Prolonged calcium influx after termination of light-induced calcium release in invertebrate photoreceptors

© The Authors, 2009 . This article is distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported License. The definitive version was published in Journal of General Physiology 134 (2009): 177-189, doi:10.1085/jgp.200910214.In microvillar photoreceptors, light stimulates the phospholipase C cascade and triggers an elevation of cytosolic Ca2+ that is essential for the regulation of both visual excitation and sensory adaptation. In some organisms, influx through light-activated ion channels contributes to the Ca2+ increase. In contrast, in other species, such as Lima, Ca2+ is initially only released from an intracellular pool, as the light-sensitive conductance is negligibly permeable to calcium ions. As a consequence, coping with sustained stimulation poses a challenge, requiring an alternative pathway for further calcium mobilization. We observed that after bright or prolonged illumination, the receptor potential of Lima photoreceptors is followed by the gradual development of an after-depolarization that decays in 1–4 minutes. Under voltage clamp, a graded, slow inward current (Islow) can be reproducibly elicited by flashes that saturate the photocurrent, and can reach a peak amplitude in excess of 200 pA. Islow obtains after replacing extracellular Na+ with Li+, guanidinium, or N-methyl-D-glucamine, indicating that it does not reflect the activation of an electrogenic Na/Ca exchange mechanism. An increase in membrane conductance accompanies the slow current. Islow is impervious to anion replacements and can be measured with extracellular Ca2+ as the sole permeant species; Ba can substitute for Ca2+ but Mg2+ cannot. A persistent Ca2+ elevation parallels Islow, when no further internal release takes place. Thus, this slow current could contribute to sustained Ca2+ mobilization and the concomitant regulation of the phototransduction machinery. Although reminiscent of the classical store depletion–operated calcium influx described in other cells, Islow appears to diverge in some significant aspects, such as its large size and insensitivity to SKF96365 and lanthanum; therefore, it may reflect an alternative mechanism for prolonged increase of cytosolic calcium in photoreceptors.This work was supported by National Science Foundation grant 0639774

Fiscal Year 2007

This annual report documents the locations, magnitudes, and geologic interpretations of earthquakes recorded for the Hanford monitoring region of south-central Washington in fiscal year 2007 (October 2006 through September 2007). The report provides summaries of seismic events recorded during the first three quarters of fiscal year 2007 and contains a more comprehensive discussion of seismic events for the fourth quarter of the fiscal year

Third Quarter Hanford Seismic Report for Fiscal Year 2008

The Hanford Seismic Assessment Program (HSAP) provides an uninterrupted collection of high-quality raw and processed seismic data from the Hanford Seismic Network for the U.S. Department of Energy and its contractors. The Hanford Seismic Assessment Team locates and identifies sources of seismic activity and monitors changes in the historical pattern of seismic activity at the Hanford Site. The data are compiled, archived, and published for use by the Hanford Site for waste management, natural phenomena hazards assessments, and engineering design and construction. In addition, the seismic monitoring organization works with the Hanford Site Emergency Services Organization to provide assistance in the event of a significant earthquake on the Hanford Site. The Hanford Seismic Network and the Eastern Washington Regional Network consist of 44 individual sensor sites and 15 radio relay sites maintained by the Hanford Seismic Assessment Team. For the Hanford Seismic Network, fourteen local earthquakes were recorded during the third quarter of fiscal year 2008. The largest event recorded by the network during the third quarter (May 18, 2008 - magnitude 3.7 Mc) was located approximately 17 km east of Prosser at a depth of 20.5 km. With regard to the depth distribution, five earthquakes occurred at shallow depths (less than 4 km, most likely in the Columbia River basalts), six earthquakes at intermediate depths (between 4 and 9 km, most likely in the pre-basalt sediments), and three earthquakes were located at depths greater than 9 km, within the crystalline basement. Geographically, eight earthquakes occurred in swarm areas and six earthquakes were classified as random events. The largest event recorded by the network during the third quarter occurred on May 18 (magnitude 3.7 Mc) and was located approximately 17 km east of Prosser at a depth of 20.5 km. This earthquake was the highest magnitude event recorded in the 46-47 N. latitude / 119-120 W. longitude sector since 1975. The May 18 event, not reported as being felt on the Hanford site or causing any damage, was communicated to the PNNL Operations Center per HSAP communications procedures. The event is not considered to be significant with regard to site safety and not unprecedented given the site’s seismic history. The Hanford strong motion accelerometer (SMA) stations at the 200 East Area, 300 Area, and the 400 Area were triggered by the May 18 event. The reportable action level of 2% g for Hanford facilities is approximately 12 times larger than the peak acceleration (0.17%) observed at the 300 Area SMA station and no action was required

Protective effect of the isoflavone equol against DNA damage induced by ultraviolet radiation to hairless mouse skin

Equol, an isoflavonoid metabolite produced from the dietary isoflavone daidzein by the gut microflora in mammals, has been found to protect not only against ultraviolet (UV) radiation-induced cutaneous inflammation and photoimmune suppression, but also have anti-photocarcinogenic properties in mice. Because the state of DNA damage has been correlated with suppression of the immune system and photocarcinogenesis, we have therefore examined the potential of equol to offer protection from solar-simulated UV (SSUV) radiation-induced DNA damage in hairless mice by the immunohistochemical approach using monoclonal antibody specific for cyclobutane pyrimidine dimers (CPDs; H3 antibody). Topical application of 20 µM equol lotion, which was applied both before and after SSUV significantly reduced the number of CPDs. This reduction was evident immediately after SSUV exposure, at 1 h after exposure, and at 24 h after exposure, revealing 54%, 50%, and 26% reduction in CPDs, respectively. When the same concentration was applied for 5 consecutive days after SSUV exposure, there was no significant difference in the reduction of CPDs immediately after SSUV irradiation or at 1 hour afterwards, but there were significant reductions of 23% and 42% at 24 and 48 h after SSUV exposure, respectively. Despite apparently reducing the number of CPDs post-SSUV, topically applied equol did not appear to increase the rate of dimer removal. To conclude, equol applied topically prior to SSUV irradiation offers protection against CPD formation in hairless mice, possibly by acting as a suncreen and thus inhibiting DNA photodamage

Third Quater Seismic Report for Fiscal Year 2007

The Hanford Seismic Assessment Program (HSAP) provides an uninterrupted collection of high-quality raw and processed seismic data from the Hanford Seismic Network for the U.S. Department of Energy and its contractors. The Hanford Seismic Assessment Team locates and identifies sources of seismic activity and monitors changes in the historical pattern of seismic activity at the Hanford Site. The data are compiled, archived, and published for use by the Hanford Site for waste management, Natural Phenomena Hazards assessments, and engineering design and construction. In addition, the seismic monitoring organization works with the Hanford Site Emergency Services Organization to provide assistance in the event of a significant earthquake on the Hanford Site. The Hanford Seismic Network and the Eastern Washington Regional Network consist of 41 individual sensor sites and 15 radio relay sites maintained by the Hanford Seismic Assessment Team. For the Hanford Seismic Network, 16 local earthquakes were recorded during the third quarter of fiscal year 2007. The largest event (magnitude 2.0) occurred on April 16, 2007 and was located 4 km southwest of the 400 Area in the Columbia River basalts at a depth of approximately 3 km. Stratigraphically, 7 earthquakes occurred in the Columbia River basalts (approximately 0-5 km depth), 1 earthquake in the pre-basalt sediments (approximately 5-10 km depth), and 8 earthquakes in the crystalline basement (approximately 10-25 km depth). Geographically, 8 earthquakes occurred in swarm areas, and 8 earthquakes were classified as random events. The Hanford SMA network was triggered on the 300 Area and the 400 Area SMA by the 2.0 Mc seismic event that occurred on April 16, 2007. The maximum vertical acceleration was 0.07 % g and the maximum horizontal acceleration was 0.05% g at the 300 Area SMA, 13.5 km from the event. At the 400 Area SMA, only 5.2 km from the event, the maximum vertical acceleration was 0.25 % g and the maximum horizontal acceleration was 0.23% g. These are the first recordings of a small local earthquake on the SMA network. The reportable action level of 2% g for Hanford facilities is approximately 8 times larger than the peak accelerations observed at the 400 Area and no action was required

Fiscal Year 2008

The Hanford Seismic Assessment Program (HSAP) provides an uninterrupted collection of high-quality raw and processed seismic data from the Hanford Seismic Network for the U.S. Department of Energy and its contractors. The HSAP is responsible for locating and identifying sources of seismic activity and monitoring changes in the historical pattern of seismic activity at the Hanford Site. The data are compiled, archived, and published for use by the Hanford Site for waste management, natural phenomena hazards assessments, and engineering design and construction. In addition, the HSAP works with the Hanford Site Emergency Services Organization to provide assistance in the event of a significant earthquake on the Hanford Site. The Hanford Seismic Network and the Eastern Washington Regional Network consist of 44 individual sensor sites and 15 radio relay sites maintained by the Hanford Seismic Assessment Team. During fiscal year 2008, the Hanford Seismic Network recorded 1431 triggers on the seismometer system, which included 112 seismic events in the southeast Washington area and an additional 422 regional and teleseismic events. There were 74 events determined to be local earthquakes relevant to the Hanford Site. The highest-magnitude event (3.7 Mc) occurred on May 18, 2008, and was located approximately 17 km east of Prosser at a depth of 20.5 km. With regard to the depth distribution, 13 earthquakes were located at shallow depths (less than 4 km, most likely in the Columbia River basalts), 45 earthquakes were located at intermediate depths (between 4 and 9 km, most likely in the pre-basalt sediments), and 16 earthquakes were located at depths greater than 9 km, within the crystalline basement. Geographically, 54 earthquakes were located in swarm areas and 20 earthquakes were classified as random events. The May 18 earthquake was the highest magnitude event recorded since 1975 in the vicinity of the Hanford Site (between 46 degrees and 47 degrees north latitude and 119 degrees and 120 degrees west longitude). The event was not reported as being felt on the Hanford Site or causing any damage and was communicated to the Pacific Northwest National Laboratory Operations Center per HSAP communi¬cations procedures. The event is not considered to be significant with regard to site safety and not unprecedented given the site’s seismic history. The Hanford strong motion accelerometer (SMA) stations at the 200 East Area, 300 Area, and 400 Area were triggered by the May 18 event. The maximum acceleration recorded at the SMA stations (0.17% at the 300 Area) was 12 times smaller than the reportable action level (2% g) for Hanford Site facilities

Effect of Geochemical and Physical Heterogeneity on the Hanford 100D Area In Situ Redox Manipulation Barrier Longevity

The purpose of this study was to quantify the influence of physical and/or geochemical heterogeneities in the Hanford 100D area In Situ Redox Manipulation (ISRM) barrier, which may be contributing to the discontinuous chromate breakthrough locations along the 65-well (2,300 ft long) barrier. Possible causes of chromate breakthrough that were investigated during this study include: (1) high hydraulic conductivity zones; (2) zones of low reducible iron; and (3) high hydraulic conductivity zones with low reducible iron. This laboratory-scale investigation utilized geochemical and physical characterization data collected on 0.5 to 1 foot intervals from four borehole locations. Results of this laboratory study did not provide definitive support any of the proposed hypotheses for explaining chromate breakthrough at the Hanford 100-D Area ISRM barrier. While site characterization data indicate a significant degree of vertical variability in both physical and geochemical properties in the four boreholes investigated, lateral continuity of high conductivity/low reductive capacity zones was not observed. The one exception was at the water table, where low reductive capacity and high-K zones were observed in 3 of four boreholes. Laterally continuous high permeability zones that contain oxic sediment near the water table is the most likely explanation for high concentration chromium breakthrough responses observed at various locations along the barrier. A mechanism that could explain partial chromate breakthrough in the ISRM barrier is the relationship between the field reductive capacity and the rate of chromate oxidation. Subsurface zones with low reductive capacity still have sufficient ferrous iron mass to reduce considerable chromate, but the rate of chromate reduction slows by 1 to 2 orders of magnitude relative to sediments with moderate to high reductive capacity. The original barrier longevity estimate of 160 pore volumes for homogeneous reduced sediment, or approximately 20 years, (with 5 mg/L dissolved oxygen and 2 ppm chromate) is reduced to 85 pore volumes (10 years) when the wide spread 60 ppm nitrate plume is included in the calculation. However, this reduction in barrier lifetime is not as great for high permeability channels, as there is insufficient time to reduce nitrate (and consume ferrous iron). If the cause of laterally discontinuous breakthrough of chromate along the ISRM barrier is due to oxic transport of chromate near the water table, additional dithionite treatment in these zones will not be effective. Treatment near the water table with a technology that emplaces considerable reductive capacity is needed, such as injectable zero valent iron

Disruption of Microtubules Sensitizes the DNA Damage-induced Apoptosis Through Inhibiting Nuclear Factor κB (NF-κB) DNA-binding Activity

The massive reorganization of microtubule network involves in transcriptional regulation of several genes by controlling transcriptional factor, nuclear factor-kappa B (NF-κB) activity. The exact molecular mechanism by which microtubule rearrangement leads to NF-κB activation largely remains to be identified. However microtubule disrupting agents may possibly act in synergy or antagonism against apoptotic cell death in response to conventional chemotherapy targeting DNA damage such as adriamycin or comptothecin in cancer cells. Interestingly pretreatment of microtubule disrupting agents (colchicine, vinblastine and nocodazole) was observed to lead to paradoxical suppression of DNA damage-induced NF-κB binding activity, even though these could enhance NF-κB signaling in the absence of other stimuli. Moreover this suppressed NF-κB binding activity subsequently resulted in synergic apoptotic response, as evident by the combination with Adr and low doses of microtubule disrupting agents was able to potentiate the cytotoxic action through caspase-dependent pathway. Taken together, these results suggested that inhibition of microtubule network chemosensitizes the cancer cells to die by apoptosis through suppressing NF-κB DNA binding activity. Therefore, our study provided a possible anti-cancer mechanism of microtubule disrupting agent to overcome resistance against to chemotherapy such as DNA damaging agent

A direct signaling role for phosphatidylinositol 4,5-bisphosphate (PIP2) in the visual excitation process of microvillar receptors

Author Posting. © American Society for Biochemistry and Molecular Biology, 2005. This article is posted here by permission of American Society for Biochemistry and Molecular Biology for personal use, not for redistribution. The definitive version was published in Journal of Biological Chemistry 280 (2005): 16784-16789, doi:10.1074/jbc.M414538200.In microvillar photoreceptors the pivotal role of phospholipase C in light transduction is undisputed, but previous attempts to account for the photoresponse solely in terms of downstream products of phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis have proved wanting. In other systems PIP2 has been shown to possess signaling functions of its own, rather than simply serving as a precursor molecule. Because illumination of microvillar photoreceptors cells leads to PIP2 break-down, a potential role for this phospholipid in phototransduction would be to help maintain some element(s) of the transduction cascade in the inactive state. We tested the effect of intracellular dialysis of PIP2 on voltage-clamped molluscan photoreceptors and found a marked reduction in the amplitude of the photocurrent; by contrast, depolarization-activated calcium and potassium currents were unaffected, thus supporting the notion of a specific effect on light signaling. In the dark, PIP2 caused a gradual outward shift of the holding current; this change was due to a decrease in membrane conductance and may reflect the suppression of basal openings of the light-sensitive conductance. The consequences of depleting PIP2 were examined in patches of light-sensitive microvillar membrane screened for the exclusive presence of light-activated ion channels. After excision, superfusion with anti-PIP2 antibodies induced the appearance of single-channel currents. Replenishment of PIP2 by exogenous application reverted the effect. These data support the notion that PIP2, in addition to being the source of inositol trisphosphate and diacylglycerol, two messengers of visual excitation, may also participate in a direct fashion in the control of the light-sensitive conductanceThis work was supported by National Institutes of Health Grant EY07559