A disulfide-bonded short chain collagen synthesized by degenerative and calcifying zones of bovine growth plate cartilage

Studies of collagen synthesis by specific sections of individual fetal bovine costochondral junction growth plates were conducted and histologically related to the zones from which the sections were derived. Sections were metabolically labeled in organ culture to examine the synthesis of collagen and its precursors. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that Type II collagen was the major species synthesized in all tissue sections; 1 alpha, 2 alpha, 3 alpha collagen chains were synthesized in all growth plate sections and to a small extent in the fetal structural cartilage. A short chain collagen was synthesized predominantly in the zones of degeneration and provisional calcification and accounted for 8-12% of the radioactivity in this section. This short chain collagen has 63-kDa subunits which are converted to 46-kDa species by limited proteolysis with pepsin. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated that both the pepsin- and non-pepsin-treated forms of short chain collagen are disulfide-bonded. Digestion with bacterial collagenase showed that the 46-kDa and a major portion of the 63-kDa forms are collagenous. Pulse-chase studies in organ culture did not demonstrate an obvious precursor to the 63-kDa form, and there was no conversion to the 46-kDa after 20 h. Synthesis of short chain collagen appears to be specific to the process of endochondral ossification in the growth plate; its appearance may be critical to this transition process.     

Seasonal Fluctuations of Globodera tabacum solanacearum as Estimated by Two Soil Extraction Techniques

Two soil extraction methods were compared to determine their efficiency in recovering cysts and juveniles of a tobacco cyst nematode, Globodera tabacum solanacearum. The methods were equally efficient when extracting nematodes from soil samples seeded in the laboratory; however, there was a significant extraction method × month interaction when the methods were used to estimate field soil populations over 2 years. The centrifugal sugar flotation method recovered greater numbers of cysts when densities were near 400 cysts/100 cm³ soil and greater numbers of juveniles in all samples. The sugar flotation method recovered greater numbers of cysts during months when densities were less than 400 cysts/100 cm³ soil. Numbers of cysts and juveniles were lowest in June and July following land tillage in May. A soil freeze in January 1982 may have been responsible for unusually high numbers of recovered cysts in February and March 1982, a pattern that did not occur in 1983.     

Endocrine and metabolic responses of the collared peccary (Tayassu tajacu) to immobilization with ketamine hydrochloride

Serial physiological responses were examined for 150 min from captive collared peccaries during immobilization with ketamine hydrochloride. Rectal temperatures decreased significantly (P less than 0.01) during anesthesia. Serum concentrations of total proteins, albumin, cholesterol, alanine aminotransferase, and calcium declined significantly (P less than 0.05) during the first 45 min post-immobilization before stabilizing. Concentrations of lactate dehydrogenase and alkaline phosphatase in sera showed similar but nonsignificant (P greater than 0.05) trends. Inorganic phosphorus and aspartate aminotransferase concentrations increased significantly (P less than 0.05) throughout the trial. Concentrations of serum glucose and glucocorticoid during the immobilization period were highly variable between individuals. Serum electrolytes, urea nitrogen, creatinine, gammaglutamyl transferase and progesterone were not significantly (P greater than 0.05) affected by immobilization. Elevations in serum testosterone were noted. Results indicated appropriate sampling times relative to immobilization for assay of particular serum biochemicals and steroid hormones during investigations of the physiology of the collared peccary.     

Observations on 6-mercaptopurine activity in Saccharomyces cerevisiae

Abstract Hydrogen fluoride treatment of [¹⁴C-glycerol]lipoteichoic acid synthesized by growing Streptococcus faecium ATCC9790 in the presence of 1,3[¹⁴C]glycerol produced five radioactive, water-soluble products which were identified by chromatographic and analytical techniques to be tetraglucosyl glycerol, triglucosyl glycerol, diglucosyl glycerol, monoglucosyl glycerol and unsubstituted glycerol. The percent composition of each varied modestly from culture to culture and ranged between 7 and 8% for the tetra-, 20.5 and 31.2% for the tri-, 11.3 to 23.5% for the di-, 20.9 to 26.8% for the mono-, and 23.1 to 34.8% for the unsubstituted glycerol. The same glucosylated glycerol compounds could be obtained in an in vitro reaction in which a 30 000 × g particulate enzyme catalyzed the incorporation of [³H]glucose from UDP [³H]glucose into lipoteichoic acid.     

Sulphate reduction in oxic and sub-oxic North-East Atlantic sediments

Abstract Oxic and sub-oxic N.-E. Atlantic sediments were examined for sulphate-reducing activity. Oxygen and/or nitrate reduction are probably the dominant mineralisation processes in the abyssal plain sediment studied. A low rate of sulphate reduction (0.1 nmol SO²⁻₄/ml/day) was recorded in the surface 5 cm of the continental slope sediment, together with the presence of a range of sulphate-reducing bacteria (SRB). A higher activity of sulphate reduction (2.2 nmol SO²⁻₄/ml/day) occurred in the continental shelf sediment which led to a small decrease in pore water sulphate and an increase in titration alkalinity. This sediment contained approx. 10²–10³ acetate, lactate and propionate oxidising SRB/ml. No low- M _r organic acids were detected in these sediments. However, amendment with 75 μM acetate stimulated sulphate-reducing activity in the shelf sediment.     

Domes,arches and urchins: The skeletal architecture of echinoids (Echinodermata)

Summary A combination of simple membrane theory and statical analysis has been used to determine how stresses are carried in echinoid skeletons. Sutures oriented circumferentially are subject principally to compression. Those forming radial zig-zags are subject to compression near the apex and tension near the ambitus. Radial and circumferential sutures in Eucidaris are equally bound with collagen fibers but in Diadema, Tripneustes, Psammechinus, Arbacia and other regular echinoids, most radial sutures are more heavily bound, and thus stronger in tension. Psammechinus, Tripneustes and several other echinoids have radial sutures thickened by ribs which increase the area of interlocking trabeculae. Ribs also increase flexural stiffness and carry a greater proportion of the stress. Further, ribs effectively draw stress from weaker areas pierced by podial pores, and increase the total load which can be sustained.Allometry indicates that regular echinoids become relatively higher at the apex as size increases, thus reducing ambital stresses. Some spatangoids with very high domes (eg Agassizia) maintain isometry, but others (eg Meoma) become flatter with size. Both holectypoids (Echinoneus) and cassiduloids (Apatopygus) maintain a constant height to diameter relationship. Flattening, and consequently ambital tensile stress, is greatest in the clypeasteroids. In this group the formation of internal buttresses which preferentially carry stress, reaches maximum development. A notable exception, however, is the high domed Clypeaster rosaceus.In this analysis it was assumed that local buckling or bending does not occur. The test of some echinoids (e.g. Diadematoida) have relatively wide sutures swathed in collagen, which allows local deformation. Others (e.g. Arbacia) have rigid sutures with reduced collagen. In Psammechinus and other members of the Order Echinoida, in addition to rib formation, inner and outer surface trabeculae are thickened so that the individual plates are stiffened. Some spatangoids (Meoma, Paleopneustes) have extensive sutural collagen, but the cassiduloid Apatopygus has collagen confined to junctions of sutures, and elsewhere the joints are strengthened and stiffened by fusion of trabeculae. Fusion of surface trabeculae is almost complete in the holectypoid, Echinoneus, and the sutures are obscured.     

Heterogeneity of alkaline small subunits of ribulose 1,5 bisphosphate carboxylase-oxygenase from Medicago sativa and M. falcata

The isolated leaf proteins of lucerne (Medicago sativa L. and M. falcata L.) were fractionated by Sepharose 6B column chromatography. Analysis of fractionated proteins indicated that the 2nd peak component was almost entirely ribulose 1,5 bisphosphate carboxylase-oxygenase (Rubisco) which represented 57% of the total recovered protein.Rubisco yielded one large subunit (LSU) and one small subunit (SSU) polypeptide after SDS gel electrophoresis.Isoelectric focusing of the SSU of Rubisco from genotypes of M. sativa cv. Hunter River (HR), Hairy Peruvian (HP) and of M. falcata (MF) showed two SSU components for HR and HP, and three components for MF. Most components of genotypes were located in the alkaline region of the gel. While the pIs of the SSU components of HR and HP were identical they differed from those of the SSU of MF thus demonstrating heterogeneity for SSU in Medicago.It is suggested that the alkaline nature of SSU may have some adaptive physiological significance.Abbreviations Rubisco ribulose bisphosphate 1,5-carboxylase-oxygenase - LSU large subunit - SSU small subunit - HR Hunter River - HP Hairy Peruvian - MF Medicago falcata - SDS Sodium dodecyl sulphate - TCA trichloracetic acid     

Effects of phorbol myristate acetate,phorbol dibutyrate,ethanol, dimethylsulfoxide,phenol, and seven metabolities of phenol on metabolic cooperation between chinese hamster v79 lung fibroblasts

The effect of phorbol myristate acetate, phorbol dibutyrate, ethanol, dimethylsulfoxide, phenol, and seven metabolites of phenol on metabolic cooperation were assessed as a function of mutant cell recovery from populations of cocultivated hypoxanthine-guanine phosphoribosyl transferase-deficient mutant (HGPRT–) and wild-type (HGPRT+) Chinese hamster V79 lung fibroblasts. Phorbol myristate acetate and phorbol diputyrate, two established tumor promoters, were potent inhibitors of metabolic cooperation. Ethanol and dimethylsulfoxide, solvents commonly used to prepare chemicals for testing, weakly inhibited metabolic cooperation. Phenol and phenylglucuronide had no effect on metabolic cooperation. Four oxidative metabolites (1,4-benzoquinone, catechol, hydroxyquinol and quinol) inhibited metabolic cooperation. Phenylsulfate weakly inhibited metabolic cooperation. Conversely, 2-methoxyphenol, a methylated derivative of catechol, appeared to enhance metabolic cooperation. These results generallyAbbreviations CAS Chemical Abstracts Service - DMSO dimethylsulfoxide - ETOH ethanol - HGPRT hypoxanthine-guanine phosphoribosyl transferase - HGPRT+ HGPRT-competent - HGPRT– HGPRT-te]deficient - MC metabolic cooperation - MC+ metabolic cooperation-competent - MC– metabolic cooperation-deficient - MEM minimum essential medium - PDBu phorbol dibutyrate - PMA phorbol myristate acetate - 6TG 6-thioguanine - 6TG^r 6-thioguanine-resistant - 6TG^s 6-thioguanine-sensitive - V79/MC assay Chinese hamster V79 lung fibroblast assay for metabolic cooperation     

Leakage from Seedling Roots, Inoculated with Phytophthora cinnamomi

A sequential study of electrolyte leakage from roots inoculated with Phytophthora cinnamomi demonstrated changes in leakage in response to infection. These changes may be characterised in terms of susceptibility and resistance. Field resistant species showed two types of response (i) rapid leakage 2—4 h after inoculation, such as in Eucalyptus calophylla, E. maculata and Gahnia radula, (ii) no significant increase in leakage with infection, for example in cereals and in juncus bufonius. Susceptible species, such as Xanthorrhoea australis, E. sieberi and E. marginata showed slow but continually increasing leakage after inoculation, and usually lost significantly more electrolyte than field resistant species. Both electrolyte leakage and susceptibility of two Eucalyptus species varied with root temperature. Roots of both the susceptible and field resistant Eucalyptus species grown at 14°C showed similar leakage patterns to those grown at 24°C although no lesions formed at 14°C. Both amount of leakage and lesion length increased at 28°C. E. marginata (susceptible) lost significantly greater quantities of electrolyte than E. calophylla at each temperature tested. Leakage from artificially wounded roots did not vary with either temperature or with host susceptibility. No significant changes in conductivity were recorded with saprophytic colonization of roots, or with incubation in either low molecular weight culture filtrate or β-glucan solutions. Vacuum infiltration with the culture filtrate or the β-glucan slightly increased initial electrolyte loss in comparison with that of the controls. The increased leakage from infected roots may be toxin mediated or due to enzymie degradation of host plasma membranes, and occurred within the region of the limited lesion in the case of field resistant species, compared with the larger zone of extending necrosis characteristic of susceptible species.