Anti-predator adaptations in a neustonic insect (Velia caprai)

Summary The risk of a Velia caprai (Heteroptera: Veliidae) individual to fall victim to brown trout (Salmo trutta) was demonstrated experimentally to be markedly lower when several bugs were simultaneously exposed to the trout. V. caprai was found distasteful to brown trout, and a high proportion was ejected alive after being captured. We suggest that school formation in V. caprai lowers the risk of predation because frequent predator-prey encounters may assist in retention of the avoidance learned by the predator. Individual behavioural responses to attacks were variable and the frequency of expansion skating and thanatosis was temperature dependent. Thanatosis postures were either symmetric with the legs pressed to the body or irregular.     

Increased acidic phospholipids in rat brain membranes after chronic ethanol administration

Two types of plasma membranes isolated from rat brain cortex were used to study the membrane-perturbing properties of ethanol. Rats administered ethanol in the form of a liquid diet showed an increase in levels of phosphatidylserines, phosphatidylinositols and phosphatidic acids as compared to controls. The results present evidence that chronic ethanol treatment results in an increase in the acidic phospholipids in brain membranes. This type of membrane modification may have important implications for the function of membrane transport enzymes such as (Na+, K+)-ATPase, which also increases in activity upon chronic ethanol administration.     

Influence of steady-state and fluctuating salinities on the oxygen consumption and activity of some brackish water shrimps and fishes

Oxygen consumption, locomotory activity and, in some cases, osmoregulatory responses of different populations of Palaemon adspersus (Rathke) and Pomatoschistus microps (Krøyer) from the Isefjord ($\text{S 19‰}$) and Karrebaek Fjord ($\text{S 12‰}$) in Denmark and the Barther Bodden ($\text{S 6‰}$) in the G.D.R. to short-term salinity fluctuations, and after long-term adaptation, were tested. The same tests were performed on populations of Gasterosteus aculeatus (L.), Palaemonetes varions (Leach) (both from Barther Bodden, G.D.R.) and Palaemon elegans (Rathke) (Black Sea, Bulgaria, $\text{S 18‰}$). The steady-state experiments showed that the standard metabolic rates of P. adspersus and Pomatoschistus microps reach their lowest levels at mean biotope salinities at both 10 and 20°C. In contrast, the routine metabolic rates of both species are independent of salinity in the ecological salinity range.All Palaemon adspersus and Pomatoschistus microps populations responded to sudden changes in salinity with increased locomotory activity and respiration regardless of the direction of stressing. Metabolic adaptation in these euryhaline species, which is not synchronous with osmotic readjustment, takes from 5 to 12 h, depending on the salinity gradient.The polystenohaline Palaemon elegans from the Black Sea and the holeuryhaline Palaemonetes varians from the Barther Bodden exhibit similar short adaptation times (≈ 2 h) to identical salinity gradients but in different salinity zones.     

Comparison of peptidase activities in some fungi pathogenic to arthropods

Peptidases, highly specific toward several synthetic chromogenic peptides, were found in the mycelia of four arthropod pathogenic fungi: Aphanomyces astaci, Beauveria bassiana, Metarrhizium anisopliae, and Paecilomyces farinosus. A. astaci peptidases had high hydrolyzing activities toward most of the peptides, especially those with arginine in the P1 position, while those of B. bassiana and P. farinosus readily hydrolyzed peptides with valine and arginine, as well as proline and tyrosine in the P2 and P1 positions, respectively. The hydrolyzing capacities of M. anisopliae peptidases were similar to A. astaci, but showed lower specific activities. Casein or azocoll was only hydrolyzed by A. astaci peptidases. B. bassiana and M. anisopliae had a very low hydrolyzing capacity toward casein and could not degrade azocoll. P. farinosus had no hydrolyzing activity toward casein or azocoll. Only peptidases from the crayfish pathogen A. astaci could degrade the crayfish cuticle. The peptidase preparations of A. astaci and B. bassiana hydrolyzing MeO-Suc-Arg-Pro-Tyr-pNA or Bz-Phe-Val-Arg-pNA were of the serine type. The possible importance of peptidase activity of arthropod pathogenic fungi in the infection process is discussed.     

Effect of gossypol on testicular blood flow and testosterone production in rats

Rats were treated with highly purified gossypol acetic acid at doses of 15 or 30 mg/kg day-1 for 6 weeks to produce an effect on spermatogenesis as shown by reduced sperm motility and increased sperm malformation rates. The treated rats did not differ from the controls in the body weight growth curves and reproductive organ weights. When stimulated with hCG, testicular blood flow was increased in the low dose group; the testosterone concentrations in peripheral and testicular venous blood were also increased to a greater extent than those of the control group. No difference was found between the high dose and control groups in testicular blood flow or testosterone concentrations. The morphology of the Leydig cells was apparently normal, although some degenerative changes in the germinal epithelium were observed in the high dose group. Therefore, there is no evidence in our experiment to show any anti-androgenic effect following 6-week treatment of gossypol in rats, even at the dose of 30 mg/kg day-1.     

The role of birds as definitive hosts and intermediate hosts of heteroxenous coccidians

Sarcocystis-like oocysts-sporocysts were found in four species of owls (Asio otus, Bubo bubo, Strix aluco, and Tyto alba) and in five species of predatory birds (Accipiter gentilis, Accipiter nisus, Buteo buteo, Circus aeruginosus, Falco tinnunculus). In addition, the muscles of 15 of 41 (36.5%) pheasants (Phasianus colchicus) and one of two jays (Garrulus glandarius) were found to harbor three types of Sarcocystis. Three of 15 (20%) infected pheasants had type I cystozoites (6-8 X 2 microns) in muscle homogenates, but sarcocysts were not seen whereas the other 12 infected pheasants had type II cystozoites (16 X 2-3 microns) and sarcocysts (90 X 600 microns) in their muscles. The one infected jay had type III cystozoites (8-10.5 X 2.5-3 microns) and sarcocysts (35-40 X greater than 770 microns) in its muscles.     

Two-dimensional gel analysis of zymogen-activating factors in the small intestine of the mouse

Zymogen-activating factors in the mouse were investigated by two-dimensional electrophoresis. Mouse pancreatic zymogens--trypsinogen-I group (Try G-I group), trypsinogen-II (Try G-II), and chymotrypsinogen (Chy G)--were purified using DEAE-cellulose column chromatography. Analysis by two-dimensional electrophoresis, using the purified zymogens as substrates, revealed enterokinase isozymes and chymotrypsinogen-activating factors in both the intestinal extract and luminal fluid. Mouse enterokinase was separated into at least two bands in the first-dimensional gel, each able to activate both trypsinogens Try G-I group and Try G-II. Chymotrypsinogen-activating factors were separated into several bands in the first-dimensional gel. Some activating factors showed mobilities similar to those of mouse enterokinase isozymes. Moreover, other activating factors that can activate chymotrypsinogen were present only in the more anodal area of the first-dimensional gel. These findings indicate that at least two enterokinases and several chymotrypsinogen-activating factors play an important role in the process of activating digestive enzymes.     

The fibrin-binding site of human plasminogen. Arginines 32 and 34 are essential for fibrin affinity of the kringle 1 domain

Kringle 1 (Tyr 79/Leu 80-His 167 and Tyr 79/Leu 80-Tyr 173), a chymotryptic fragment of human plasminogen that has high affinity for fibrin and omega-aminocarboxylic acids, has been subjected to modification with 1,2-cyclohexanedione to identify arginine residues essential for ligand binding. Reaction of 1,2-cyclohexanedione with kringle 1 was found to rapidly abolish the fibrin-Sepharose affinity of the fragment, whereas the affinity for lysine-Sepharose was lost at a significantly slower rate. Successive affinity chromatography of modified kringle 1 on fibrin- and lysine-Sepharose was used to separate kringle 1 that lost affinity for fibrin-, but retained affinity for lysine-Sepharose from kringle 1 that lost affinity for both affinants. The modified proteins were subjected to structural studies in order to locate the labeled arginine residues in kringle 1. These studies have revealed that modification of Arg 34 leads to the loss of both the fibrin- and lysine-Sepharose affinities of kringle 1, whereas reaction of Arg 32 abolishes fibrin affinity but leaves lysine-Sepharose affinity unaltered. The results suggest that Arg 32 and Arg 34 are both involved in fibrin binding and that Arg 34 is also involved in binding omega-aminocarboxylic acids. Previous NMR studies on kringles have indeed shown that the segment containing residue 34 is in the proximity of and interacts with the omega-aminocarboxylic acid-binding site. This interaction may explain the influence of omega-aminocarboxylic acids on fibrin binding by kringle 1.     

Residual hemolytic and proteolytic activity expressed by Bb after decay-dissociation of C3b,Bb

Bb (Mr = 63,000) is the catalytic site-bearing subunit of the C3 convertase of the alternative complement pathway, C3b,Bb, which is dissociated from the complex upon decay of the enzyme. Because purified Bb induced certain leukocyte activities, we examined whether it expresses residual hemolytic or proteolytic activity. Hemolytic activity of Bb was tested by using Factor B- or Factor D-depleted normal human serum and rabbit or sheep erythrocytes. Proteolytic activity of Bb was assessed by using purified C3 or C5 as substrates and SDS-PAGE to detect protein cleavage. Bb expressed metal-dependent hemolytic activity that was approximately 100-fold lower than that of Factor B. This activity could be inhibited by Factor H and enhanced by properdin. Low but statistically significant binding of 125I-labeled Bb to C3b on erythrocytes was demonstrated. Monoclonal antibodies that bind to Bb but not to intact Factor B inhibited the Bb hemolytic activity. Purified Bb cleaved C3 to C3a and C3b, as evidenced by the appearance of the alpha'-chain of C3b. It also cleaved C5 to C5a and C5b when cobra venom factor was present in the reaction mixture. Metal ions were required for expression of proteolytic activity, and Ni supported the activity better than Mg. These results indicate that decayed Bb has residual C3 and C5 cleaving activity and hemolytic activity, expression of which appears to require its association with C3b, C3(H2O), or cobra venom factor. These observations may aid in explaining the mechanism of action of Bb on leukocytes.     

Lipid/myelin basic protein multilayers. A model for the cytoplasmic space in central nervous system myelin

A multilayered complex forms when a solution of myelin basic protein is added to single-bilayer vesicles formed by sonicating myelin lipids. Vesicles and multilayers have been studied by electron microscopy, biochemical analysis, and X-ray diffraction. Freeze-fracture electron microscopy shows well-separated vesicles before myelin basic protein is added, but afterward there are aggregated, possibly multilayered, vesicles and extensive planar multilayers. The vesicles aggregate and fuse within seconds after the protein is added, and the multilayers form within minutes. No intra-bilayer particles are seen, with or without the protein. Some myelin basic protein, but no lipid, remains in the supernatant after the protein is added and the complex sedimented for X-ray diffraction. A rather variable proportion of the protein is bound. X-ray diffraction patterns show that the vesicles are stable in the absence of myelin basic protein, even under high g-forces. After the protein is added, however, lipid/myelin basic protein multilayers predominate over single-bilayer vesicles. The protein is in every space between lipid bilayers. Thus the vesicles are torn open by strong interaction with myelin basic protein. The inter-bilayer spaces in the multilayers are comparable to the cytoplasmic spaces in central nervous system myelins . The diffraction indicates the same lipid bilayer thickness in vesicles and multilayers, to within 1 A. By comparing electron-density profiles of vesicles and multilayers, most of the myelin basic protein is located in the inter-bilayer space while up to one-third may be inserted between lipid headgroups. When cytochrome c is added in place of myelin basic protein, multilayers also form. In this case the protein is located entirely outside the unchanged bilayer. Comparison of the various profiles emphasizes the close and extensive apposition of myelin basic protein to the lipid bilayer. Numerous bonds may form between myelin basic protein and lipids. Cholesterol may enhance binding by opening gaps between diacyl-lipid headgroups.