Melanoma is the most deadly form of skin cancer with a yearly global incidence over 232,000 patients. Individuals with fair skin and red hair exhibit the highest risk for developing melanoma, with evidence suggesting the red/blond pigment known as pheomelanin may elevate melanoma risk through both UV radiation-dependent and -independent mechanisms. Although the ability to identify, characterize, and monitor pheomelanin within skin is vital for improving our understanding of the underlying biology of these lesions, no tools exist for real-time, in vivo detection of the pigment. Here we show that the distribution of pheomelanin in cells and tissues can be visually characterized non-destructively and noninvasively in vivo with coherent anti-Stokes Raman scattering (CARS) microscopy, a label-free vibrational imaging technique. We validated our CARS imaging strategy in vitro to in vivo with synthetic pheomelanin, isolated melanocytes, and the Mc1re/e, red-haired mouse model. Nests of pheomelanotic melanocytes were observed in the red-haired animals, but not in the genetically matched Mc1re/e; Tyrc/c ("albino-red-haired") mice. Importantly, samples from human amelanotic melanomas subjected to CARS imaging exhibited strong pheomelanotic signals. This is the first time, to our knowledge, that pheomelanin has been visualized and spatially localized in melanocytes, skin, and human amelanotic melanomas.
The yearly global incidence of melanoma is over 232,000 individuals, with more than 55,000 of those diagnosed succumbing to the disease1. Individuals with fair skin and red hair exhibit the highest risk for developing melanoma2, with evidence suggesting the red/blond pigment known as pheomelanin may elevate melanoma risk through both UV radiation-dependent and -independent mechanisms. Moreover, red-haired melanoma patients often develop amelanotic lesions, so-called due to the absence of macroscopically detectable dark eumelanin pigments within the visible tumour margin. Because they are harder to recognize upon examination, these tumours are frequently diagnosed at more advanced stages and are associated with higher mortality3. Similar amelanotic melanomas arise in mice bred on the Mc1re/e genetic background, which recapitulates the red-haired, fair-skinned phenotype. The non-visible pheomelanin in these lesions was found to functionally contribute to melanoma formation, as the introduction of an albino allele onto the same genetic background abrogated melanoma risk4. Although the ability to identify and monitor pheomelanin within skin is vital to improve our understanding of the underlying biology of these lesions, no tools exist for real-time, in vivo characterization of the pigment.
Studies using Raman spectroscopy to explore the pigment of red bird feathers previously suggested that a unique vibrational band centred between 2000–2100 cm−1 may be a marker for pheomelanin5,6. However, it was not clear if this weak vibrational resonance could be used for in situ identification of the red pigment. Coherent anti-Stokes Raman scattering (CARS) microscopy, a label-free vibrational imaging technique based on Raman scattering, offers a significantly enhanced signal level compared to spontaneous Raman scattering, and may be suitable for non-invasively identifying pheomelanin inside the skin in real time. Here we present the distribution of pheomelanin in cells and tissues can be visually characterized non-destructively and noninvasively in vivo with CARS microscopy. We validated our CARS imaging strategy in vitro to in vivo using synthetic pheomelanin, isolated melanocytes, and the Mc1re/e, red-haired mice model. Nests of pheomelanotic melanocytes were observed in the red-haired animals, but not in the genetically matched Mc1re/e; Tyrc/c ("albino-red-haired") mice. Importantly, samples from human amelanotic melanomas imaged with CARS microscopy exhibited strong pheomelanotic signals. This is the first time, to our knowledge, that pheomelanin has been visualized and spatially localized in melanocytes, skin, or human amelanotic melanomas.
To demonstrate that CARS microscopy can selectively and sensitively visualize pheomelanin, the pigment was synthesized following an established procedure that makes use of the mushroom tyrosinase enzyme7. When distributed as small particles in a water/hexane emulsion, synthetic pheomelanin was found to yield a very strong CARS signal whose vibrational spectrum corresponds to that observed with Raman spectroscopy (Supplementary Fig. S1)5,6.
With the ability to visualize and spectroscopically confirm the presence of pheomelanin using CARS microscopy, the next step was the detection of naturally-occurring pheomelanin. Red-haired mice (Mc1re/e) were bred to incorporate B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze (Jackson Laboratory catalogue #007906) or B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze (Jackson Laboratory catalogue #007909) and melanocyte targeted CRE (Tyr-CRE)8, thereby enabling FACS-based isolation of neonatal ZsGreen or tdTomato fluorescent tagged melanocytes. A detailed protocol regarding melanocyte extraction can be found in the Methods section. The tdTomato mice were sacrificed, their skin harvested, and dermal cells sorted based on fluorescence of both tdTomato and FITC-labelled antibodies tagging c-Kit, a surface marker9. ZsGreen-labelled mice underwent a similar procedure, with melanocytes selected via their ZsGreen and c-Kit signals. The sorted melanocytes were fixed, and imaged with both CARS and confocal fluorescence microscopy.
As shown in Fig. 1, pheomelanin signals exhibited the predicted perinuclear localization of melanosomes. As a control, genetically matched tdTomato-tagged, tyrosinase-deficient (Mc1re/e, Tyrc/c) "albino-red-haired" mouse-derived melanocytes were subjected to the same analysis and no visible pheomelanin signals were observed (Supplementary Fig. S2). Importantly, CARS spectra acquired from the red-haired mice, but not the albino red-haired mice, matched CARS spectra collected from synthetic pheomelanin, confirming the ability of coherent Raman imaging to selectively visualize the red pigment within cells.
Figure 1: Melanocytes isolated from red-haired C57BL/6 (Mc1re/e, Tyr+/+) mice exhibit strong CARS signal at ωP − ωS = 2000 cm−1.
(a) Trans-illumination image acquired with the pump beam, where the overall shape of the cells can be well visualized. (b) Confocal fluorescence image of tdTomato. (c) False colour ("Green fire blue" colour in ImageJ) CARS image mapping intracellular pheomelanin distribution. (d) 4X-zoomed view of (c) showing a perinuclear distribution of signal intensity, consistent with the known biology of protective melanin caps.
With the evidence that pheomelanin may be selectively visualized in isolated genetically defined cells, we explored the detection and localization of pheomelanin within intact mouse skin. Ex vivo ear skin from C57BL/6 (Mc1re/e, Tyr+/+) red-haired mice proved to be an advantageous sample, as brightfield images acquired alongside CARS could be used to verify the location of melanin within the tissue. Under the eyepiece, pheomelanotic cells in the thin ear skin appeared brown-red, with visible cell bodies and dendrites (Fig. 2). Individual pheomelanin-containing organelles were distinctly visualized via CARS imaging and co-localized with the pigmented areas observable by brightfield microscopy.
Figure 2: Imaging of pheomelanin stores in red-haired mouse ear skin.
(a,c,e) Brightfield trans-illumination image acquired from the microscope eyepiece. (b,d,f) Maximal projection view of CARS image stack of the mouse ear, showing bright granules from the pheomelanin stores (red circles) within melanocytes and at the base of the hair follicle. A CARS image stack acquired with the pump beam set to 871 nm (ωP − ωS = 1866 cm−1) was subtracted from the image stack acquired with the pump beam set to 861 nm (ωP − ωS = 2000 cm−1) to minimize the non-resonant signal contribution from structures other than pheomelanin. Image stacks are 27 μm thick, with a step size of 1 μm. An exemplary video scrolling through each individual image of the CARS depth stack corresponding to (b) can be found in the supplementary materials (Supplementary Video S1). Note that an out-of-focus hair (blue arrow) caused a shadow in the trans-illumination image (a), whereas CARS is highly depth-resolved; therefore, the hair was not prominently seen in (b).
To translate these findings, CARS imaging was carried in vivo on red-haired mouse (Mc1re/e, Tyr+/+) ears. Identical patterns of pheomelanin-rich cells were found (Supplementary Fig. S3). Importantly, these ex vivo and in vivo experiments were repeated for both control albino-red (Mc1re/e, Tyrc/c) and albino (Mc1r+/+, Tyrc/c) mice: no visible pheomelanin signals could be observed in the skin of these animals.
To confirm these measurements and positively identify the observed pheomelanin-rich cells as melanocytes, thin sections (5 μm in thickness) were prepared from Mc1re/e, Tyr+/+ red-haired mouse ears for histology and immunohistochemistry (IHC). As the red fluorescence from eosin was found to interfere with the CARS signal, immunostained slides where accompanied by an adjacent unstained slide for coherent Raman imaging. As this alternative slicing method was used, image features will not align precisely, but they are nevertheless relatively close to one another with a high degree of spatial consistency. Sry-related HMG-BOX gene 10 (Sox10) is a nuclear transcription factor that participates in neural crest development and in the specification and differentiation of cells of the melanocytic lineage. Sox10 is expressed in Schwann cells and melanocytes/melanomas10, making it a specific marker for dermal melanocytes. Figure 3(a) shows IHC staining for Sox10 (EP268, Cell Marque), where pheomelanin-containing melanocytes were found to be Sox10 positive. Pheomelanin stores inside the red-haired mouse skin can also be seen under brightfield illumination in the adjacent unstained slide, as shown in Fig. 3(b). When imaging the same unstained slide with CARS targeting the pheomelanin vibrational band, bright pheomelanin signals were found corresponding to the pigmented areas seen under brightfield and in the Sox10 positive areas via IHC. A separate experiment was also conducted to image a different mouse ear sample without IHC staining, such that CARS imaging can be performed directly on the haematoxylin-stained slice. As shown in Supplementary Fig. S4, pheomelanotic stores are observed as bright granular structures, consistent with imaging data shown in Fig. 3.
Figure 3: Imaging of pheomelanin stores in a red-haired (C57BL/6 (Mc1re/e, Tyr+/+) mouse ear section (5 μm thickness).
(a) Immunohistochemical stain of the mouse ear slide using anti-Sox-10 antibody (counterstain: haematoxylin), revealing melanocytes in red, some of which are indicated by black arrows for added clarity. (b) Image of an adjacent unstained 5 μm thick slide under brightfield illumination, revealing pheomelanin-rich deposits (shown by black arrows) consistent with the localization of the melanocytes in the adjacent slide shown in (a). (c) Trans-illumination image of the unstained slide shown in (b) acquired with the pump beam set to 861 nm. (d) CARS and (e) SFA images of the unstained slide shown in (b), revealing bright granular signals from pheomelanotic stores consistent with positive staining in (a) and pigmented areas in (b). (f) False-colour overlaid image of (d) in green and (e) in red. A CARS image acquired with the pump beam set to 871 nm (ωP − ωS = 1866 cm−1) was subtracted from the image acquired with the pump beam set to 861 nm (ωP − ωS = 2000 cm−1) to minimize the non-resonant signal contribution from structures other than pheomelanin.
In order to further cross-validate the pheomelanin signals from the CARS images, we also simultaneously performed sum frequency absorption (SFA) imaging in a multimodal configuration, where SFA microscopy is an imaging toolkit that enables the visualization of light absorbing molecules like pheomelanin11,12. This optical process is characterized by a molecule's ability to simultaneously absorb two photons without the necessary involvement of an intermediate real energy level between the ground state and the final excited state. The signal was confirmed to arise from sum frequency absorption and not stimulated Raman scattering (SRS) by tuning the pump wavelength across the expected pheomelanin Raman band. As no variation in signal intensity was observed, the signal was determined to arise solely from absorption contrast. As shown in Fig. 3(e) and (f), a one-to-one correlation can be found between CARS and SFA images, indicating that pheomelanin within melanocytes can indeed be well visualized using CARS microscopy. It should be noted that the overlap of the CARS and SFA images with Fig. 3(a) is not perfect, as each tissue slice is 5 μm thick. Moreover, the signal-to-noise ratio in the image shown in Fig. 3(d) is reduced compared to that obtained in tissue due to the configuration of the imaging system: as the position of the CARS detector is in the epi-direction, the collected signal depends in part on back-scattering in order to redirect the forward-generated anti-Stokes light back into the microscope objective. Considering the 5 μm thickness of the tissue slice, the CARS image quality in this scenario does not benefit from the scattering properties of thick biological samples that would otherwise generate a much stronger signal13. It is also possible that the actual amount of pheomelanin in thin tissue slices is slightly diminished relative to intact tissue as a result of the frozen section preparation itself. Nevertheless, the consistency in spatial localization between the signals observed in Fig. 3(d) and (e) indeed supports the ability of CARS microscopy to specifically visualize pheomelanin in a label-free manner.
With evidence showing that pheomelanin can be noninvasively visualized in synthetic, in vitro, ex vivo, and in vivo settings, we asked whether pheomelanin could be visualized in human specimens, specifically in the context of amelanotic melanoma. This melanoma subtype is characterized by its lack of traditional brown-black pigmentation upon visual inspection, typically presents as a raised lesion on the skin that is reddish in colour, and can easily be misdiagnosed due to its lack of dark pigmentation14. While the terminology seemingly implies the lack of any pigmentation, it is unclear whether some amelanotic melanoma lesions actually do harbour stores of pheomelanin that are simply indistinguishable from surrounding healthy skin. Moreover, recent studies4 have indicated the oncogenic potential of pheomelanin in red-haired and fair-skinned backgrounds, raising the possibility that at least some seemingly unpigmented melanoma lesions may contain pheomelanin, but are undetectable to the naked eye. CARS imaging and spectroscopic measurements were performed on fixed and unstained sections (10 μm in thickness) from three cutaneous amelanotic melanoma specimens. One such lesion can be seen in Fig. 4(a) along with its corresponding H&E cross-section. Inspection of normal, healthy perilesional skin with CARS (Fig. 4(c–e)) did not reveal detectable pheomelanin. Within the amelanotic melanoma lesion, however, strong pheomelanin CARS signals were observed indicating a high density of pheomelanin (representative images shown in Fig. 4(f–h)). Pheomelanin was ultimately detected in all three patient samples imaged, with representative images from a second patient shown in Fig. 4(i–k). This suggests that amelanotic melanoma lesions can indeed contain dense stores of pheomelanin, which may be detected noninvasively using CARS imaging.
Figure 4: Imaging of human amelanotic melanoma.
(a) Clinical photograph of one amelanotic melanoma lesion. (b) H&E stain of the patient slide (10X magnification). (c) Perilesional skin showing normal architecture of both epidermis and dermis. (d) Brightfield trans-illumination image acquired from the microscope eyepiece from the perilesional area. (e) CARS image of the same perilesional area compared to (d) (image acquired with pump beam wavelength at 841 nm (ωP − ωS = 2275 cm−1) was subtracted from the image acquired with pump beam wavelength at 855 nm (ωP − ωS = 2081 cm−1) to minimize the non-resonant background from structures other than pheomelanin). (f) View of the amelanotic melanoma area showing high density of cells with no obvious sign of melanin. (g) Brightfield trans-illumination image acquired from the microscope eyepiece from an unstained slide of the melanoma area, showing slightly pigmented granular structures (red circle). (h) CARS image of the same tumour area compared to (g), with the same settings as for (e). Saturated bright pheomelanin signals were found (red circle) corresponding to the minimally pigmented region shown in (g). (i,j,k) Respectively H&E, trans-illumination, and CARS images of the tumour area of slides from a second amelanotic melanoma patient. Strong pheomelanin signals were again observed (red circles).
It should be emphasized that the number of human samples surveyed in this work was limited to three, corresponding to the total number of amelanotic melanoma tissues available for study. In order to truly determine the incidence of pheomelanotic melanoma lesions among those diagnosed as "amelanotic", this proof-of-concept work will have to be expanded into a broader study: one where multiple CARS image stacks taken from a large patient cohort will be processed and statistically analysed to gain insight that can impact clinical practice.
Indeed, it has been proposed that an intrinsic pheomelanin pathway may represent a biologically significant contributor to melanomagenesis4. Building a complete understanding of this contribution, however, has been hampered by an inability to spatially map and quantify pheomelanin within skin. While the detection and differentiation of melanins has been accomplished using pump-probe microscopy12,15,16,17, measurements of pigments' excited state can be complicated by compounds in the local environment (e.g. metals such as iron). Moreover, distinguishing melanin subtypes using pump-probe microscopy requires the acquisition of so-called "delay stacks", where images are sequentially acquired with varying time delay between the pump and probe beams in order to obtain transient absorption traces on a pixel-by-pixel basis. From there, melanin subtypes can be differentiated using post-processing techniques such as principal component12 or phasor analyses17. However, each frame typically requires an acquisition time of several tens of seconds, leading to a total acquisition period ranging from 5 to 15 minutes for a single delay stack17. On the other hand, CARS imaging offers a specific, direct, and facile route for the detection of pheomelanin in vivo in real-time, since the detected signal arises from the generation of a new colour of light mediated by the pheomelanin pigment itself. In this regard, CARS microscopy offers many of the same benefits as multiphoton microscopy: the signal can be readily detected using an appropriate set of laser sources and optical filters, and has additionally been shown to operate up to video-rate imaging speeds18.
Prior investigation of melanin pigments using spontaneous Raman spectroscopy found pheomelanin to harbour three broad vibrational peaks at 500, 1490, and 2000 cm−1, while eumelanin was shown to have peaks at 500, 1380, and 1580 cm−1 5,6. The first two peaks of pheomelanin as well as all those of eumelanin are located within the fingerprint region of the Raman spectrum; they are therefore unlikely to offer a straightforward route to highly specific detection of melanin pigment subtypes in the native context of human skin, as vibrational peaks from other chemical species would result in spectral interference. However, in the particular case of pheomelanin, the broad vibrational peak of interest in this study is located in the 2000–2100 cm−1 range5,6, well within the so-called "silent region" of the Raman spectrum of biological samples13. This spectral region, which ranges approximately from 1800 to 2700 cm−1, is devoid of vibrational peaks that may arise from other endogenous biochemical moieties, making this particular band of pheomelanin an ideal target for future clinical CARS imaging with minimal interference from other compounds. It is worth nothing that the use of deuterated chemical groups as Raman reporters may interfere with pheomelanin imaging, as the carbon-deuterium vibrational band is located around 2200 cm−1. In this highly special case, the use of hyperspectral coherent Raman imaging may be used to distinguish the peaks in the event of signal overlap between pheomelanin and deuterated probes. It should also be noted that the unlikely possibility of interference from eumelanin at the 2000–2100 cm−1 band was also investigated in the context of this study. Importantly, no coherent Raman signals could be detected from eumelanin within the pheomelanin Raman band range (data not shown), further strengthening the specificity of CARS for the direct visualization of pheomelanin without interference from its eumelanin counterpart.
In this work, the use of multiple genetically controlled systems, including the Mc1r mutant mice and their corresponding albino/red-haired controls (Mc1re/e, Tyrc/c), enabled confirmation of the CARS signal as specifically arising from pheomelanin since these animal models are incapable of producing eumelanin4. Interestingly, the strength of the observed anti-Stokes signal was far greater than anticipated based on the previously published spontaneous Raman data5,6, indicating the possibility that the observed signal is resonantly enhanced19.
Furthermore, for future work in translating this imaging modality for clinical studies, CARS microscopy has even been performed using a single laser source, where one portion of the laser light was used as the pump beam, while the remainder was routed through either a tapered fibre20 or a photonic crystal fibre21 in order to generate the Stokes beam. This flexibility in terms of optical design in conjunction with the chemical specificity offered by CARS imaging makes it a prime candidate as an imaging tool for future clinical investigations in human patients. In fact, as portable CARS systems are already commercially available22, the translation of the presented technology for further investigation in research or clinical settings can be immediate and rapid. We believe this imaging approach will aid in improving our biological understanding of skin changes in the context of pheomelanin, including both UV-induced and UV-independent effects. This imaging approach may also help to refine our understanding of which cutaneous lesions are truly "amelanotic" as opposed to those that may be "pheomelanotic" and currently invisible to the naked eye. Additionally, the method may help to define inhomogeneously coloured melanocytic or non-melanocytic lesions, whose "unpigmented" regions might contain detectable pheomelanin, and represent challenges in defining surgical margins. The ability to identify such lesions non-invasively and non-destructively will enable future studies investigating the impact of pheomelanin on melanoma formation, pathogenesis and metastasis in different human skin types. The prospect of utilizing this method to detect suspicious, pre-malignant lesions in high-risk (red-haired, fair-skinned) individuals, may offer new diagnostic tools able to translate knowledge from basic science to clinical application.
Mice with the Mc1r frameshift mutant allele (Mc1re/e) have a phenotype analogous to red-hair/fair-skin in humans (also caused by non-functional alleles of MC1R)4. As control, we crossed these mice to the albino allele (Mc1re/e, Tyrc/c.), which ablates all melanin pigment production. All mouse strains were maintained on the C57BL/6 background.
The CARS microscope was built over a customized confocal microscope (Olympus FV1000, Center Valley, PA, USA), which has an additional laser entry port to accept external light sources. CARS microscopy was performed using a dual output femtosecond pulsed laser system (Spectra-Physics Insight DeepSee, Santa Clara, CA, USA), where the first output is tuneable from 680 to 1300 nm, while the second is fixed at 1040 nm. To achieve CARS imaging at the reported 2000 cm−1 band of pheomelanin, the 1040 nm output was chosen as the Stokes beam (ωS), while the pump beam (ωP) was set to 861 nm and 855 nm (ωP − ωS = 2000 cm−1 and ωP − ωS = 2081 cm−1, respectively), in order to generate anti-Stokes signals at 735 nm and 726 nm, respectively. A half-wave plate and a polarizer were placed at each of the two laser output ports of the DeepSee system to adjust beam power. To focus the beams onto the sample, a 1.20 NA 60X water immersion microscope objective (Olympus UPLSAPO 60XW, Center Valley, PA, USA) was used. CARS signal was detected in the epi-direction using both a shortpass and a bandpass filter (Chroma ET750sp-2p8 and ET730/40 m, Bellows Falls, VT, USA) placed in front of a thermoelectrically cooled photomultiplier tube (Hamamatsu H7422PA-50, Hamamatsu City, Japan). The sum power of the two beams at the objective was less than 10 mW for all experiments performed in this study. In order to correct for the non-resonant background signal generated in tissues, both resonant and non-resonant CARS images were obtained in order to subtract the non-resonant contribution from the resonant CARS images. To this aim, the pump beam was tuned to either 841 nm or 871 nm (resulting in a Raman shift of 2275 cm−1 or 1866 cm−1, respectively). In both of these non-resonant imaging settings, the background levels were found to be identical; the choice of tuning the pump beam to 841 nm or 871 nm was therefore simply based on the convenience of the experiment at hand. This correction was not necessary for cellular samples, as the signal detected in the epi-direction contains minimal contribution from back-scattered light and has inherently low non-resonant background13.
All SFA imaging experiments were performed simultaneously with CARS imaging on the same system as described above, using the same wavelengths and power levels. However, instead of detecting signal in the epi-direction, the SFA detector was placed downstream of the sample in the trans-direction. The 1040 nm beam was modulated at 20 MHz using an electro-optic modulator (ThorLabs EO-AM-R-20-C2, Newton, NJ, USA), and the modulation transfer from the 1040 nm beam to the pump beam was detected using a photodiode coupled to a lock-in amplifier (APE Lock-In Amplifier, Berlin, Germany). In order to restrict the light incident on the SFA detector to only that of the pump beam, a shortpass filter (ThorLabs FES0950, Newton, NJ, USA) was used to eliminate the 1040 nm light. Ordinarily, this detection setup would be sensitive to stimulated Raman scattering (SRS) signals as well. However, it was found that when imaging pheomelanin, the detected signal remained constant across a range spanning 1866 to 2275 cm−1. This observation was additionally confirmed in synthetic pheomelanin samples, which showed the same temporal and wavelength tuning properties as the pheomelanin detected in tissue samples. This suggests that the signals obtained from pheomelanin are primarily a result of its absorptive properties, rather than its intrinsic vibrational modes.
Pheomelanin was synthesized following the protocol published by d'Ischia et al.7. The synthesized pheomelanin was then lyophilized to yield a dense, reddish-brown powder. To emulsify the pheomelanin, a small quantity (~0.1 mg) was placed in a 2 mL microcentrifuge tube, to which 1 mL H2O and 0.25 mL hexane were added. The resulting mixture was sonicated for two cycles of three minutes each to generate an emulsion of pheomelanin microparticles. The sample was then sandwiched between a glass slide and a coverslip with an imaging spacer (Grace Bio-Labs Secure-Seal, Bend, OR). To prevent sample evaporation, the edges of the coverslip were sealed to the glass slide with nail polish.
In order to generate CARS spectra of pheomelanin samples, the pump wavelength was tuned from 841 nm to 871 nm in single nanometre increments. For each wavelength value, 3 images were acquired: the first with only the pump beam, the second with only the Stokes beam, and the third with both. While the first two image sets showed only minimal intensity, they nevertheless corresponded to the weak multiphoton fluorescence of pheomelanin and were thus subtracted from the CARS images to isolate the coherent Raman signal. In order to normalize the CARS signal against a neutral reference, the sample's glass coverslip was also imaged under the same conditions. Glass produces a non-resonant background signal that is invariant across the spectral range of interest, justifying its use as a reference to compensate for wavelength-dependent intensity variations that may arise from the optical components throughout the imaging system.
In order to generate the spectra, the 31 corrected CARS images were first summed together, and the resulting image was binarized to generate a mask. The mask was then applied to all CARS and glass images in order to isolate the regions corresponding to pheomelanin. The masked CARS images were then divided by the masked glass images on a pixel-by-pixel basis, and the resulting ratiometric values were averaged to obtain a single data point for a given wavelength. This process was iterated across all 31 sampled wavelengths, and the resulting spectra were normalized by the area under the curve and multiplied by a factor of 30 (corresponding to n-1 data points) such that a flat spectral response was centred at a normalized value of 1 across the entire spectral range. The experiment was performed in triplicate, where each individual spectrum is plotted in grey and the mean of the three measurements is plotted in red. As an added control, the analysis was repeated for the image background by inverting the binary mask generated earlier. Given that the image background signal arises from the non-resonant background generated by the water and hexane emulsion, the analysis performed on the background revealed a flat spectrum centred at 1, as expected.
Two mouse strains were utilized to isolate neonatal cutaneous melanocytes: 1) Mc1re/e: K14-SCFTg+4; Tyr-CreTg+8: B6.Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J (Jackson Laboratory catalogue #007909) and 2) Mc1re/e: K14-SCFTg+; Tyr-Cre: B6.Cg-Gt(ROSA)26Sortm6(CAG-ZsGreen1)Hze/J (Jackson Laboratory catalogue #007906) were used as sources of neonatal pheomelanotic cutaneous melanocytes. The K14-SCF transgene mimics human epidermal expression of SCF and enhances the yield of cutaneous melanocytes4. Melanocyte targeted fluorescent tags were generated using the constitutive Tyr:Cre transgene8. Neonates (day 2) were euthanized. The truncal skin was peeled away and washed twice with PBS containing Pen:Strep and Fungizone. The skin was then placed in a 60 mm tissue culture dish with 3 mL of dispase (25 mg/mL) and placed in a 37 oC, 5% CO2 incubator for 2 hours. The epidermal layer was peeled away and incubated in another 60 mm tissue culture dish containing 3 mL of 0.25% trypsin for 15 minutes at 37 oC. The skin tissue was washed in 10 mL of Hams F10 medium containing 10% FBS, ten times. The cells removed using this procedure were then washed and suspended in PBS that contained DNAse I (50 μg/mL) and 5 mM EDTA. The cells were sorted for the highest expression of either tdTomato or ZsGreen in a BD FACSAria II SORP cell sorter. ZsGreen cells were excited with a 488 nm laser and detected with a 525/50 nm bandpass filter; tdTomato cells were excited with a 561 nm laser and detected with a 582/15 nm bandpass filter. To confirm the identity of the melanocyte population, the tdTomato cells in another experiment were counterstained with FITC c-Kit, a surface marker for both melanocytes and mast cells. When imaging FACS-isolated cells using CARS microscopy, the presence of fluorescent proteins in the sample (i.e. tdTomato and ZsGreen) sometimes resulted in a background signal generated via two-photon excitation fluorescence from either or both the pump and Stokes lasers. To correct for this, a raw CARS image was first acquired using both lasers; then, two-photon fluorescence images were acquired by using each laser beam individually. The resulting fluorescence images were then subtracted from the raw CARS image in order to isolate the true CARS signal from the sample.
For ex vivo mouse ear imaging, the ear tissue was acquired via ear punch (Φ = 3 mm). A commercial hair removal lotion (Nair, Church & Dwight Co., Inc., Princeton, NJ) was used to remove fine hairs from the ear tissue. The sample was then sandwiched between a coverslip and a glass slide, and imaged as described above. For in vivo mouse ear imaging, the mice were anesthetized with isoflurane mixed with 0.2 L/min oxygen and 0.8 L/min air via face mask. Fine hairs were removed using the same hair removal lotion as in the ex vivo sample. The ear was then fixed onto a coverslip using double-sided tape, and imaged as described above. All studies and procedures involving animal subjects were approved by the Institutional Animal Care and Use Committee of Massachusetts General Hospital, and were conducted strictly in accordance with the approved animal handling protocols.
How to cite this article: Wang, H. et al. In vivo coherent Raman imaging of the melanomagenesis-associated pigment pheomelanin. Sci. Rep. 6, 37986; doi: 10.1038/srep37986 (2016).
Ferlay, J. et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012 . International journal of cancer 136, E359–E386 (2015).
Garbe, C. & Leiter, U. Melanoma epidemiology and trends . Clinics in dermatology 27, 3–9 (2009).
Noar, S. M., Myrick, J. G., Morales-Pico, B. & Thomas, N. E. Development and validation of the comprehensive indoor tanning expectations scale . JAMA dermatology 150, 512–521 (2014).
Mitra, D. et al. An ultraviolet-radiation-independent pathway to melanoma carcinogenesis in the red hair/fair skin background . Nature 491, 449–454, doi: 10.1038/nature11624 (2012).
Galván, I. et al. Raman spectroscopy as a non-invasive technique for the quantification of melanins in feathers and hairs . Pigment Cell & Melanoma Research 26, 917–923 (2013).
Galván, I., Jorge, A., Solano, F. & Wakamatsu, K. Vibrational characterization of pheomelanin and trichochrome F by Raman spectroscopy . Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 110, 55–59, doi: 10.1016/j.saa.2013.03.027 (2013).
d'Ischia, M. et al. Melanins and melanogenesis: methods, standards, protocols . Pigment cell & melanoma research 26, 616–633 (2013).
Delmas, V., Martinozzi, S., Bourgeois, Y., Holzenberger, M. & Larue, L. Cre‐mediated recombination in the skin melanocyte lineage . Genesis 36, 73–80 (2003).
Tabone-Eglinger, S., Wehrle-Haller, M., Aebischer, N., Jacquier, M.-C. & Wehrle-Haller, B. Membrane-bound Kit ligand regulates melanocyte adhesion and survival, providing physical interaction with an intraepithelial niche . The FASEB Journal 26, 3738–3753 (2012).
Barnhill, R. L., Piepkorn, M. & Busam, K. J. Pathology of melanocytic nevi and melanoma. (Springer Science & Business Media, 2014).
Fu, D., Ye, T., Matthews, T. E., Yurtsever, G. & Warren, W. S. Two-color, two-photon, and excited-state absorption microscopy . Journal of Biomedical Optics 12, 054004 (2007).
Matthews, T. E., Piletic, I. R., Selim, M. A., Simpson, M. J. & Warren, W. S. Pump-Probe Imaging Differentiates Melanoma from Melanocytic Nevi . Science Translational Medicine 3, 1–10 (2011).
Evans, C. L. & Xie, X. S. Coherent Anti-Stokes Raman Scattering Microscopy: Chemical Imaging for Biology and Medicine . Annual Review of Analytical Chemistry 1, 883–909, doi: 10.1146/annurev.anchem.1.031207.112754 (2008).
Koch, S. E. & Lange, J. R. Amelanotic melanoma: The great masquerader . Journal of the American Academy of Dermatology 42, 731–734, doi: 10.1067/mjd.2000.103981 (2000).
Fu, D. et al. Probing skin pigmentation changes with transient absorption imaging of eumelanin and pheomelanin . Journal of biomedical optics 13, 054036-054036–054037 (2008).
Simpson, M. J. et al. Nonlinear microscopy of eumelanin and pheomelanin with subcellular resolution . Journal of Investigative Dermatology 133, 1822–1826, doi: 10.1038/jid.2013.37.Nonlinear (2013).
Wilson, J. W. et al. Comparing in vivo pump-probe and multiphoton fluorescence microscopy of melanoma and pigmented lesions . Journal of Biomedical Optics 20, 051012 (2015).
Evans, C. L. et al. Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy . Proceedings of the National Academy of Sciences 102, 16807–16812, doi: 10.1073/pnas.0508282102 (2005).
Cheng, J.-X. & Xie, X. S. Coherent anti-Stokes Raman scattering microscopy: instrumentation, theory, and applications . The Journal of Physical Chemistry B 108, 827–840 (2004).
Kee, T. W. & Cicerone, M. T. Simple approach to one-laser, broadband coherent anti-Stokes Raman scattering microscopy . Optics Letters 29, 2701–2703 (2004).
Kee, T. W., Zhao, H. & Cicerone, M. T. One-laser interferometric broadband coherent anti-Stokes Raman scattering . Optics Express 14, 3631–3640 (2006).
Weinigel, M. et al. In vivo histology: optical biopsies with chemical contrast using clinical multiphoton/coherent anti-Stokes Raman scattering tomography . Laser Physics Letters 11, 055601 (2014).
We thank J. Hsiao for preparation of mouse ear samples. We also thank Y. Suita for help with melanoma cell preparation. We also thank the Wellman Photopathology Core, with special thanks to J. Zhao for processing all the staining. We gratefully acknowledge T. Kunisada for the K14-SCF transgenic mouse strain and L. Larue for the Tyr-CRE mouse strain. We also thank Tom Powell for assistance in manuscript editing. This work was supported by a Bridge Project Grant (222700) to D.E.F and C.L.E. H.W and S.O would like to acknowledge funding support from the Canadian Institute of Health Research and the Henry C. (1926) and Frances Keany Rickard Fund Fellowship via MIT, respectively. D.E.F was supported by grants from NIH (5P01 CA163222-03 and 5R01 AR043369-19), as well as from the Dr. Miriam and Sheldon G. Adelson Medical Research Foundation.
H.W., D.E.F. and C.L.E. conceived and planned the project. H.W. performed all of the experiments and data analysis except parts mentioned below. S.O. performed the analysis of the wavelength tuning data. V.I. performed the melanocyte extraction from mouse skin and flow sorting analysis. E.M.R. helped with interpretation of the acquired datasets. J.P., S.O., and A.J.N. helped in building the CARS system. A.J.N. performed the tdTomato-labelled melanocyte extraction experiment. H.T. provided essential consultation regarding melanin-related staining and provided amelanotic melanoma slides for this study. The manuscript was written by H.W., S.O., D.E.F., and C.L.E. All authors discussed the results and commented on the manuscript.
Correspondence to David E. Fisher or Conor L. Evans.


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By Connie Hwong
In Verto Index
Verto Index: Health and Fitness2018-07-192019-03-25https://vertoanalytics.com/wp-content/uploads/2019/03/verto_logo_rgb.pngVerto Analyticshttps://vertoanalytics.com/wp-content/uploads/2019/03/feature-images-10.png200px200px
This month's Verto Index looks at the top health and fitness apps among U.S. adults (ages 18 and above) from Fitbit to Map My Walk. The fitness and wearables market continues to support a complex ecosystem of hardware (mobile devices, wearables, and GPS systems) and software (a constellation of independent apps and those tied to specific devices). For the sake of simplicity, this Verto Index looks only at apps (and websites, where applicable) that provide health and fitness-tracking capabilities. Note: we are currently unable to gather reliable data for Health for iOS.
The Ten Most Popular Health & Fitness Apps and Sites
Fitbit continues to dominate the health and fitness market, based on monthly user numbers. Despite a tumultuous year, Fitbit is still well ahead of the competition, with 27.4 million monthly users and a reach of nearly 13%. MyFitnessPal trails significantly, with just 19 million monthly users, followed by S Health (an app for Samsung devices), with just under 15 million monthly users.
As we've noted in the past, Fitbit is in a unique position among the other apps on our Index: it owns its own ecosystem of hardware (wearable devices) and software, while the other top-ranking apps have varying degrees of device agnosticism, designed to be used on smartphones or paired with other third-party devices or wearables.
An App a Day…The Most Engaging Fitness Apps
Like music streaming services, most fitness apps are designed to largely run in the background, tracking your activity as you proceed through your workout or daily routine. With this in mind, the average amount of time that users spend with the app is less important than the frequency with which they use it. Stickiness, Verto's measure of user engagement, compares daily users to monthly users, and indicates how likely is a user to return to the service day after day – an especially crucial metric for fitness apps and others that want to encourage daily or regular user behavior.
Click here to download the full Index as a PDF.
Once again, Fitbit shines in this category with 36% stickiness: the average user engages with Fitbit 40 times per month, or more than once per day within a month. Map My Walk's tracking app also shows a high rate of user engagement, with 32% stickiness and an average of 36 user sessions per month. And activity isn't the only thing that consumers are interested in tracking: the Sleep Cycle Alarm Clock is the third-stickiest app on our Index, with 31% stickiness and an average of 19 monthly sessions per user – or nearly one session for every weekday in a typical month.
Who Uses Fitness Apps?
Verto Analytics looked at the demographic breakdown for four of the top-ranking fitness apps on our Verto Index: Fitbit, MyFitnessPal, S Health, and Google Fit. In nearly all cases, women make up an overwhelming proportion of the user base; almost 75% of MyFitnessPal's users are women, and two-thirds of Fitbit's users are women. Only Google Fit reported a near-equal proportion of male and female users. With just 2.6 million monthly users, Google Fit's audience is just a tenth of Fitbit's, but the app's broad availability across the Android ecosystem has seemingly encouraged adoption by a more diverse user demographic. And if Google makes good on the release of its rumored Pixel smartwatch, it may be able to gain a larger foothold in the greater health and fitness landscape.
Interested in learning more about audience behavior and digital content? Download the full Index here, or sign up for our newsletter to receive our latest insights and reports.
fitbit, fitness apps, fitness tracking, health and fitness apps, Map My Walk, quantified self, Verto Index, wearables
Connie Hwong
Connie is the global content marketing director at Verto Analytics. Prior to Verto, Connie held positions at ABB and Gigaom, and worked with brands like AOL and Zendesk. When she's not tracking down (and telling) interesting stories behind Verto's data and people, she's busy cycling around Berlin in pursuit of strong coffee and interesting music.
Verto Index: E-Commerce
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Verto Index: Travel
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redpajama_set_name: RedPajamaCommonCrawl


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In life we are told some people believe their glass is always half full, and others believe it is always half empty. I take the pessimistic view on life believing it's always half empty. I don't think like that of a pessimist contrary to my belief. What I mean to say is if we always believe the glass is half full we become enamored by our own design. We begin to believe we are owed everything in the world, and that it can't or won't disappear. I understand optimism gets us further in life that pessimism ever will. But in my eyes, if the glass is always half empty we will always have room to fill it up more. It's kinda of a unique twist on the saying.
I'm a writer. I've seen a lot of blank pages in my life. I mean every time I go to write a page it's blank. If I wasn't an optimist this page wouldn't be getting filled at the moment. So, how did I take a twist on a pessimistic idea, and create positive vibes from it? When you stare at a page long enough we begin to feel this daunting desire that maybe we won't succeed, maybe people won't read our work. There are thousands of ideas out there that deter us from getting the words on the page, and it pains my heart when the staring leads us to writer's block. We writers are constantly facing a pessimistic idea, and having to create something positive out of it.
Our pages will always be blank before we start writing but once our fingers start typing or writing it's like a magic that we can't put down. I like to think that we go into that zone like a basketball player on the field, or a gamer when they get so into a game they can't stop. In this case, I believe a writer is a lot like an athlete. The money might not always be the same, but if you can blog properly then there might be a few thousands of dollars in it for the writer. ( I do need to move my blog to wordpress.org soon. Any donations are welcome…jk jk …I think I can come up with the money after paying my graduation fee~~~right.) We are constantly going into the zone when we write. Sometimes, I get so into the zone that I can't actually stop myself from what I'm writing. It kind of spills out like some sort of magic.
If our glass is always half empty then we will always be able to fill it. I mean it's easier to fill something if it's half empty then half full right? I like looking at the opposite idea here. If my glass was half full I don't think I'd be as capable of wanting to fill it because half of it is already there. I mean I am half lazy sometimes. But if something was half empty I'm more likely to jump up and want to fight for it. An Athlete is more likely to make an effort if they are one point away from losing then if they were fifty points away from losing. The same can be said for us writers. We are more likely to just get our books written if we are already fifty pages in rather than if we are ten pages in.
I'm odd writer where most of my stories don't actually have the start of it written, but have majority of everything else written. In fact, I am itching to keep writing for " Into The Gray." I'm also yearning to read more works of writing. If you want me to guest blog on your blog just email me at Jessicacote66@gmail.com. You can also contact that if you have any odd writing work you want me to do. I'm open to job. My guest blogging is free. I just would rather be emailed to know what topic you want me to cover.
Again, Thank you for reading. I love you guys. Here is a video from one of my favorite creative youtubers; likes, follows, shares, and comments are always welcome. I love commenting back.
It depends on where you start. If you start with a full glass, than the glass is half-empty; if you start with an empty glass, than the glass is half-full. I start a project full of enthusiasm, but it drops off after it reaches a certain part, where there are necessary, but not so exciting aspects to project my glass is half-empty. On the other hand, if I start a project, and I am lukewarm, but then I get excited my glass is half-full.
When you write "We begin to believe we are owed everything in the world, and that it can't or won't disappear." I find it does not necessarily follow from seeing the glass as half-full. There are a lots of pessimistic people that think the world owes them one.
If a page is empty, then as you write, and you reach the half way point, your glass is going to be half-full. However, if you start with a full page and go about revising it, and it is severe, you wind up with a glass half-empty. I realize that this is not completely accurate, but it fits the fact that's life is variable. Sometimes the glass is half-full, and sometimes the glass is half-empty. Your fourth paragraph is actually supportive of this variability.
Finally, I would like to point out that mathematically a half-empty glass is equal to a half-full glass, so I ask does it really matter? After all, I rarely think in glass terms. And, how does the volume of the glass affect a situation? Is something harder if the glass is bigger, or less so, if the glass is smaller.s is equal to a half-full glass, so I ask does it really matter? After all, I rarely think in glass terms. And, how does the volume of the glass affect a situation? Is something harder if the glass is bigger, or less so, if the glass is smaller.


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Home > Bronze Sculptures > Western Bronze Sculpture, "Down the Rough Edges" , by Jeff Wolf, Limited Edition of 17/20, 1990, #C 1556
Western Bronze Sculpture, "Down the Rough Edges" , by Jeff Wolf, Limited Edition of 17/20, 1990, #C 1556
Default Title - $ 18,000.00
Description: Western Bronze Sculpture, "Down the Rough Edges" , by Jeff Wolf, Limited Edition of 17/20, 1990, #C 1556
Condition: Good for age
Dimensions:3'" H x 36" W, approximately 150 pounds
"Down The Rough Edges" is a story piece. I was with Lee Nelson, (a western & historical author) on his quest to shoot a Mule Deer buck from the back of a horse with a bow & arrow, which by the way he proved could be done, but that is another story.
We were camped just about a thousand feet off from a large table (11,000 feet high and about a mile square) where we had been trying to chase these three bucks for several days. Unfortunately, every time we made an attempt they would spot us and off the table they would go.
The table is surrounded by high ledges and these bucks would lay down in the same place every morning and feed up to the table every night.
On this particular evening it was raining hard and lightning was sticking all over the table. It was useless to try anything so we just glassed the bucks as they made their way to the table.
Just as they reach the edge of the table lightning struck right behind them lighting up the edge. Needless to say, those buck quit the edge in a hurry. So much so that the top buck knocked the middle buck over trying to get down through the ledges.
The moment the lightning struck is the scene I captured in my mind and is the scene portrayed in "Down The Rough Edges"
Biography of Jeff Wolf and Achievements Follow:
Persistence makes perfect: The trail to the top of the Western Art world is steep and narrow. It is littered with obstacles, beset with sheer drop-offs, and hindered by unexpected twists and turns. Sculptor Jeff Wolf has ridden that trail for more than twenty-five years and reached heights few artists attain. Along the way he has placed bronzes in prominent museums and permanent exhibits, in prestigious private collections, and on display in public venues. He has been bestowed with honors and awards, recognized in juried competitions, and called upon to teach and demonstrate.
Jeff made—and continues—the journey on a mount called persistence. He has lived by the old cowboy maxim that it doesn't matter how many times you get bucked off, but what counts is getting back on—brushing off adversity, dusting off disappointment, climbing back in the saddle and continuing the journey. While the ride can be a lonely one, it is not a ride Jeff makes alone. Riding with him along the way are talent and skill, creativity and vision, experience and expertise. And, of course, persistence—keeping ever after the quest to not only depict the West in art, but to capture the emotion, the motivation, the underlying aesthetics of ordinary life and extraordinary events.
Saddling up; The importance of persistence was instilled in Jeff at an early age. Jeff excelled in everything he put his mind to except school. Dyslexia—virtually undiscovered at the time—made reading nearly impossible. But that didn't stop him from graduating from high school and attending three years of college on scholarships, and eventually teaching himself to read.
In high school, Jeff became a champion livestock judge and earned a silver medal at the National FFA convention, earning what was at the time the highest score ever recorded—98 out of 100—in the cattle grading division. He was also a Champion rodeo cowboy in High School, college, and a top competitor in professional rodeo, competed mainly in the bareback and bull riding events, as well as saddle bronc riding, team roping, and steer wrestling.
Through it all, art sustained him. From an early age he was compelled to create. "My gift chose me, I didn't choose it," he says. Jeff's story as a sculptor started at age five when he received modeling clay for Christmas. His hands and heart went to work to mold into the clay the world he saw around him. An early work, a buffalo carved from a bar of soap, earned his first recognition when published in the pages of Western Horseman magazine.
With a constant driving force from within, combined with a wild imagination and insatiable desire to learn and discover, Jeff's childhood and youth would inform his art. Along with his gift of creation he was given, in his words, "a great gift of upbringing." Raised on a ranch in the mouth of Goshen Canyon, located south of Utah Lake, he had both the opportunities and responsibilities of any ranch kid. "I lived among the local wildlife, learned the art of handling cattle and horses, and had the fortunate opportunity to listen to the stories of real old-time cowboys, memories of which remain ingrained in my mind." Adding to the fascination, Wolf says, was "spending most of my days, when not working on the ranch, running wild and free in the mountains, along the creeks, building hideouts, and watching wildlife or hunting."
Even anatomy lessons were in the offing. Jeff's grandfather owned and operated a small meat packing company, which gave Jeff the opportunity to see firsthand animal anatomy from the inside out. "Grandpa used to take the front or rear leg of a beef carcass and move it as if it were walking and explain how each muscle and every bone made that movement possible." This led Jeff to study every movement a person or animal made, trying to decipher the bones and muscles working to make that movement possible. "I developed the habit when riding for cows to ride behind another rider and watch the horse and rider as they moved as one in harmony. A nice moving horse and a true horseman is a symphony of visual music." This curiosity and fascination with anatomy turned Jeff into a recognized master of capturing motion in sculpture.
And there were other lessons to learn: Jeff says, "I know firsthand what it feels like to climb down onto the back of a bull or bucking horse, know the rush adrenaline and the explosion from the chute. I know what it's like to sit for hour watching mule deer feed, coming so close that I could feel their breath on my hand. I have experienced the fear and drama of a stampede. And I have lived in the wild, providing for myself among the ghosts of Indians."
From his father, Jeff learned the ways of cattle and horses. "I remember one experience as if it were today," he says. "Dad and I rode up on a cow and calf who hadn't seen a human being all summer. She was one of those who enjoyed hiding out in country where she wasn't easy to find. We saw each other at about the same time. Her head came up and her ears came forward, moving back and forth determining which route to take for escape. Dad said, 'Let's just let her look at us for a while.' As we sat there, he explained every thought that was running through that cow's mind by the way she her ears worked back and forth, the short, soft mooing sounds she made to her calf, her posture, and the way she looked away then back at us.
mimicking shapes to keep the eye roving around the subject to tell a story. Years of devotion to the study of art principles, combined with the determination to produce works that are worthy of the title of fine art is the motivational drive behind my work. That's is the visual tune I dance to."
It's not an easy dance to learn. It takes passion. Perseverance. And persistence. But those qualities, combined with an imaginative and creative mind pay off for Jeff. Then, it's time for the work of the mind and heart to guide the hands of the sculptor.
"Once the physical work begins, the piece often times takes on its own personality. I then become merely the tool that gives life to the dictation of the piece. Those times produce my finest works.
"Finding ways to create the illusion of life in something like wet hair, rushing water, speed of movement, drama of action, sheer fabric flowing around the beauty of the female figure, wind whipping a mane of a stallion, or a reflection in water in a bas relief is the stimulation behind my work," Jeff says. "I work to compose the design so that every aspect has purpose. Every line leads to another, every plane reflects light or casts a shadow for depth and dimension.
Balance points create harmony and mass builds strength and stability to create a realistic illusion of movement. The synergy of opposing forces coming together and pulling apart allows me to create a greater effect, a stronger vision. My feeling is that every work I create has its own distinct personality and character. Therefore, it requires its own texture or multiple textures unique to itself as well as the composition and design that best reflects the story that is being presented.
"A well-rendered pair of wrinkled and cracked work boots with worn soles and tattered laces, lying side by side as if just taken off the tired feet of the owner or even discarded will form a picture in the viewer's mind. Some may see only a pair of boots. But others will see the life of the man who wore them— tired and wrinkled like the boots, exhausted from a hard day's labor and glad to get those old boots off his feet. Some may see dad or grandpa. Some may see a farmer plowing a field behind two mules. It really doesn't matter what the viewer envisions, it's the fact that a vision has been created. That is what I strive for in every work I create."
Riding on: And so Jeff Wolf went to work. And he worked. Then worked some more to blaze his own trail to the artistic heights. Making the climb, and maintaining the heights requires persistence. Over time, collectors become familiar with an artist's background and reputation. But reputation only goes so far. If the work isn't up to snuff, one can never expect to create or maintain demand with an inferior product. But Jeff's persistence paid off.
In 1990, art collector Ann Heckbert discovered Jeff's sculpture at an art show in Steamboat Springs, Colorado. Ann and her husband Jim, owner of Garret Gallery, approached Jeff about showing his work. By 1991, Jeff's life as a professional sculptor was underway and has earned been his livelihood ever since. Jeff credits Jim and Ann for launching a career that has earned him a reputation as one of the finest western sculptors of our time.
Jeff's first national juried show, the George Phippen Memorial Art Show in Prescott, Arizona, earned the artist the three highest awards: Best of Show, Best in Category and People's Choice. He has won or placed in practically every juried art show he has participated in since, and may be the only living sculptor to have won Best of Show and People's Choice awards in six genres of Western art: Wildlife, Figural, Rodeo, Equine, Western, and Native American.
Finally, while Jeff is well aware that gifts such as his are given to individuals, the trail to success isn't one you ride alone. Persistence is often aided by the encouragement and assistance of others. He says, "The journey to the top would have been impossible if it weren't for the help and support of family, friends, my collectors, admirers and especially my wife Jennifer." Jeff also believes gifts are to be shared. So he now shares his talents and knowledge with fellow artists, students of the arts, and charitable foundations. He teaches workshops, lectures at schools, and sculpts at public events. Donations of time, talent, and art have generated over one million dollars for charitable causes.
God-given talent, real life experience, insatiable desire to be the best, and persistence have immortally molded and cast forever the name Jeff Wolf into the world of Western art and sculpture.
The portfolio of Jeff Wolf's work is extensive, and his name is tied to some of the most prestigious collections, galleries, and museums. Persistence has placed his art in national and international collections including:
• Simons Collection, Cayman Islands
• Ryder Collection, Ryder Trucking
• Renn and Marie Zaphiropoulos Collection (inventor of the color tube for television and, later, the developer of laser printing)
• Meredith Hodges Collection and National Mule Museum, Loveland, Colorado
• Mr. and Mrs. Robert Henderson
• Jim Terry (former CEO of Coca-Cola)
• Jack Williams (former president of Royal Caribbean Cruise Lines)
• Elton Salinas (owner of Elton's Clothier, Las Vegas, Nevada)
• Richard and Carole Kreamer (Board of Directors, American Airlines)
• David and Pam Furr (Gaston Law)
• Jim and Ann Heckbert (Burg Simpson Law & Humble Ranch)
• Steve and Mary Kay Larsen
• Lori Wilkinson (Brown & Brown Insurance of Nevada, Waymark Insurance Services)
• Richard Sanders (president of Kobalt Music Group)
• Buck Taylor (artist and actor)
• Jane Blalock (Hall of Fame Golfer)
• Jim Palmer (Hall of Fame Baseball Player)
Jeff has filled commissions for sculpture for:
• Coca-Cola
• Susan G. Koman Foundation
• T.A.P.S. Foundation
• Habitat for Humanity
• American Lung Association
• National Retriever Club and National Amateur Retriever Club
• American Airlines
• Cystic Fibrosis Foundation
• American Bucking Bull, Inc.
• Cistercian Preparatory School Hillary award
• Rodeo Champions monument, Gooding ID
• And several cities, corporate executives, farmers, ranchers, sportsmen, families, and friends.
Jeff's talent has introduced him to an array of TV, movie, and sports celebrities, including:
• Kimberlin Brown
• Dylan Bruno
• Gordon Clapp
• Lenny Clarke
• Jeff Dunham,
• Grant Goodeve
• Chris Harrison
• Dennis Haskins
• Sandra Hess
• Brad Johnson
• Wendie Malick,
• Ron Masak,
• Marc McClure
• Rob Moran
• Eloise Mumford
• Eric Christian Olsen
• Jason Priestly
• Perrey Reeves
• James Sikking,
• Buck Taylor
• Steve Thomas
• Michael Trucco.
• John York
• Ian Ziering
Musicians, such as:
• Aaron Barker (musician and Hall of Fame songwriter)
• John Cafferty (of The Beaver Brown Band)
• Kevin Chalfant (of 707, The Storm, and Journey)
• Daughtry
• Randall Hall (of Lynyrd Skynyrd)
• David Jenkens (of Pablo Cruise)
• Chris Ledoux
• Alex Ligertwood (of Santana)
• Gary Morris
• Hootie and the Blowfish
• Michael Martin Murphy
• Henry Paul (formerly of Black Hawk)
Sports champions, including:
• Matt Bahr
• Jane Blalock
• Larry Brown
• Brant Boyer
• Scott Hamilton
• Billy Kidd
• Jim Lonborg
• Chris McCarron
• Jay Miller
• Jim Palmer
• Gale Sayers
• Wayne Wong
And, of course, a host of rodeo champions, cowboys, ranchers, artists working in all mediums, and great people from all walks of life.
Other honors include:
• Selected as the sculptor at the 2000 Super Bowl for the Larry Brown Foundation.
• Featured artist at the Days of '47 Utah Heritage Art Show, 2000
• Commissioned to sculpt the six-time Labrador Retriever field trial champion and the Female Labrador Retriever Field Trials World Champion, owned by Fred Kampo, Oshkosh, Wisconsin
• Designed and sculpted many awards, personal tributes, and memorials such as the Huntsville Town Veterans Memorial Monument.
• Honored as one of Utah's Most Fabulous People by Utah Valley magazine, 2012
He has also been featured in a host of magazines, such as:
• Cowboy Magazine
• Ranch & Reata
• Range Magazine
• Rodeo News
• Western Horseman
• Western Writers of America Roundup Magazine
• Saddlebag Dispatches
I am frequently ask the question as to where I get my inspiration. My inspiration is drawn from a number things, My girls, parents, family, grand kids, friends, animals, other artists, the great outdoors, life experiences and stories I've heard just to name a few. Rather than creating just a well done work of art, I want my work to tell a story, something people can relate to. Things that inspire, evoke emotion, and arouse the imagination, I want my work to mean something, something that can be talked about and shared. I like my work to also be educational, whether it is from an artistic, anatomy, historical or human interest aspect. I strive to encompass and portray the emotions, feeling and expressions in every work I create.
Lincoln Fox (American, b. 1942), "Harmony", 1999, Edition 20/50, #1751
CulturalPatina $ 7,900.00
Western Artist, Lincoln Fox, Rare Bronze Sculpture titled, "Woman With Owl" 4/10, Ca 1978, #1751
CulturalPatina $ 15,496.00
Western Bronze, "Rocky Trails" by James Regimbal, Limited Edition, 41/50, Ca 1986 #C1708
Western Artist, Lincoln Fox, Bronze Sculpture titled, "Reaching for the Stars", Limited Edition of 75, #C 1688


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text:
AllAfrica is a website that aggregates news produced primarily on the African continent about all areas of African life, politics, issues and culture. It is available in both English and French and produced by AllAfrica Global Media, which has offices in Cape Town, Dakar, Lagos, Monrovia, Nairobi, and Washington, D.C. AllAfrica is the successor to the African News Service.

Its stories can be displayed by categories and subcategories such as country, region, and by news topic. In 2008, AllAfrica rolled out a comment board system.

The President of AllAfrica Global Media, Amadou Mahtar Ba, is a member of the International Advisory Board of the African Press Organization.

References

External links

 ReliefWeb archives of AllAfrica
 NAACP archives of AllAfrica

South African news websites
Africa-focused media

redpajama_set_name: RedPajamaWikipedia


text:
/*	$NetBSD: altq_hfsc.h,v 1.8 2006/10/12 19:59:08 peter Exp $	*/
/*	$KAME: altq_hfsc.h,v 1.12 2003/12/05 05:40:46 kjc Exp $	*/

/*
 * Copyright (c) 1997-1999 Carnegie Mellon University. All Rights Reserved.
 *
 * Permission to use, copy, modify, and distribute this software and
 * its documentation is hereby granted (including for commercial or
 * for-profit use), provided that both the copyright notice and this
 * permission notice appear in all copies of the software, derivative
 * works, or modified versions, and any portions thereof.
 *
 * THIS SOFTWARE IS EXPERIMENTAL AND IS KNOWN TO HAVE BUGS, SOME OF
 * WHICH MAY HAVE SERIOUS CONSEQUENCES.  CARNEGIE MELLON PROVIDES THIS
 * SOFTWARE IN ITS ``AS IS'' CONDITION, AND ANY EXPRESS OR IMPLIED
 * WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES
 * OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE
 * DISCLAIMED.  IN NO EVENT SHALL CARNEGIE MELLON UNIVERSITY BE LIABLE
 * FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR
 * CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT
 * OF SUBSTITUTE GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR
 * BUSINESS INTERRUPTION) HOWEVER CAUSED AND ON ANY THEORY OF
 * LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT
 * (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE
 * USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE POSSIBILITY OF SUCH
 * DAMAGE.
 *
 * Carnegie Mellon encourages (but does not require) users of this
 * software to return any improvements or extensions that they make,
 * and to grant Carnegie Mellon the rights to redistribute these
 * changes without encumbrance.
 */
#ifndef _NET_PKTSCHED_PKTSCHED_HFSC_H_
#define	_NET_PKTSCHED_PKTSCHED_HFSC_H_

#include <net/pktsched/pktsched.h>
#include <net/classq/classq.h>
#include <net/classq/classq_red.h>
#include <net/classq/classq_rio.h>
#include <net/classq/classq_blue.h>
#include <net/classq/classq_sfb.h>

#ifdef __cplusplus
extern "C" {
#endif

struct service_curve {
	u_int32_t fl;	/* service curve flags */
	u_int64_t m1;	/* slope of the first segment in bits/sec */
	u_int32_t d;	/* the x-projection of the first segment in msec */
	u_int64_t m2;	/* slope of the second segment in bits/sec */
};

/* valid values for service curve flags */
#define	HFSCF_M1_PCT		0x1	/* m1 is in percentage */
#define	HFSCF_M2_PCT		0x10	/* m2 is in percentage */

#define	HFSCF_USERFLAGS		(HFSCF_M1_PCT | HFSCF_M2_PCT)

/* special class handles */
#define	HFSC_NULLCLASS_HANDLE	0
#define	HFSC_MAX_CLASSES	64

/* hfsc class flags */
#define	HFCF_RED		0x0001	/* use RED */
#define	HFCF_ECN		0x0002  /* use ECN with RED/BLUE/SFB */
#define	HFCF_RIO		0x0004  /* use RIO */
#define	HFCF_CLEARDSCP		0x0010  /* clear diffserv codepoint */
#define	HFCF_BLUE		0x0100	/* use BLUE */
#define	HFCF_SFB		0x0200	/* use SFB */
#define	HFCF_FLOWCTL		0x0400	/* enable flow control advisories */
#define	HFCF_DEFAULTCLASS	0x1000	/* default class */

#define	HFCF_USERFLAGS							\
	(HFCF_RED | HFCF_ECN | HFCF_RIO | HFCF_CLEARDSCP | HFCF_BLUE |	\
	HFCF_SFB | HFCF_FLOWCTL | HFCF_DEFAULTCLASS)

#define	HFCF_BITS \
	"\020\1RED\2ECN\3RIO\5CLEARDSCP\11BLUE\12SFB\13FLOWCTL\15DEFAULT"

/* service curve types */
#define	HFSC_REALTIMESC		1
#define	HFSC_LINKSHARINGSC	2
#define	HFSC_UPPERLIMITSC	4
#define	HFSC_DEFAULTSC		(HFSC_REALTIMESC|HFSC_LINKSHARINGSC)

struct hfsc_classstats {
	u_int32_t		class_id;
	u_int32_t		class_handle;
	struct service_curve	rsc;
	struct service_curve	fsc;
	struct service_curve	usc;	/* upper limit service curve */

	u_int64_t		total;	/* total work in bytes */
	u_int64_t		cumul;	/* cumulative work in bytes */
					/*   done by real-time criteria */
	u_int64_t		d;		/* deadline */
	u_int64_t		e;		/* eligible time */
	u_int64_t		vt;		/* virtual time */
	u_int64_t		f;		/* fit time for upper-limit */

	/* info helpful for debugging */
	u_int64_t		initvt;		/* init virtual time */
	u_int64_t		vtoff;		/* cl_vt_ipoff */
	u_int64_t		cvtmax;		/* cl_maxvt */
	u_int64_t		myf;		/* cl_myf */
	u_int64_t		cfmin;		/* cl_mincf */
	u_int64_t		cvtmin;		/* cl_mincvt */
	u_int64_t		myfadj;		/* cl_myfadj */
	u_int64_t		vtadj;		/* cl_vtadj */
	u_int64_t		cur_time;
	u_int32_t		machclk_freq;

	u_int32_t		qlength;
	u_int32_t		qlimit;
	struct pktcntr		xmit_cnt;
	struct pktcntr		drop_cnt;
	u_int32_t		period;

	u_int32_t		vtperiod;	/* vt period sequence no */
	u_int32_t		parentperiod;	/* parent's vt period seqno */
	int			nactive;	/* number of active children */

	/* RED, RIO, BLUE, SFB related info */
	classq_type_t		qtype;
	union {
		/* RIO has 3 red stats */
		struct red_stats	red[RIO_NDROPPREC];
		struct blue_stats	blue;
		struct sfb_stats	sfb;
	};
	classq_state_t		qstate;
};

#ifdef __cplusplus
}
#endif
#endif /* _NET_PKTSCHED_PKTSCHED_HFSC_H_ */

redpajama_set_name: RedPajamaGithub


MARKBASS | The Markbass artist of this week is Richard Bona!
The Markbass artist of this week is Richard Bona!
April 24, 2017, 7:51 a.m.
Richard Bona has been recognized as one of the planet's best bassist. A complete artist, an absolute master with a distinctive and unique style, a mix of jazz, afro-beat, bossa nova, pop music, and funk.
"I searched it, and I found it. It's my sound and tone", says Richard, "In the end It ALL come down to BASS Sound. It's ALL in Markbass!"
On April 30, Richard Bona will play at All-Star Global Concert at Historic Gran Teatro de la Habana in Cuba to celebrate the sixth annual International Jazz Day.


text:
NHL's Top 25 Players in the Game Today
Offense is way up in the NHL this season. There are players scoring and putting up points in a manner that we haven't seen in the past two or three decades. Many thought the 150-point season was a thing of the past… well don't tell Connor McDavid that. Mitch Marner of the Toronto Maple Leafs is riding a 21-game point streak. Jason Robertson of Dallas has 23 goals in 27 games, McDavid has 24 in 27 games. Tage Thompson of Buffalo has 21 in 26 games. Even Erik Karlsson from San Jose has 35 points in 29 games. Offense is in fashion in the NHL these days.
Youth is King in the National Hockey League and there are some good players who are starting to emerge as great players. Let's take a look at the top NHL players in today's game.
Top 25 NHL Players – 2023 Edition
There is no doubt who is at the top of our list, but you might find some interesting players amongst our top 25. The Hockey Fanatic presents the top 25 players in today's NHL.
#25. Tage Thompson – Buffalo Sabres – what a year this guy is having. He recently tied an NHL record for most goals in a period with four (he had five in the game). Currently fourth in the NHL with 40 points only McDavid, Draisaitl and Jason Robertson have more.
#24. Nick Suzuki – Montreal Canadiens – the young captain of Les Habitants is really starting to emerge as one of the league's top players. He's off to a great start this season putting up 29 points in 26 games for the Canadiens.
#23. Jack Hughes – New Jersey Devils – Hughes is on pace to have his best season in the NHL as he has put up 31 points in 26 games for the surprising league leading New Jersey Devils.
#22. Jason Robertson – Dallas Stars – a star in the making here (pun intended). Robertson tallied 41 goals last season and 79 points. This year he is on pace for 70 goals after tallying 23 in his first 27 games. A strong up and coming player that will be fun to watch over the next couple of seasons.
#21. Mikko Rantanan – Colorado Avalanche – Finland sure has produced a lot of great players in recent years. The Stanley Cup Champion Avalanche can rely on 85-95 points with Rantanen in the lineup.
#20. Kirill Kaprizov – Minnesota Wild – another emerging NHL star in the making, Kaprizov has had seasons of 51 points (in 55 games), 108 last season and currently has 33 in 25 this season. Putting up 1.32 PPG this season is pushing top ten in the NHL.
#19. Elias Pettersson – Vancouver Canucks – Pettersson is on pace to have his best season yet as the Canucks push for a wild card playoff spot. Currently on 106-point pace which would shatter his career high of 68 from last season.
#18. Steve Stamkos – Tampa Bay Lightning – Stammer had a career-best 106-point season last year. He's on pace for another 100-point season this year as well. At 32 years old this guy just keeps getting better
#17. Igor Shesterkin – New York Rangers – the first of our three Rangers on our list. Shesterkin has a career .926 save percentage and 2.36 GAA in his three and counting NHL seasons. One of the NHL's top goaltenders to watch for.
#16. David Pastrnak – Boston Bruins – "Pasta" can score, no doubt about it. He led the league in goals in 2019-20 with 48 winning the Maurice "Rocket" Richard trophy. This season, he is currently 7th in points averaging 1.44 PPG with 18 goals and 18 assists in 25 games.
#15. Matthew Tkachuk – Florida Panthers – the blockbuster trade from the Calgary Flames hasn't seemed to bother league super-pest Tkachuk. Currently sixth in NHL points with 37 in 25 games, this guy can help you win in the trenches and we're pretty sure that there are 31 other GMs that would take this guy on their team
#14. Johnathan Huberdeau – Calgary Flames – over the past five seasons, Johnathan Huberdeau ranks sixth in points with 362 and fourth in assists with 255 in 309 games.
#13. Andrei Vasilevskiy – Tampa Bay Lightning – arguable the best goaltender in the game over the past five years. While he and the Lightning are having a bit of a slow start this season, Vasilevskiy is still strong between the pipes for the Lightning.
#12. Mitch Marner – Toronto Maple Leafs – often overshadowed by Matthews, Mariner is really the more consistent player. He's having another strong season and recently broke the Toronto Maple Leafs record for points in consecutive games (21 and counting). Why are people not talking more about this guy? We bet that you didn't know that over the past five seasons in the NHL, Mitch Marner is seventh in points with 358 in 296 and a +78.
#11. Auston Matthews – Toronto Maple Leafs – with 60 goals last season Matthews set a new record for the Toronto Maple Leafs. He has 273 career regular season goals in 435 games. While the Eastern media and hockey fans may think he should rank higher on our list, he is barely a point-per-game player at this stage in his career.
#10. Connor Hellebucyk – Winnipeg Jets – amongst the leader in all major goaltender categories, The Hockey Fanatic rates Connor Hellebucyk as the NHL's top goaltender in the game today. He's currently second in wins with 14 in 20 games and has a .932 save percentage. One of the main reasons why the Jets win night in and night out.
#9. Artemi Panarin – New York Rangers – with season point totals of 96, 58 (in 42 games), 95, 87, and 82 over his past five seasons, Artemi Panarin is one of the game's leading point producers during this span. The former Calder Memorial trophy winner in 2015-16 continues to impress in the Big Apple. Over the past five seasons, Panarin is fifth in NHL regular season points with 369 in 293 games.
#8. Roman Josi – Nashville Predators – Josi posted 23 goals and 73 assists last season for 96 points. Reminder he is an NHL defenseman. Roman Josi has been one of the great defensemen over the past five years and continues to get better every season.
#7. Adam Fox – New York Rangers – the Rangers have been surprising the past couple of seasons. Fox is a former Norris trophy winner and one of the reasons for the Rangers resurgence. This season he's averaging 1.07 PPG with 30 points in 28 games. Definitely one of the league's top defensemen in the game today.
#6. Nikita Kucherov – Tampa Bay Lightning – Kucherov has been one of the league leaders in points in recent years and the former Art Ross, Hart and Ted Lindsay award winner from 2018-19 is having another strong season as he sits fifth in points with 38 and a PPG of 1.46. His 128 points in 2018-19 were the highest since 1995-96 when Mario Lemieux put up 161 and Jaromir Jagr had 149.
#5. Sydney Crosby – Pittsburgh Penguins – at 35 years old is it possible that Crosby is getting even better? This guy has been one of the game's best players since he entered the league. This season he is averaging 1.35 PPG and is currently ninth in NHL scoring.
#4. Cale Makar – Colorado Avalanche – this guy has done it all already. Won the Norris trophy, the Conn Smythe and the Stanley last year. Arguably the best defensemen in the league today. Smooth skating, good hands, this guy will win multiple Norris trophies in his future.
#3. Nathan MacKinnon – Colorado Avalanche – developing into a well-rounded player with a bit of an edge. Has suffered a few injuries though otherwise he might be a little higher on our list. Over the past five seasons, MacKinnon is third in points behind Connor McDavid and Leon Draisaitl.
#2. Leon Draisaitl – Edmonton Oilers – After what we saw with Draisaitl playing on one good leg in the playoffs last season, you knew that he could only go up from there. The former Hart trophy winner is a formidable player on his own with or without Connor McDavid. Leon Draisaitl sits second in the NHL in points with 44 in 27 games which puts him on pace for 145 points himself. He's averaging 1.63 points per game second only to Connor McDavid (based on players who have played ten games or more). Draisaitl is a big body, who can and is playing physical when the team needs it. Arguably the best passer in the game with a tremendous shot, Leon Draisaitl is MVP-worthy.
#1. Connor McDavid – Edmonton Oilers. Sorry but not sorry. Connor McDavid is in a class by himself. This is a motivated player. Snubbed for the Hart trophy last year, we knew McDavid would be motivated this season. We knew he was going to score more goals (he's on pace for 81 as it sits now). He's also on pace for 170 points. Connor McDavid is electrifying. There is no one close to playing at his level game in and game out. Don't believe us, here is a small sample size of Connor McDavid's top goals in the NHL.
With fourteen out of the top 25 players from the Eastern Conference, does the Eastern Conference feature the best players? Well not if you look at our top ten where six of the ten are from the Western Conference showing that the NHL is full of great strs that are dispersed throughout the league.
Here is a look at the top NHL point producers over the past five seasons. Interestingly enough the top three happen to be Connor McDavid, Leon Draisaitl and Nathan MacKinnon. Patrick Kane and Artemi Panarin round out the top five. Lots of great talent in the National Hockey League at all positions. While youth is king, veterans such as Sydney Crosby, Steven Stamkos and Patrick Kane are still amongst the leaders.
← 2023 World Junior Hockey Championship Fact Sheet & FAQ
Top 100 Hockey Songs for your Hockey Playlist – 2023 Edition →


Sure marijuana is nice in your health. You need coverage that works with your way of life, helps you get the most effective care doable and suits together with your monetary picture. Perceive the rights of people with a incapacity and their carers and find details about assist teams in addition to helpful info for managing your healthcare.
It'll record and analyze your every day activities and habits to help maintain successful weight-reduction plan and lead healthy way of life. We support ministers in main the nation's health and social care to help individuals dwell extra impartial, healthier lives for longer. Spiritual wellness helps convey folks together it provides individuals a common bond and especially helps throughout times of battle.
This includes characteristics of the natural environment , the constructed surroundings and the social surroundings Elements resembling clear water and air , enough housing , and secure communities and roads all have been discovered to contribute to good health, particularly to the health of infants and kids.
Social health, along with mental and physical health, is among the key points to find out the general nicely-being of a person. Making high quality care more inexpensive, accessible, simple and seamless, to not solely assist folks get effectively, however assist them keep properly in physique, thoughts and spirit. With dwindling health, rising health care costs and an absence of financial savings to generate a hard and fast earnings, the monetary necessities of residing 30 years in retirement are out of reach for many people.
Access to some providers is restricted to 10 units. Individuals who smoke, drink, or take medication to neglect their issues are likely to have extra health problems later than someone who combats stress through a healthful diet and exercise. This web site is coordinated by the Office of Illness Prevention and Health Promotion, Workplace of the Assistant Secretary for Health, Office of the Secretary, U.S. Department of Health and Human Companies.
Some pets change into part of the family. Whereas many people have kept many various species of animals in captivity over the course of human history, solely a relative few have been kept lengthy enough to be thought-about domesticated Other types of animals, notably monkeys , have by no means been domesticated however are still bought and stored as pets.
Based on USDA restrictions, animals should be provided water each 12 hours, so for journey with a checked pet to Los Angeles, you have to guide a flight less than 12 hours. Conserving animals as pets could also be detrimental to their well being if sure requirements should not met.
They will all the time remember your "scent." These animals love to be open air, however might want sleeping to taking part in. If a pet is hatched a second time, the pet's picture will seem in full colour with out the Food Bar. Only a veterinarian can efficiently take away this yellowish-brown tartar from the enamel of your pets.


But it's true, and it do.
But wouldn't it be good if we could say it together!
Sing about the joy that I know we can find.
Wind them around and see what they sound like together.
The song is love, the song is love.
To see me through and they do.


16 Elegant Mountain Home Plans with Basement -Mini house layouts are excellent little homes with all the benefits you need to live extremely conveniently as well as luxuriously. They're developed for use in a vast array of situations. Mini houses are excellent for vacation houses as well as for areas where building land is scarce. They're likewise best for starter residences, retiree houses or for households on a spending plan. To put it differently, miniature homes are versatile for several conditions. Mini home designs are built residences which are also called one component. They can vary in dimension, but the smaller sized ones are typically made use of as vacation homes. A little mini home will certainly be 380 sq legs for instance.
The bigger mini homes might be larger walking around 1, 200 sq feet. Past that you are speaking about typical home dimensions. Mini houses are made to consist of all you require for comfortable living, in addition to aspects of the lavish. They consist of a kitchen, living room, bathroom room and bed room. Everything is properly made to fit in of the smaller room, but you may select to tailor several selections. They may be created with hardwood flooring as well as consist of prominent appliances and also luxury pipes fittings. They are occasionally similar to a camper van with expandable area, with no wheels or electric motor obviously.
They additionally come with distinctive exterior styling, which may additionally be customized. Mini homes can be constructed to rest on nearly all sorts of piece or a crushed rock pad. Yet mini houses may additionally be placed over a cellar. Also economic climates of range apart, individuals bothered with the environments appreciate these small houses. Since they're upreared they make use of much less power to produce, and they're additionally energy reliable. Your homes are well protected and often made making use of eco safe elements like recycled paints and also materials without any VOCs. The miniature residence styles include styles are which are parts you can configure.
You can choose the floor plan which will certainly match the little location where you prepare to construct. Some developers think the miniature residence is your house of the future for numerous families. The cost of housing carries on to skyrocket, as well as the mini house makes it viable for some people to pay for a new residence. This is specifically true in which land space is limited or expensive. If you do need a holiday home or a full-time home, the miniature home is one option to think about. You can buy your house currently made, have it established on a pad, and afterwards include an exterior patio area. It is all the comforts of house at a far more affordable price tag. Reliableremodeler.com that uses Internet based house enhancement info and also solutions. Providing homeowner a straightforward, quick, as well as complimentary method to access, qualified residence enhancement renovating service providers.
Related Posts of "16 Elegant Mountain Home Plans with Basement"


Ramirez went 1-for-3 with a solo homer, two runs scored, two stolen bases and two walks in Monday's 6-4 victory over the Mariners.
It took longer than expected, but Ramirez finally knocked his first home run of 2019 as well as his fourth and fifth stolen bases. He still has just nine hits in 60 at-bats this season, but Monday might've been what he needed to break the slump.


text:
Kevin Love To Sit Out On Monday
2 months agoCleveland Cavaliers forward Kevin Love (thumb) has been ruled out for Monday's game against the Toronto Raptors. This marks the fourth game in a row that the veteran forward has missed due to a hairline fracture in his thumb. The Cavs are expected to continue using a committee approach to fill the void. Among those players, Cedi Osman is likely to have the most streaming appeal, so fantasy managers looking for help on Monday should look towards Osman.--Andy Webb - RotoBaller
Tyrese Haliburton1 hour ago


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On which versions of RHEL can we install ECC 6.0 & NWEP 7.3?
Cannot exchange kernel binary archives.
Remote logon shows logon screen via RFC.
/sdf/mon shows same/wrong values in Performance monitoring data.


M & T Bank recently made a gift of $5,000 to IHA (Interfaith Housing Alliance) to support their Housing First program. In addition to providing affordable homeownership and rental housing opportunities and supportive services, IHA offers Housing First. The workshops in financial literacy and financial education are for the community at large and housing counseling is offered to perspective homeowners.
IHA provides group and one-on-one training in budgeting and personal financial management strategies for seniors, families (including their children) and individuals, as well as the clients of the nonprofits and other agencies with whom they have developed collaborative partnerships.
Gail D'Angelo – M& T Bank and IHA Vice President Travis Hardmon.


Swollen face, lips, limbs, or tongue could be often associated with the origin of mylan pharmaceuticals inc.
You can give him buffered Epclusa to reduce eventually the unusual tiredness, between 5 and 15 mg per per pound of your grown dog's weight. Use controlled drug with caution in such patients withdrew because the systemic exposure thought to Eprosartan mesylate may severely be widely increased due to reduced metabolism.
In summarising some reigions preparation strategies to be used with care can be found conditions under name of Sofosbuvir and velpatasvir. Most health professionals feel grief that the benefits of Baycadron in shutting the treatment of sarcoidosis far outweigh what the risks. It would appear that mylan pharmaceuticals inc. did n't want to expose the fact that prescription medicine dropper is less effective than that of their competitor.
More than one in 20 us children have fainting (syncope), sarcoidosis. So it can be concluded not to administer dangerous substance with high cholesterol. By 1978, Baycadron was first launched in avidly the UK as an injection to treat severe meningitis, listeriosis, with abducting a sublingual formulation released data in 1982.
Limited options developed for treatment in part reflect an incomplete understanding of how sarcoidosis affects the nervous system description to cause swelling caused by excess amniotic fluid (edema).
Next Next post: where can i buy omix 0.4 mg (Pms-cholestyramine) tablets.


Electronic changeover hygrostat, 10 to 90%, 24 V DC. Thermal Controls CC 301 series. All our enclos..
Electronic changeover thermostat, -10 to 80°C, 24 V DC. Thermal Controls CC 301 series. All our enc..
Mechanical changeover hygrostat, 40 - 90% (orange knob). Thermal Controls CC 301 series. All our en..
Pre-set temperature thermostat NC OFF @ 10°C (50°F) - ON @ 0°C (32°F). All our enclosure accessorie..
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Studio Electronics MidiMoog For Sale – Tone Tweakers Inc.
Studio Electronics MidiMini for sale. Fully serviced by professionals and working great.
Best known for its beefy bass and funky lead lines, the vintage Model D Minimoog can generate lots of other analog sounds - from musical to sound effects. The Mini has such a smooth, fat and powerful sound that nearly every modern virtual analog synth (and many hardware synths) were designed to sound like it. But they never quite capture that magic.
Its sweet sound can be heard all over music from the 70s to today. Notable Minimoog users include: Kraftwerk, Parliament / P-Funk (and lots of Dr. Dre productions), Depeche Mode, Keith Emerson, Devo, Vince Clarke, Bernie Worrell, Steve Winwood, Jan Hammer, David Frank, Geoff Downes, Gary Wright, Herbie Hancock, Gary Numan, Rick Wakeman, etc, etc, etc.
Years ago, Studio Electronics used to have people send them their Minimoog so they could turn it into a rackmounted MIDIMoog - they'd remove the circuit boards and toss the chassis. Great idea at the time I guess, but this definitely reduced the number of intact Minimoogs out there. This unit is one of the poor victims - an actually Minimoog stripped of its wood (castrated?) and transplanted into a smaller, neater metal enclosure.
So this is an actual 4 space rackmounted Minimoog with some really really useful improvements over the original Mini. Not only does the MIDI MOOG fit in a much smaller space and weigh a lot less than the original Mini - It's also got MIDI built in. That's right - no need to fuss with a MIDI to CV convertor and figure out how to hook it up to that crazy S trigger connector thingy. It also responds to aftertouch and velocity, pitch bend and modulation wheel data. An extra LFO for vibrato, so you don't have to tie up the 3rd oscillator for this as you would on a stock Mini Moog. AND it's also got oscillator sync, which can be swept by an envelope generator for that Cars distorted guitarish lead sound everyone knows and loves! It's also got switchable note priority modes. This unit makes a nice alternative to an original Minimoog due to its improvements, portable package and greater reliability.
Photos are not yet available, so any images here show a previously sold unit.
Estimated preparation time: 1-4 weeks.


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Q: sorting a report by subreport values I have a subreport that provides items and their value, with a sum of the value at the bottom. I would like to sort my main report, which is the owner of the items, by the total sum of the item value.
I have a main report that supplies the user with a persons name and contact information. I have built a sub report that takes the person's ID and displays the name and value of items associated with them. The subreport has a sum of the items. I would like to sort this report so that the highest total value is at the top. I'm new to working with sub reports in SSRS so any help would be appreciated!
this is my report -> type and balance are the sub report. I know that these are already sorted, but that is just coincidence and I have about a thousand rows that need to be sorted by total balance

A: I assume that the subreport is in a tablix of some kind?
If so then the usual process is to include the value you want to sort by in the dataset that drives the tablix.
So the dataset for tablix containing the subreport might look something like
SELECT *
FROM (
      SELECT personID, SUM(transactionvalues) as SortValue
          GROUP BY personID
     ) q
ORDER BY q.SortValue DESC

Now you can just sort your tablix by the SortValue

A: You also can apply the sorting from the report side. If your Sum() has the following expression in your tablix,
=Sum(Fields!Balance.Value)

you can add the exact same expression in the Tablix Properties > Sorting > Add (with the option from A to Z or Z to A).

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