Cerebral blood flow (CBF) is known to be dysregulated in persons with human immunodeficiency virus 1 (HIV-1), for uncertain reasons. This is an important issue because impaired vasoreactivity has been associated with increased risk of ischemic stroke, elevated overall cardiovascular risk and cognitive impairment.
To test whether dysregulation of CBF might be due to virally-induced neuroinflammation, we used a well-defined animal model (GFAP-driven, doxycycline-inducible HIV-1 Tat transgenic (Tat-tg) mice). We then exposed the mice to a brief hypercapnic stimulus, and assessed cerebrovascular reactivity by measuring 1) changes in cerebral blood flow, using laser Doppler flowmetry and 2) changes in vascular dilation, using in vivo two-photon imaging.
Exposure to brief hypercapnia revealed an underlying cerebrovascular pathology in Tat-tg mice. In control animals, brief hypercapnia induced a brisk increase in cortical flow (20.8% above baseline) and vascular dilation, as measured by laser Doppler flowmetry and in vivo two-photon microscopy. These responses were significantly attenuated in Tat-tg mice (11.6% above baseline), but cortical microvascular morphology and capillary density were unaltered, suggesting that the functional pathology was not secondary to vascular remodeling. To examine the mechanistic basis for the diminished cerebrovascular response to brief hypercapnia, Tat-tg mice were treated with 1) gisadenafil, a phosphodiesterase 5 (PDE5) inhibitor and 2) tetrahydrobiopterin (BH4). Gisadenafil largely restored the normal increase in cortical flow following hypercapnia in Tat-tg mice (17.5% above baseline), whereas BH4 had little effect. Gisadenafil also restored the dilation of small (<25 μm) arterioles following hypercapnia (19.1% versus 20.6% diameter increase in control and Tat-tg plus gisadenafil, respectively), although it failed to restore full dilation of larger (>25 μm) vessels.
Taken together, these data show that HIV-associated neuroinflammation can cause cerebrovascular pathology through effects on cyclic guanosine monophosphate (cGMP) metabolism and possibly on PDE5 metabolism.
HIV-associated neurocognitive disorder (HAND) is characterized by sensory, motor and cognitive dysfunctions that result from HIV infection . HAND remains a major clinical concern despite the widespread use of combination antiretroviral therapy, and a recent study showed that HAND was detected in 52% of HIV-infected persons enrolled in a large multisite patient cohort . HAND represents a continuum of symptoms that may reflect a slowly progressing, multifactorial, degenerative process [3, 4]. While these symptoms are thought to be the result of HIV-induced neuroinflammation, the overall pathogenesis of this disease remains incompletely understood.
The effects of HIV-induced neuroinflammation on neuronal structure and function have been extensively studied in both in vitro and in vivo experimental model systems. In contrast, cerebral blood flow (CBF) was initially recognized to be dysregulated in persons with HIV-associated neurologic disease more than 20 years ago [5–10], but remains poorly understood at the mechanistic level. Recently, Ances and colleagues showed that the resting CBF of HIV-infected individuals is significantly decreased in both the lenticular nuclei and the visual cortex . Follow-up studies revealed that HIV infection and aging independently affect functional and resting flow to cortical structures , and have shown that resting CBF in persons with HIV infection is reduced to a level equivalent to that of HIV-1 negative persons who are 15 to 20 years older . Not only is resting CBF reduced in the setting of HIV-1 infection [11–13], but cerebrovascular responses to metabolic demand are also perturbed . This may have important implications for neurocognitive function.
To test whether dysregulation of CBF might be due to virally induced neuroinflammation, we used a well-defined animal model in which the pro-inflammatory viral Tat protein is expressed exclusively within the central nervous system (GFAP-driven, doxycycline-inducible HIV-1 Tat transgenic (Tat-tg) mice ). To confirm our findings, we also used a second experimental animal model, in which wild-type mice were exposed acutely to HIV-1 Tat by direct intracerebral injection (Tat-ICI; ).
We explored the cerebrovascular response of these animals to a defined hypercapnic stimulus, by measuring changes in CBF over the somatosensory cortex using laser Doppler flowmetry, in combination with two-photon in vivo imaging of cortical vessels. We found a significant loss of normal responsiveness to hypercapnia in Tat-exposed mice, relative to controls. This pathology was initially revealed by a long (5 minute) exposure to moderate hypercapnia (6% inspired CO2) which was then recapitulated during brief exposure (30 seconds); this transient hypercapnic challenge was selected for subsequent experiments so as to avoid hemodynamic changes as a result of acidosis [16, 17], which can occur following longer challenges .
In both our chronic (Tat-tg) and acute (Tat-ICI) mouse models for HIV-1 neuroinflammation, the vascular response to brief, moderate hypercapnia was significantly attenuated when compared to control animals (that is, wild-type littermates in the case of Tat-tg mice, or mice injected with saline in the case of Tat-ICI mice). These responses occurred in the absence of changes in cortical microvascular morphology and capillary density in the Tat-tg mice, suggesting that the functional pathology could not be attributed to vascular remodeling.
To explore the mechanistic basis for this pathology, we examined the possible dysregulation of the nitric oxide-cyclic guanosine monophosphate (NO-cGMP) axis within the neurovascular unit, which regulates the hypercapnic dilatory response [19–23]. We next performed experiments to directly address the mechanistic role of the NO-cGMP axis in contributing to the diminished vascular response to brief hypercapnia in mice with HIV-1 associated neuroinflammation. To do this, we exposed Tat-tg mice to hypercapnia in the presence or absence of 1) gisadenafil, a phosphodiesterase 5 (PDE5) inhibitor that prevents degradation of cGMP [24, 25], and 2) tetrahydrobiopterin (BH4) which is a limiting cofactor necessary for NO production [26, 27]. Treatment with BH4 had little effect on the cerebrovascular response to brief hypercapnia, whereas the PDE5 inhibitor largely restored the normal increase in cortical flow following hypercapnia in Tat-tg mice. Gisadenafil also restored the dilation of small (<25 μm) arterioles following hypercapnia, although it failed to restore full dilation of larger (>25 μm) vessels. This suggests that normalization of flow resulting from (PDE5) inhibition was predominantly determined by the functional recovery of smaller arterioles (<25 μm) within the cortex.
All animal procedures were approved by the University Committee on Animal Research. Adult (10 to 12 weeks old) C57BL/6 male mice were obtained from Charles River Laboratory and housed with a 12-hour light and 12-hour dark photoperiod. Food and water were provided ad libitum. Acute Tat-induced neuroinflammation was produced by stereotactic intracranial injection (ICI) to the right somatosensory cortex, essentially as described . Briefly, a 10-μl microvolume syringe (NanoFil, World Precision Instruments, Sarasota, FL, USA) and 35-gauge needle were silanized (Sigmacote, Sigma, St. Louis, MO, USA) to prevent Tat adhesion. Anesthesia was induced with isofluorane at a rate of 3.5 L/min and maintained at 2.0 L/min in a 50% oxygen and 50% nitrogen gas mixture. A small craniotomy (0.5 mm) was made 1 mm lateral to the sagittal suture at Bregma level −0.5 using a rotary hand tool. Syringe and needle were mounted in an Ultramicropump III syringe pump (World Precision Instruments, Sarasota, FL, USA) that was fixed to a three-axis micromanipulator. The needle tip was then maneuvered into the craniotomy and advanced to a depth of approximately 500–700 μm below cortical surface. Three μl of Tat (recombinant Tat1-72, 1 mg/ml, produced and purified from E. coli) dissolved in sterile saline (0.9% NaCl), was delivered at a rate of 80 nL/min. This dosage has been shown to elicit a strong neuroinflammatory response in mice following intracerebral injection . As controls, we used 1) sterile saline, 2) Tat inactivated by heating at 95°C for 10 min and 3) 3 μg of recombinant oligomeric HIV-1 Env (HIV-1YU2 gp140 ). The needle was then removed and the craniotomy was filled with bone wax (Ethicon, Raleigh, NC, USA). Three simple interrupted sutures were used to close the scalp, which was then treated with topical antibiotic. Rimadyl (5 mg/kg) was administered to provide post-surgical analgesia. Forty-eight hours post ICI, cerebrovascular reactivity (CVR) to carbon dioxide was examined.
Tat-transgenic (Tat-tg) mice were a generous gift from Drs. Pamela Knapp and Kurt Hauser (Virginia Commonwealth University) . At 8 weeks old, male mice were fed pellets infused with 6 mg/kg doxycycline (Harlan Laboratories, South Easton, MA, USA), ad libitum, for 3 weeks prior to use in experiments. Non-transgenic littermates (WT) were used as controls, and were also fed doxycycline-infused food. The expression of Tat mRNA in cortices of Tat-tg mice exposed to doxycycline was tested by RT-PCR, as described . Figure 1E shows that treatment with doxycycline dramatically increased the level of Tat expression as expected [30, 31]. Animals were housed with a 12-hour light and 12-hour dark photoperiod.
BH4 (15 mg/kg; Sigma, St. Louis, MO, USA) and gisadenafil besylate (UK-369003, 2 mg/kg; Tocris Bioscience, Bristol, UK) were suspended in a 5% DMSO, 95% physiologic saline solution and was administered by intraperitoneal (IP) injection to mice 2 hours before exposure to hypercapnia; control animals received vehicle alone.
Cerebral blood flow was measured using either bilateral or unilateral laser Doppler flowmetry (LDF) (BLF 21 D, Transonic Systems Inc., Ithaca, NY, USA) over the somatosensory cortex, as described . Physiological parameters were measured as described . Briefly, mean arterial pressure (MAP) was monitored via a femoral artery catheter. Arterial blood gases (ABG) and pH were measured with a blood gas analyzer (Siemens, Rapidlab 248, Erlangen, Bavaria, Germany) in 40 μl microsamples of blood from the femoral artery; blood samples were obtained prior to recording baseline, and after 30 second or 5 minute CO2 exposure was complete. The oxygen saturation and heart rate (HR) were continuously monitored using MouseOx (Harvard Apparatus, Starr Life Science Corporation, Holliston, MA, USA). Body temperature was monitored with a rectal probe.
Male mice (25 to 28 g) acutely or chronically exposed to Tat were anesthetized with a urethane/xylazine (1 g/kg, 2 mg/kg, respectively) combination via IP injection. This regimen was chosen to avoid the perturbation of cerebral blood flow that is observed with volatile anesthetics . Animals were placed on a heating pad (37°C) to prevent anesthesia-induced hypothermia, and temperature was monitored rectally for the duration of experiment. Bilateral LDF was used to examine CBF in the acute model of HIV-induced neuroinflammation (in this case, the uninjected left hemisphere served as a control for the injected right hemisphere). Unilateral LDF measurements were recorded for Tat-tg mice. In all cases, initial cortical baseline blood flow was recorded for 1 minute after which 6% CO2 was added to a 21% O2 air mixture for either 5 minutes or 30 seconds; CBF was then recorded for a total of 9 or 5 minutes, respectively.
Urethane/xylazine anesthetized mice were securely placed into a stereotactic frame and a thin skull window was created as described in . To visualize blood vessels, Texas Red-dextran (MW 70 kDa, Life Technologies, Invitrogen, Grand Island, NY, USA) was injected intravenously (10 mg/kg, in 0.1 ml of saline) into the right femoral vein. Arterioles were identified by the intrinsic auto-fluorescence of smooth muscle (as visualized using a 480/20 bandpass emission filter), branching patterns and blood flow direction. Imaging was performed on a Spectra PhysicsMaiTai HP DeepSee/Olympus Fluoview FV1000 multiphoton imaging setup with a 25× NA 1.05 water immersion microscope objective, and recorded by FluoView1000 software (Olympus America, Center Valley, PA, USA). The excitation wavelength was 880 nm, with the laser power at the sample set below 10 mW. The Texas Red fluorescence was detected using a 607/36 bandpass emission filter. Vessel diameter was extracted from two-photon images of pial arterioles using ImageJ software (NIH). Diameter was measured at two time points - at baseline and again at 30 seconds following exposure to 6% CO2.
Tat-tg and WT animals were injected with Texas Red-dextran as described above. Dye was allowed to circulate for 5 minutes, and anesthetized animals were then sacrificed by cervical dislocation and decapitated. Intact brains were removed, washed with cold artificial cerebrospinal fluid (aCSF, Harvard Apparatus, Starr Life Science Corporation, Holliston, MA, USA) and placed into a brain slicing matrix (Zivic Instruments, Pittsburgh, PA, USA). Tissue was cut into 2 mm coronal slices and placed onto glass slides with a shallow depression containing aCSF. A glass cover slip was placed over the sectioned tissue and a drop of aCSF was added to the cover slip to provide a liquid interface for the water immersion microscope objective. Images were taken from Bregma level −1.0, interaural 3.10, approximately 1.5 mm from midline. A 25x, 100-μm Z-stack was imaged through the cortex which was then skeletonized and analyzed in three dimensions using AMIRA software (Visage Imaging, San Diego, CA, USA).
The specificity of the PDE5 inhibitor gisadenafil besylate on PDE5 was evaluated by PDE activity assay on recombinant PDE5A and PDE1A proteins expressed in Cos7 cells as described in . The cGMP PDE activity of Cos7 cell lysates were assayed in buffer containing 20 mM Tris–HCl (pH 7.5), 3 mM MgCl, 15 mM magnesium acetate, 1 μM cGMP, 3H]cGMP (100,000 cpm/tube), either 400 mM EGTA (for PDE5A lysates) or 200 mM CaCl and 4 g/ml of CaM (for PDE1A lysates), and indicated concentrations of PDE5 inhibitor gisadenafil. All PDE assay reactions were started by adding the substrate into premixed other components. Reactions were incubated at 30°C for 15 minutes, and then terminated by boiling for 1 minute. After cooling, 2.5 mg/mL snake venom (Sigma, St. Louis, MO, USA) (with 5'-nucleotidase activity) was added to each reaction, and reactions were incubated at 30°C for 10 minutes. Hydrolyzed products were then separated by DEAE-sepharose anionic exchange columns, eluted from columns, and measured via liquid scintillation counter. Enzymatic activity was calculated as percentage total radioactivity minus background, and was established in a linear range prior to initiation of each experiment.
Given the relatively small number of subjects (mice) in each group, it's difficult to justify the normality assumption of the data distribution. Therefore we used the nonparametric permutation test to compare maximum CBF values reached after exposure to CO2 in different groups. To compare arteriole diameters before and after exposure to CO2, a two-tailed t-test was used.
The brain vasculature is exquisitely sensitive to changes in tissue and blood levels of carbon dioxide (pCO2) . Increased pCO2, or hypercapnia, induces a potent global vasodilation in the surface pial arterioles of the neocortex without the need for sensory or motor stimulation . Therefore, we used this dilatory responsiveness as a model to determine whether Tat exposure induces pathology within the vessel itself.
We initially examined responses in Tat-tg mice. Cerebral blood flow was measured by laser Doppler flowmetry, with unilateral placement of the laser Doppler probe (since both hemispheres expressed Tat). Baseline was established for 1 minute after which animals were exposed to 6% CO2 for 5 minutes (Figure 1A, the shaded area on the time line shows the period during which animals were exposed to CO2). The peak vasodilatory response to hypercapnia was significantly attenuated (P = 0.01; nonparametric permutation test) in Tat-tg (21.5% increase in CBF) compared to WT mice (40.3% increase in CBF) (Figure 1B).
Since long hypercapnic challenges have the potential to cause acidosis  and resulting hemodynamic changes [16, 17], we measured physiologic parameters before and after the 5-minute exposure to 6% CO2 (Table 1). There were no differences between Tat-tg, non-tg littermates (WT) or Tat-tg mice not induced with DOX (Tat-tg UI). However, after a 5 minute exposure to 6% CO2, mice developed moderate acidosis. We therefore next assessed whether a more transient exposure to CO2 would produce a similar CBF response without causing significant physiological changes. Table 1 shows that short (30 seconds) exposure minimized the magnitude of acidosis, while still eliciting a robust increase in cortical flow (Figure 1C). Comparison of the peak response of Tat-tg (11.6% increase in CBF, above baseline) and WT (20.8% increase in CBF, above baseline) mice to a brief hypercapnic challenge (Figure 1D) revealed a statistically significant difference (P = 0.01; nonparametric permutation test). The peak response of Tat-tg UI mice to a brief hypercapnic challenge (20.7% increase in CBF, above baseline) was not different from response of WT mice.
PaO2, partial arterial pressure of O2; UI, Uninduced. Data shown as mean ± SEM.
To determine whether the vascular response to hypercapnia was altered in an acute model of HIV-induced neuroinflammation , c57BL/6 mice age 8to 12 weeks, were anesthetized and administered one of the following by stereotactic intracerebral injection to the cortex of the right hemisphere (RH): 1) 3 μl of saline (0.9% NaCl), 2) 3 μl (1 mg/ml) of recombinant HIV-1 Tat in saline, 3) the same amount of heat-inactivated Tat in saline, or 4) 3 μl (1 mg/ml) of recombinant oligomeric HIV-1 Env in saline (HIV-1YU2 gp140). The cortex of the left hemisphere (LH) was not injected. Two days later, the response to hypercapnia was evaluated using flowmetry with bilaterally placed laser Doppler probes. Each animal served as its own control, by comparing flow on the manipulated right hemisphere (RH) to that on the unmanipulated LH. Figure 2A shows data for c57BL/6 mice injected with saline (RH), challenged with 6% CO2 for 30 seconds and recorded over a 5- minute period. This brief exposure to hypercapnia led to a brisk increase in CBF. The peak increase in CBF was not statistically different in the unmanipulated LH (30.6%) and saline-injected RH (21.3%) of these animals (Figure 2B) (P = 0.3; nonparametric permutation test). Figure 2C shows data for mice injected with Tat (RH) and challenged for 30 seconds with 6% CO2. The magnitude of the induced change in CBF was significantly lower in the Tat-injected RH (7.3%) versus the non-injected LH (26.3%) (P = 0.01; nonparametric permutation test) (Figure 2D). Figure 2E shows data for control mice injected with heat-inactivated Tat (RH) and challenged for 30 seconds with 6% CO2. The magnitude of the induced change in CBF was similar in the RH (24.6%) and the unmanipulated LH (29.5%) (Figure 2F). Finally, Figure 2G shows data for mice injected with oligomeric HIV-1 Env (RH) and challenged for 30 seconds with 6% CO2. The magnitude of the induced change in CBF was similar in the RH (29.3%) and the unmanipulated LH (32.1%) (Figure 2H).
The data presented in Figure 2 show that, consistent with what was observed in the chronic model, animals acutely exposed to Tat also had a diminished increase in CBF, following exposure to brief hypercapnia. Physiological parameters remained consistent during the 30 second administration of CO2 between the WT and Tat-tg mice (Table 1), suggesting that observed increases from baseline CBF (Figure 2) were a result of cerebrovascular responses rather than peripheral hemodynamic accommodations such as increased mean arterial pressure (MAP).
It is generally accepted that cortical blood flow is governed by surface pial arterioles , which run along the surface of the cortex, and then dive deeply into the parenchyma of the brain where they form an extensive cortical capillary network. Intracranial delivery of HIV-1 Tat is known to upregulate the release of inflammatory cytokines such as tumor necrosis factor α, interleukin 1β and interleukin-6 within the CNS [39–41]. By doing so, Tat creates a highly inflammatory, and potentially pro-angiogenic, environment (reviewed in [42, 43]). This suggests the possibility that chronic HIV-associated neuroinflammation might lead to increased microvascular density and/or increased vessel tortuosity in Tat-exposed mice, which could potentially modulate CVR, and therefore confound the interpretation of our data.
To assess whether perturbations of cerebrovascular reactivity might be due to changes in microvessel morphology and density (resulting in impaired flow), we imaged the cortical microvasculature using multiphoton microscopy in Tat-tg, and WT animals. Three- dimensional (500 μm x 500 μm x 100 μm) image stacks were then quantitatively analyzed using Amira software. For each image stack (Figure 3A) a three-dimensional vascular 'skeleton' was created by manually tracing vessels in three-dimensional space using the Amira filament tool. Then vessel length, radius, volume and branching parameters (including the numbers of nodes and segments) were extracted. Vessel branching was defined as segments between two nodes where a node is either an endpoint of a vessel or point at which multiple segments arise (Figure 3B). There was no statistically significant difference in the number of nodes and segments between WT and Tat-tg animals (Figure 3C; P = 0.120 and P = 0.483, respectively; t-test). Mean segment length (Figure 3D) was also found to be statistically indistinguishable in the two groups (P = 0.279; t-test). To determine whether vessel density within the cortex was altered in Tat-tg animals, mean radius (Figure 3E) and total vessel volume (Figure 3F) were calculated. No significant difference between WT and Tat-tg mice was detected (P = 0.490, and P = 0.431, respectively; t-test).
Hypercapnia is thought to induce vasodilation through the NO-cGMP pathway between perivascular neurons and/or endothelial cells and vascular smooth muscle [19–23]. To test whether this pathway might be dysregulated in the context of HIV-associated neuroinflammation, we conducted hypercapnic challenge experiments in Tat-tg mice that were treated with either 1) tetrahydrobiopterin (BH4), an essential and potentially limiting cofactor in the production of NO [26, 27] or 2) an inhibitor of PDE5, which regulates cellular cGMP levels . Physiological parameters were not changed by these treatments (Table 1).
Since some PDE5 inhibitors can also interact with PDE1 isotypes found within the cerebral vasculature, we first confirmed the specificity of gisadenafil for PDE5. This was directly tested with recombinant PDE5A and PDE1A overexpressed in COS-7 cells (Figure 4A). Using this approach, we found the IC50 of gisadenafil for PDE5A to be 3.6 nM, similar to its reported IC50 of 1.23 nM . In contrast, we found the IC50 of gisadenafil for PDE1A to be 9.1 μM, an approximately 2500-fold difference in specificity. Thus, gisadenafil at the concentrations used in this study should be specific to PDE5.
Tat-tg animals were treated with BH4 or gisadenafil besylate (UK-369,003; [24, 25]) by intraperitoneal injection 2 hours before measurement of the cerebrovascular response to brief hypercapnia. BH4 supplementation did not restore the normal cerebrovascular response to brief hypercapnic challenge (Figure 4B). In contrast, Tat-tg animals treated with the PDE5 inhibitor gisadenafil showed a marked restoration of the normal cerebrovascular response to 30 second hypercapnic challenge (17.5% peak increase in CBF), relative to untreated Tat-tg mice (11.6% peak increase in CBF) (Figure 4B). Combination treatment with both BH4 and gisadenafil had a slightly lesser (15.7%) effect compared to gisadenafil alone (Figure 4B). Longer (5 minute) exposures to CO2 were also performed in animals treated with gisadenafil to determine if these changes to CVR were repro ducible under extended hypercapnia. Tat-tg animals treated with gisadenafil showed marked improvement (P = 0.003, nonparametric permutation test) in CVR (42.9% peak increase in CBF) compared to non-treated Tat-tg animals (21.5% peak increase in CBF) (Figure 4C).
Since the morphology and gross structure of pial arterioles was unchanged in our model for HIV-associated neuroinflammation, we reasoned that the physical response of pial arterioles to pCO2 could be changed due to vessel pathology, and underlie the attenuated cerebrovascular reactivity in Tat-tg mice. We therefore measured changes in the diameter of these vessels in response to brief hypercapnia, by performing in vivo two-photon microscopy in both WT and Tat-tg mice, during CO2 administration.
A thinned skull window was prepared over the right somatosensory cortex , and Texas Red-dextran was then administered intravenously. Each animal was imaged for 1-minute baseline, then 6% CO2 was then delivered for 30 seconds via nose cone and imaging was continued for a total of 5 minutes. Representative images for the peak responses of WT and Tat-tg mice to intermittent hypercapnia are shown in Figure 5. In WT mice, cerebral arterioles, of all sizes dilated as expected in response to brief hypercapnia (Figure 5A,B), whereas in Tat-tg mice, this dilatory response was more attenuated (Figure 5C,D) in larger vessels (>25 μm). Treatment of Tat-tg mice with gisadenafil largely restored the normal vasodilatory response to hypercapnia (Figure 5E,F) by restoring small vessel (<25 μm) dilatory capacity.
These findings were quantitated by measuring the percent change of vessel diameter after exposure to CO2, as compared to baseline, using ImageJ software (NIH). Increases in pial vessel diameters (1 to 50 μm) were significantly different (P <0.0001; t-test) in WT (19.6%) and Tat-tg (10.0%) animals, showing compromise of the normal vasodilatory response to hypercapnia in Tat-tg model (Figure 5G). Interestingly, greater attenuation was observed in the dilation of larger caliber vessels (Figure 5I; >25 μm) (20.5% in WT mice, versus 6.7% in Tat-tg mice; P <0.0001; t-test) compared to smaller caliber vessels (Figure 5H; <25 μm) (19.1% in WT mice, versus 13.8% in Tat-tg mice; P = 0.0251; t-test).
We next examined individual arterioles in Tat-tg mice that were treated with gisadenafil. Comparison of all vessels (Figure 5G; 1 to 50 μm diameter) from untreated Tat-tg (10.0% peak increase in vessel diameter) and Tat-tg animals treated with gisadenafil (14.3% peak increase in vessel diameter), showed a significant improvement in dilatory capacity (P = 0.0135; t-test). However, vessel dilation failed to achieve levels observed for WT mice (19.6% increase in vessel diameter; P = 0.0069, t-test). Interestingly, smaller arterioles (1 to 25 μm diameter) in Tat-tg mice treated with the PDE5 inhibitor exhibited a robust dilatory response (20.6% increase in vessel diameter) to brief hypercapnia (Figure 5H), which was statistically equivalent (P = 0.5447; t-test) to the dilatory response of comparably sized vessels in WT animals (19.1% increase in vessel diameter). The dilation of larger arterioles (26 to 50 μm diameter) in Tat-tg mice was also enhanced by treatment with the PDE5 inhibitor (10.6% increase in vessel diameter), compared to untreated Tat-tg mice (6.7% increase in vessel diameter) (P = 0.0187; t-test). However, the dilatory response of these treated larger vessels was not fully restored when compared to WT mice (20.5%) (Figure 7I; P <0.0001; t-test).
We used hypercapnia as an experimental tool to examine the regulation of cerebrovascular reactivity in the context of HIV-induced neuroinflammation. Our initial experiments used a relatively extended (5 minute) exposure to moderate hypercapnia, consistent with previous studies in other mouse and rat models of cerebrovascular reactivity (6% CO2 stimulus with 9-minute evaluation of CBF) [45–47]. These studies demonstrated a significant loss of normal vascular responsiveness to hypercapnia in Tat-exposed mice, relative to controls (Figure 1). However, we noted that there was considerable acidemia in the animals at the end of this CO2 administration period (Table 1). Respiratory acidosis of this kind  can stimulate systemic hemodynamic change through β-adrenergic signaling [16, 17]. To avoid this, we therefore explored the administration of a 30-second CO2 challenge. This shorter hypercapnic exposure recapitulated our initial long exposure findings (compare Figures 1A with 1C), while minimizing the potential confounder of moderate respiratory acidosis (Table 1).
We proceeded to use this hypercapnic challenge model to examine the regulation of CBF in the context of two experimental animal models for HIV-induced neuroinflammation. In both models, mice were exposed to HIV-1 Tat (either by direct intracranial injection or through enforced transgene expression within the CNS), in order to induce a neuroinflammatory state mimicking that found in persons with HIV-associated neurocognitive disorders. In both models, we detected significant reductions in cerebrovascular reactivity to brief hypercapnia, suggesting an underlying cerebrovascular pathology that was induced by exposure to HIV-1 Tat. Mice exposed acutely to intracranial Tat showed a more profound depression in the cerebrovascular response to hypercapnia compared to Tat-tg mice. This difference might be explained by a robust inflammatory response to acutely injected Tat, compared to a more slowly developing, chronic inflammation produced in the Tat-tg mouse during three weeks of DOX-induced Tat expression. Since HIV-associated neuroinflammation is also a chronic disease, all remaining experiments were conducted in the Tat-tg model.
Tat-tg mice showed a 50% attenuation of the normal increase in CBF, following exposure to hypercapnia. This suggests that the dysregulation of CBF in persons living with HIV-1 may be a direct result of virally-induced neurovascular inflammation, and not secondary to peripheral inflammation or peripheral vascular disease associated with HIV-1 infection. Additionally, these findings suggest that decreased resting CBF in persons living with HIV  might be explained not only as a result of reduced neuronal demand (due to neuronal damage), but also as the consequence of an HIV-1 induced vascular dysfunction. Clinically, the failure to regulate CBF in the face of changing metabolic demand, such as increased CO2 levels, may in turn lead to transient episodes of cerebral ischemia, further perpetuating neuronal damage and inflammation.
In addition to examining regional cerebral blood flow using laser Doppler flowmetry, we also conducted experiments to look at the response of individual vessels to brief hypercapnia. To do this, we conducted in vivo two-photon imaging of cerebral blood vessels, using a thin-skull window (intact cranium). This allowed preservation of normal intracranial pressure, CSF dynamics and hemodynamics. We found that Tat-induced neuroinflammation reduced the overall magnitude of CO2-induced vasodilation, with an especially profound effect on larger vessels (>25 μm diameter) (Figure 5).
Cerebrovascular responses to hypercapnia are believed to occur through an NO-cGMP pathway between perivascular neurons and smooth muscle cells [19–23]. This pathway is highly susceptible to dysregulation by reactive oxygen species produced during inflammation [48, 49]. Moreover, HIV-1 Tat has also been shown to significantly decrease endothelial NOS (eNOS) expression in porcine coronary arteries . We therefore conducted experiments to examine the role of the NO-cGMP axis in contributing to the diminished vascular response to brief hypercapnia in mice with HIV-1 associated neuroinflammation.
First, we tested whether increasing the supply of NO might correct the vascular response to CO2. NO production by NOS is dependent on its cofactor tetrahydrobiopterin (BH4) [26, 27]. When BH4 levels are inadequate, the reduction of O2 by NOS is no longer coupled to L-arginine oxidation, resulting in generation of superoxide rather than NO. To test whether this uncoupling of NOS was responsible for impaired CVR in our model, we supplemented Tat-tg and WT mice with BH4 and then exposed them to brief hypercapnia. BH4 had no effect on the cerebrovascular responses to brief hypercapnia, which remained attenuated in the Tat-tg mice. In contrast, when Tat-tg mice were treated with a PDE5 inhibitor (gisadenafil), expected to prevent degradation of cGMP in arterial smooth muscle cells [24, 25], their cerebrovascular response to brief hypercapnia was normalized. This indicates that CO2 chemosensing was not impaired in the Tat-tg mice, and suggests that HIV-associated neuroinflammation may cause cerebrovascular pathology, through effects on cGMP metabolism and possibly PDE5. Interestingly, while gisadenafil produced significant recovery of the normal CBF response to brief hypercapnia in both small and large diameter vessels (Figure 5H, I, respectively), it showed selectivity for smaller vessels (<25 μm). One possible explanation is that the dilatation of the larger vessels is more reliant on NO [51, 52], which may not have been adequately replenished with BH4 supplementation. An alternative explanation is that PDE5 expression may be limited to the smaller vessels, whereas other PDE family members exist within the larger arterioles. Future studies will need to address the large vessel responsiveness to different PDE inhibitors. More importantly, however, inhibition of PDE5 was sufficient to restore the normal increase in CVR. This suggests that responses in the smaller vasculature may be of greater importance, compared to larger arterioles (>25 μm), in overall control of CBF following brief hypercapnic stimulation. Additionally, it is possible that already available PDE5 inhibitors could be used to limit further neuroinflammation initiated by transient episodes of tissue ischemia, due to decreased CVR to CO2 or other CBF regulators signaling through the NO-cGMP pathway.
Overall, this study reports three major findings. First, HIV-associated neuroinflammation (induced by either acute or chronic exposure to HIV-1 Tat) produced a cerebrovascular pathology, that was uncovered upon brief exposure to moderate hypercapnia. Second, this cerebrovascular pathology may selectively affect larger vessels (>25 μm) and finally, inhibition of PDE5 by gisadenafil besylate restored the normal cerebrovascular response to transient hypercapnia in mice with underlying HIV-associated neuroinflammation by fully restoring the dilatory capacity of the smaller pial arterioles. These findings may have important implications for persons living with HIV-1 especially because impaired vasoreactivity to hypercapnia has been associated with increased risk of ischemic stroke, elevated overall cardiovascular risk and cognitive impairment [53–58].
This study shows that HIV-associated neuroinflammation can directly result in cerebrovascular pathology. This may explain the dysregulation of cerebral blood flow and cerebrovascular reactivity in persons living with HIV. Our findings also implicate PDE5 as a key mediator of this process, and suggest that pharmacologic inhibition of this enzyme may restore normal cerebrovascular responses. This could have important therapeutic implications for the treatment of HIV-associated neurocognitive disorders.
All animal experiments were approved by the University of Rochester Committee on Animal Research, and were conducted in accordance with institutional, federal and local animal use regulations.
This work was supported by the following grants from the National Institutes of Health (NIH): T32GM007356 (JNS), T32DA007232 (FO) and R01DA026325. JNS was also supported in part by a HHMI "Med-into-Grad" Fellowship in Cardiovascular Science, and TL was supported by the Summer Research with NIDA Program. This research was also supported in part by the University of Rochester Developmental Center for AIDS Research (NIH P30AI078498).
We thank Drs. Kurt Hauser and Pam Knapp (Virginia Commonwealth University) for generously providing HIV-1 Tat transgenic mice, Dr. Phil Ray (University of Kentucky) for providing recombinant HIV-1 Tat protein, and Dr. Linda Callahan and Maria Jepson of the University of Rochester Two-Photon Core Facility for technical assistance and advice. Finally, we also thank Drs. Handy Gelbard, Charles Lowenstein, Craig Morrell (University of Rochester), Beau Ances (Washington University), James Duffin (University of Toronto) and Ariane Jay (Cornell Veterinary College) for helpful discussions.
JNS carried out in vivo imaging and laser Doppler flowmetry (LDF), helped to conceive the study, and drafted the manuscript. OP conducted in vivo imaging and LDF, and helped to conceive the study. WK conducted experiments to analyze the specificity of gisadenafil for PDE5 versus PDE1. JTZ assisted with surgeries for blood gas and MAP data. MG carried out genotyping and breeding of Tat-tg mice. TL carried out LDF experiments. FO analyzed cGMP levels. CF performed the statistical analysis. CY contributed to the design and conception of experiments to examine the NO/cGMP axis. KAK participated in the design of the study, the performance of in vivo imaging studies, and the analysis of in vivo imaging data. SD conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.


text:
Q: how to transfer value to another page with link?
Hello i'm stil learning, using Codeigniter can someone tell me, or give example code?
what i need is in Round Id we have 111 i want give it link and search database with value 111 how to do that? here the code i tried but still not right

                Round Id
                Player Id
                Bet Place
                Total Bet
                Win
                Lose

            <?php foreach ($tbl_bet_spot as $data) {?>

                    <a href="<?php echo site_url('account/detail_round_id?select=111');?>"><?php echo $data->round_id;?>
                    <?php echo $data->id;?>
                    <?php echo $data->bet;?>
                    <?php echo $data->total_bet;?>
                    <?php echo $data->win;?>
                    <?php echo $data->lose;?>

            <?php } ?>

controller
public function detail_round_id(){
        $select = $_GET['select'];

        $data['tbl_bet_spot'] = $this->login_model->selectRoundId_by_round($select)->result();

        print_r ($data);
    }

i just try with my code and it work now, but it's static in here
<a href="<?php echo site_url('account/detail_round_id?select=111');?>"><?php echo $data->round_id;?>

how i can send this value <?php echo $data->round_id;?> properly into controller? thanks a lot.

A: Use this code
<a href="<?php echo site_url()?>/account/detail_round_id/<?php echo $data->round_id;?>"><?php echo $data->round_id;?>

controller
public function detail_round_id(){
        $select = $this->uri->segment(3);

        $data['tbl_bet_spot'] = $this->login_model->selectRoundId_by_round($select)->result();

        print_r ($data);
    }

A: Try this may help you,
In view make link like this,
<a href="<?php echo site_url('account/detail_round_id/'.$data->round_id);?>"><?php echo $data->round_id;?>

And in controller add parameter like this,
public function detail_round_id($id){
    $data['tbl_bet_spot'] = $this->login_model->selectRoundId_by_round($id)->result();
    print_r ($data);
}

A: view page you pass value like this
  <a href="<?php echo base_url(); ?>account/detail_round_id/select/<?=$data->round_id?>"><?php echo $data->round_id;?>

In controller get value like this
 $select=$this->uri->segment(4);

hope this will help

redpajama_set_name: RedPajamaStackExchange


Iyanya has released a new single titled 'Iyanu'. Iyanu (Holy Water) is a Yoruba word that means miracle. The 'Iyanu' song was released globally for streaming and broadcast on Tuesday, November 7th, 2017.
Meanwhile, an official video for the song which was shot in Italy by European Director, Omar will be released on Tuesday, November 14th.


text:
Trade Policy and Moral Leadership
David Warsh says that if the US wants to reestablish itself as a world leader in trade policy, it must "regain its traditional leadership in human rights." That means the next president must acknowledge the "grim, incontrovertible truth" that US "officials committed what a dispassionate jury would recognize as a series of serious crimes under American law in the years after 2001:
On the Significance of Geneva, by David Warsh: It's August. Is anyone surprised that trade talks in Geneva collapsed last week… As the Financial Times put it, "…the annual breakdown of the Doha round of trade talks is becoming a summer ritual. … It is time to be brave, swallow hard and accept that the Doha round in its present form has failed.†Indeed. …
What's going on in the world of international trade? … To think genuinely august thoughts about what might come next in global trade, step back and ponder events on a grand scale. A couple of recent books come in handy here. …
Power and Plenty: Trade, War and the Global Economy in the Second Millennium, by Ronald Findlay … and Kevin O'Rourke … is the result of a collaboration between a political economist and a historical economist. A Splendid Exchange: How Trade Shaped the World, by William Bernstein, provides the movie version of the story. … You can get the flavor of the man and his … book from his well-attended website, Efficient Frontier.
It is Power and Plenty that has captured my attention. … I am not … going to summarize a thousand years of international trade, except to say the authors organize their account around what they say are the three great events of the last thousand years: the Black Death of the fourteenth century; the "discovery†and incorporation of the New World into the Old starting in the sixteenth century; and the Industrial Revolution, beginning in the nineteenth century. Nor should you spend much more of your time here. …
I will say that the dustjacket … conveys the book's takeaway message. The authors write, "The greatest expansions of world trade have tended to come, not from the bloodless tântonnement of some fictional Walrasian auctioneer but from the barrel of a Maxim gun, the edge of a scimitar, or the ferocity of nomadic horsemen.â€
Thus successive epochs in global trade have, for the most part, been demarcated by the outbreaks of major wars or imperial expansions: Pax Mongolica, Pax Manchurica, Pax Mughalica, Pax Britannica, Pax Americana. "Each era can be seen as one in which trade is conducted within a geopolitical framework established by the previous major war or conflict, that is in turn altered by the outbreak of the next war, setting the stage for the next epoch, and so on. It is natural to suspect that the accumulating economic and geopolitical tensions unleashed in the course of each period of peace, prosperity and trade culminate in successive rounds of conflict, so that wars, rather than being exogenous or external shocks to the world system, have been inherent in its very nature as it has evolved over the past millennium.â€
That's very far from the concerns for farmers' shares that scuttled the talks in at World Trade Organization headquarters in Geneva last week. Nobody will go to war over India's attempt to protect itself from cheap agricultural exports from Brazil. What, then, are the accumulating tensions that engage the attention of Professors Findlay and O'Rourke? It's central Asia and the Caucasus that interest them, and the various routes from their petroleum riches to Western Europe and the Indian Ocean. …
For in the end, a book about trade is a book about geography – about how the human-built world grew up around straits, harbors, rivers, roads, and natural resources. Writers less well-grounded in the interdependence between trade and politics would have been astonished to find the twenty-first century dawn with the United States embroiled in a two wars along the route that Alexander the Great took to India 2300 years before, but not Findlay and O'Rourke. The US occupation of Iraq in 2003 inflicted a "devastating blow†to American prestige and its claims of moral leadership, they say. It may yet turn out that common threats emanating from the Eurasian heartland will force Japan and Western Europe to make common cause with their US ally once again.
The moral of Power and Plenty is that, instead of worrying about the collapse of the Doha Round, friends of trade should be thinking hard about what comes next. The fifty years of the Cold War otherwise had little to recommend it, but at least it imposed a collective discipline on the two major blocs and their client states. In a world of many different blocs, it appears that Pax Americana will be replaced by some new set of stabilizing rules. But what will they be?
That is what makes the next election so interesting. There seems to be widespread agreement that, in the first years of the new century, the leadership of the United States lost its way and behaved in uncharacteristic ways. The essence of the departure, however, is in much dispute.
I don't mean the decision to invade Iraq. That f
alls somewhere along a continuum that runs from Woodrow Wilson's decision to intervene in Europe during World War I through Harry Truman's decision to go to war in Korea in the summer of 1950 to Lyndon Johnson's deceptive escalation of the war in Vietnam. Reasonable people can disagree.
I mean the assault on the principle that there exist basic human rights that must be observed and protected, even in time of war.
Jane Mayer is the former Wall Street Journal reporter who, in thirteen articles in the New Yorker since 2001, has documented the most important details of the behind-the-scenes American response to the 9/11 attacks. In her new book, The Dark Side: The Inside Story of How the War on Terror Turned into a War on American Ideals, she concludes by comparing the Bush administration's panic to President Roosevelt's internment of Japanese-Americans during World War II.
To me, that similitude seems too charitable. From Mayer's account, it appears clear that US leaders in the office of the Vice President and the Justice Department engaged in a fundamental repudiation of the rules for the treatment of non-combatants and prisoners of war hammered out in treaty negotiations over 150 years and collectively known as the Geneva Conventions.
On the strength of reporting by Mayer and others, there is good reason to suspect that that these officials committed what a dispassionate jury would recognize as a series of serious crimes under American law in the years after 2001 – in Iraq, in the "dark prisons†in Europe, in the detention center in Guantanamo Bay, Cuba.
I don't know whether or not it is necessary that these officials be prosecuted. Probably not: these matters are incredibly divisive. But if the US is to regain its traditional leadership in human rights – meaning if it is once again to become the world leader in trade policy, among many other things – its leaders, including both John McCain and Barack Obama, sooner or later must acknowledge this grim, incontrovertible truth.
I disagree on prosecution. Justice alone is enough to compel prosecution – people should be responsible for their actions – but the moral hazard issue is also worth worrying about. What's to stop another administration from doing the same or worse? I don't expect it will happen, but I don't care what their title is or who might get upset about it, people who broke the law as it relates to torture or other crimes need to be prosecuted for it. Human rights begin at home.
[Let me add one more thought. In financial markets, the argument for ignoring moral hazard and letting some people escape responsibility for bad choices is that the punishment – letting banks fail – would potentially cause excessive harm to the the innocent by bringing down the larger economy. If the harm can be confined to those who caused the problem, or if the harm is minimal (there's always some social cost to prosecution), then there is no reason to intervene, and good reasons to let banks fail.
I suppose a similar argument can be made here. Prosecuting for these crimes would create so much divisiveness that it would endanger the system by bringing it down now, or by doing long-term harm to fundamental institutions. Obviously I don't buy that argument, I don't think there's much of a chance it would bring down our system of government, and I think the long-term danger to our important institutions is larger if we don't prosecute.]
Posted by Mark Thoma on Tuesday, August 5, 2008 at 02:07 AM in Economics, International Trade
Permalink TrackBack (0) Comments (31)
The Anticommons
Can "too much ownership" cause gridlock and the "underuse and waste" of resourses?:
The Permission Problem, by James Surowiecki: In the second decade of the twentieth century, it was almost impossible to build an airplane in the United States. That was the result of a chaotic legal battle among the dozens of companies—including one owned by Orville Wright—that held patents on the various components that made a plane go. No one could manufacture aircraft without fear of being hauled into court. The First World War got the industry started again, because Congress realized that something needed to be done to get planes in the air. It created a "patent pool,†putting all the aircraft patents under the control of a new association and letting manufacturers license them for a fee. Had Congress not stepped in, we might still be flying around in blimps.
The situation that grounded the U.S. aircraft industry is an example of what the Columbia law professor Michael Heller, in his new book, "The Gridlock Economy,†calls the "anticommons.†We hear a lot about the "tragedy of the commonsâ€: if a valuable asset (a grazing field, say) is held in common, each individual will try to exploit as much of it as possible. Villagers will send all their cows out to graze at the same time, and soon the field will be useless. When there's no ownership, the pursuit of individual self-interest can make everyone worse off. But Heller shows that having too much ownership creates its own problems. If too many people own individual parts of a valuable asset, it's easy to end up with gridlock, since any one person can simply veto the use of the asset.
The commons leads to overuse and destruction; the anticommons leads to underuse and waste. … Even divided land ownership can have unforeseen consequences. Wind power, for instance, could reliably supply up to twenty per cent of America's energy needs—but only if new transmission lines were built, allowing the efficient movement of power… Don't count on that happening anytime soon. Most of the land that the grid would pass through is owned by individuals, and nobody wants power lines running through his back yard.
The point isn't that private property is a bad thing, or that the state should be able to run roughshod over the rights of individual owners. Property rights (including patents) are essential… But property rights need to be limited to be effective. The more we divide common resources like science and culture into small, fenced-off lots, Heller shows, the more difficult we make it for people to do business and to build something new. Innovation, investment, and growth end up being stifled. …
In theory, one should be able to break a gridlock by striking a deal that would leave all sides better off. Sometimes that happens. … [But…] One reason deals founder is that there are simply too many interested parties. If, in order to create a new drug, you have to strike bargains with thirty or forty other companies… often things go awry because owners won't make a deal at a reasonable price…
Recent experimental work by the psychologist Sven Vanneste and the legal scholar Ben Depoorter helps explain why. When something you own is necessary to the success of a venture, even if its contribution is small, you'll tend to ask for an amount close to the full value of the venture. And since everyone in your position also thinks he deserves a huge sum, the venture quickly becomes unviable. So the next time we start handing out new ownership rights—whether via patents or copyright or privatization schemes—we'd better try to weigh all the good things that won't happen as a result. Otherwise, we won't know what we've been missing.
Trade Policy and Moral Leadership added by Mark Thoma on August 26, 2008

redpajama_set_name: RedPajamaCommonCrawl


text:
const project = require('../config/project.config')
const server = require('../server/main')
const debug = require('debug')('app:bin:server')

server.listen(project.server_port)
debug(`Server is now running at http://localhost:${project.server_port}.`)

redpajama_set_name: RedPajamaGithub


text:
Just Garden Storage helps you tidy the clutter, transforming your patio or garden into a neat, clutter free zone.
We have updated our Privacy and Cookie Policy as shown below to comply with the General Data Protection Regulation (GDPR), which came into effect on May 25 2018.
Just Garden Storage is a trading name of Westcountry Brands Ltd. Registered in England: 07959428.
About this Privacy and Cookie Policy
By accessing our website www.justgardenstorage.co.uk you agree that our Privacy and Cookie Policy will apply whenever you access this website on any device. For privacy related questions please write to our Data Protection Officer at the address below.
Whilst most updates and amendments to this Privacy and Cookie Policy will be minor, Just Garden Storage may change this policy from time to time, and at Just Garden Storage sole discretion to meet the requirements of our business and in order to comply with changes to the General Data Protection Regulation (GDPR). Changes will be updated on this website www.justgardenstorage.co.uk
Access to your information and correction
You have the right to request a copy of the information that we may hold about you. If you would like a copy of some or all of your personal information, please write to our Data Protection Officer at the address below.
We aim to make sure that your personal information is accurate and up to date. You may ask us to correct or remove information you think is inaccurate. We take the security of your personal data seriously and have adopted a policy of compliance with the data protection laws of the United Kingdom to take reasonable care to prevent any unauthorised access to personal data.
Cookies
Like other websites we use cookies on www.justgardenstorage.co.uk Cookies are small text files stored on your computer by your browser e.g Chrome, Safari, Firefox, Internet Explorer.
Cookies contain information about the use of your computer but do not include personal information about you (they do not capture your name, for example). They are used for many things, such as remembering whether you have visited our site before, so that you remain logged in, or to help us work out how many new website visitors we receive each month.
Modern internet browsers accept cookies automatically but you can change the settings of your browser to erase cookies or prevent automatic acceptance if you prefer.
The following links explain how you can control cookies in your browser. Please note that if you turn off cookies in your browser, the changes you make will apply to all websites not just www.justgardenstorage.co.uk:
Internet Explorer: https://support.microsoft.com/en-gb/help/17442/windows-internet-explorer-delete-manage-cookies
Chrome: https://support.google.com/chrome/answer/95647?co=GENIE.Platform%3DDesktop&hl=en
Safari: https://support.apple.com/en-gb/guide/safari/manage-cookies-and-website-data-sfri11471/mac
Social Media and Third Party Cookies
Links to social media websites that we use are run by other companies. These companies place cookies on your computer when you use them on our site or if you are already logged in to them. You can find out more about these third party companies Privacy and Cookie Policies here:
Facebook (Facebook's data policy): https://www.facebook.com/privacy/explanation
You Tube (YouTube video player cookie policy): https://support.google.com/youtube/answer/2801895?hl=en-GB
Twitter (Twitter's privacy policy): https://twitter.com/privacy
Google Analytics
We use Google Analytics on our website www.justgardenstorage.co.uk to help us understand how visitors interact with our website and for aggregating information about the traffic to our site. We do not enable Data Collection for Advertising Features or Remarketing purposes, nor do we run Google AdSense advertising on our website.
We also anonymize your IP address as standard to protect your privacy.
Google's privacy policy can be found here: https://policies.google.com/privacy
You can also opt out of this if you want with this extension: https://tools.google.com/dlpage/gaoptout
www.justgardenstorage.co.uk features links to selected travel partners and third party websites. Our privacy policy only applies to Just Garden Storage so when you click on an external link to one of these third party websites or use our handy booking widget you should always take the time to read their own, specific privacy policy.
We earn money when you go on to make a purchase with one of our partners, having used www.justgardenstorage.co.uk. When you click through to a third party website from one of our links and make a purchase, the third party company pay us a fee, which does not impact on the price you pay!
justgardenstorage.co.uk © 2020 is a participant in the Amazon EU Associates Programme, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to amazon.co.uk
At no stage in the process do we collect personal data on you or your purchase, this is handled by the third party website you buy from.
We do not give paid advertisers personal data from Just Garden Storage. Should you chose to visit their website your contract will be with them. Again, no personal data is taken by Just Garden Storage.
We also serve a number of external adverts on our website together with sponsored links. These external links include clickable text, banner and / or image links that will take you to specific third party websites. These third party websites will typically have detailed privacy policies in place, relating directly to the adverts they serve.
We do not gather personal data about the adverts and sponsored links you click on. Any cookie placed on your computer will be done so by the third party website.
Contact Forms
Our contact form asks you to read this Privacy and Cookie Policy and tick the box to agree to the use of the information you provide being used to contact you. Data is not used for any other purpose other than to answer your enquiry.
If you have questions about this Privacy and Cookie Policy, please write to: Westcountry Brands Ltd, 20-22 Wenlock Road, London N1 7GU.


Metaglossary.com - Definitions for "clock"
A machine for measuring time, indicating the hour and other divisions; in ordinary mechanical clocks for domestic or office use the time is indicated on a typically circular face or dial plate containing two hands, pointing to numbers engraved on the periphery of the face, thus showing the hours and minutes. The works of a mechanical clock are moved by a weight or a spring, and it is often so constructed as to tell the hour by the stroke of a hammer on a bell. In electrical or electronic clocks, the time may be indicated, as on a mechanical clock, by hands, but may also be indicated by direct digital readout, with the hours and minutes in normal Arabic numerals. The readout using hands is often called analog to distinguish it from the digital readout. Some clocks also indicate the seconds. Clocks are not adapted, like the watch, to be carried on the person. Specialized clocks, such as atomic clocks, may be constructed on different principles, and may have a very high precision for use in scientific observations.
A steady pulse from a generator which is used for synchronizing sequencers, drum machines, etc. Common sequencer timing clock rates are 24, 48, 96,120,240,384,or 480 pulses-per-quarter note. MIDI timing clocks run at a rate of 24 ppqn. Also called the resolution, PPQ, ticks, etc of a sequencer, ie Logic Audio uses 980 pulses-per-quarter note to control timing of MIDI events.
The device used to time the 25-minute-per-game allotment per player. Electronic timers are preferred followed by analog timers, such as those used in chess.
A device providing signals used in a transmission system to control the timing of certain functions such as the duration of signal elements or the sampling rate. A device that generates periodic, accurately spaced signals used for such purposes as timing, regulation of the operations of a processor, or generation of interrupts.
A timing reference signal, e.g., on an MVIP bus.
An oscillator-generated signal that provides a timing reference for a transmission link; used to control the timing of functions such as sampling interval, signaling rate, and duration of signal elements.
A device or instrument for maintaining and displaying time.
A steady pulse from a generator which is used for synchronizing sequencers, drum machines, etc. Common sequencer timing clock rates are 24, 48, or 96 pulses-per-quarter note. MIDI timing clocks run at a rate of 24 ppqn. The EIII clock runs at 96 ppqn.
Equipment that provides a timing signal.
In serious TOURNAMENTS and MATCHES each player has a fixed amount of time to play either a certain number of moves or the whole game. A player who exceeds the time limit loses as long as his opponent has enough MATERIAL left to get CHECKMATE. A chess clock has two races. On making a move the player presses the button on top of his clock to start his opponent's clock ticking. Digital clocks are now being used in many TOURNAMENTS.
Most churches have them, not all are working. Where churches have clocks and bells, they are usually linked together so that the bells will sound the hours, and usually quarters as well.
A chip on the motherboard that regulates the timing of events in the computer.
Digital signal in the form of a rectangular pulse train or a square wave.
An electronic circuit that generates evenly spaced pulses at speeds of millions of cycles per second. The pulses are used to synchronize the flow of information through the computers internal communication channels.
A signal defined for the purpose of synchronizing the actions of a system component, so that the behavior is reliable, repeatable, and stable. Clocks controling the data path vs. the control path of a design thread are represented separately in flowHDL.
A special circuit that sends pulses of current to the CPU and other computer components.
The electronic circuit in a computer that generates a steady stream of timing pulses - the digital signals that synchronize every operation. The system clock signal is precisely set by a quartz crystal, typically at a specific frequency between 1 and 50 megahertz. The clock rate of a computer is one of the prime determinants of its overall processing speed, and it can go as high as the other components of the computer allow.
The device used for timing total time taken by each player during a game. Generally, any fraction of a minute over time costs the player 10 points PLUS each further minute over time attracts an additional 10-point penalty. ANALOGUE CLOCK Has two clock dials plus a flag on each side which drops when the minute hand exactly reaches the number 12. DIGITAL CLOCK Has a digital display on each side which will display a negative number when a player has gone over time.
Devices which produce a clock pulse used to ensure the correct timing of other devices on the board / schematic.
On computers the system clock is an electronic circuit, which keeps a constant stream of pulses. These pulses help to synchronize the computer operation. Generally it is a quartz crystal, which is operating between 1-50 Mega Hertz (MHz). The clock speed helps to determine the overall processing speed of the computer system.
A timing source for charge transfer functions within CCD chips.
The time-of-day Clock appears on the screen whenever you change channels or press the DISPLAY key.
an instrument for time measuring, in seconds, minutes and hours, as analog or digital formats.
Any electronic oscillator used to generate a timing reference signal. Its purpose is to synchronise the data being taken from a disc by a CD or DVD player.
Paired clocks used in all official tournaments and in club games. After a player moves, he depresses a lever that stops his clock and starts his opponent's. Each clock, therefore, registers only the elapsed time for one player. If a player exceeds the time limit set on his clock, a flag falls and he loses the game, even if he has a clear winning position.
An oscillator producing timing pulses to synchronize various elements of a system. In switching mode power supplies, a clock is used to produce the power pulses that are modulated to control power transfer. In digital interfaces that communicate on a bus (such as the IEEE-488) a clock is used to synchronize the data transfer and commands.
A Front Panel control that displays the local time.
A square wave used to synchronize several different circuit elements.
a source of timing information, especially important when two or more devices must be synchronized. See synchronization.
The source of a computer= s timing signals. It synchronizes every operation of the CPU. Some newer CPU= s are able to perform more than one task per clock cycle.
A device that generates accurate, periodic signals for synchronisation purposes.
n 1: The oscillator that generates the timing pulses that coordinate the parts of and enable the flow of data within a digital device. 2: A circuit powered by battery that keeps track of the date and time for human and various system requirements.
A square waveform used for synchronizing and timing of several circuits.
The timing circuit for a microprocessor.
The clock not only keeps track and displays the system date and time, its most important function is to regulate the timing and speed of all the functions performed by your computer.
a circuit or device that generates a periodic signal used for timing or synchronizing signals.
(Horloge) : Function allowing the computer to maintain date and time, and to calculate the elapsed time between two events.
A timing device that generates the basic periodic signal used as a source of synchronizing signals in digital equipment.
Regular low frequency pulse or square wave used for driving sequencers arpeggiators etc.
A clock signal (in electronic music) is a regular series of electronic pulses sent from one sequencer to another in order to keep the two units properly synchronized (a system which is mainly associated with drum machines). In MIDI, the clock signal is a regular series of MIDI clock messages, rather than just a simple series of pulses.
Timing signals used in synchronous transmission. More generally: the source(s) of timing signals sequencing electronic events.
The internal system which provides the timing or cycle time constraint which controls the allowed bus transit time for data. The time window under which data and instructions can be internally processed in a computer. This directly relates to the speed of the internal computer.
The clock, in cryptology, was a method devised by Polish mathematician-cryptologist Jerzy Różycki, at the Polish General Staff's Cipher Bureau, to facilitate decrypting German Enigma messages. This method sometimes made it possible to determine which of the Enigma machine's rotors was at the far right, that is, in the position where the rotor always revolved at every depression of a key.
A figure or figured work on the ankle or side of a stocking.
To ornament with figured work, as the side of a stocking.
A knitted or embroidered ornament on a stocking, usually running up the side.
An ornament on the side (usually over the ankle) of a stocking.
also known as a molecular clock. One of the most important assumptions that is often made using molecular data is the existence of a clock. In other words, there is sometimes an assumption being made either implicitly or explicitly that the rate of change of residues in molecules is approximately constant. Making this assumption is fraught with dangers, but a great many empirical studies have shown that (with some errors) the assumption of a clock often holds true. If there are some good reference points (say, fossils or significant geological events) it is sometimes possible to use molecular data to estimate the time of events for which there is no other alternative.
A watch, esp. one that strikes.
The striking of a clock.
A large beetle, esp. the European dung beetle (Scarabæus stercorarius).
Clock was primarily Stu Allen and Pete Pritchard, who were the nucleus of this house, turned eurodance, turned disco act. In 1994, they hailed themselves as the British "2 Unlimited" when they recruited rapper Marcus Thomas, and vocalist Lorna Saunders to front the act. Marcus left in 1998, to join the band Tzant, to be replaced by Ché-gun Peters.
Clock is the name of a Swedish hamburger restaurant chain originally founded by an off-shoot from the US based Carrols. The chain suffered from mismanagement and declared bankruptcy; as the government had acquired the forfeited company, it was incorporated into the state-owned restaurant chain SARA.
To call, as a hen. See Cluck.
A media object that defines a transformation on a TimeBase.
Clock is when the disc is spinning clockwise, i.e in the same direction of the clock.
The completion of one cycle of a frequency.
A continuous variable that changes with a constant rate.
The code used to identify the college or main department from which an employee is paid.


Visiup company has a resort for sale Vietnam for foreigners in the Vietnamese territory.
Beautiful resort for sale in Vietnam in Phan Thiet. It is luxurious, spacious and relaxing. It also has a restaurant, pool, spa, suite, sauna and green space... The resort is located in front of the sea in a area paradise. This resort can attract, seduce and retain all customers.


Yes! You can use either an email client such as Outlook to manage Alumni Email for Life, or simply use the web interface. Client configuration instructions can be found at http://help.outlook.com. Please remember that your username in this case will be username@tulane.edu.
Office 365 is hosted by Microsoft and therefore you will manage your Alumni Email for Life password through Alumni Email for Life as opposed to the Tulane University Password Portal. For instructions on changing your password please click here.


text:
Request DemoClient Login
Reputation Intelligence
RepTrak Platform
What is RepTrak
Reputation Insights
RepTrak Difference
Leading and Lagging: How the 100 Most Reputable Companies are (and aren't) meeting environmental expectations
Corporate Responsibility/ESG
Reputation Risk
Higher Purpose
28 Apr 2021Corporate Responsibility/ESG
Time is running out for climate action — and for businesses to contribute to the solution.
As the Leaders' Summit on Climate concludes and Earth Day passes, it is a race against time to aggressively reduce greenhouse gas emissions, reduce plastic, and protect marine ecosystems. Governments are deepening their national strategies and targets, while investor expectations are mounting for businesses to deliver against their climate pledges. This isn't just a call for government action. The public is specifically looking for corporations to step up.
And the public is paying attention.
Corporate environmental promises aren't necessarily new, but the expectation for companies to take accountability for their environmental impact and take action is growing. The making and breaking of these promises influences public perception, boosting and damaging brand power and reputation.
We turned to our 2021 Global RepTrak 100 Most Reputable Companies to study the power of environmental action on reputation and perception. RepTrak's annual study is the definitive ranking of corporate reputation for the world's leading companies, showcasing how people feel, think, and act towards brands – all based on RepTrak's proprietary reputation analytics and data.
Read the full Global RepTrak 100 report here.
The Top 100 are determined by 7 key drivers: Performance, Products/Services, Leadership, Citizenship, Governance, Workplace, and Innovation.
RepTrak also measures an ESG (Environmental – Social –Governance) Score, reflecting public perception of performance against 17 factors. These include environmental considerations like: acts responsibly to protect the environment, reduces its environmental footprint, manages energy efficiently, and is a responsible user of natural resources, among others.
In the sink-or-swim year of 2020, the Top 100 rose to the occasion and achieved a highest ever score of 74.9.
Although RepTrak's Top 100 are exemplary in their key reputation drivers, many receive average marks across the board for "acting responsibly to protect the environment."
In fact, ESG Scores (Environmental - Social - Governance) were lower than the average Reputation Scores, with significant variation across the individual Environmental, Social, and Governance Scores. Globally, the overall ESG Score was an "average" 69.5, with Social receiving the highest perception score, followed by Governance and then Environment.
RepTrak data reveals a low ESG score results in a 20% willingness to buy, while a high ESG score results in a 60% willingness.
This leaves room for improvement.
In July 2020, BlackRock Inc. issued a report that called out 244 companies for failing to prepare their operations for climate change or inform their investors of the risks.
And the Top 100 are working on it. Sixty percent of the Top 100 have established Science-Based Targets (SBTi) for their carbon emissions reduction strategies, while 1 out of 3 have set a target to use 100% Renewable Energy.
More than one-third have communicated a goal of Net Zero Emissions by 2050 (if not sooner), with many signing onto the Climate Pledge to achieve carbon neutrality by 2040. While this is higher than the estimated 20% of the Fortune 2000 who have set Net Zero Emissions targets, many of these commitments have been made within the past year and may not be known to the general public.
Some Top 100 companies have even more ambitious pledges, including:
#10 Microsoft: Net Zero by 2030, with an additional goal of removing all carbon emitted since it was founded in 1975
#36 IKEA: "Climate positive" by 2030
#46 Apple: Net Zero by 2030
#92 Amazon: co-founded The Climate Pledge in 2019, aiming to be Net Zero by 2040 and now over 100 companies have signed on (e.g., #50 Colgate-Palmolive, #26 Visa).
Environmental perception is just one factor to influence reputation, but as the environmental conversation heats up (literally), its importance to reputation will only grow.
And consumers are noticing what is – and isn't – said about environmental efforts.
O2, a UK-based telecommunications company, recently launched their O2 vs CO2 campaign. This proactive multi-channel campaign communicates their initiative to buy 100% renewable electricity to power their network. The desire to engage a mass-audience and spread awareness of O2's environmental efforts was influenced, in part, by their RepTrak insights.
O2 is the only mobile network operator to publicly commit to net zero carbon by 2025, and the only network to receive the highest certification for carbon reduction in their supply chain.
Dedicated to listening to their audience and community, RepTrak's data highlighted that while the brand has strong environmental credentials, O2's sustainability commitments were not widely known or understood. These insights encouraged O2 to build a creative and impactful campaign to raise awareness of their environmental track record and ambitious Net Zero commitments, while also strengthening citizenship and ESG perceptions amongst key audiences.
As corporate expectations rise, O2 has taken a proactive approach, demonstrating their commitment to their customers and to tackling climate change – and then communicating it deliberately.
Even for the Top 100, the commitment to sustainability initiatives is very loud and shiny, with updates and setbacks quiet. Advertising and communications are crucial in maintaining transparency, accountability, and positively building reputation.
As companies tackle their environmental promises, they won't just have to make and accomplish serious environmental goals – they'll have to communicate their efforts and results very clearly.
Curious about how the public perceives your ESG efforts? Request a demo.
Related <b>{category}</b> stories
VIDEO: RepTrak in 60 seconds
RepTrak provides reputation management software, measuring and monitoring corporate reputation in near real time. We break it down in 60 seconds.
FAST FACTS: 2021 Airline Reputation
Throughout 2020 and 2021, airline reputation ebbed and flowed with COVID anxieties and restrictions. We took a deep dive into our proprietary reputation data to understand their unique reputation journey and compiled the Fast Facts.
Home for the holidays: How airline reputation faired in 2021
Amid the swirling chaos of travel restrictions and vaccine hesitancy and demands, air travel became a peculiar stage to act out our frustrations and anxieties. The flights are nonstop and so is our corporate reputation tracking software, so we took to the friendly skies for a closer look at how airline reputation faired from 2020 – 2021.
Show all stories about Corporate Responsibility/ESG →
RepTrak
Boston Global HQ 399 Boylston Street, Floor 7, Boston, MA 02116 +1 617 758 0955, info@reptrak.com


text:
Vintage & Antique Engagement Rings
Ogham Jewellery
Pre-Loved Jewellery
Remodelling Jewellery
Established over 30 years ago after Carol returned from Thailand where she had lived for many years training in goldsmithing and studying gemstones. Over the years, Carol continued to study and train in gemstones and today she is Irelands first and only female member of The Institute of Registered Jewellery Valuers in London and Irelands first female Fellow of the institute in London. Carol is a full graduate Gemmologist from the Gemological Institute of America and Fellow of the National association of Goldsmiths of Great Britain.
For valuations www.dublinjewelleryvaluations.ie
Specialising in handmade engagement rings and jewellery to suit your budget
Carol Clarke Jewellers is a family run business specialising in diamond engagement rings and unusual gold jewellery for thirty years. We are situated in 7 Royal Hibernian Way Dawson Street Dublin 2. The business was first started by Carol Clarke over twenty years ago.
Carol, FIRV., FNAG., G.G (GIA), AJP (GIA), P. J. Val Dip, R.J Dip, BJIG, trained as a goldsmith in Bangkok, Thailand, while studying jewellery design at the Bangkok Jewelry Institute. Carol also studied at the Asian Institute of Gemological Sciences in Bangkok. After her training, Carol came back to Ireland and started her own business designing and making unique pieces of jewellery in gold, platinum and silver. Carol also studied with the National Association of Goldsmiths of Great Britain and holds the Retail Jewellers Diploma with distinction. Carol is also the only person in Ireland to have completed the Gemological Institute of America Insurance Replacement Appraisal examination which is just one of a number of gemological examinations that Carol has achieved.
Carol FIRV., FNAG., G.G (GIA), AJP (GIA), P. J. Val Dip, R.J Dip, BJIG works with her husband Eamon who also studied art in Dublin and who has won numerous awards over the years for his designs and workmanship as a goldsmith.Colette also holds a Retail Jewellers Diploma from the National Association of Goldsmiths of Great Britain. Ann, Marie and Sandra also hold various jewellery qualifications from various jewellery associations, so together we make a group of very qualified jewellers! We feel that we offer you a very high and informative standard of service.
We are sure you will find something of interest on our site from our Claddagh rings, Celtic engagement rings, Ogham rings, Claddagh pendants, Celtic wedding bands, etc.
Carol Clarke
7 Royal Hibernian Way,
Dawson Street,
info@carolclarke.ie
© 2023 Carol Clarke - All Rights Reserved. Web Development by Pro Web Design


This year we had excelent place in Los Lances beach. The riders had fight in Freestyle and big air. 7 days of different weather conditions has shown us beutifull tricks and unforgetable show of the strongest riders in the World!


Artists in Community Partnerships – This program invests in Ventura County by partnering qualified, accomplished artists from all disciplines with public and private organizations and businesses to address problems and opportunities through creative and artistic means. Applications may be submitted by the project artist or the partnering entity. The applications are reviewed by a panel created by the Arts Council for consistency with the objectives of grant program. The funding from the Arts Council varies from year to year based on contributions from supporting foundations. Each project is generally funded on a three-part basis: one third from the partnering entity, one third by the Arts Council (up to a prescribed limit), and one third by the artist raising funds from the community at large. The at-large contributions are often raised through such on-line fund-raising sites as KickStarter.
The program is announced through the media and the Council's own contact list (a good reason to be "Registered" as an Affiliate with the Council). Interested parties are invited to an information session where the program is explained and examples of past projects are presented. Once the winners have been announced, a limited amount of time is allowed to collect the required funds to complete the project. Most projects must be completed within a year. Plays based on interviews with war veterans have been written (and later staged), public art installed, and fire memorials created (among other projects) as a part of this program.
Images of past projects Is this a good way to insert images?


text:
Home :: Health
Effective vaccine against HIV/AIDS comes closer to reality
Washington, Thu, 08 Mar 2012 ANI
Washington, March 8 (ANI): Scientists including one of an Indian origin may be on the verge of developing a vaccine that will provide long-lasting protection against repeated exposures to human immunodeficiency virus (HIV).
The team at Emory University and GeoVax Labs, Inc. has developed a vaccine that can protect nonhuman primates against multiple exposures to simian immunodeficiency virus (SIV) given in three clusters over more than three years. SIV is the nonhuman primate version of HIV.
Harriet L. Robinson, PhD, chief scientific officer at GeoVax Labs, Inc., and former director of the division of microbiology and immunology at Yerkes National Primate Research Center, has been leading the research team with Rama Rao Amara, PhD, associate professor of microbiology and immunology, Yerkes National Primate Research Center and the Emory Vaccine Center.
The vaccine regimen included a DNA prime vaccine that co-expressed HIV proteins and granulocyte-macrophage colony-stimulating factor (GM-CSF). GM-CSF is a normal protein that promotes the initiation of immune responses and thus enhances the ability of the vaccine to elicit blocking antibodies for the SIV virus before it enters cells.
Vaccination consisted of two DNA inoculations at months 0 and 2 to prime the vaccine response and then two booster inoculations at months 4 and 6. The booster vaccine was MVA, an attenuated poxvirus expressing HIV proteins. Six months after the last vaccination, both vaccinated and unvaccinated animals were exposed to SIV through 12 weekly exposures, resulting in an 87 percent per exposure efficacy and 70 percent overall protection.
Over the next two years uninfected animals were exposed multiple times in two more series, resulting in an 82 percent per exposure efficacy during the second series and an 84 percent per exposure efficacy during the third series.
"Repeated challenges in animals are used to mimic sexual transmission. The hope is that the results in the nonhuman primate models will translate into vaccine-induced prevention in humans," said Robinson.
"It is impressive to note that protection could be observed against both neutralization sensitive and neutralization resistant viruses," said Amara.
Neutralization is the process by which some antibodies can block virus infection.
A first generation GeoVax DNA/MVA vaccine that does not co-express GM-CSF has shown excellent safety and reproducible vaccine responses in Phase 1 and 2a clinical trials in more than 400 uninfected people.
These trials, supported and conducted by the National Institutes of Health HIV Vaccine Trials Network, have set the stage for the second-generation GM-CSF co-expressing vaccine to move from its initial Phase 1 safety testing slated to start in March of this year to a Phase 2b efficacy trial in participants who are at high risk of exposure to HIV. The vaccine is designed for a version of the virus prevalent in the Americas.
The research was presented Wednesday at the Conference on Retroviruses and Opportunistic Infections (CROI) in Seattle, Wash. (ANI)
Read More: Khyala Phase - I | Khyala Phase - Ii | Okhla Industrial Area Phase-i | Dakshinpuri Phase-i | Dakshinpuri Phase-ii | Dakshinpuri Phase-iii | East Of Kailash Phase-i | Roop Wash | Roop Wash Pachgain | Amara | N.v.colony | H.b.colony Alwar | 3 Eme Center | W.r.s.colony | R.s.p.colony Gangrel | Spinning Mills Center | Jp Nagar Iii Phase | T.k.colony | Indian Research Po | Mankundu Govt.colony


text:
© ONDINE COHANE
Author & Journalist
La Bandita
Eat, Stay, Buy Locally: Treading Lightly on the Road (The New York Times)
by Ondine Cohane
Between the intricate shrines of Angkor Wat, the diverse landscapes of the countryside and the kindness of its people — and despite the horrors of its recent history — Cambodia knocks a lot of people off their feet. It certainly had that effect on me when I first visited a few years ago and then …
Following Dylan Thomas in Wales (The New York Times)
Climbing along a steep coastal path through a forest in southern Wales, with russet red and tawny brown autumn leaves crunching beneath my feet, I reached a crest where the trailhead looked back onto a long estuary lined with salt flats. The River Taf ran through the headlands before me, its glacier-cut course unmistakable …
Finding a Setting That Captures a Scene (The New York Times, Feb 2014)
Wes Anderson's films are best known for their eccentric characters and rigorously whimsical tone. But just as distinctively captured are their settings. Think of the colors and textures of the Indian landscapes in "The Darjeeling Limited," for example, or the lovingly rendered Mediterranean locations in "The Life Aquatic With Steve Zissou." The director's latest …
CohaneTravels:Update Bangkok
As many of you know, Bangkok has become one of my very favorite cities in the world. And I am not alone in my desire to visit it frequently: according to the new Global Destination Index it is now the most visited city on the globe, even more than spots like New York and London. …
Ondine Cohane
After Ondine Cohane edited Conde Nast Traveler's Word of Mouth section for a couple of years, she got her dream job, as a contributing editor at the magazine.
In Norway, the Journey Is the Destination (The New York Times)
Always Italy
By FRANCES MAYES and ONDINE COHANE(National Geographic)
The Innkeeper's Diaries (Journey)
CohaneTravels
Daily Traveler Blog
A writer finds emotional solace on some of…
When Ondine Cohane and her husband opened a…
The Aeolian Islands: Beautiful Italian Countryside, Few Tourists (Conde Nast Traveler)
© 2012 - 2021 - ondinecohane.com - ondine@ondinecohane.com