I—IV feeding on rotifers were examined in the laboratory. Prey used in direct observations of predatory behaviour included a variety of rotifers (Symhacta pectlnata, S. ohUmga, Polyarthra remata, Asplanchna girodi, Keratella crassa, spined and unspined forms of Keratella cochlearis) and two crustaceans (Bosmitia longirostris, Mesocyclops edax nauplii. 2. In general, strike efficiencies (percentage of strikes resulting in inges- tion) increased in successive instars I—III. Early instar (I and II) strike efficiencies were low when compared with other invertebrate predators. For a given instar. mean prey handling times varied among prey species more than strike efficiencies. Mean handling times for small, soft-bodied rotifers were lowest and those for wide, hard-bodied prey were highest. 3. Instar I exhibited significantly greater selectivity for the small, soft- bodied S. obUmga than for the larger S. pectinata, hard-bodied K. crassa, and spined and unspined forms of K. cochlearis. Instars II—IV positively selected both the large and small Symhaeta species over all Keratella species. The relationship between Chaobortts selectivity and prey value (weight of prey per unit handling time) can be described by a power function. Ingestion rates of rotifers by older instars (III and IV) are among the highest reported for invertebrate predators. 4. Rotifer vulnerability to Chaoborus predation probably depended on rotifer cuticle texture, body width, and hydrodynamic disturbances. Spined rotifers were not necessarily protected from Chaoborus predation because Chaohorus can manipulate and swallow them. Giguere et al.'s 1982) encounter rate model must be modified to predict encounter rates of slow-moving rotifer prey with Chaohorus.     

Short-term movements of chalk-stream invertebrates

SUMMARY 1. Trays of gravel were implanted for 24 h periods in several chalk streams. The trays were variously modified to monitor colonization from different sources, particularly drift (D), other downstream movements (DSM), upstream movements (USM), and all sources combined (ALL).
2. The abundance of invertebrates in the streams was high (maximum of 717 per 0.04 m²) and colonization of the trays was extensive, especially by Gammarus pulex , the predominant species in all but one of the streams (maximum number of individuals of G. pulex per tray: for D, 473; for DSM, 462; tor USM, 286; and tor ALL, 639). In some cases benthic densities were achieved in the trays in the 24 h exposure period, and in most cases any one source of colonization could provide benthic densities in a few days.
3. Patterns of movement differed between species, between size classes of G. pulex , and between sites for particular species. No one source of colonization could be regarded as generally predominant although downstream movements in total (D + DSM) usually far exceeded USM. DSM was usually an important source of colonization.
4. The extent and direction of movement appeared to be determined by local factors and were explained in terms of the high densities of invertebrates and the current and substratum characteristics of each site.
5. Estimates of the relative importance of different sources of colonization varied greatly according to the criteria used in determining proportions.     

Tracheostomy Tube Failure in Obstructive Sleep Apnea

In our institutions we routinely do posttracheostomy sleep studies on patients being treated for obstructive sleep apnea. We have identified several patients who failed to show objective evidence of improvement after tracheostomy. From our studies we have found that both mechanical obstruction and concomitant respiratory control dysfunction caused this failure. A unique tracheostomy tube was constructed to treat the subset of patients with internal collapse of the tracheostomy tube.     

An abnormal terminal X-Y interchange accounts for most but not all cases of human XX maleness

To determine if human XX maleness results from an abnormal chromosomal X-Y interchange, we studied the inheritance of the paternal pseudoautosomal region in nine patients. Those six patients in whom Y-specific DNA was found (Y(+)) inherited the entire pseudoautosomal region from the paternal Y chromosome and lost that of the paternal X chromosome. Moreover, in three Y(+) cases, we observed the deletion of a paternal Xp locus tightly linked to the pseudoautosomal region. These results definitively show that an abnormal and terminal X-Y interchange during paternal meiosis causes Y(+)XX maleness. In contrast, no abnormal X-Y interchange was observed in any of the three Y(-) cases analyzed, suggesting that maleness can occur in the absence of any Y-specific DNA.     

Startle responding and context conditioning. Naloxone pretreatment and stimulus intensity

The possibility that acoustic startle stimuli could support a conditional response (freezing) to contextual stimuli was investigated. Rats were exposed to three acoustic startle stimuli on the first day, and one on the second day. On day 1, 20 rats received naloxone pretreatment and another 20 received saline (placebo) pretreatment. Half of each group received a high-intensity acoustic stimulus, the other half a low-intensity acoustic stimulus. Both the higher stimulus intensity and the naloxone pretreatment led to greater freezing behavior during the 3-minute test period before the single startle stimulus on day 2. These findings support the notion that increased actual or perceived intensity of the acoustic startle stimulus increases conditioning to contextual stimuli as indexed by freezing behavior.     

Early Pavlovian studies on the effects of starvation. A contribution to the history of neurodietetics

This communication is based on the pioneering studies performed by I. P. Pavlov's associates, Yu. P. Frolov (in 1918) and I. S. Rozental' (in 1918/1919) at the Institute of Experimental Medicine, in Petrograd. The changes in the "independent variable"--the dogs' diet--were not planned but were the consequences of severe shortages of food for man and beast. The principal generalization concerns the order in which different forms of "complex nervous activity" were impaired: the order is opposite to the order in which they emerge in the process of ontogenesis. First to suffer was "internal inhibition," as documented by the failure of stimulus differentiation. This was followed by the decrease in the magnitude of well-established conditional responses (CRs). As a result of a more severe impairment of the excitatory processes, it became difficult or impossible to establish new CRs. In time, previously established CRs to artificial stimuli, visual and acoustic, disappeared totally. The CRs to natural conditional stimuli (CSs) were maintained fairly well but, eventually, they too decreased markedly. In the terminal phase of starvation, the unconditional salivary reflexes continued to function, although their magnitude was depressed.