Arginase,an s-phase enzyme in a human cell line

Suspension cultures of ‘Chang liver’ cells were synchronized by preincubation in a glutamine-deficient medium or by thymidine blockade. Specific arginase activity varied in the synchronized cultures, being high when the number of S-phase cells was maximal. A relationship between high arginase activity and a high percentage of (S+G₂) cells was also found when unsynchronized cells were separated by velocity sedimentation. The increase in arginase activity near the G₁/S border was totally inhibited in the presence of cycloheximide. The rate of decrease in activity after addition of the drug indicated that the variations in the rate of synthesis of the enzyme, while the rate of degradation was more or less constant, corresponding to 4–6% per h. The role of arginase in cells lacking a urea cycle and the regulation of arginase activity in ‘Chang liver’ cells is discussed.     

Endorphin mediated increase in pain threshold induced by long-lasting exercise in rats

Rats were trained to run spontaneously, without stress, in running wheels. The running activity increased gradually and could reach a plateau of 7 km/night after 3–4 weeks. During the first hour of running in the dark phase the squeak threshold increased significantly and remained high in the morning. The degree of increased threshold was correlated to the amount of running activity. The squeak threshold declined during the following 6 hours of inactivity. A rapid decrease in threshold occurred after naloxone (1–2 mg/kg i.p.). It is suggested that long-lasting muscle exercise (e.g. jogging), acupuncture, and low frequency electrical stimulation of afferent nerve fibres produce discharges in muscle afferents which influence central endorphin mechanics giving analgetic effects.     

Compartmentalisation of nitrogenase in a non-heterocystous cyanobacterium: Trichodesmium contortum

Abstract In Trichodesmium contortum , nitrogenase was detected in only a limited number (about 10%) of microscopically distinguishable, consecutively arranged cells in central regions of the trichomes. Cells with nitrogenase also contained the photosystem II associated pigment phycoerythrin. These cells were not distinguishable from other cells on a structural basis, but were clearly visible at low magnification microscopy as all in the zone were more compact and shorter than those on either side. The compartmentalisation of nitrogenase into a chain of cells and in a possibly photosynthetic environment represents a previously undescribed phenomenon. The nitrogenase containing cells apparently perform the O₂ protective function of heterocysts yet are different in several aspects.     

Steroidal glycoalkaloids in cell and shoot teratoma cultures ofSolanum dulcamara

Bittersweet (Solanum dulcamara L., Solanaceae) is of interest as a source of steroidal alkaloids for the commercial production of hormones. Since glycoalkaloid production is positively correlated to differentiation, tumor and teratoma cultures of the soladulcidine chemotype were established by transformation withAgrobacterium tumefaciens. A newly developed HPLC-system, which allowed separation and sensitive quantitation of the glycoalkaloids soladulcidine-tetraoside, solamargine and solasonine, was used to analyse glycoalkaloid profiles in plants and cultures. Tumors and teratoma were charcterized by a shift in their alkaloid pattern from soladulcidine tetraoside to the solasodine glycosides solamargine and solasonine. Shoot teratoma showed a total glycoalkaloid content of 1% dw, which is about fivefold higher than in the source plant. A regenerated plant retained the altered alkaloid spectrum; the levels, however, equalled those of the source plant. From the alteration of alkaloid pattern in the transformed cultures suggestions can be made concerning the biosynthetic pathway. Completion of the biosynthesis of the aglycone is likely to be complete before glycosylation occurs.     

Estimation of desmosponge (Porifera,Demospongiae) larval settlement rates from short-term recruitment rates: Preliminary experiments

During a study of the spatial and temporal patterns of desmosponge (Porifera, Demospongiae) recruitment on rocky and coral reef habitats of Santa Marta, Colombian Caribbean Sea, preliminary attempts were made to estimate actual settlement rates from short-term (1 to a few days) recruitment censuses. Short-term recruitment rates on black, acrylic plastic plates attached to open, non-cryptic substratum by anchor screws were low and variable (0–5 recruits/plate in 1–2 days, sets of n=5–10 plates), but reflected the depth and seasonal trends found using mid-term (1 to a few months) censusing intervals. Moreover, mortality of recruits during 1–2 day intervals was low (0–12%). Thus, short-term censusing intervals can be used to estimate actual settlement rates. To be able to make statistical comparisons, however, it is necessary to increase the number of recruits per census by pooling data of n plates per set, and to have more than one set per site or treatment.     

Hemoglobins,XLVIIII

Summary Hagfish hemoglobin has three main components, one of which is Hb III. It is monomeric and consists of 148 amino acid residues (M = 17 350). Its complete primary structure, previously published, is discussed here. The proximal amino acid (F8) of the heme linkage is histidine as always in the hemoglobins, but the regularly expected distal histidine E7 is substituted by glutamine. This substitution, leading to a new kind of heme linkage, has hitherto only been demonstrated in opossum hemoglobin. It is suggested that E7, Gln, is directed out of the heme pocket, and that the adjacent Ell, Ile, is directed toward the inside of the pocket, giving the distal heme contact instead of histidine.Myxine Hb III has an additional tail of 9 amino acid residues at its N-terminal end, as has the hemoglobin ofLampetra fluviatilis. The genetic codes ofMyxine andLampetra hemoglobins show 117 differences, in spite of many morphological resemblances between hagfish and lamprey. Their primary hemoglobin structures show differences substantial enough to bo compatible with the divergence of the two families some 400–500 million years ago.     

MHC restriction of male-antigen-specific T helper cells collaborating in antibody responses

By priming female C57BL/6 mice with syngeneic male spleen cells and enriching inguinal and paraaortic lymph node cells in long-term culture (LTC) by repeated restimulations, H-Y-specific T helper cells can be produced. In response to male spleen cells carrying I-Ab antigens these cells activate antigenexpressing B cells to secrete polyclonal antibody. Before the end of the second week in LTC it was impossible to detect any helper activity. Induction of plaque-forming cells (PFC) also requires simultaneous recognition of antigen and I-A-encoded determinants in the stimulator-responder spleen-cell population. The testing of spleen cells fromH-2 recombinant strains as stimulator-responders to anti-H-Y helper T cells of C57BL/6 origin also revealed that other genes, telomeric toI-A, control the magnitude of both specific T-cell proliferation and helper-dependent B-cell activation.     

Differentiation-related changes of cytokeratin expression in cultured keratinocytes and in fetal, newborn, and adult epidermis

Cytokeratin expression in differentiating cultured foreskin keratinocytes was studied using chain-specific anti-cytokeratin monoclonal antibodies directed against cytokeratins 4, 8, 10, 13, 18, and 19, respectively. Keratinocytes were cultured at low Ca2+ concentration (0.06 mM) to repress differentiation. At confluency, the cells were switched to high Ca2+ concentration (1.6 mM) to induce differentiation. Cells were harvested 0, 3, 8, 16, 24, 48, and 72 h after the switch. Keratinocytes cultured throughout at high Ca2+ concentration were also harvested. Immunoblots of cytokeratin preparations isolated from these cultures showed that cytokeratins 4, 13, and 19 were not present in nondifferentiating keratinocytes but could be detected from about 16 h after the Ca2+ switch. Immunohistochemical studies were performed on frozen sections of cell sheets incubated with anti-cytokeratin and anti-vimentin. Expression of cytokeratins 4, 13, and 19 was seen in superficial cells. Cytokeratin 10 was locally present in suprabasal and superficial cells. Vimentin was present in 40-70% of the basal cells and in only a few differentiating keratinocytes. Expression of cytokeratins 8 and 18 could not be detected. The same antibodies were also used to stain sections from fetal (15, 20, and 29 weeks), newborn (40 weeks), and mature (5 and 75 years) epidermis. In the 15-week-old epidermis, basal cells were positive for cytokeratins 8 and 19 and locally for cytokeratin 4; intermediate cells expressed cytokeratins 4, 10, 13, and 19; and the periderm contained cytokeratins 4, 8, 13, 18, and 19. In the 20-week-old epidermis, cytokeratin 4 had disappeared from the basal cell layer and cytokeratin 19 was present only locally; in the intermediate cell layer, cytokeratins 4 and 19 had disappeared; and in the periderm, the expression of the cytokeratins studied was the same as that in the 15-week-old epidermis. The basal cells of the 29-week-old fetal epidermis, the newborn epidermis, and the mature epidermis are negative with all antibodies tested, except for some scattered cells in the fetal and newborn skin, presumably Merkel cells, that were positive for cytokeratins 8, 18, and 19. Suprabasal cells in all specimens were positive only for cytokeratin 10. With respect to the cytokeratins studied, our results show that cultured differentiating keratinocytes resemble the suprabasal cells of early fetal epidermis. Basal cells of cultured keratinocytes resemble the basal cells of late fetal, newborn, and adult epidermis and therefore support previous observations.     

Practically relevant concentrations of deoxynivalenol in diets for growing-finishing pigs offered as mash or pellets

A complete 2 x 3 two factorial design was applied to investigate the effects of Fusarium-infected wheat (2.5 mg DON/kg, 0, 25 and 50% of the diets), feed processing (mash and pellets) and the interactions thereof on fattening pigs (96, n= 16/group). Feed-to-gain ratio was significantly increased by contaminated wheat (2.65; 2.62 and 2.73 kg/kg for diets containing 0, 25 and 50% Fusarium-infected wheat, respectively) while digestibility of nutrients and metabolizable energy were not affected by the wheat batch. The feed processing also resulted in significant differences in feed-to-gain ratio but was accompanied by significant effects on the digestibility of organic matter and crude fat and on the metabolizable energy. Clinical chemical parameters were not significantly altered by the inclusion of the infected wheat. The lymphocyte proliferation capacity was not significantly affected by any of the experimental factors. A contribution of the feed processing to the variation of the deoxynivalenol (DON) effect may not be deduced from the present results.