Naphthalene oxygenase fromCorynebacterium renale: Characterisation and mechanism of oxygenation

The formation ofcis-l,2,-dihydroxy-l,2,-dihydronaphthalene from naphthalene by naphthalene oxygenase, purified fromCorynebacterium renale ATCC 15075, was demonstrated to involve oxidation of a mol NADH and consumption of one mol oxygen. The enzyme contains one g-atom Fe²⁺ and one FAD. Catalase inhibited product formation and H2O2 could substitute for NADH in the reaction. Superoxide dismutase inhibited enzyme activity when either NADH or H₂O₂ was present; the generation of superoxide anion on addition of NADH to the enzyme, in the absence of naphthalene, was detected by the nitro blue tetrazolium reduction method. Hydroxyl radical scavengers, ethanol, mannitol and sodium benzoate, inhibited product formation when either NADH or H₂O₂ was present. Electron spin resonance studies, under aerobic conditions, indicated that iron of the enzyme underwent valence changes during the course of the reaction     

Initial steps in the degradation of n-alkane-1-sulphonates by Pseudomonas

The primary reaction in the degradation of n-alkane-1-sulfonates by Pseudomonas is the hydroxylation of the carbon atom bearing the sulfonate group. The 1-hydroxy-n-alkane-1-sulfonate (aldehyde-bisulfite adduct) formed easily hydrolyses to give the corresponding aldehyde and bisulfite. The enzyme catalysing the hydroxylation reaction depends for its action on the presence of molecular oxygen and NADH. The kinetics of this reaction and the substrate specificity of the enzyme were studied using a crude enzyme extract and a spectrophotometric assay method based on co-oxidation of NADH with the sulfonate.     

Immune response in human subjects at high altitude

Levels of immunoglobulins were determined in persons exposed to high altitude. The individuals studied included high altitude natives, sea level residents at high altitude for 2 years, and recent arrivals at high altitude. Increased IgG and IgA levels were found in high altitude natives and sea level residents at high altitude for 2 years when compared with sea level residents. In recent arrivals marked increase of IgG and IgG levels and slight rise in IgM was seen. Recent arrivals who suffered from high altitude pulmonary oedema showed marked elevation of IgG, IgM, and IgA. Immunoglobulin responses to both primary and secondary TAB inoculation were of a higher magnitude and more sustained at high altitude than at sea level.     

Fluorescence investigations of coenzyme and substrate interactions in mannitol-1-phosphate dehydrogenase ofEscherichia coli

Coenzyme and substrate interactions with mannitol-1-phosphate dehydrogenase fromEscherichia coli (a dimer of MW 45,000) have been studied by fluorescence spectroscopy. NAD⁺ quenches the fluorescence emission of the protein tryptophan residues; shifting the excitation wavelength from 280 to 290 nm results in an increase in this quenching and a red shift in the emission maximum. NAD⁺ also quenches the fluorescence of covalently attached pyridoxyl phosphate, and this quenching is accompanied by a spectral broadening above 425 nm. Fructose-6-phosphate increases the binding of NAD⁺, but causes a slight reduction in the quenching of the tryptophan fluorescence observed at saturating levels of coenzyme, and reverses the NAD⁺-induced broadening in the pyridoxyl phosphate emission spectrum. NADH quenches the protein emission much less than NAD⁺; this quenching is not changed by shifting the excitation wavelength and is not affected by the presence of bound mannitol-1-phosphate. Titrations monitoring the quenching by NADH indicate a single class of NADH binding sites, while titrations monitoring NADH fluorescence suggest that coenzyme fluorescence is more enhanced when NADH is bound to less than half of the total enzyme subunits, with the emission per NADH molecule bound decreasing as the number of NADH molecules bound increases. In the absence of coenzyme, neither fructose-6-phosphate nor mannitol-1-phosphate have any effect on the protein tryptophan emission; however, both substrates induce specific changes in the emission spectrum of covalently attached pyridoxyl phosphate. These results suggest that the different coenzymes and substrates cause specific conformational changes in mannitol-1-phosphate dehydrogenase.     

NaturalMycoplasma arthritidis infection in mice

B6C3F₁ mice from a hybrid production colony frequently were serologically positive by enzyme-linked immunosorbent assay (ELISA) and consistently negative by culture forMycoplasma pulmonis. Subsequently, 162 mice were obtained and intensively studied using an expanded group of cultural procedures, ELISA, and histopathology. Lesions attributable to mycoplasma infection were not found, butMycoplasma arthritidis was isolated from 20 mice. TheM. pulmonis ELISA was positive (IgM, IgG, or both) in 113 mice. Selected sera were tested simultaneously in both theM. pulmonis ELISA and in an ELISA usingM. arthritidis antigen, and were found to be positive in both the IgM and IgG classes in both ELISAs. Thus, cross-reacting antibody was produced in mice naturally infected withM. arthritidis, confirming previous observations based on experimental infections. To our knowledge, this is the first report of naturalM. arthritidis infection in laboratory mice.     

The role of homoeologous group 1 chromosomes in the control of rRNA genes in wheat

The presence of rRNA genes on homoeologous chromosomes 1A, 1B, and 1D of hexaploid wheat was investigated by rRNA/DNA hybridization, using DNA purified from aneuploid and substitution line derivatives of the variety Chinese Spring. Doubling the number of 1B chromosomes increased the number of rRNA genes by 31–49% but deleting the 1B chromosomes decreased the number by only 15–23%. This suggests that changes may occur in rRNA gene multiplicity at other nucleolar organizer sites to partially compensate for a deficiency of rRNA genes. There was no unequivocal evidence of rRNA genes on Chinese Spring chromosome 1A or 1D, but other varieties were shown to have rRNA genes on chromosome 1A. These results are consistent with the cytological observations that chromosomes 1A and 1B but not 1D possess nucleolar organizers.     

MiR-199a is overexpressed in plasma of type 2 diabetes patients which contributes to type 2 diabetes by targeting GLUT4

Decreased GLUT4 expression and impaired GLUT4 cell membrane translocation are involved in type 2 diabetes mellitus (T2DM) pathogenesis so the factors impacting GLUT4 expression may be associated with T2DM. In this study, we identified four miRNAs: miR-31, miR-93, miR-146a, and miR-199a which suppress GLUT4 expression in HEK293T cells. Subsequently, we determined expression of these four miRNAs in plasma samples of T2DM patients, T2DM susceptible individuals, and healthy controls and found miR-199a was overexpressed in patients’ plasma compared with healthy control. Because the miR-199a binding site in GLUT4 3′UTR is highly conserved among vertebrates, we detected the glucose uptake in rat L6 myoblast cells through gain- and loss-of-function of miR-199a. We found that miR-199a can repress glucose uptake in L6 cells, which was rescued by GLUT4 overexpression. These results indicate that T2DM patients may have a high level miR-199a that reduce GLUT4 expression and contribute to the insulin resistance. Hence, miR-199a may be a novel biomarker for risk estimation and classification in T2DM patients.     

The genes for ribosomal RNA in diploid and polytene chromosomes of Drosophila melanogaster

The proportion of the Drosophila genome coding for ribosomal RNA was examined in DNA from both diploid and polytene tissues of Drosophila melanogaster by rRNA-DNA hybridization. Measurements were made on larvae with one, two, three and four nucleolus organizer regions per genome. In DNA from diploid tissues the percent rDNA (coding for 28S and 18S ribosomal DNA) was found to be in proportion to the number of nucleolus organizers present. The number of rRNA genes within a nucleolus organizer therefore does not vary in response to changes in the number of nucleolus organizers. On the other hand, in DNA from cells with polytene chromosomes the percent rDNA remained at a level of about 0.1% (two to six times lower than the diploid values), regardless of either the number of nucleolus organizers per genome or whether the nucleolus organizers were carried by the X or Y chromosomes. This independence of polytene rDNA content from the number of nucleolus organizers is presumably due to the autonomous polytenization of this region of the chromosome. When the rDNA content of DNA from whole flies is examined, both the rDNA additivity of the diploid cells and the rDNA independence of polytene cells will affect the results. This is a possible explanation for the relative rDNA increase known to occur in X0 flies, but probably not for the phenomenon of rDNA magnification. — In further studies on DNA from larval diploid tissues, the following findings were made: 1) the Ybb-chromosome carries no rDNA; 2) flies carrying four nucleolus organizers do not tend to lose rDNA, even after eleven generations, and 3) the nucleolus organizer on the wild type Y chromosome may have significantly less rDNA than does that on the corresponding X chromosome.     

Isolation and characterisation of immunoglobulins in the serum of precolostral piglets

The presence of actively synthetized immunoglobulin in the serum of newborn precolostral germfree piglets was confirmed. This immunoglobulin, being of IgG antigenic type, carries determinants of typical IgG heavy and light chains and has a sedimentation constant of 4S. The first antibodies formed in germfree piglets after immunization with sheep red blood cells are of macroglobulinemic (IgM) nature. They are followed by formation of more slowly sedimenting antibodies of the IgG type. No fast sedimenting antibodies of the IgG type were detected.     

Biological activity of 125iodine labelled globin mRNA in an Ehrlich ascites cell-free system

Globin mRNA was isolated from avian erythroblasts and labelled with ¹²⁵iodine to a specific activity of 4.5×10⁶ dpm μg^-1 mRNA. The labelling procedure was modified as compared to common techniques in order to protect the mRNA against degradation. When this RNA is translated in an Ehrlich ascites cell-free system globin chains are synthesized as is demonstrated by the analysis of the in vitro synthesized products. The biological activity of ¹²⁵iodine labelled mRNA has proven to be 30% of that of unlabelled mRNA.     

Gill Monogenea of deepwater and surface fish in southeastern Australia

The faunas of gill Monogenea of marine teleost fishes in deep and surface waters of southeastern Australia are compared, based on extensive surveys: 1563 fish (66 or 67 species, 35 families, 15 orders) in deepwater; 1862 fish (46 species, 26 families, 7 orders) in surface water. Relative species diversity (number of species of Monogenea/all fish species examined) is approximately five times greater in surface waters. There is a similarly low relative species diversity of Monogenea in the northwestern Pacific and northwestern Atlantic deepwater. Deepwater Monogenea in all seas belong mainly to the Diclidophoroidea (13 of 19 species in southeastern Australia, 14 of 17 species in the northwestern Pacific, at least 9 of 12 (?) species in the northwestern Atlantic) predominantly Diclidophoridae. Important groups of surface Monogenea in southeastern Australia are the Microcotylidae (34 of 83 species), Dactylogyridae Ancyrocephalinae (15 species) and Capsaloidea (12 species); only 10 species belong to the Diclidophoroidea and 2 of those to the Diclidophoridae. It is concluded that deepsea Monogenea in southeastern Australia show no or very little relationship with surface Monogenea of the same geographical area, but close relationship with Monogenea in the deepsea of other geographical areas. Some deepsea Monogenea have a wide geographical distribution in the Atlantic and Pacific Oceans. Arctic and Antarctic Monogenea also are not related to deepwater forms. The main group of deepsea Monogenea is considered to be archaic.