Mitochondrial metabolism reveals a functional architecture in intact islets of Langerhans from normal and diabetic Psammomys obesus

The cells within the intact islet of Langerhans function as a metabolic syncytium, secreting insulin in a coordinated and oscillatory manner in response to external fuel. With increased glucose, the oscillatory amplitude is enhanced, leading to the hypothesis that cells within the islet are secreting with greater synchronization. Consequently, non-insulin-dependent diabetes mellitus (NIDDM; type 2 diabetes)-induced irregularities in insulin secretion oscillations may be attributed to decreased intercellular coordination. The purpose of the present study was to determine whether the degree of metabolic coordination within the intact islet was enhanced by increased glucose and compromised by NIDDM. Experiments were performed with isolated islets from normal and diabetic Psammomys obesus. Using confocal microscopy and the mitochondrial potentiometric dye rhodamine 123, we measured mitochondrial membrane potential oscillations in individual cells within intact islets. When mitochondrial membrane potential was averaged from all the cells in a single islet, the resultant waveform demonstrated clear sinusoidal oscillations. Cells within islets were heterogeneous in terms of cellular synchronicity (similarity in phase and period), sinusoidal regularity, and frequency of oscillation. Cells within normal islets oscillated with greater synchronicity compared with cells within diabetic islets. The range of oscillatory frequencies was unchanged by glucose or diabetes. Cells within diabetic (but not normal) islets increased oscillatory regularity in response to glucose. These data support the hypothesis that glucose enhances metabolic coupling in normal islets and that the dampening of oscillatory insulin secretion in NIDDM may result from disrupted metabolic coupling. <br/

Monitoring Cl- movement in single cells exposed to hypotonic solution

Self-referencing ion--selective electrodes (ISEs), made with Chloride Ionophore I-Cocktail A (Fluka), were positioned 1-3 microm from human embryonic kidney cells (tsA201a) and used to record chloride flux during a sustained hyposmotic challenge. The ISE response was close to Nernstian when comparing potentials (VN) measured in 100 and 10 mM NaCl (deltaVN = 57 +/- 2 mV), but was slightly greater than ideal when comparing 1 and 10 mM NaCl (deltaVN = 70 +/- 3 mV). The response was also linear in the presence of 1 mM glutamate, gluconate, or acetate, 10 microM tamoxifen, or 0.1, 1, or 10 mM HEPES at pH 7.0. The ISE was approximately 3 orders of magnitude more selective for Cl- over glutamate or gluconate but less than 2 orders of magnitude move selective for Clover bicarbonate, acetate, citrate or thiosulfate. As a result this ISE is best described as an anion sensor. The ISE was 'poisoned' by 50 microM 5-nitro-2-(3phenylpropyl-amino)-benzoic acid (NPPB), but not by tamoxifen. An outward anion efflux was recorded from cells challenged with hypotonic (250 +/- 5 mOsm) solution. The increase in efflux peaked 7-8 min before decreasing, consistent with regulatory volume decreases observed in separate experiments using a similar osmotic protocol. This anion efflux was blocked by 10 microM tamoxifen. These results establish the feasibility of using the modulation of electrochemical, anion-selective, electrodes to monitor anions and, in this case, chloride movement during volume regulatory events. The approach provides a real-time measure of anion movement during regulated volume decrease at the single-cell leve

A recombinant rubella virus E1 glycoprotein as a rubella vaccine candidate.

A recombinant rubella virus E1 (rE1) glycoprotein was produced and some of its chemical and immunological features were characterized. Two animal models were then used to establish that the rE1 glycoprotein and rubella virus particles shared antigenic and immunogenic properties. In the first one, sera from rE1 glycoprotein-immunized BALB/c mice neutralized in vitro rubella virus infection. In the second model, severe combined immune deficient (SCID) mice implanted with tonsil fragments from rubella immune donors and immunized with rE1 glycoprotein produced human anti-rubella virus antibodies. Altogether, these results showed that immunization with rE1 glycoprotein elicited neutralizing anti-rubella virus antibodies. This study thus indicated that the rE1 glycoprotein could constitute a non-replicating rubella vaccine

A 22,000 14C year BP sediment and pollen record of climate change from Laguna Miscanti (23°S), northern Chile

Lake sediments and pollen, spores and algae from the high-elevation endorheic Laguna Miscanti (22°45′S, 67°45′W, 4140 m a.s.l., 13.5 km2 water surface, 10 m deep) in the Atacama Desert of northern Chile provide information about abrupt and high amplitude changes in effective moisture. Although the lack of terrestrial organic macrofossils and the presence of a significant 14C reservoir effect make radiocarbon dating of lake sediments very difficult, we propose the following palaeoenvironmental history. An initial shallow freshwater lake (ca. 22,000 14C years BP) disappeared during the extremely dry conditions of the Last Glacial Maximum (LGM; 18,000 14C years BP). That section is devoid of pollen. The late-glacial lake transgression started around 12,000 14C years BP, peaked in two phases between ca. 11,000 and <9000 14C years BP, and terminated around 8000 14C years BP. Effective moisture increased more than three times compared to modern conditions (∼200 mm precipitation), and a relatively dense terrestrial vegetation was established. Very shallow hypersaline lacustrine conditions prevailed during the mid-Holocene until ca. 3600 14C years BP. However, numerous drying and wetting cycles suggest frequent changes in moisture, maybe even individual storms during the mid-Holocene. After several humid spells, modern conditions were reached at ca. 3000 14C years BP. Comparison between limnogeological data and pollen of terrestrial plants suggest century-scale response lags. Relatively constant concentrations of long-distance transported pollen from lowlands east of the Andes suggest similar atmospheric circulation patterns (mainly tropical summer rainfall) throughout the entire period of time. These findings compare favorably with other regional paleoenvironmental data