Dynamic Observation of Retinal Response to Pressure Elevation in a Microfluidic Chamber.

Dynamic observation of cell and tissue responses to elevated pressure could help our understanding of important physiological and pathological processes related to pressure-induced injury. Here, we report on a microfluidic platform capable of maintaining a wide range of stable operating pressures (30 to 200 mmHg) while using a low flowrate (2-14 µL/h) to limit shear stress. This is achieved by forcing flow through a porous resistance matrix composed of agarose gel downstream of a microfluidic chamber. The flow characteristics were investigated and the permeabilities of the agarose with four different concentrations were extracted, agreeing well with results found in the literature. To demonstrate the capability of the device, we measured the change in intracellular Ca2+ levels of retinal ganglion cells in whole mouse retina in response to pressure. The onset of enhanced pressure results in, on average, an immediate 119.16% increase in the intracellular Ca2+ levels of retinal ganglion cells. The demonstrated microfluidic platform could be widely used to probe cell and tissue responses to elevated pressure.

Probing Light-Stimulated Activities in the Retina via Transparent Graphene Electrodes.

Graphene has triggered tremendous research due to its superior properties. In particular, the intrinsic high light transmission illustrates the unique advantage in neural biosensing. Here, we combine perforated flexible graphene electrodes with microfluidic platforms to explore real-time extracellular electrical activities of retinal ganglion cells (RGCs). Under light stimulation, the transparent graphene electrodes have demonstrated the capability of recording the electrical activities of stimulated RGCs in direct contact. Different types of RGCs have shown three distinct light induced patterns, ON, OFF, and ON-OFF, which are primarily operated by cone photoreceptors. Moreover, the observed spiking waveforms can be divided into two groups: the biphasic waveform usually occurs at contacts with soma, while the triphasic waveform is likely related to the axon. Under high K+ stimulation, the graphene electrodes exhibit higher electrical sensitivity than gold counterparts with an average 2.5-fold enhancement in spiking amplitude. Furthermore, a strong response has been observed with the firing rate first increasing and then ceasing, which could be due to the potassium-induced neural depolarization. These results show that graphene electrodes can be a promising candidate in the electrophysiology studies of retina and offer a route to engineering future two-dimensional materials-based biosensors.

Enhanced photocurrent response speed in charge-density-wave phase of TiSe2-metal junctions.

Group IVB transition metal dichalcogenides (TMDCs) have attracted significant attention due to their predicted high charge carrier mobility, large sheet current density, and enhanced thermoelectric power. Here, we investigate the electrical and optoelectronic properties of few-layer titanium diselenide (TiSe2)-metal junctions through spatial-, wavelength-, temperature-, power- and temporal-dependent scanning photocurrent measurements. Strong photocurrent responses have been detected at TiSe2-metal junctions, which is likely attributed to both photovoltaic and photothermoelectric effects. A fast response time of 31 µs has been achieved, which is two orders of magnitude better than HfSe2 based devices. More importantly, our experimental results reveal a significant enhancement in the response speed upon cooling to the charge-density-wave (CDW) phase transition temperature (TCDW = 206 K), which may result from dramatic reduction in carrier scattering that occurs as a result of the switching between the normal and CDW phases of TiSe2. Additionally, the photoresponsivity at 145 K is up to an order of magnitude higher than that obtained at room temperature. These fundamental studies not only offer insight for the photocurrent generation mechanisms of group IVB TMDC materials, but also provide a route to engineering future temperature-dependent, two-dimensional, fast electronic and optoelectronic devices.

Ultrafast Photocurrent Response and High Detectivity in Two-Dimensional MoSe2-based Heterojunctions.

Two-dimensional (2D) transition metal dichalcogenide (TMDC) materials have garnered great attention on account of their novel properties and potential to advance modern technology. Recent studies have demonstrated that TMDCs can be utilized to create high-performing heterostructures with combined functionality of the individual layers and new phenomena at these interfaces. Here, we report an ultrafast photoresponse within MoSe2-based heterostructures in which heavily p-doped WSe2 and MoS2 flakes share an undoped MoSe2 channel, allowing us to directly compare the optoelectronic properties of MoSe2-based heterojunctions with different 2D materials. Strong photocurrent signals have been observed in both MoSe2-WSe2 and MoSe2-MoS2 heterojunctions with a photoresponse time constant of ∼16 µs, surmounting previous MoSe2-based devices by three orders of magnitude. Further studies have shown that the fast response is independent of the integrated 2D materials (WSe2 or MoS2) but is likely attributed to the high carrier mobility of 260 cm2 V-1 s-1 in the undoped MoSe2 channel as well as the greatly reduced Schottky barriers and near absence of interface states at MoSe2-WSe2/MoS2 heterojunctions, which lead to reduced carrier transit time and thus short photocurrent response time. Lastly, a high detectivity on the order of ∼1014 Jones has been achieved in MoSe2-based heterojunctions, which supersedes current industry standards. These fundamental studies not only shed light on photocurrent generation mechanisms in MoSe2-based heterojunctions but also open up new avenues for engineering future high-performance 2D optoelectronic devices.

In situ monitoring of electrical and optoelectronic properties of suspended graphene ribbons during laser-induced morphological changes.

Exploring ways to tune and improve the performance of graphene is of paramount importance in creating functional graphene-based electronic and optoelectronic devices. Recent advancements have shown that altering the morphology of graphene can have a pronounced effect on its properties. Here, we present a practical and facile method to manipulate the morphology of a suspended graphene ribbon using a laser to locally induce heating while monitoring its electrical and optoelectronic properties in situ. Electrical measurements reveal that the conductance of suspended graphene transistors can be tuned by modifying its morphology. Additionally, scanning photocurrent measurements show that laser-induced folded graphene ribbons display significantly enhanced localized photocurrent responses in comparison with their flat counterparts. Moreover, the localization of the laser-induced heating allows for a series of folds to be induced along the entire graphene ribbon, creating targeted photocurrent enhancement. Through further investigations, it is revealed that the photo-thermoelectric effect is the primary mechanism for the increased photocurrent response of the device. Our experimental results explore the mechanisms and consequences of the folding process as well as provide a strategy to manipulate morphology and physical properties of graphene for future engineering of electronics and optoelectronics.

Near-infrared optical transitions in PdSe2 phototransistors.

We investigate electronic and optoelectronic properties of few-layer palladium diselenide (PdSe2) phototransistors through spatially-resolved photocurrent measurements. A strong photocurrent resonance peak is observed at 1060 nm (1.17 eV), likely attributed to indirect optical transitions in few-layer PdSe2. More interestingly, when the thickness of PdSe2 flakes increases, more and more photocurrent resonance peaks appear in the near-infrared region, suggesting strong interlayer interactions in few-layer PdSe2 help open up more optical transitions between the conduction and valence bands of PdSe2. Moreover, gate-dependent measurements indicate that remarkable photocurrent responses at the junctions between PdSe2 and metal electrodes primarily result from the photovoltaic effect when a PdSe2 phototransistor is in the off-state and are partially attributed to the photothermoelectric effect when the device turns on. We also demonstrate PdSe2 devices with a Seebeck coefficient as high as 74 µV K-1 at room temperature, which is comparable with recent theoretical predications. Additionally, we find that the rise and decay time constants of PdSe2 phototransistors are ∼156 µs and ∼163 µs, respectively, which are more than three orders of magnitude faster than previous PdSe2 work and two orders of magnitude over other noble metal dichalcogenide phototransistors, offering new avenues for engineering future optoelectronics.

Association of childhood adiposity measures with adulthood knee cartilage defects and bone marrow lesions: a 25-year cohort study.

OBJECTIVE: To describe the associations between childhood adiposity measures and adulthood knee cartilage defects and bone marrow lesions (BMLs) measured 25 years later. METHODS: 327 participants from the Australian Schools Health and Fitness Survey (ASHFS) of 1985 (aged 7-15 years) were followed up 25 years later (aged 31-41 years). Childhood measures (weight, height and skinfolds) were collected in 1985. Body mass index (BMI), overweight status and fat mass were calculated. Participants underwent 1.5 T knee magnetic resonance imaging (MRI) during 2008-2010, and cartilage defects and BMLs were scored from knee MRI scans. Log binomial regressions were used to examine the associations. RESULTS: Among 327 participants (47.1% females), 21 (6.4%) were overweight in childhood. Childhood adiposity measures were associated with the increased risk of adulthood patellar cartilage defects (Weight relative risk (RR) 1.05/kg, 95% confidence interval (CI) 1.01-1.09; BMI 1.10/kg/m2, 1.01-1.19; Overweight 2.22/yes, 1.21-4.08; fat mass 1.11/kg, 1.01-1.22), but not tibiofemoral cartilage defects. Childhood adiposity measures were not significantly associated with adulthood knee BMLs except for the association between childhood overweight status and adulthood patellar BMLs (RR 2.87/yes, 95% CI 1.10-7.53). These significant associations persisted after adjustment for corresponding adulthood adiposity measure. CONCLUSION: Childhood adiposity measures were associated with the increased risk of adulthood patellar cartilage defects and, to a lesser extent, BMLs, independent of adulthood adiposity measures. These results suggest that adiposity in childhood has long-term effects on patellar structural abnormalities in young adults.

A phase I, first in human study of FP-1039 (GSK3052230), a novel FGF ligand trap, in patients with advanced solid tumors.

BACKGROUND: Fibroblast growth factors (FGFs) play important roles in multiple cancers by supporting tumor growth and angiogenesis. FP-1039 (GSK3052230) is a FGF ligand trap consisting of the extracellular domain of FGF receptor 1 (FGFR1) fused with the Fc region of IgG1. FP-1039 binds and neutralizes multiple FGFs that normally bind FGFR1. The primary objective of this phase I study was to evaluate the safety and tolerability of FP-1039. PATIENTS AND METHODS: Eligible patients with metastatic or locally advanced solid tumors for which standard treatments were ineffective were treated with weekly doses of FP-1039 for 4 weeks, followed by 2 weeks observation. RESULTS: Thirty-nine subjects received a mean of 6 infusions of FP-1039 at doses ranging from 0.5 to 16 mg/kg weekly, with no maximally tolerated dose identified. Grade 3 or greater treatment emergent adverse events were uncommon. Four dose-limiting toxicities were reported at doses of 0.75 mg/kg (urticaria), 1 mg/kg (intestinal perforation and neutropenia), and 16 mg/kg (muscular weakness). Drug exposure was dose proportional, and the terminal elimination half-life was 2.6-3.9 days following a single dose. Target engagement as measured by low free plasma FGF2 levels was achieved. FGF pathway dysregulation was uncommon. No objective responses were observed. CONCLUSION: In nonselected cancer patients with advanced disease, treatment with FP-1039 was well tolerated and toxicities associated with small molecule drugs that inhibit FGFR tyrosine kinases, including hyperphosphatemia, were not observed. Further studies of FP-1039 in patients selected for FGF pathway dysregulation, who are most likely to benefit, are now underway.

Evidence for horizontal and vertical transmission in Campylobacter passage from hen to her progeny.

Campylobacter is an important human pathogen, and consumption of undercooked poultry has been linked to significant human illnesses. To reduce human illness, intervention strategies targeting Campylobacter reduction in poultry are in development. For more than a decade, there has been an ongoing national and international controversy about whether Campylobacter can pass from one generation of poultry to the next via the fertile egg. We recognize that there are numerous sources of Campylobacter entry into flocks of commercial poultry (including egg transmission), yet the environment is often cited as the only source. There has been an abundance of published research globally that refutes this contention, and this article lists and discusses many of them, along with other studies that support environment as the sole or primary source. One must remember that egg passage can mean more than vertical, transovarian transmission. Fecal bacteria, including Campylobacter, can contaminate the shell, shell membranes, and albumen of freshly laid fertile eggs. This contamination is drawn through the shell by temperature differential, aided by the presence of moisture (the "sweating" of the egg); then, when the chick emerges from the egg, it can ingest bacteria such as Campylobacter, become colonized, and spread this contamination to flock mates in the grow house. Improvements in cultural laboratory methods continue to advance our knowledge of the ecology of Campylobacter, and in the not-so-distant future, egg passage will not be a subject continuously debated but will be embraced, thus allowing the development and implementation of more effective intervention strategies.

Subtype selective NMDA receptor antagonists induce recovery of synapses lost following exposure to HIV-1 Tat.

BACKGROUND AND PURPOSE: Neurocognitive disorders afflict approximately 20% of HIV-infected patients. HIV-1-infected cells in the brain shed viral proteins such as transactivator of transcription (Tat). Tat elicits cell death and synapse loss via processes initiated by NMDA receptor activation but mediated by separate downstream signalling pathways. Subunit selective NMDA receptor antagonists may differentially modulate survival relative to synaptic changes. EXPERIMENTAL APPROACH: Tat-evoked cell death was quantified by measuring propidium iodide uptake into rat hippocampal neurons in culture. The effects of Tat on synaptic changes were measured using an imaging-based assay that quantified clusters of the scaffolding protein postsynaptic density 95 fused to green fluorescent protein. KEY RESULTS: Dizocilpine, a non-competitive NMDA receptor antagonist, inhibited Tat-induced synapse loss, subsequent synapse recovery and Tat-induced cell death with comparable potencies. Memantine (10 µM) and ifenprodil (10 µM), which preferentially inhibit GluN2B-containing NMDA receptors, protected from Tat-induced cell death with no effect on synapse loss. Surprisingly, memantine and ifenprodil induced synapse recovery in the presence of Tat. In contrast, the GluN2A-prefering antagonist TCN201 prevented synapse loss and recovery with no effect on cell death. CONCLUSIONS AND IMPLICATIONS: Synapse loss is a protective mechanism that enables the cell to cope with excess excitatory input. Thus, memantine and ifenprodil are promising neuroprotective drugs because they spare synaptic changes and promote survival. These GluN2B-preferring drugs induced recovery from Tat-evoked synapse loss, suggesting that synaptic pharmacology changed during the neurotoxic process. NMDA receptor subtypes differentially participate in the adaptation and death induced by excitotoxic insult.

Effect of vaccinating breeder chickens with a killed Salmonella vaccine on Salmonella prevalences and loads in breeder and broiler chicken flocks.

The objective of this study was to evaluate the effect of vaccination of breeder chickens on Salmonella prevalences and loads in breeder and broiler chicken flocks. Chickens housed on six commercial breeder farms were vaccinated with a killed Salmonella vaccine containing Salmonella Typhimurium, Salmonella Enteritidis, and Salmonella Kentucky. Unvaccinated breeders placed on six additional farms served as controls. Eggs from vaccinated and unvaccinated breeder flocks were kept separately in the hatchery, and the resulting chicks were used to populate 58 commercial broiler flock houses by using a pair-matched design. Vaccinated breeder flocks had significantly higher Salmonella-specific antibody titers than did the unvaccinated breeder flocks, although they did not differ significantly with respect to environmental Salmonella prevalences or loads. Broiler flocks that were the progeny of vaccinated breeders had significantly lower Salmonella prevalences and loads than broiler flocks that were the progeny of unvaccinated breeders. After adjusting for sample type and clustering at the farm level, the odds of detecting Salmonella in samples collected from broiler flocks originating from vaccinated breeders were 62% lower (odds ratio [95% confidence interval] = 0.38 [0.21, 0.68]) than in flocks from unvaccinated breeders. In addition, the mean load of culture-positive samples was lower in broilers from vaccinated breeders by 0.30 log most probable number per sample (95% confidence interval of -0.51, -0.09; P = 0.004), corresponding to a 50% decrease in Salmonella loads. In summary, vaccination of broiler breeder pullets increased humoral immunity in the breeders and reduced Salmonella prevalences and loads in their broiler progeny, but did not significantly decrease Salmonella in the breeder farm environment.

CCL5 evokes calcium signals in microglia through a kinase-, phosphoinositide-, and nucleotide-dependent mechanism.

Microglia, the resident macrophages of the CNS, are responsible for the innate immune response in the brain and participate in the pathogenesis of certain neurodegenerative disorders. Chemokines initiate activation and migration of microglia. The beta-chemokine CCL5 induces an elevation in intracellular calcium concentration ([Ca(2+)](i)) in human microglia. Here, we examined the signal transduction pathway linking activation of chemokine receptor CCR5 to an elevation in [Ca(2+)](i) in cultured microglia by using pharmacological approaches in combination with Fura-2-based digital imaging. The CCL5-induced response required Janus kinase (Jak) activity and the stimulation of an inhibitory G protein. Multiple downstream signaling pathways were involved, including phosphatidylinositol 3-kinase (PI3K), Bruton's tyrosine kinase (Btk), and phospholipase C (PLC)-mediated release of Ca(2+) from inositol 1,4,5-trisphosphate (IP(3))-sensitive stores. Activation of both the kinase and the lipase pathways was required for eliciting the Ca(2+) response. However, the majority of the [Ca(2+)](i) increase was derived from sources activated by NAD metabolites. Cyclic ADP-ribose (cADPR) evoked Ca(2+) release from intracellular stores, and ADPR evoked Ca(2+) influx via a nimodipine-sensitive channel. Thus, a multistep cascade couples CCR5 activation to Ca(2+) increases in human microglia. Because changes in [Ca(2+)](i) affect chemotaxis, secretion, and gene expression, pharmacologic modulation of this pathway may alter inflammatory and degenerative processes in the CNS.

Degradation and inactivation of plasma tumor necrosis factor-alpha by pancreatic proteases in experimental acute pancreatitis.

BACKGROUND: Release of TNFalpha is thought to play an important role in mediating systemic effects in acute pancreatitis (AP). We have been unable to find an elevation of plasma TNFalpha in AP and hypothesize that it is susceptible to catabolism by circulating pancreatic proteases. METHODS: (1) AP was induced in Sprague-Dawley rats by cerulein hyperstimulation preceded by intraductal infusion of saline (mild) or glycodeoxycholic acid (severe). Healthy and sham-operated animals served as controls. Severity of pancreatitis was confirmed by histology. Plasma TNFalpha levels were measured at various time points after induction of AP with competitive ELISA. (2) Recombinant rat TNFalpha (rrTNFalpha) was incubated with trypsin, elastase, chymotrypsin and pepsin. Western Blot was performed to visualize TNF degradation. (3) RrTNFalpha was incubated in a concentration and time-dependant manner with proteases and TNF bioactivity was evaluated with a cytotoxicity assay. RESULTS: (1) Plasma TNFalpha levels in severe pancreatitis were significantly lower than in sham-operated controls after 0.5 and 6 h. (2) Incubation with proteases showed degradation in the presence of trypsin, elastase and chymotrypsin and no effect of pepsin. (3) There was a concentration dependent inactivation of rrTNFalpha in the presence of pancreatic proteases and a complete time-dependent inactivation in the presence of trypsin. CONCLUSION: Plasma TNFalpha does not rise in experimental AP, and levels are significantly lower in severe pancreatitis compared to sham-operated controls. Our study demonstrates degradation and inactivation of TNFalpha by pancreatic proteases, suggesting that it is unlikely it plays an important role in the development of distant organ failure.

Diazepam inhibits HIV-1 Tat-induced migration of human microglia.

During HIV-1 encephalitis, the chemotaxis-inducing activity of Tat may enhance the viral life cycle through recruitment of additional susceptible microglial cells to foci of infection. Benzodiazepines (BDZs) readily penetrate the blood-brain barrier and are known to possess anti-inflammatory properties. Pretreatment of human microglial cells with peripheral (Ro5-4864) and mixed (diazepam), but not central (clonazepam), benzodiazepine receptor ligands was found to potently suppress HIV-1 Tat-induced chemotaxis. Application of Tat to microglial cells evokes an increase in intracellular calcium concentration ([Ca(2+)]i) that rapidly desensitizes the cells. Diazepam's inhibitory effect was associated with its ability to block Tat-induced [Ca(2+)]i mobilization. These data support the notion that through their effects on microglia, peripheral BDZ receptor ligands could alter the neuropathogenesis of HIV-1.

Plasma membrane calcium ATPase plays a role in reducing Ca(2+)-mediated cytotoxicity in PC12 cells.

In many cell types, cell death induced by a variety of insults is accompanied by an increase in intracellular calcium. The Ca(2+) homeostatic mechanisms affected by such insults, however, have not been fully determined. Recent evidence indicates that kainic acid-induced seizures alter plasma membrane calcium ATPase mRNA expression within vulnerable hippocampal cell populations before the onset of cell death. We examined the effects of altering plasma membrane calcium ATPase expression on cell vulnerability in rat pheochromocytoma 12 cells. Pheochromocytoma 12 cells are vulnerable to Ca(2+) overload induced by the Ca(2+) ionophore A23187. Reverse transcriptase-PCR and Western blot data indicated that plasma membrane calcium ATPase isoform 4b constitutes a major calcium pump isoform in the pheochromocytoma 12 cells. Therefore, permanently transfected pheochromocytoma 12-derived cell lines were established that either over-expressed plasma membrane calcium ATPase isoform 4b, or suppressed the expression of the endogenous plasma membrane calcium ATPase isoform 4. Over-expressing clones were less vulnerable to Ca(2+)-mediated cell death induced by A23187 whereas "antisense" clones were considerably more susceptible. These data indicate that regulation of plasma membrane calcium ATPase expression may be critical to cellular survival when cells are exposed to pathological increases in intracellular calcium.

Cannabinoids inhibit the formation of new synapses between hippocampal neurons in culture.

The principal psychoactive ingredient in marijuana, Delta(9)-tetrahydrocannabinol, has been shown to inhibit adenylyl cyclase activity in vitro and can lead to impairment of memory in vivo. cAMP-induced changes in synaptic plasticity are thought to underlie memory formation. We examined the effects of cannabinoid receptor agonists on forskolin-induced formation of new synapses between rat hippocampal neurons in culture. Functional synaptic boutons were identified with FM1-43-based digital imaging. Cannabimimetic drugs prevented the recruitment of new synapses by inhibiting the formation of cAMP. The inhibition produced by Win55212-2, a synthetic cannabinoid receptor agonist, was stereoselective and was reversed by a selective CB1 receptor antagonist. Both Delta(9)-tetrahydrocannabinol and the endogenous ligand, anandamide, inhibited the formation of new synapses. Win55212-2 blocked the formation of new synapses induced by forskolin, but not those evoked by a membrane permeant cAMP analog. Thus, activation of cannabinoid receptors can modulate synaptic plasticity independent of direct effects on neurotransmitter release. Preventing the formation of new synapses may contribute to the impairment of memory produced by cannabinoids.

Use of home hemoglobin A1c test kits to monitor the effectiveness of diabetes care.

BACKGROUND: Periodic measurement of glycated hemoglobin (HbA1c) is highly recommended for people with diabetes to determine whether their blood glucose is adequately controlled. Quality improvement programs initiated by health plans often focus on ensuring that HbA1c is being monitored in members with diabetes. To focus improvement efforts on members with poor blood glucose control, health plans need to know which members have high HbA1c levels. Recent development of home test kits provides another opportunity for health plans to help members measure their HbA1c and to identify members with high levels. METHODS: A sample of members from two health plans who were sent HbA1c self-test kits in January 2000 participated in a telephone interview. To understand why members did or did not use self-test kits sent by their health plans, the survey focused on perceived ease of use, outcomes, and normative beliefs. RESULTS: In the group of 380 members who were interviewed, 170 (45%) used the kit. HbA1c values were > 8 mg/dl in 43%. Among the 170 who used the kit, 160 said that they would use the kit. Their most common reason for using the kit was to find out how well their blood glucose was being controlled (48%). Convenience (12%) was the next most frequent reason for using the kit. Among the 210 members who did not use the kit, 81 members said that they would not or were not sure if they would when interviewed. Their most frequent reason for not using the kit was duplication of tests done by physicians (34%). Others were too busy (12%), wanted to talk with their physician (11%), or had difficulty using the kit (11%). CONCLUSIONS: Because the majority of health plan members did not use the kit and the majority who did use the kit had HbA1c levels < 8 mg/dl, sending home test kits to members did not result in a high yield of members with elevated HbA1c levels. Physicians' support for use of the kits and efforts to make kits easier to use might increase use. Efforts to avoid duplication of physicians' measurements could make this strategy to identify members with poorly controlled levels of blood glucose more cost-effective, although health plans would not know which monitored members might benefit most from programs to improve care of diabetes.

Differentiation induces up-regulation of plasma membrane Ca(2+)-ATPase and concomitant increase in Ca(2+) efflux in human neuroblastoma cell line IMR-32.

Precise regulation of intracellular Ca(2+) concentration ([Ca(2+)](i)) is achieved by the coordinated function of Ca(2+) channels and Ca(2+) buffers. Neuronal differentiation induces up-regulation of Ca(2+) channels. However, little is known about the effects of differentiation on the expression of the plasma membrane Ca(2+)-ATPase (PMCA), the principal Ca(2+) extrusion mechanism in neurons. In this study, we examined the regulation of PMCA expression during differentiation of the human neuroblastoma cell line IMR-32. [Ca(2+)](i) was monitored in single cells using indo-1 microfluorimetry. When the Ca(2+)-ATPase of the endoplasmic reticulum was blocked by cyclopiazonic acid, [Ca(2+)](i) recovery after small depolarization-induced Ca(2+) loads was governed primarily by PMCAs. [Ca(2+)](i) returned to baseline by a process described by a monoexponential function in undifferentiated cells (tau = 52 +/- 4 s; n = 25). After differentiation for 12-16 days, the [Ca(2+)](i) recovery rate increased by more than threefold (tau = 17 +/- 1 s; n = 31). Western blots showed a pronounced increase in expression of three major PMCA isoforms in IMR-32 cells during differentiation, including PMCA2, PMCA3 and PMCA4. These results demonstrate up-regulation of PMCAs on the functional and protein level during neuronal differentiation in vitro. Parallel amplification of Ca(2+) influx and efflux pathways may enable differentiated neurons to precisely localize Ca(2+) signals in time and space.

Assessing photosynthetic downregulation in sunflower stands with an optically-based model.

Using a simple light-use efficiency model based on optical measurements, we explored spatial patterns of photosynthetic activity in fertilized and unfertilized sunflower stands. The model had two components: (1) absorbed photosynthetically active radiation (APAR), and (2) radiation-use efficiency. APAR was the product of photosynthetic photon flux density (PPFD) and leaf absorptance, which was derived from leaf reflectance. Radiation-use efficiency was either assumed to be constant or allowed to vary linearly with the photochemical reflectance index (PRI), a measure of xanthophyll cycle pigment activity. When efficiency was assumed to be constant, the model overestimated photosynthetic rates in upper canopy layers exposed to direct PPFD, particularly in the unfertilized canopy due to the greater photosynthetic downregulation associated with higher levels of photoprotective (de-epoxidized) xanthophyll cycle pigments in these conditions. When efficiency was allowed to vary according to the PRI, modeled photosynthetic rates closely matched measured rates for all canopy layers in both treatments. These results illustrate the importance of considering reduced radiation-use efficiency due to photosynthetic downregulation when modeling photosynthesis from reflectance, and illustrate the potential for detecting radiation-use efficiency through leaf optical properties. At least under the conditions of this study, these results also suggest that xanthophyll cycle pigment activity and net carbon uptake are coordinately regulated, allowing assays of Photosystem II activity to reveal changing rates of net assimilation. Because the optical methods in this study are adaptable to multiple spatial scales (leaf to landscape), this approach may provide a scalable model for estimating photosynthetic rates independently from flux measurements.

Dynamics of Salmonella contamination in a commercial quail operation.

Control of carcass contamination requires knowledge of the source and dynamics of spread of Salmonella in commercial poultry production. We examined Salmonella contamination at a U.S. commercial quail operation. Pulsed-field gel electrophoresis (PFGE) was used to type isolates in order to trace Salmonella throughout this production environment. During a 6-mo survey, Salmonella serotypes hadar, typhimurium, typhimurium variant Copenhagen, and paratyphi were encountered within this poultry operation. Ninety-four percent of the Salmonella isolated from breeder and production houses and from carcass rinses belonged to Salmonella serotypes typhimurium variant Copenhagen and hadar. There were six distinct S. typhimurium variant Copenhagen genetic types, as identified by PFGE, present within this particular poultry operation. Seventy-nine percent of S. typhimurium variant Copenhagen identified from the environment of the breeder and production houses produced the same PFGE pattern. Thirty-eight percent of S. typhimurium Copenhagen isolated from carcass rinses and the breeder house shared the same PFGE DNA pattern. This study demonstrates the transmission of salmonellae throughout this production environment, from the breeders to their progeny and to the birds ultimately processed for human consumption.