Deoxyribonucleic acid relatedness inYersinia enterocolitica andYersinia enterocolitica-like organisms

Yersinia enterocolitica andY. enterocolitica-like strains were characterized by DNA relatedness. These strains formed four distinct DNA relatedness groups: (i) the 5 classical biotypes ofY. enterocolitica sensu stricto as designated by Wauters; (ii) strains that are rhamnose positive and also positive in tests for melibiose, α-methyl-d-glucoside, raffinose, and Simmons' citrate; (iii) strains that are rhamnose positive but negative in tests for melibiose, α-methyl-d-glucoside, and raffinose; (iv) sucrose-negative, Voges-Proskauer-negative, trehalose-positive strains.     

Prediction of effluent pollution level in activated sludge systems II. The effect of recirculation and recirculation ratio

The paper describes substrate removal and biomass production in activated sludge wastewater treatment systems with special attention to the effect of recirculation. Applicability of the models presented is verified experimentally.     

Seasonal fluctuation of zooplankton populations in lower Delaware Bay

Quantitative zooplankton samples were obtained monthly or bi-monthly 15 times from June 1974 to May 1975 at three stations in lower Delaware Bay. Two 12-hour cruises were also conducted at one of the stations.Arthropods dominated the samples in terms of number of species and number of individuals. The number of zooplankton from surface samples ranged from 58/m³ in August to 21,092/ m³ in June, while bottom samples varied from 259/m³ in August to 30,395/ m³ in October. In general, larger concentrations of individuals were found in bottom samples.Only on three occasions did meroplankton exceed the holoplankton, and these occurred at the shallow water stations. Meroplankton comprised a larger percentage of the bottom samples than surface samples. Zoeae of Neopanope sayi and Uca sp. contributed mainly to the large proportion of meroplankton in July 1974, veligers of Mytilus edulis in January 1975, and nauplii of Balanus sp. in May 1975.Copepods were the largest component of the population throughout most of the year. At all stations and depths, Arctica tonsa dominated most of the summer samples. In the spring of 1975, A. tonsa was replaced by Centropages hamatus, Temora longicornis, and Pseudocalanus minutus.During the 12-hour cruises there were higher numbers of individuals in the bottom waters in the day with migration to surface waters in the afternoon and evening. Based on cluster analysis, five time-related assemblages were discerned: June, July–August, September–November, December, January–May. Comparison of Delaware Bay zooplankton with other estuarine systems indicated that the densities obtained locally were most similar to those reported in the York River, Virginia.     

Quantitation of submandibular proteins resolved from normal individuals and children with cystic fibrosis

Submandibular secretions collected from children with cystic fibrosis (CF) showed increased protein concentration (milligrams/milliliter) and increased amylase specific activity (units/milligram of protein) relative to normal secretions. These differences between normal (N) and CF secretions were as follows: protein, 1.25 ± 0.51 (N), 1.75 ± 0.35 (CF) (P < 0.02); and amylase, 58 ± 18 (N), 80 ± 19 (CF) (P < 0.001). To determine the basis for elevated protein in CF saliva, several major proteins resolved by polyacrylamide disc gel electrophoresis were quantitated by densitometry. These included four phosphoproteins (PP), serum albumin, an acid phosphatase-containing fraction, amylase, and an unidentified protein referred to as PI-7.1. Together, these proteins comprise greater than 75% of the total protein in the secretion. Differences in individual protein concentrations (milligrams/milliliter) resolved from normal and CF secretions, respectively, were as follows: PP₂, 0.02 ± 0.01, 0.03 ± 0.02 (NS, not significant); PP₃, 0.06 ± 0.04, 0.05 ± 0.03 (NS); acid phosphatase fraction, 0.06 ± 0.04, 0.12 ± 0.07 (P < 0.05); amylase, 0.09 ± 0.04, 0.27 ± 0.16 (P < 0.01); and pI-7.1, 0.04 ± 0.02, 0.13 ± 0.08 (P < 0.02). Amylase, the most significant contributor to the elevated protein, comprised 26% of the total protein of normal secretions and 38% of the total protein of CF secretions. Thus, our results do not support the concept of a generalized increase in all organic components in CF submandibular secretions but, rather, increases in specific proteins, namely amylase, component pI-7.1, and an acid phosphatase-containing fraction.     

Partial characterization of prostaglandin synthetase in the reproductive tract of the male house cricket, acheta domesticus

An active prostaglandin (PG) synthetase was found in the 12100 g pellet of reproductive tract homogenates of the male house cricket, $\text{Acheta domesticus}$. Comparatively, the 12100 g supernatant and the microsomal fractions were inactive. The PG synthetase in the pellet fraction was characterized in terms of cofactor, temperature, pH, and incubation time requirements. Indomethacin, a known inhibitor of mammalian PG synthetase, was not inhibitory to the cricket synthetase. The procedure and findings are relevant to PG synthetase studies of any organism or tissue.     

Sunshine and the geographical distribution of the alleles of the Gc system of plasma proteins

Summary Following the discovery by Daiger et al. (1975) that the Gc proteins of human plasma act as the carriers of vitamin D, the authors have plotted on a world map all available data on the frequency of the allele Gc ², and compared the distribution with that of sunlight. With some exceptions high frequencies of Gc ² correspond to low levels of sunlight and vice versa. Similar comparisons within Ireland show no such relation. The results are discussed in relation to natural selection and the incidence of rickets, due to vitamin D deficiency.     

Symmetry, orientation and rotational mobility in the a3 heme of cytochrome c oxidase in the inner membrane of mitochondria

The photoinduced linear dichroism of absorption changes resulting from photolysis of the complex between heme a₃ of the cytochrome oxidase and CO is studied. The experiments started from isotropic solutions or suspensions of the enzyme both in its isolated form and in mitochondria. The anisotropy responsible for the linear dichroism was induced by excitation with a flash of linearly polarized light. The dichroic ratios observed with various systems; polymerized enzyme in solution, enzyme in mitochondria and in submitochondrial particles (at 20 °C as well as at liquid N₂-temperature) all approached a value of 4/3 which characterizes a chromophore which is circularly degenerate. Therefrom we conclude that the interaction of heme a₃ with its microenvironment within the protein does not break its four-fold symmetry.

The experiments with mitochondria and submitochondrial particles suspended in aqueous buffer revealed similarly high dichroic ratios without any dichroic relaxation other than a rather slow one which could be attributed to the rotation of the whole organelle in the suspending medium. Therefrom we conclude that the cytochrome oxidase either is totally immobilized in the membrane, or that it carries out only limited rotational diffusion around a single axis coinciding with the symmetry axis of heme a₃. In the light of independent evidence for a transmembrane arrangement of the oxidase and for the general fluidity of the inner mitochondrial membrane we consider anisotropic mobility of the cytochrome oxidase around an axis normal to the plane of the membrane as the most likely interpretation. Then our experimental results imply that the plane of heme a₃ is coplanar to the membrane.     

Active human erythropoietin expressed in insect cells using a baculovirus vector: A role for N-linked oligosaccharide

Biologically active recombinant human erythropoietin has been expressed at high levels in an insect cell background. Expression involved the preparation of a human erythropoietin cDNA, the transfer of this cDNA to the Autographa californica nuclear polyhedrosis virus (AcNPV) genome under the polyhedrin gene promoter, and the subsequent infection of Spodoptera frugiperda cells with recombinant AcNPV. Erythropoietin cDNA was prepared through the expression of the human erythropoietin gene in COS cells using pSV2 and the construction of a COS cell cDNA library in bacteriophage Lambda GT10. Prior to transfer to the AcNPV genome, erythropoietin cDNA isolated from this library was modified at the 3′-terminus in order to replace genomic erythropoietin for SV40 cDNA derived from pSV2. Transfer of this cDNA to AcNPV and the infection of S. frugiperda cells with cloned recombinant virus led to the secretion of erythropoietin: based on bioassay, rates of hormone secretion (over 40 U/ml per h) were 50-fold greater than observed for COS cells. The purified recombinant product possessed full biological activity (at least 200000 U/mg), but was of lower M_r (23000) than human erythropoietin produced in COS cells (30000) or purified from urine (30000 to 38000). This difference was attributed to the glycosylation of erythropoietin in S. frugiperda cells with oligosaccharides of only limited size. Further removal of N-linked oligosac-charides from this M_r 23000 hormone using N-Glycanase yielded an apo-erythropoietin (M_r 18000) which possessed substantially reduced biological activity. These results indicate that glycosylation, but not the normal processing of oligosaccharides to complex types, is required for the full hormonal activity of human erythropoietin during red cell development.     

Particulate matter exposure induces persistent lung inflammation and endothelial dysfunction

Epidemiologic and animal studies have shown that exposure to particulate matter air pollution (PM) is a risk factor for the development of atherosclerosis. Whether PM-induced lung and systemic inflammation is involved in this process is not clear. We hypothesized that PM exposure causes lung and systemic inflammation, which in turn leads to vascular endothelial dysfunction, a key step in the initiation and progression of atherosclerosis. New Zealand White rabbits were exposed for 5 days (acute, total dose 8 mg) and 4 wk (chronic, total dose 16 mg) to either PM smaller than 10 mum (PM(10)) or saline intratracheally. Lung inflammation was quantified by morphometry; systemic inflammation was assessed by white blood cell and platelet counts and serum interleukin (IL)-6, nitric oxide, and endothelin levels. Endothelial dysfunction was assessed by vascular response to acetylcholine (ACh) and sodium nitroprusside (SNP). PM(10) exposure increased lung macrophages (P<0.02), macrophages containing particles (P<0.001), and activated macrophages (P<0.006). PM(10) increased serum IL-6 levels in the first 2 wk of exposure (P<0.05) but not in weeks 3 or 4. PM(10) exposure reduced ACh-related relaxation of the carotid artery with both acute and chronic exposure, with no effect on SNP-induced vasodilatation. Serum IL-6 levels correlated with macrophages containing particles (P=0.043) and ACh-induced vasodilatation (P=0.014 at week 1, P=0.021 at week 2). Exposure to PM(10) caused lung and systemic inflammation that were both associated with vascular endothelial dysfunction. This suggests that PM-induced lung and systemic inflammatory responses contribute to the adverse vascular events associated with exposure to air pollution.