Mammalian prestin is a weak Cl⁻/HCO3⁻ electrogenic antiporter.

The lateral membrane of mammalian cochlear outer hair cells contains prestin, a protein which can act as a fast voltage-driven actuator responsible for electromotility and enhanced sensitivity to sound. The protein belongs to the SLC26 family of transporters whose members are characterised as able to exchange halides for SO(4)(2-) or HCO(3)(-) yet previous analyses of mammalian prestin have suggested that such exchange functions were minimal. Here anion transport is investigated both in guinea-pig outer hair cells (OHCs) and in an expression system where we employ a sensitive intracellular pH (pH(i)) probe, pHluorin, to report HCO(3)(-) transport and to monitor the small pH(i) changes observable in the cells. In the presence of extracellular HCO(3)(-), pH(i) recovered from an acid load 4 times faster in prestin-transfected cells. The acceleration required a chloride gradient established by reducing extracellular chloride to 2 mm. Similar results were also shown using BCECF as an alternative pH(i) sensor, but with recovery only found in those cells expressing prestin. Simultaneous electrophysiological recording of the transfected cells during bicarbonate exposure produced a shift in the reversal potential to more negative potentials, consistent with electrogenic transport. These data therefore suggest that prestin can act as a weak Cl(-)/HCO(3)(-) antiporter and it is proposed that, in addition to participating in wide band cochlear sound amplification, prestin may also be involved in the slow time scale (>10 s) phenomena where changes in cell stiffness and internal pressure have been implicated. The results show the importance of considering the effects of the endogenous bicarbonate buffering system in evaluating the function of prestin in cochlear outer hair cells.

Omega-3 fatty acids modulate collagen signaling in human platelets.

Dietary intake of the omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) results in cardioprotective benefits. However, the cellular and physiological bases for these benefits remain unclear. We hypothesized that EPA and DHA treatments would interfere with collagen-mediated platelet signaling. Thirty healthy volunteers received 28 days of 3.4 g/d EPA+DHA with and without a single dose of aspirin. Clinical hematologic parameters were then measured along with assays of collagen-stimulated platelet activation and protein phosphorylation. Omega-3 therapy led to a small but significant reduction in platelets (6.3%) and red blood cells (1.7%), but did not impair clinical time-to-closure assays. However, collagen-mediated platelet signaling events of integrin activation, α-granule secretion, and phosphatidylserine exposure were all reduced by roughly 50% after omega-3 incorporation, and collagen-induced tyrosine phosphorylation was significantly impaired. The diminished platelet response to collagen may account for some of the cardioprotective benefits provided by DHA and EPA.

The remarkable cochlear amplifier.

This composite article is intended to give the experts in the field of cochlear mechanics an opportunity to voice their personal opinion on the one mechanism they believe dominates cochlear amplification in mammals. A collection of these ideas are presented here for the auditory community and others interested in the cochlear amplifier. Each expert has given their own personal view on the topic and at the end of their commentary they have suggested several experiments that would be required for the decisive mechanism underlying the cochlear amplifier. These experiments are presently lacking but if successfully performed would have an enormous impact on our understanding of the cochlear amplifier.

Incomplete data on the Canadian cohort may have affected the results of the study by the International Agency for Research on Cancer on the radiogenic cancer risk among nuclear industry workers in 15 countries.

In 1995 the International Agency for Research on Cancer (IARC) completed a study that involved nuclear workers from facilities in the USA, UK and Canada. The only significant, though weak, dose-related associations found were for leukaemia and multiple myeloma. The results for the Canadian cohort, which comprised workers from the facilities of Atomic Energy of Canada Limited (AECL), were compatible with those for the other national cohorts. In 2005, IARC completed a further study, involving nuclear workers from 15 countries, including Canada. In these results, the dose-related risk for leukaemia was not significant but the prominent finding was a statistically significant excess relative risk per sievert (ERR Sv(-1)) for 'all cancers excluding leukaemia'. Surprisingly, the risk ascribed to the Canadian cohort for all cancers excluding leukaemia, driven by the AECL sub-cohort, was significantly higher than the risk estimate for the 15-country cohort as a whole. We have attempted to identify why the results for the AECL cohort were so discrepant and had such a remarkable influence on the 15-country risk estimate. When considering the issues associated with data on the AECL cohorts and their handling, we noted a striking feature: a major change in outcome of studies that involved Canadian nuclear workers occurred concomitantly with the shift to when data from the National Dose Registry (NDR) of Canada were used directly rather than data from records at AECL. We concluded that an important contributor to the considerable upward shift in apparent risk in the 15-country and other Canadian studies that have been based on the NDR probably relates to pre-1971 data and, in particular, the absence from the NDR of the person-years of workers who had zero doses in the calendar years 1956 to 1970. Our recommendation was for there to be a comprehensive evaluation of the risks from radiation in nuclear industry workers in Canada, organisation by organisation, in which some of the anomalies that we have identified might be addressed.

Atom detection and photon production in a scalable, open, optical microcavity.

A microfabricated Fabry-Perot optical resonator has been used for atom detection and photon production with less than 1 atom on average in the cavity mode. Our cavity design combines the intrinsic scalability of microfabrication processes with direct coupling of the cavity field to single-mode optical waveguides or fibers. The presence of the atom is seen through changes in both the intensity and the noise characteristics of probe light reflected from the cavity input mirror. An excitation laser passing transversely through the cavity triggers photon emission into the cavity mode and hence into the single-mode fiber. These are first steps toward building an optical microcavity network on an atom chip for applications in quantum information processing.

Biologically based analysis of lung cancer incidence in a large Canadian occupational cohort with low-dose ionizing radiation exposure, and comparison with Japanese atomic bomb survivors.

Lung cancer incidence is analyzed in a large Canadian National Dose Registry (CNDR) cohort with individual annual dosimetry for low-dose occupational exposure to gamma and tritium radiation using the two-stage clonal expansion model (TSCE) and extensions of the model with up to 10 initiation steps. Models with clonal expansion turned off provide very poor fits and are rejected. Characteristic and distinct temporal patterns of excess relative risk (ERR) are found for dose response affecting early, middle, or late stages of carcinogenesis, that is, initiation with one or more stages, clonal expansion, or malignant conversion. Both fixed lag and lag distributions are used to model time from first malignant cell to incidence. Background rates are adjusted for gender and birth cohort. Lacking individual smoking data, surrogate annual smoking doses based on U.S. annual per capita cigarette consumption appear to account for much of the birth cohort effect, leaving radiation dose response relatively unchanged. The mean cumulative exposure for males receiving nonzero cumulative doses of gamma and tritium radiation was 18.2 mSv. The males have a significant dose response with 33 out of a total of 322 lung cancer cases attributable to radiation. There were 78 incident lung cancer among females, (with mean cumulative exposure of 3.8 mSv among females with nonzero exposure). The dose response for females appears smaller than for males but does not differ significantly from zero or from the male dose response. Findings for males include significant dose-response relationships for promotion and malignant conversion, but not initiation, and a protraction effect (sometimes called an inverse-dose-rate effect, where risk increases with protraction of a given dose). The dose response predicted by our analysis appears consistent with the risk for lung cancer incidence in the Japanese atomic bomb survivors cohort, provided that proper adjustments are made for duration of exposure and differences in background rate parameters.

Investigating time patterns of variation in radiation cancer associations.

AIMS: In occupational settings, carcinogenic exposures are often repeated or protracted over time. The time pattern of exposure accrual may influence subsequent temporal patterns of cancer risk. The authors present several simple models that may be used to evaluate the influence of time since exposure or age at exposure on cancer incidence or mortality in an occupational cohort. METHODS: A cohort of 40,415 nuclear industry workers was identified via the Canadian National Dose Registry. Vital status and cause of death were ascertained through 1994. Associations between ionising radiation and mortality due to lung cancer, leukaemia, and cancers other than lung and leukaemia were quantified using conditional logistic regression models with risk sets constructed by incidence density sampling. A step function, a bilinear function, and a sigmoid function were used to evaluate temporal variation in exposure effects. RESULTS: Step and sigmoid functions were used to explore latency and morbidity periods. For analyses of lung cancer, leukaemia, and other cancers the best fitting models were obtained when exposure assignment was lagged by 13, 0, and 5 years, respectively. A bilinear function was used to evaluate whether exposure effects diminished with time since exposure. In analyses of lung cancer and leukaemia, there was evidence that radiation effects attenuated with protracted time since exposure. In analyses of age at exposure, there was evidence of variation in radiation mortality associations for analyses of lung cancer and leukaemia; discounting radiation doses accrued at younger ages (for example, 15-35 years) led to significant improvements in model fit. CONCLUSIONS: This paper illustrates empirical approaches to evaluating temporal variation in the effect of a protracted exposure on disease risk.

Cavitation in a lubrication flow between a moving sphere and a boundary.

A heavy sphere is free to move inside a rotating horizontal cylinder filled with viscous liquid. The steady motion is essentially Stokesian, and the sphere rotates at a fixed location with a lubrication layer between the ball and the wall. The symmetry of the flow field suggests there will be no force to balance the normal component of the ball's weight. However, we show that a normal force can arise when a cavitation bubble is present. The bubble size was measured as a function of the cylinder rotation rate and agrees well with a model which uses the force and torque balances on the sphere.

Regulation of the voltage-gated potassium channel KCNQ4 in the auditory pathway.

The potassium channel KCNQ4, expressed in the mammalian cochlea, has been associated tentatively with an outer hair cell (OHC) potassium current, I(K,n), a current distinguished by an activation curve shifted to exceptionally negative potentials. Using CHO cells as a mammalian expression system, we have examined the properties of KCNQ4 channels under different phosphorylation conditions. The expressed current showed the typical KCNQ4 voltage-dependence, with a voltage for half-maximal activation (V(1/2)) of -25 mV, and was blocked almost completely by 200 microM linopirdine. Application of 8-bromo-cAMP or the catalytic sub-unit of PKA shifted V(1/2) by approximately -10 and -20 mV, respectively. Co-expression of KCNQ4 and prestin, the OHC motor protein, altered the voltage activation by a further -15 mV. Currents recorded with less than 1 nM Ca(2+) in the pipette ran down slowly (12% over 5 min). Buffering the pipette Ca(2+) to 100 nM increased the run-down rate sevenfold. Exogenous PKA in the pipette prevented the effect of elevated [Ca(2+)](i) on run-down. Inhibition of the calcium binding proteins calmodulin or calcineurin by W-7 or cyclosporin A, respectively, also prevented the calcium-dependent rapid run-down. We suggest that KCNQ4 phosphorylation via PKA and coupling to a complex that may include prestin can lead to the negative activation and the negative resting potential found in adult OHCs.

Decreases in occupational exposure to ionizing radiation among Canadian dental workers.

OBJECTIVE: To describe doses of ionizing radiation and their possible associations with mortality rates and cancer incidence among Canadian dental workers. METHODS: The National Dose Registry (NDR) of Canada was used to assess occupational dose of ionizing radiation received by dental workers. The NDR cohort includes 42,175 people classified as dental workers. Subjects in the NDR were linked to both the Canadian Mortality Database and the Canadian Cancer Database to ascertain cause of death and cancer incidence, respectively. RESULTS: The cohort consisted of 9,051 male and 33,124 female dental workers. A total of 656 incident cases of cancer and 558 deaths were observed. The standardized mortality ratio associated with all-cause mortality was 0.53 (90% confidence interval [CI] 0.49-0.57). The incidence of cancer among dental workers was lower than that for the Canadian population for all cancers except melanoma of the skin (for melanoma, the standardized incidence ratio was 1.46 [90% CI 1.14-1.85]). Occupational doses of ionizing radiation among dentists and dental workers have decreased markedly since the 1950s. CONCLUSIONS: Dental workers receive very low doses of ionizing radiation, and these doses do not appear to be associated with any increase in cancer incidence; the increased incidence of melanoma is more likely related to other risk factors such as exposure to ultraviolet radiation from sunlight.

Apical endocytosis in outer hair cells of the mammalian cochlea.

Outer hair cells (OHCs), the sensory-motor cells of the mammalian cochlea, contain an endocytic tubulovesicular compartment below their apical stereocilia. We have used two-photon imaging of FM1-43 in the intact epithelium to show that these cells take up membrane in a Ca(2+)-dependent manner from a distinct apical site. The uptake rate was 0.8 microm(2)/s and internalized membrane was trafficked rapidly to a compartment along the lateral wall and distinct intracellular compartments. Double labelling with FM1-43 and DiOC(6), an endoplasmic reticulum (ER) marker, showed that these compartments are part of the tubulovesicular endoplasmic reticulum of OHCs. Labelling with a lysosomal marker showed that OHC lysosomes are restricted to the apex. Using the protein marker wheat germ agglutinin (WGA-FITC) we demonstrate that apical protein internalization and trafficking is about eight times slower than membrane internalization. Using double labelling with FM1-43 and WGA-FITC, we show that membrane and protein internalization are apically colocalized but that patterns of protein and membrane traffic differ. Protein was targeted only to the most apical third of the lateral wall. In control conditions, OHCs displayed only weak WGA-FITC surface labelling at the site of endocytosis. Lowering the rate of apical endocytosis increased this surface signal. The results suggest that OHCs endocytose membrane and membrane proteins with a high turnover rate and that these cells may use apical endocytosis to sort proteins via an indirect pathway to the lateral membrane.

Analysis of mortality among Canadian nuclear power industry workers after chronic low-dose exposure to ionizing radiation.

Studies of radiation-associated risks among workers chronically exposed to low doses of radiation are important, both to estimate risks directly and to assess the adequacy of extrapolations of risk estimates from high-dose studies. This paper presents results based on a cohort of 45,468 nuclear power industry workers from the Canadian National Dose Registry monitored for more than 1 year for chronic low-dose whole-body ionizing radiation exposures sometime between 1957 and 1994 (mean duration of monitoring = 7.4 years, mean cumulative equivalent dose = 13.5 mSv). The excess relative risks for leukemia [excluding chronic lymphocytic leukemia (CLL)] and for all solid cancers were 52.5 [95% confidence interval (CI): 0.205, 291] and 2.80 (95% CI: -0.038, 7.13) per sievert, respectively, both associations having P values close to 0.05. Relative risks by dose categories increased monotonically for leukemia excluding CLL but were less consistent for all solid cancers combined. Although the point estimates are higher than those found in other studies of whole-body irradiation, the difference could well be due to chance. Further follow-up of this cohort or the combination of results from multiple worker studies will produce more stable estimates and thus complement the risk estimates from higher-dose studies.

Flow and fracture in drying nanoparticle suspensions.

Drying aqueous suspensions of monodisperse silica nanoparticles can fracture in remarkable patterns. As the material solidifies, evenly spaced cracks invade from the drying surface, with individual cracks undergoing intermittent motion. We show that the growth of cracks is limited by the advancement of the compaction front, which is governed by a balance of evaporation and flow of fluid at the drying surface. Surprisingly, the macroscopic dynamics of drying show signatures of molecular-scale fluid effects.

Sugar transport by mammalian members of the SLC26 superfamily of anion-bicarbonate exchangers.

The mammalian cochlea contains a population of outer hair cells (OHCs) whose electromotility depends on an assembly of 'motor' molecules in the basolateral membrane of the cell. Named 'prestin', the molecule is a member of the SLC26 anion transporter superfamily. We show both directly and indirectly that SLC26A5, rat prestin, takes up hexoses when expressed in several cell lines. Direct measurements of labelled fructose transport into COS-7 cells expressing prestin are reported here. Indirect measurements, using imaging techniques, show that transfected HEK-293 or CHO-Ki cells undergo reversible volume changes when exposed to isosmotic glucose-fructose exchange. The observations are consistent with the sugar transport. A similar transport was observed using a C-terminal green fluorescent protein (GFP)-tagged pendrin (SLC26A4) construct. Cells transfected with GFP alone did not respond to sugars. The data are consistent with fructose being transported by prestin with an apparent Km=24 nm. From the voltage-dependent capacitance of transfected cells, we estimate that 250,000 prestin molecules were present and hence that the single transport rate is not more than 3000 fructose molecules s(-1). Comparison of the transfected cell swelling rates induced by fructose and by osmotic steps indicates that water was co-transported with sugar. We suggest that the structure of SLC26 family members allows them to act as neutral substrate transporters and may explain observed properties of cochlear hair cells.

Can you still see the cochlea for the molecules?

It is now established that the mammalian cochlea uses active amplification of incoming sound to achieve sensitivity. Cellular details are emerging slowly. Recent studies of sensory hair cells have highlighted the possible molecular bases for amplification and the components of sensitivity regulation within the cochlea: a synthesis is likely to depend on effective and physiologically informed modelling.

Purinergic control of intercellular communication between Hensen's cells of the guinea-pig cochlea.

1. Hensen's cells in the isolated cochlea were stimulated by extracellular adenosine 5'-triphosphate (ATP) applied to their endolymphatic surface while changes in membrane current and intracellular calcium concentration ([Ca2+]i) were measured simultaneously. The response consisted of (i) an initial rapid inward current accompanied by elevation of the [Ca2+]i, (ii) a more slowly rising inward current accompanied by a rise of the [Ca2+]i and (iii) a slowly developing reduction of input conductance. 2. The slower responses were maintained in the absence of extracellular Ca2+. Similar responses were produced by increasing the [Ca2+]i via UV flash photolysis of intracellular D-myo-inositol 1,4,5-trisphosphate, P4(5)-(1-(2-nitrophenyl)ethyl) ester (caged InsP3) loaded at pipette concentrations of 8-16 microM. 3. The slow inward current, reversing around 0 mV, was blocked by 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). 4. Bath application of U-73122 (1 microM), a phospholipase C inhibitor, eliminated the slow Ca2+-release component of the response to ATP. It is proposed that the effects of ATP are mediated by the co-activation of ionotropic P2X and metabotropic P2Y receptors. 5. Immunohistochemistry using light and electron microscopy revealed that inositol 1,4,5-trisphosphate (InsP3) receptors delineate a network within the cells. 6. The coupling ratio (CR) between cell pairs measured in dual patch-clamp recordings was 0.356 +/- 0.024. The coupling reversibly decreased to 51 % of the control within 2 min of applying 100 microM ATP. Flash photolysis of 32 microM intracellular caged InsP3 and 1 mM caged Ca2+ reduced CR to 42 and 62 % of the control, respectively. 7. We propose that endolymphatic ATP via P2X and P2Y receptors can control intercellular communication amongst Hensen's cells by reducing gap junction conductance in a Ca2+- and InsP3-dependent manner.

First analysis of cancer incidence and occupational radiation exposure based on the National Dose Registry of Canada.

A cohort study was conducted to investigate the relation between cancer incidence and occupational exposure to ionizing radiation. Records containing dose information from 1951 to 1988 for 191,333 persons were extracted from the National Dose Registry of Canada. The records were linked to the Canadian Cancer Data Base, with incidence data from 1969 to 1988. Standardized incidence ratios were calculated using Canadian cancer incidence rates stratified by age, sex, and calendar year. Excess relative risks were obtained from internally based dose-response analyses. The following significant results were found for males and females combined: a deficit in the standardized incidence ratio for all cancers combined; elevated standardized incidence ratios for thyroid cancer and melanoma; and elevated excess relative risks for rectum, leukemia, lung, all cancers combined, all except lung, and all except leukemia. For males, cancers of the colon, pancreas, and testis also showed significantly elevated excess relative risks. The specific cancer types listed above have been implicated in previous studies on occupational exposure to ionizing radiation, except for testis, colon, and melanoma, while the findings on thyroid cancer from previous studies are inconclusive. The thyroid standardized incidence ratios in this study are highly significant, but further investigation is needed to assess the possibility of association with occupational radiation exposure.