Production,cross reactivity,and epitope analysis of monoclonal antibodies against rat kidney gamma glutamyltransferase

Eighteen IgGl monoclonal antibodies (blabs) have been produced against gamma-glutamyl transferase (GGT) from rat kidney. They were specific to the light subunit of the enzyme with affinity constants ranging from 0.3 to 7.5 10⁸ M^–1, while they did not react with GGT from other sources i.e. human and pig kidney, rat and guinea pig liver, suggesting species and organ specificity. Two of the blabs (N° 11 and 21) lost their immunoreactivities towards rat kidney GGT in the presence of N-acetyl-neuraminic acid, while immunoreactivities of the other blabs were unchanged. Furthermore, Mabs No 11 and 21 did not react with desialylated rat kidney GGT. These findings suggest that N-acetyl-neuraminic acid is involved in the epitopes recognized by these two Mabs.Abbreviations ELISA enzyme linked immunosorbent assay - GGT gamma-glutamyltransferase - Mab monoclonal antibody - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis     

Hepatic mitochondrial cytochrome P-450 system. Purification and characterization of two distinct forms of mitochondrial cytochrome P-450 from beta-naphthoflavone-induced rat liver

We have purified two distinct isoforms of mitochondrial cytochrome P-450 from beta-naphthoflavone (beta-NF)-induced rat liver to greater than 85% homogeneity and characterized their molecular and catalytic properties. One of these isoforms showing an apparent molecular mass of 52 kDa is termed P-450mt1 and the second isoform with 54-kDa molecular mass is termed P-450mt2. Cytochrome P-450mt2 comigrates with similarly induced microsomal P-450c (the major beta-NF-inducible form) on sodium dodecyl sulfate-polyacrylamide gels and cross-reacts with polyclonal antibody monospecific for cytochrome P-450c. Cytochrome P-450mt2, however, represents a distinct molecular species since it failed to react with a monoclonal antibody to P-450c and produced V8 protease fingerprints different from P-450c. Cytochrome P-450mt1, on the other hand, did not show any immunochemical homology with P-450c or P-450mt2 as well as partially purified P-450 from control mitochondria. Electrophoretic comparisons and Western blot analysis show that both P-450mt1 and P-450mt2 are induced forms not present in detectable levels in control liver mitochondria. A distinctive property of mitochondrial P-450mt1 and P-450mt2 was that their catalytic activities could be reconstituted with both NADPH-cytochrome P-450 reductase as well as mitochondrial specific ferredoxin and ferredoxin reductase electron transfer systems, while P-450c showed exclusive requirement for NADPH-cytochrome P-450 reductase. Cytochromes P-450mt1 and P-450mt2 were able to metabolize xenobiotics like benzo(a)pyrene and dimethyl benzanthracene at rates only one-tenth with cytochrome P-450c. Furthermore, P-450mt1, P-450mt2, as well as partially purified P-450 from control liver, but not P-450c, showed varying activities for 25- and 26-hydroxylation of cholesterol and 25-hydroxylation of vitamin D3. These results provide evidence for the presence of at least two distinct forms of beta-NF-inducible cytochrome P-450 in rat hepatic mitochondria.     

Type A behavior and job performance: some suggestive findings

This study examined Type A and Type B differences in job performance, psychosomatic complaints, and career progression among white-collar employees (N = 218) in a field setting. Significant differences were found between Type A's and Type B's in quality of performance, effort exerted at the job, and psychosomatic complaints. Employees' cultural background and sex moderated some of the relationships observed in the study. Implications of the findings for future research on the topic are discussed.     

Phosphorus depletion in the rhizosphere as influenced by soil moisture

To study the influence of soil moisture on phosphorus (P) depletion in the rhizosphere, maize (Zea mays cv. Trak) was pre-grown in vermiculite filled-PVC tubes for 9 days and then the plants with the tubes were transplanted into soil columns maintained at two soil moisture levels () of 0.14 and 0.20 cm³ cm^–3 for 10 days. The soil columns were separated at 1 cm depth by a nylon screen of 53 m inner mesh size, into 1 cm soil layer above and 3 cm soil column below screen. A root mat developed over the screen, but root hairs only could penetrate it. Regardless of the soil moisture level in the columns, and adequate and equal water and nutrients supply was maintained via wicks from an external nutrient solution to the plant roots in vermiculite. After 10 days, the soil columns were separated from the root mats, quickly frozen in liquid nitrogen and sliced into thin layers (0.2mm) using a refrigerated microtome to give soil samples at defined distances from the root mats for analyses. Lower soil moisture (=0.14) resulted in narrower and steeper depletion profile of 0.5 M NaHCO₃ extractable P (NaHCO₃-P_i) as compared to higher soil moisture (=0.20). Depletion of P in soil solution in the immediate vicinity of root mats did not differ much but the extension of the depletion zones was 0.10 cm at =0.14 and 0.20 cm at =0.20. The depletion up to 0.05cm with =0.14 and up to 0.07 cm with =0.20 was uniform, and may be attributed to the depletion in the root hair zone. Beyond the root hair zones, the theory of diffusion and mass flow was able to explain the observed differences in shape and extent of the P depletion profiles at the two soil moisture levels.     

The role of infectious agents in sudden infant death syndrome

Abstract Epidemiological factors associated with susceptibility to respiratory infections are similar to those associated with Sudden Infant Death Syndrome. Here we review the evidence that respiratory pathogens might be involved in some cases of Sudden Infact Death Syndrome in the context of factors identified in epidemiological studies of cot deaths: the age range affected; mother's smoking; respiratory viral infections; immunisation status. Both laboratory and epidimiological evidence suggests that vulnerability of infants to infectious agents depends on interactions between genetic, developmental and environmental factors that contribute to colonisation by microorganisms, the inflammatory and specific immune responses and the infants' physiological responses to inflammatory mediators. A model is proposed to explain how microorganisms might trigger a series of events resulting in some of these unexpected deaths and discusses how the present recommendations regarding child care practices might help reduce the numbers of Sudden Infant Death Syndrome cases associated with infectious agents.     

Evidence for the role of glycoprotein G of respiratory syncytial virus in binding of Neisseria meningitidis to HEp-2 cells

Abstract Viral glycoproteins G and F are expressed on the surface of cells infected with respiratory syncytial virus (RSV). We investigated the role of these proteins in the previously reported enhanced binding of Neisseria meningitidis to RSV-infected HEp-2 cells. Virus particles attached to bacteria were detected by immunofluorescence with flow cytometry. Binding of FITC-labelled bacteria to RSV-infected cells was significantly inhibited by monoclonal antibody against glycoprotein G. Unlabelled bacteria interfered with binding of the anti-G monoclonal antibody to these cells. These interactions were not found with a monoclonal antibody against glycoprotein F. We propose that glycoprotein G of RSV expressed on the surface of infected cells might act as an additional receptor for meningococci.     

Effect of exogeneous methyl oleate on the time course of some parameters of Streptomyces hygroscopicus NRRL B-1865 culture

Addition of methyl oleate to a Streptomyces hygroscopicus NRRL B-1865 culture modified the metabolic properties of this strain. This addition decreased the pH of the medium, increased the valine uptake of the cells and reduced their consumption of glucose until the beginning of antibiotic biosynthesis, which was delayed. At the same time, an increase in growth (× 1.8) and a marked improvement in antibiotic production (× 20) could be observed. The use of labelled methyl oleate showed that methyl oleate was not a precursor of antibiotics produced by S. hygroscopicus NRRL B-1865. It is suggested that methyl oleate addition may cause some alteration in membrane permeability, inducing an increase in H⁺ extrusion and stimulating the accumulation of branched amino acids, known to be direct precursors of polyether antibiotics. Correspondence to: L. David     

Silicon and Nitric Oxide-Mediated Regulation of Growth Attributes,Metabolites and Antioxidant Defense System of Radish (<i>Raphanus sativus</i> L.) under Arsenic Stress

Arsenic (As) contaminated food chains have emerged as a serious public concern for humans and animals and are known to affect the cultivation of edible crops throughout the world. Therefore, the present study was designed to investigate the individual as well as the combined effects of exogenous silicon (Si) and sodium nitroprusside (SNP), a nitric oxide (NO) donor, on plant growth, metabolites, and antioxidant defense systems of radish (Raphanus sativus L.) plants under three different concentrations of As stress, i.e., 0.3, 0.5, and 0.7 mM in a pot experiment. The results showed that As stress reduced the growth parameters of radish plants by increasing the level of oxidative stress markers, i.e., malondialdehyde and hydrogen peroxide. However, foliar application of Si (2 mM) and pretreatment with SNP (100 µM) alone as well as in combination with Si improved the plant growth parameters, i.e., root length, fresh and dry weight of plants under As stress. Furthermore, As stress also reduced protein, and metabolites contents (flavonoids, phenolic and anthocyanin). Activities of antioxidative enzymes such as catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (POD), and polyphenol oxidase (PPO), as well as the content of non-enzymatic antioxidants (glutathione and ascorbic acid) decreased under As stress. In most of the parameters in radish, As III concentration showed maximum reduction, as compared to As I and II concentrations. However, the individual and combined application of Si and NO significantly alleviated the As-mediated oxidative stress in radish plants by increasing the protein, and metabolites content. Enhancement in the activities of CAT, APX, POD and PPO enzymes were recorded. Contents of glutathione and ascorbic acid were also enhanced in response to co-application of Si and NO under As stress. Results obtained were more pronounced when Si and NO were applied in combination under As stress, as compared to their individual application. In short, the current study highlights that Si and NO synergistically regulate plant growth through lowering the As-mediated oxidative stress by upregulating the metabolites content, activity of antioxidative enzymes and non-enzymatic antioxidants in radish plants.