Production of poly(3-hydroxyalkanoates) from α, ω-alkanedioic acids and hydroxylated fatty acids by Alcaligenes sp.

Summary Poly(3-hydroxybutyrate) (P(3HB)) was produced from a series of , -alkanedioic acids of both even and odd carbon numbers by the Alcaligenes sp. AK201. In contrast, copolymers of 3HB and 3-hydroxyvalerate(P(3HB-co-3HV)) were produced from hydroxylated fatty acids of even carbon numbers such as 12-hydroxystearate and 2-hydroxyoctanoate. The biosynthetic pathways to poly(3-hydroxyalkanoates)(P(3HA)) are discussed.     

Mating of the fission yeast occurs independently of pmd1 + gene product,a structural homologue of budding yeast STE6 and mammalian P-glycoproteins

The pmd1 ⁺, a multidrug resistance gene of the fission yeast Schizosaccharomyces pombe, encodes a protein similar to the budding yeast Saccharomyces cerevisiae STE6 gene product and mammalian P-glycoproteins. The STE6 protein is a membrane transporter of a-factor, a mating pheromone of a-type S. cerevisiae, which is structurally related to M-factor of the fission yeast. However, heterothallic or homothallic pmd1 null mutant cells of S. pombe, which were constructed by means of gene disruption, showed no significant decrease in the mating abilities. On the other hand, the multidrug resistance conferred by the pmd1 ⁺ was overcome by the treatment with verapamil, a typical inhibitor of mammalian P-glycoproteins. These results indicate that the pmd1 ⁺ gene product is functionally similar to mammalian P-glycoproteins, rather than to the budding yeast STE6.     

Whole cell K and Cl currents in dissociated eccrine secretory coil cells during stimulation

Using the whole-cell voltage clamp (to determine the membrane current) and current clamp (to determine membrane potential) methods in conjunction with the nystatin-perforation technique, we studied the effect of methacholine (MCh) and other secretagogues on whole cell K and Cl currents in dissociated rhesus palm eccrine sweat clear cells. Application of MCh by local superfusion induced a net outward current (at a holding potential of ?60 mV and a clamp voltage of 0 mV), and a transient hyperpolarization by 5.6 mV, suggesting the stimulation of K currents. The net outward current gradually changed to the inward (presumably Cl) currents over the next 1 to 2 min of continuous MCh stimulation. During this time the membrane potential also changed from hyperpolarization to depolarization. The inward currents were increasingly more activated than outward (presumably K) currents during repeated MCh stimulations so that a net inward current (at ?60 mV) was observed after the fourth or fifth MCh stimulation. Ionomycin (10 μm) also activated both inward and outward current. The observed effect of MCh was abolished by reducing extracellular [Ca] to below 1 nm (Ca-free + 1 mm EGTA in the bath). MCh-activated outward currents were inhibited by 5 mm Ba and by 0.1 mm quinidine, although these agents also suppressed the inward currents. Bi-ionic potential measurements indicated that the contribution of Na to the membrane potential was negligible both before and after MCh or ISO (isoproterenol) stimulations and that the observed membrane current was carried mainly by K and Cl. MCh increased the bi-ionic potential by step changes in external K and Cl concentrations, further supporting that MCh-induced outward and inward currents represent K and Cl currents, respectively. Stimulation with ISO or FK (forskolin) resulted in a depolarization by about 55 mV and a net inward (most likely Cl) current independent of external Ca. CT-cAMP mimicked the effects of FK and ISO. The bi-ionic potential, produced by step changes in the external Cl concentration, increased during ISO stimulation, whereas that of K decreased. This indicates that the ISO-induced inward current is due to Cl current and that K currents were unchanged or slightly decreased during stimulation with ISO or 10 μm FK. Both myoepithelial and dark cells responded only to MCh (but not to FK) with a marked depolarization of the membrane potential due to activation of Cl, but not K, currents. We conclude that MCh stimulates Ca-dependent K and Cl currents, whereas ISO stimulates cAMP-dependent Cl currents in eccrine clear cells.     

Effects of zinc,selenium, and calcium on the nephrotoxicity of cadmium in primary cultures of rat renal proximal epithelial cells

The influence of essential metals, like zinc, selenium, and calcium, on the nephrotoxicity of cadmium was studied in primary cultures of rat proximal tubular cells. Damage to kidney cells was assessed by measuring the release of lactate dehydrogenase (LDH), γ-glutamyltranspeptidase (GTP), and β-N-acetylglucosaminidase (NAG) from cells into the medium and the cellular concentration of protein. Incubation with 200 μM cadmium in the presence of equivalent molar or lower concentrations of zinc and selenium showed greater release of LDH and NAG than cadmium alone, indicating an enhanced effect. However, metallothionein (MT) induced by pretreatment with a nontoxic concentration of zinc decreased significantly the release of enzyme from cells and elevated cellular protein levels in response to MT levels. MT provided partial protection against the nephrotoxicity of cadmium. Decreased calcium levels in the incubation medium also resulted in markedly increased release of LDH and NAG from cells exposed to cadmium and reduced cellular protein levels. These findings suggest that variations in zinc and calcium intake may affect the development of cadmium-induced renal dysfunction.     

Vaginal smear cycle in 4-day cyclic Chinese hamsters, Cricetulus griseus]

The contents of vaginal smear of 4-day cyclic Chinese hamster (Cricetulus griseus) was investigated every 3 hours for 5 days. A light-dark cycle of 14--10 hr was used with the lights turned on at 6 : 00 a.m. Estrous cycle of the Chinese hamster determined by vaginal smears can be divided into 6 periods. The proestrous phase started at about 0 : 00 of day 1, the day of the proestrous phase was designated as day 1 of the estrous cycle. In the afternoon of the same day 1, nucleated epithelial cells gradually increased in number (proestrus : I), and the vaginal contents became to consist solely of nucleated epithelial cells at about 18 : 00 to 21 : 00 (estrus : II). At about 0 : 00 of day 2, however, nucleated epithelial cells were superseded suddenly by cornified epithelial cells, and this phase lasted for 9 to 12 hr (metestrus I : III). Towards the end of the cornified stage, nucleated cells appeared in short duration (metestrus II : IV). And then, in the evening of day 2, leucocytes gradually increased in number with degeneration of nucleated cells (diestrus I : V-1). On day 3, vaginal smear contained a large amount of mucus as well as degenerated nucleated cells and leucocytes (diestrus II : V-2). At about 21 : 000 of day 4, some cornified epithelial cells were seen and then proestrous stage was returned. The females were mated with 3 to 5 males in the evening of day 1, copulation was confirmed in 83.7% females in the next morning,thus the copulation in the Chinese hamster may be thought to occur during the vaginal smear stage of nucleated epithelial cells (estrous phase), i.e. about 18 : 00 to 24 : 00 of day 1.     

Population studies ofCavelerius saccharivorus Okajima (Heteroptera: Lygaeidae): Adult dispersal in relation to the density

Two adjacent stations (H and L) were set up to study movement of C. saccharivorus adults in sugarcane fields. At the beginning of the study the density of C. saccharivorus including all stages of development was quite different between the two. The density of the first generation adult on Station H was about 5 times that on Station L. The number of C. saccharivorus on both the stations became almost the same one month after the beginning of the study. At the beginning of the study macropterous adults were more numerous in Station H than in Station L. However percentage of macropterous adults on Station L increased after one month whereas that on Station H declined. About 2,000 marked adults were released on each station during the early period of the emergence of the first generation adult. Marked insects were recaptured on both the stations one month after the release. The adults released on the dense population (H) tended to disperse more actively than those on the scarce population (L). Marked macropterous adults moved more actively than brachypterous ones. The density related dispersal of adults was considered to be an important factor to regulate the population density.     

Stimulation by subsynaptosomal fractions of transmitter efflux from plain synaptic vesicle fraction

The effects were investigated of purified subsynaptic fractions on the efflux of radioactivity from a plain synaptic vesicle fraction which had incorporated [³H]dopamine. About 50% of the radioactivity incorporated into the plain vesicles (120 g protein) was liberated on exposure to purified synaptic membranes (30 g protein). The synaptic membrane-dependent efflux appeared to depend on both adenosine triphosphate and divalent cations, especially Ca²⁺. Of the subcellular fractions used, the heavy microsomal fraction showed the same effects as the synaptic membrane fraction. Purified synaptic junctions exhibited the strongest stimulating effects: the efflux was 2 times greater than that observed with synaptic membranes. The stimulating effects of myelin were less than oneseventh of those of synaptic junctional fraction. These observations may indicate that the transmitters are liberated by interaction of vesicle membrane with synaptic membrane in the presence of ATP and divalent cations.Abbreviations EDTA ethylenediamine tetraacetic acid - EGTA ethyleneglycol bis-(-aminoethylether)-N,N-tetraacetic acid - AMP-PNP adenyl imidodiphosphate     

Immobilization of Glucose Isomerase-Containing Streptomyces phaeochromogenes Cells in Fine-Particle Form

A new preparation method for immobilizing Streptomyces phaeochromogenes cells in fine-particle form was investigated using radiation-induced polymerization at low temperatures with previously salted out hydrophilic monomers. Using this method, it was found that the glucose isomerase activity of the immobilized cell particles was markedly higher than that of immobilized cells in block form obtained without salting out of the monomer. The diameter of the particles was varied by changing the irradiation temperature or the concentrations of monomer and salt. The magnitude of the enzymatic activity increased with decreasing particle diameter. K_m values of the immobilized cell particles were close to that of the intact cell. These facts suggested that the cells were trapped on the surface of the particle.