The Sodium/Proton Exchanger NHE8 Regulates Late Endosomal Morphology and Function

The pH and lumenal environment of intracellular organelles is considered essential for protein sorting and trafficking through the cell. We provide the first evidence that a mammalian NHE sodium (potassium)/proton exchanger, NHE8, plays a key role in the control of protein trafficking and endosome morphology. At steady state, the majority of epitope-tagged NHE8 was found in the trans-Golgi network of HeLa M-cells, but a proportion was also localized to multivesicular bodies (MVBs). Depletion of NHE8 in HeLa M-cells with siRNA resulted in the perturbation of MVB protein sorting, as shown by an increase in epidermal growth factor degradation. Additionally, NHE8-depleted cells displayed striking perinuclear clustering of endosomes and lysosomes, and there was a ninefold increase in the cellular volume taken up by LAMP1/LBPA-positive, dense MVBs. Our data points to a role for the ion exchange activity of NHE8 being required to maintain endosome morphology, as overexpression of a nonfunctional point mutant protein (NHE8 E225Q) resulted in phenotypes similar to those seen after siRNA depletion of endogenous NHE8. Interestingly, we found that depletion of NHE8, despite its function as a sodium (potassium)/proton antiporter, did not affect the overall pH inside dense MVBs.     

A statistical method for correlating tRNA sequence with amino acid specificity.

A statistical method for finding the nucleotide positions in tRNA sequences that correlate with amino acid specificity has been developed. The procedure involves finding the subset of nucleotide positions and groups of positions where the marginal density of one amino acid tRNA class does not overlap that of any other amino acid class. The procedure is an application of a statistical method known as the Expectation Maximization algorithm.     

Mysterious bio-duck sound attributed to the Antarctic minke whale (Balaenoptera bonaerensis)

For decades, the bio-duck sound has been recorded in the Southern Ocean, but the animal producing it has remained a mystery. Heard mainly during austral winter in the Southern Ocean, this ubiquitous sound has been recorded in Antarctic waters and contemporaneously off the Australian west coast. Here, we present conclusive evidence that the bio-duck sound is produced by Antarctic minke whales (Balaenoptera bonaerensis). We analysed data from multi-sensor acoustic recording tags that included intense bio-duck sounds as well as singular downsweeps that have previously been attributed to this species. This finding allows the interpretation of a wealth of long-term acoustic recordings for this previously acoustically concealed species, which will improve our understanding of the distribution, abundance and behaviour of Antarctic minke whales. This is critical information for a species that inhabits a difficult to access sea-ice environment that is changing rapidly in some regions and has been the subject of contentious lethal sampling efforts and ongoing international legal action.     

Cyanobacteria passage through drinking water filters during perturbation episodes as a function of cell morphology, coagulant and initial filter loading rate

Eight pilot-scale in-line filtration trials were performed to evaluate the passage of cyanobacterial cells through drinking water filters after sudden increases in hydraulic loading rates. Trials were performed at 30 °C using two coagulant combinations (aluminum sulfate and cationic polymer or ferric chloride and cationic polymer), two initial filter loading rates (7 or 10 m/h) and two species of morphologically different cyanobacteria (Microcystis aeruginosa or Anabaena flos aquae). The filter was perturbed by instantaneously increasing the hydraulic loading rate by 50%. Filter influent and effluent water qualities were characterized by measuring turbidity, particles and chlorophyll a. The observed post-perturbation filter effluent chlorophyll a peaks were 1.6–48 times greater than the pre-perturbation averages. Chlorophyll a peaks were larger for M. aeruginosa than for A. flos aquae. Chlorophyll a peaks were also larger for the higher (10 m/h) than for the lower (7 m/h) initial filter loading rate. The post-perturbation effluent turbidity peaks were 1.4–7.2 times greater than the pre-perturbation averages. The post-perturbation effluent particle peaks were 6.5–25 times greater than the pre-perturbation averages. These results indicate that particles were a more sensitive indicator of cyanobacterial passage than turbidity.     

Enhancement of α-lactalbumin-like activity in mammary explants from pregnant rats in the absence of exogenous prolactin

Mammary explants from pregnant rats showed a progressive increase in α-lactalbumin activity during culture with insulin, hydrocortisone and prolactin. Unexpectedly, culture with only insulin and hydrocortisone produced a similar rate of increase of α-lactalbumin-like activity, but this increase commenced about 24 hr later. The delay suggests that the enhanced activity effected by insulin and hydrocortisone is not a reflection of carry-over of endogenous mammotrophic hormones. Insulin plus hydrocortisone did not stimulate casein or fatty acid synthesis by pregnancy tissue, and did not enhance α-lactalbumin-like activity in virgin rat mammary explants. Enhancement of this activity by insulin plus hydrocortisone in pregnant tissue was constant over a wide range of glucocorticoid concentrations, but was inhibited by progesterone. Available evidence indicates that the active factor in extracts from insulin-hydrocortisone-explants is a heat-stable protein which is either α-lactalbumin itself, or another molecule with similar specifier properties.     

The organization and structure of nerve and muscle in the jellyfish Cyanea capillata (coelenterata; scyphozoa)

The perirhopalial tissue and swimming muscle of Cyanea were examined with light microscopical and electron microscopical techniques. The perirhopalial tissue is a thin, triangular septum found on the subumbrellar surface of the animal. It separates part of the gastric canal system from the surrounding seawater, and is bound on two sides by radial muscle bands and on the third, the shorter side, by a rhopalium and the margin of the bell. The ectoderm of the perirhopalial tissue is composed of large, somewhat cuboidal, vacuolated, myoepithelial cells. The muscle tails of these cells form a single layer of radial, smooth muscle. Neurons of the “giant fiber nerve net” (GFNN), which form an extensive net over the perirhopalial tissue, lie at the base of the vacuolated portion of the myoepithelial cells. These neurons are visible in living tissue. The morphology of individual GFNN neurons was examined following intracellular injection of the fluorescent dye Lucifer Yellow. The neurons are usually bipolar and free of branches. At the electron microscope level, one usually finds that the GFNN neurons contain large vacuoles. The other characteristic feature of these cells is that they form symmetrical, or nonpolarized, synapses; that is, synaptic vesicles are found on both sides of the synapse. The swimming muscle is striated and composed of myoepithelial cells. Each myoepithelial cell has several muscle tails, and those of adjacent cells are linked to gether by desmosomes. The endoderm of the perirhopalial tissue also was examined. This investigation of the organization and ultrastructure of the perirhopalial tissue and surrounding muscle was undertaken to provide essential background information for an ongoing physiological study of the GFNN neurons and their synapses.     

Bacterial oxidation of polythionates: determination of tetrathionate with an ion-selective electrode.

A commercially available ion-selective electrode for nitrate was used to continuously monitor tetrathionate oxidation by Thiobacillus dentrificans. The electrode was much more sensitive to tetrathionate than to nitrate. The same electrode could also be used for the determination of trithionate.     

Testicular metamorphosis rates in European Starlings maintained under short daily photophases

Juvenile male European Starlings(Sturnus vulgaris) were maintained under discrete fixed daily photophases ranging from 1 to 11 h in duration. Treatment began on 20 December when all birds were reproductively quiescent, and continued until 14 June of the following year.In situ measurements of left testis widths at monthly intervals documented testicular width increases to levels associated with complete spermatogenesis in birds under all photoperiod regimens. Starlings maintained under the shortest and longest photoperiods required fewer days of treatment to achieve spermatogenic testes than did those under intermediate-length photoperiods. Data are consistent with the hypothesis that prolonged daily periods of darkness result in oscillations of a circadian timing system stimulating increased gonadotropin secretion and consequent testicular metamorphosis.