An increase in extracellular Ca2+ induces growth arrest and differentiation of human keratinocytes in culture. We examined possible involvement of S100C/A11 in this growth regulation. On exposure of the cells to high Ca2+, S100C/A11 was specifically phosphorylated at 10Thr and 94Ser. Phosphorylation facilitated the binding of S100C/A11 to nucleolin, resulting in nuclear translocation of S100C/A11. In nuclei, S100C/A11 liberated Sp1/3 from nucleolin. The resulting free Sp1/3 transcriptionally activated p21CIP1/WAF1, a representative negative regulator of cell growth. Introduction of anti-S100C/A11 antibody into the cells largely abolished the growth inhibition induced by Ca2+ and the induction of p21CIP1/WAF1. In the human epidermis, S100C/A11 was detected in nuclei of differentiating cells in the suprabasal layers, but not in nuclei of proliferating cells in the basal layer. These results indicate that S100C/A11 is a key mediator of the Ca2+-induced growth inhibition of human keratinocytes in culture, and that it may be possibly involved in the growth regulation in vivo as well.
Exquisite spatial and temporal control of cell growth and differentiation is a prerequisite for embryonic development and maintenance of fine tissue architecture. The epidermis is a typical tissue in which compartments of growing cells and growth-arrested terminally differentiating cells are clearly demarcated. In normal epidermis, proliferating cells are only observed in the basal layer. On moving to the upper layers, the cells stop growing, progressively follow a terminal differentiation pathway, and finally shed off from the surface. The critical step determining whether the cells withdraw from the growing cell population is regulated by a complex network involving many genes, but the precise molecular mechanisms remain largely unknown.
A number of factors have been reported to trigger differentiation of human and mouse epidermal keratinocytes in culture, i.e., increased extracellular Ca2+ concentration (Hennings et al., 1980), TGFβ (Shipley et al., 1986), forced expression of PKC (Ohba et al., 1998), and detachment of cells from the substrate (Zhu and Watt, 1996). Normal human keratinocytes (NHKs) continuously proliferate only in a culture medium with Ca2+ of <0.1 mM. An increase in the Ca2+ concentration to 1.2–2.0 mM results in termination of cell growth and induction of terminal differentiation phenotypes (Hennings et al., 1980). An increase in the extracellular Ca2+ level resulted in a sustained higher intracellular Ca2+ concentration (Sharpe et al., 1989). Some other conditions inducing differentiation of keratinocytes also lead to increased intracellular Ca2+ levels (Sharpe et al., 1989; Missero et al., 1996). An increasing gradient of Ca2+ concentration is present from the basal to the cornified layers of the epidermis in vivo (Menon et al., 1992). Mice lacking the expression of full-length extracellular Ca2+-sensing receptors showed deteriorated epidermal differentiation (Komuves et al., 2002). These results indicate that higher Ca2+ levels lead to induction of epidermal differentiation not only in culture, but also in vivo.
An elevation in the Ca2+ level triggers a number of intracellular signal transductional events, including production of inositol 1,4,5-triphosphate and 1,2-diacylglycerol; activation of calcineurin, PKCs, and Raf/MEK/ERK pathway; and tyrosine phosphorylation of p62 and fyn (Dotto, 1999; Schmidt et al., 2000). On the other hand, it has been shown that p21CIP1/WAF1 was induced 4 or 8 h after an increase in the extracellular Ca2+ level, leading to inhibition of Cdk activity and blockage of cell cycle progression (Missero et al., 1996). p21CIP1/WAF1 protein has been detected in cells of the suprabasal layers, but not in those cells of the basal layer of the human epidermis (Ponten et al., 1995). An important missing link is how the Ca2+-induced initial events lead to the induction of p21CIP1/WAF1.
In a previous work on density-dependent growth inhibition of normal human fibroblasts, we identified S100C/A11 (calgizzarin), a member of the Ca2+-binding S100 protein family, as a key mediator of growth arrest (Sakaguchi et al., 2000). In a confluent state, S100C/A11 was phosphorylated at 10Thr and translocated into nuclei, and it eventually inhibited DNA synthesis through the induction of p21CIP1/WAF1 and p16INK4a. S100C/A11 is comprised in the epidermal differentiation complex (EDC) located on chromosome 1q21 in humans. EDC encodes nearly 30 genes. About half of them are specifically expressed during Ca2+-dependent terminal differentiation of keratinocytes (e.g., profilaggrin and loricrin), and the other half are members of the S100 protein family. The S100 family proteins have been assumed to play signal transduction roles in the differentiation of epidermis and other tissues. Some of the S100 protein family members were differentially expressed in normal human skin and melanocytic lesions (Boni et al., 1997). S100C/A11 was reported to be up- or down-regulated in malignant tumors (Van Ginkel et al., 1998). These results prompted us to examine possible involvement of S100C/A11 in growth regulation of epidermal keratinocytes. Here, we show that S100C/A11 is a key mediator of the high Ca2+-induced growth arrest in human keratinocytes.
Increase in extracellular Ca2+ concentration inhibits the growth of NHK and HaCaT cells
At first, we confirmed that an increase in extracellular Ca2+ concentration from 0.03 to 1.5 mM results in inhibition of DNA synthesis of NHK cells in dose- and time-dependent manners (Fig. 1, A and B). HaCaT cells, an immortalized human keratinocyte line (Boukamp et al., 1988), were routinely cultivated in a medium with 1.5 mM Ca2+. When the concentration of Ca2+ was increased to 5 or 10 mM, DNA synthesis of HaCaT cells was inhibited (Fig. 1, A and B). Under similar conditions, Sharpe et al. (1989) and Gonczi et al. (2002) reported that there was no remarkable difference in intracellular Ca2+ level (70–100 nM) of NHK and HaCaT cells, despite different extracellular Ca2+ concentrations. Intracellular Ca2+ concentrations in NHK and HaCaT cells increased by ∼40% within 30 min after increasing the extracellular Ca2+ concentrations from 0.03 to 1.5 mM and from 1.5 to 10 mM, respectively (Fig. 1 C). Overall, the response of HaCaT cells to 10 mM Ca2+ was similar to that of NHK cells to 1.5 mM Ca2+. No immediate cytotoxicity was noted in HaCaT cells in a medium with 10 mM Ca2+ during the observation period. Therefore, throughout the present work, 0.03 and 1.5 mM Ca2+ was used as low Ca2+ media and 1.5 and 10 mM Ca2+ was used as high Ca2+ media for NHK and HaCaT cells, respectively.
S100C/A11 inhibits the growth of human keratinocytes
To examine the possible involvement of S100C/A11 in growth regulation of human keratinocytes, we transfected various constructs of S100C/A11 flanking SV40-derived NLS to exponentially growing HaCaT cells. As shown in Fig. 2 A, incorporation of BrdU was specifically inhibited in the cells transfected with wild-type S100C/A11 (indicated by arrows), whereas GFP alone did not show any effects. DNA synthesis was inhibited by ∼25% by the wild-type S100C/A11 (Fig. 2 B). This reduction is substantial, considering that the transfection efficiency was at most 40%. Introduction of recombinant S100C/A11 protein into HaCaT cells by the aid of the protein transduction domain of TAT protein (YGRKKRRQRRR; Schwarze et al., 1999) efficiently inhibited DNA synthesis (Fig. 2 C). The TAT-flanked protein was transferred to cell nuclei with an efficiency of nearly 100%.
Increase in extracellular Ca2+ concentration results in phosphorylation and nuclear translocation of S100C/A11
When extracts of NHK and HaCaT cells labeled with [32P]phosphate were analyzed by a two-dimensional gel electrophoresis, increased phosphorylation of S100C/A11 (as identified by sequencing) was noted in both cell types exposed to high Ca2+ (Fig. 3 A). Exposure to high Ca2+ increased the immunoprecipitable amount of S100C/A11 and more remarkably the level of phosphorylation in both cell types (Fig. 3 A, bottom), whereas the S100C/A11 protein levels only slightly increased by high Ca2+ (Fig. S3).
To determine the phosphorylation sites of S100C/A11, endogenous S100C/A11 protein was purified from 32P-labeled HaCaT cells and degradated to peptides. As shown in Fig. 3 B (middle and bottom), only NH2-terminal p1-1 and COOH-terminal p4 were phosphorylated in a totally high Ca2+-depending manner. When the synthesized peptides of S100C/A11 were incubated with cell extracts in vitro, NH2-terminal pepA (amino acid residues 1–23) and COOH-terminal pepF (87–105), but not four other peptides covering the middle part of S100C/A11, were significantly phosphorylated by the extracts prepared from NHK and HaCaT cells cultivated in the high Ca2+ media (unpublished data). Thr and Ser residues were phosphorylated in pepA and pepF, respectively (Fig. 3 C). Finally, phosphorylation sites in p1-1 and p4 were determined as 10Thr and 94Ser, respectively, by an amino acid sequencer (Fig. 3 D). In accordance with this, Ala-substituted forms at 10Thr of pepA and at 94Ser of pepF were not phosphorylated by the cell extracts prepared after cultivating in the high Ca2+ media (unpublished data).
Increases in Ca2+ in the media resulted in partial translocation of endogenous S100C/A11 to nuclei of NHK and HaCaT cells as demonstrated by immunostaining (Fig. 4 A) and by Western blotting and autoradiography (Fig. 4 B). Preincubation with a membrane-permeable Ca2+ chelator, BAPTA-AM, inhibited the nuclear translocation of endogenous S100C/A11 by high Ca2+ (Fig. 4 A). An S100C/A11 variant protein lacking Ca2+-binding capacity was not translocated to nuclei by high Ca2+ (Fig. 4 C). Transfection of the wild-type S100C/A11-GFP construct before the shift to high Ca2+ resulted in nuclear translocation of the protein in HaCaT cells (Fig. 4 D). This nuclear translocation was not observed when 10Thr was substituted with Ala (Fig. 4 D) or when the NH2-terminal 23 amino acids were deleted (unpublished data). 10Thr-to-Asp–substituted protein (TD) was observed in nuclei even in the low Ca2+ medium. On the contrary, 94Ser-substituted forms behaved in a manner similar to the wild-type protein. A COOH-terminal truncated form of S100C/A11 lacking the residues from 87 to 105 was transferred to nuclei by the high Ca2+ (unpublished data). These results indicate that the phosphorylation of 10Thr, but not of 94Ser, is critical for the Ca2+-induced nuclear localization of S100C/A11, and that binding of Ca2+ to S100C/A11 is necessary for the translocation.
Binding of S100C/A11 to nucleolin is essential for nuclear translocation
Because S100C/A11 lacks the canonical NLS sequence, we screened for possible interacting protein(s) using a pepA(TD)-immobilized column (pepA with Asp at the 10th residue, a phosphorylated pepA equivalent). An ∼100-kD protein was identified in extracts of HaCaT cells as showing specific binding (Fig. 5 A, left). Sequencing of the eluted protein gave an amino acid sequence of VKLAKAGKANQGD, corresponding to the NH2 terminus of human nucleolin. The binding of S100C/A11 and nucleolin was confirmed in vitro (Fig. 5 A, right; Fig. 5 B). The bound amounts of nucleolin depended on the amounts of S100C/A11 protein immobilized, and S100C/A11(TD), a 10Thr phosphorylation equivalent form, showed higher affinity to nucleolin than did the wild type (Fig. 5 B). Synthesized pepA having phosphothreonine at the 10th residue showed a higher nucleolin-binding capacity similar to pepA(TD) (Fig. S6). Only pepA, and not the other five partial peptides of S100C/A11, binds to nucleolin. Furthermore, neither GST-S100A2 nor GST-S100A6, the other members of S100 family, bound to nucleolin as assayed in vitro (unpublished data).
In the Ca2+-stimulated HaCaT cells, S100C/A11 protein that was phosphorylated and bound to nucleolin was predominantly found in the nuclei (Fig. 5 C). Among various GFP-conjugated 10Thr- or 94Ser-substituted forms of S100C/A11 transfected to HaCaT cells, only 10Thr-to-Asp–substituted forms were coprecipitated with nucleolin in the low Ca2+ medium (Fig. 5 D). In the high Ca2+ medium, phosphorylation of S100C/A11 was observed when either 10Thr or 94Ser was intact. Nucleolin was coprecipitated with 10Thr-intact forms in addition to 10Thr-to-Asp–substituted forms. BAPTA-AM inhibited the phosphorylation and interaction of wild-type S100C/A11 with nucleolin by high Ca2+ (Fig. 5 E). The S100C/A11 variant protein lacking Ca2+-binding capacity was neither phosphorylated nor coprecipitated with nucleolin by high Ca2+ (Fig. 5 E).
When small interfering RNA (siRNA) against nucleolin was applied to HaCaT cells, the protein levels were substantially reduced (Fig. 5 F, top), whereas the control siRNA against GAPDH showed no effects. Increase in Ca2+ concentration translocated endogenous S100C/A11 into nuclei in the siRNA against GAPDH-treated cells, but not in the siRNA against nucleolin-treated cells (Fig. 5 F, bottom). The necessity of nucleolin for the Ca2+-induced nuclear translocation of S100C/A11 was also confirmed in an in vitro translocation assay using permeabilized HaCaT cells by depleting nucleolin with a specific antibody (Fig. S5). These results indicate that (1) high Ca2+ treatment of HaCaT cells induces endogenous activity to phosphorylate S100C/A11 at 10Thr and 94Ser, and eventually enhances the binding of S100C/A11 to nucleolin; (2) S100C/A11 binds to nucleolin through its NH2-terminal region in a 10Thr phosphorylation–dependent manner; (3) the binding of S100C/A11 to nucleolin is essential for the translocation of S100C/A11 to nuclei; and (4) the binding of Ca2+ to S100C/A11 is necessary for the phosphorylation, binding to nucleolin, and nuclear translocation of S100C/A11.
The effector protein for growth inhibition by S100C/A11 is p21CIP1/WAF1
The next question is how nuclear S100C/A11 exerts its inhibitory action on the growth of keratinocytes. We screened a number of possible candidates and found that p21CIP1/WAF1 mRNA was remarkably induced in NHK cells either cultivated at 1.5 mM Ca2+ or exposed to TAT-S100C/A11 (Fig. 6 A). p16INK4a was also induced, but to a moderate extent. Similar induction was observed in HaCaT cells in a time-dependent manner (Fig. 6 B).
To confirm the role of S100C/A11 on the induction of p21CIP1/WAF1, anti-S100C/A11 antibody was introduced into HaCaT cells by the aid of Chariot or polyethyleneimine (PEI). The polyclonal antibody that we used was specific as shown in Fig. 6 C, and was demonstrated to recognize the NH2-terminal pepA and COOH-terminal pepF of S100C/A11 (unpublished data). Binding of the introduced antibody to endogenous S100C/A11 was demonstrated by immunoprecipitation (Fig. 6 D). Introduction of the antibody inhibited the high Ca2+-induced phosphorylation of S100C/A11 (Fig. 6 E). Fab fragments of the anti-S100C/A11 antibody also reduced the phosphorylation of S100C/A11 in HaCaT cells (Fig. S8). The induction of p21CIP1/WAF1 by TAT-S100C/A11 was dose-dependently abrogated by the antibody introduced into the cells (Fig. 6 F). The decrease in the amount of p21CIP1/WAF1 protein due to the action of anti-S100C/A11 antibody was associated with the recovery of cdk2 activity as assayed in vitro (Fig. 6 F). Inhibition of cdk2 activity by S100C/A11 introduced into HaCaT cells was dose- and time-dependent. Finally, anti-p21CIP1/WAF1 antibody (but not anti-p16INK4a antibody) completely abrogated the growth inhibition by S100C/A11 as assayed for [3H]TdR incorporation or for cdk2 activity (Fig. 6, G and H), indicating that p21CIP1/WAF1 is the principal effector protein of the S100C/A11-mediated growth inhibition of human keratinocytes. Specificity of anti-p21CIP1/WAF1 and anti-p16INK4a antibodies was confirmed as shown in Fig. 6 C.
In nuclei, S100C/A11 competitively binds to nucleolin and liberates Sp1 and Sp3, which eventually induce p21CIP1/WAF1
Next, we addressed the question of how S100C/A11 in the nuclei induces the effector p21CIP1/WAF1. Screening using Signal Transduction AntibodyArray™ (Hypromatrix) resulted in identification of Sp1 as a protein that binds to nucleolin, and the binding was inhibited by the addition of S100C/A11(TD) (Fig. 7 A). Anti-Sp2 antibody (but not anti-Sp3 antibody) was also on the membrane, but showed a negative result. Binding of Sp1 to nucleolin was confirmed by applying the extract of HaCaT cells onto GST-nucleolin beads (Fig. 7 B). Sp1 did not bind directly to S100C/A11.
To examine whether nucleolin binds to Sp1 in the cells, we performed a coprecipitation assay (Fig. 7 C). Sp1 and S100C/A11 were detected in the precipitates prepared using anti-nucleolin antibody, whereas nucleolin, but not S100C/A11, was detected in the precipitates prepared using anti-Sp1 antibody. These results indicate that binding among the proteins actually takes place in vivo, and that S100C/A11 does not bind to Sp1. In HaCaT cells cultivated in the presence of 1.5 mM Ca2+, nucleolin bound to Sp1, but not to S100C/A11. When the Ca2+ concentration was increased to 10 mM, S100C/A11 instead of Sp1 was detected in the immunoprecipitates prepared using anti-nucleolin antibody, and the bound S100C/A11 was phosphorylated. When the extract was treated with alkaline phosphatase in advance, the amount of S100C/A11 bound to nucleolin was dramatically reduced and the binding of Sp1 was recovered concomitantly. Immunoprecipitation with anti-Sp1 antibody confirmed that the stimulation with high Ca2+ liberated Sp1 from nucleolin.
Regions of nucleolin and Sp1 involved in the interaction were determined using recombinant partial proteins (Fig. 7 E). The COOH-terminal region of Sp1 (segment 3) including a zinc-finger domain was shown to be responsible for the binding to nucleolin (Fig. 7 D, top). In turn, Sp1 in the cell extract bound to segment 4 of nucleolin immobilized on a membrane (Fig. 7 D, middle). Recombinant Sp1 segment 3 also bound to the nucleolin segment 4, indicating direct interaction between the two proteins. S100C/A11 was recovered only from beads conjugated with segment 3 of nucleolin (Fig. 7 D, bottom). When segment 3 of nucleolin was introduced into HaCaT cells, the Ca2+-induced p21CIP1/WAF1 protein level was decreased and the activity of cdk2 was recovered within 8 h, despite continuous cultivation at 10 mM Ca2+ (unpublished data). This is possibly due to a dominant-negative effect of the peptide through sequestering S100C/A11 by binding. The interacting regions of the proteins are depicted in Fig. 7 E.
Binding of nucleolin in cell extracts to Sp1 (segment 3) was dose-dependently inhibited by recombinant S100C/A11 (Fig. 7 F). S100C/A11(TD) showed a stronger effect, whereas S100C/A11(TA) did not affect the binding between nucleolin and Sp1 (segment 3).
Because Sp3 is also known to activate the p21CIP1/WAF1 promoter (Prowse et al., 1997), we examined possible interaction with nucleolin and S100C/A11 and found that (1) Sp3 can bind to nucleolin segment 4 via its COOH-terminal zinc-finger domain; and (2) the binding of Sp3 to nucleolin was inhibited by S100C/A11 (unpublished data).
A gel shift assay was performed using GC-rich regions of the p21CIP1/WAF1 promoter, the well-known Sp1/Sp3-binding sites (see Materials and methods). As shown in Fig. 7 G, exposure of NHK and HaCaT cells to high Ca2+ resulted in an increase in the intensity of the shifted bands by Sp1 and Sp3, which were further retarded by the addition of anti-Sp1 or -Sp3 antibody. Under similar experimental conditions in which HaCaT cells were cultivated in a high Ca2+ medium, addition of GST-nucleolin into the incubation mixtures inhibited the extent of the band shift as compared with GST (Fig. 7 G, right). This was possibly due to sequestration of Sp1 by exogenous nucleolin. The inhibition of the band shift by GST-nucleolin was abrogated by the 10Thr phosphorylation equivalent form S100C/A11(TD), whereas wild-type S100C/A11 and S100A2 showed no effect. We found that the interaction of the proteins were dose-dependent, and addition of an excess amount of wild-type S100C/A11 showed recovering effects similar to S100C/A11(TD) (unpublished data). These results indicate the competitive nature of the binding between S100C/A11 and Sp1/Sp3 to nucleolin. We performed a series of luciferase assays using a plasmid containing the p21CIP1/WAF1 promoter region (Nakano et al., 1997) and obtained essentially the same results as those obtained by using the gel shift assay (unpublished data). Deletion of the Sp1/Sp3-binding sites in the promoter abrogated the induction of luciferase activity by S100C/A11.
Biological relevance of the S100C/A11 pathway as a mediator of the growth inhibition of keratinocytes
The findings described in the previous paragraphs indicate that S100C/A11 is involved in growth inhibition of human keratinocytes induced by high Ca2+ (Fig. 8 A). Under low Ca2+ conditions, S100C/A11 is present mostly in the cytoplasm. The unphosphorylated S100C/A11 partly binds to actin fiber via its COOH-terminal region (unpublished data). An increase in intracellular Ca2+ results in activation of an as-yet-unidentified protein kinase(s) that phosphorylates S100C/A11 at 10Thr and 94Ser. Phosphorylated S100C/A11 binds to nucleolin via its NH2-terminal region and is translocated to nuclei. In the nuclei, S100C/A11 competes with Sp1 and Sp3 for binding to nucleolin, and the resulting free Sp1 and Sp3 induce p21CIP1/WAF1.
A question that arises is to what extent the pathway is biologically relevant. To address this question, anti-S100C/A11 antibody was introduced into NHK and HaCaT cells by PEI and Chariot, respectively. The introduction efficiency was confirmed to be >70% by immunostaining. The introduced antibody inhibited the high Ca2+-induced phosphorylation of S100C/A11 (Fig. 6 E). High Ca2+ suppressed DNA synthesis to ∼17% of the control level, and the antibody recovered the DNA synthesis to 64 and 72% of the control level in NHK and HaCaT cells, respectively, indicating that S100C/A11 is a principal (if not sole) mediator of growth regulation (Fig. 8 B). The incomplete recovery may be either due to incomplete blockage of S100C/A11 function by the introduced antibody or due to the presence of some additional pathway mediating high Ca2+-induced growth inhibition.
S100C/A11 was found to be expressed at various levels in many human tissues (Fig. 8 C), among which skin showed a particularly high level of S100C/A11 expression. When normal human skin tissues were stained with anti-S100C/A11 antibody, S100C/A11 protein was detected in epidermal keratinocytes (Fig. 8 D). Nuclei of cells in the basal layer were consistently negative for S100C/A11 protein, whereas nuclei of cells in the suprabasal layers were positive. p21CIP1/WAF1 protein was not detected in the basal layer, but appeared in suprabasal cell nuclei in which S100C/A11 was present. Among 21 nuclei counted in the basal layer, 20 and 1 nuclei were negative and positive for both S100C/A11 and p21CIP1/WAF1, respectively. In the suprabasal layer, 89 nuclei were positive for S100C/A11 and p21CIP1/WAF1, whereas 11 nuclei were positive for S100C/A11, but negative for p21CIP1/WAF1. No S100C/A11-negative nuclei expressed p21CIP1/WAF1. These results indicate that S100C/A11 not only mediates high Ca2+-induced growth inhibition of human keratinocytes in culture, but most likely plays a key role in the growth regulation in vivo as well.
We have demonstrated that an S100C/A11-mediated pathway plays a key role in high Ca2+-induced growth inhibition of human keratinocytes in culture. This finding has substantial biological significance for the following reasons: (1) high Ca2+ is a physiological inducer of growth arrest and differentiation of human keratinocytes in culture, and probably in vivo as well; (2) prior introduction of anti-S100C/A11 antibody resulted in recovery of the high Ca2+-induced growth inhibition of human keratinocytes by ∼70%, indicating that S100C/A11 is a key (if not the sole) mediator of growth regulation; and (3) immunostaining of the human epidermis for S100C/A11 and p21CIP1/WAF1 showed a picture consistent with the model presented as a summary of the results of the present report using cultured human keratinocytes (Fig. 8 A).
Phosphorylation and nuclear translocation of S100C/A11
Endogenous S100C/A11 was phosphorylated at 10Thr and 94Ser in HaCaT cells exposed to high Ca2+ (Fig. 3 D), whereas only 10Thr was shown to be the major target of phosphorylation in confluent normal human fibroblasts (Sakaguchi et al., 2000). The phosphorylation of S100C/A11 at 10Thr stimulated binding to nucleolin via its NH2-terminal region, this being a prerequisite for nuclear translocation of S100C/A11 (Fig. 5, C–F). In human keratinocytes, some of the cytoplasmic S100C/A11 proteins were colocalized with actin fibers. We found that S100C/A11 binds to actin through the COOH-terminal region and that phosphorylation of 94Ser reduced the affinity (unpublished data). The liberation of S100C/A11 from actin may facilitate the association with nucleolin and the eventual nuclear translocation of S100C/A11.
Ca2+ binds to S100C/A11 and induces its conformational changes (Rety et al., 2000). The binding of Ca2+ to S100C/A11 appears to be critical for the S100C/A11-mediated growth inhibition because interference of the binding either by amino acid substitution or with a membrane-permeable Ca2+ chelator (BAPTA-AM) inhibited phosphorylation, binding to nucleolin, and nuclear translocation of S100C/A11 (Fig. 4, A and C; Fig. 5 E). Cellular kinase activity for S100C/A11 as assayed in vitro using S100C/A11 or pepA was induced by high Ca2+ in NHK and HaCaT cells. The kinase activity was detected in a cytosolic-soluble fraction, but not in an insoluble or nuclear fraction, of HaCaT cells exposed to high Ca2+. When the cell extract was applied onto a cation exchange column, the activity was eluted with 160–180 mM KCl (unpublished data). Further characterization of the kinase(s) is now in progress in our laboratory.
Interaction among nucleolin, S100C/A11, and Sp1/Sp3
Nucleolin was first identified as a protein involved in ribosomal assembly and maturation, but was later found to have diverse biological functions (Srivastava and Pollard, 1999). Although nucleolin is mainly localized in the nucleus, it shuttles between the nucleus and cytoplasm and is even present on the cell surface. When we microinjected wheat germ hemagglutinin into HaCaT cells, which is known to generally inhibit nuclear import of proteins (Finlay and Forbes, 1990), nucleolin was diffusely distributed in the cells (unpublished data). Recently, Shibata et al. (2002) reported that nucleolin mediates a signal from the cell surface to the nucleus. Nucleolin inhibited transcriptional activation by A-Myb through specific binding (Ying et al., 2000). Heat shock or irradiation was shown to mobilize nucleolin from nucleoli to promote the formation of a nucleolin–p53 complex (Daniely et al., 2002). In the present work, we showed that S100C/A11 and Sp1/Sp3 compete for binding to nucleolin. This is the first report of nucleolin providing a site for switching transcriptional activation through protein–protein interaction. Unexpectedly, it was found that S100C/A11 and Sp1/Sp3 bind to different regions of nucleolin (Fig. 7 E). Binding of S100C/A11 may change the conformation of nucleolin so that it can no longer hold Sp1 and Sp3. Although nucleolin is notoriously sticky, the interaction with Sp1/3 and S100C/A11 is unlikely to be nonspecific because (1) only the specific regions of the respective proteins show the binding capacity; and (2) other members of S100, such as S100A2 and S100A6, did not bind to nucleolin.
Transcriptional activation of p21CIP1/WAF1 by Sp1 and Sp3
Sp1/krupple-like factors act as ubiquitous and tissue-restricted transactivators in many different contexts. Sp1 and Sp3 were shown to activate the p21CIP1/WAF1 promoter (Prowse et al., 1997). In the gel shift assay, the shifted bands shown in Fig. 7 G were further retarded by incubation with anti-Sp1 and anti-Sp3 antibody. We performed a series of luciferase assays using the plasmid containing the human p21CIP1/WAF1 promoter sequence of 2.4 kb (Nakano et al., 1997). Application of TAT-S100C/A11 induced the activity of the p21CIP1/WAF1 promoter by more than 10-fold in HaCaT cells in an Sp1/Sp3 site-dependent manner (unpublished data). This induction was abrogated by pretreatment of the cells with TAT-nucleolin segment 3, but not TAT-nucleolin segment 4. These results indicate that p21CIP1/WAF1, the effector protein of S100C/A11-mediated growth inhibition (Fig. 6 G), is transcriptionally activated by Sp1 and Sp3 in human keratinocytes when exposed to high Ca2+. This induction must be p53-independent because HaCaT cells are known to have only mutated p53 genes (Lehman et al., 1993).
Biological effects of p21CIP1/WAF1 in human keratinocytes
There remains little doubt that p21CIP1/WAF1, when expressed, inhibits the growth of keratinocytes through inactivating Cdks (Gartel et al., 1996). The results of immunostaining for p21CIP1/WAF1 (Fig. 8 D) suggest that p21CIP1/WAF1 is involved in growth arrest and differentiation of human epidermal keratinocytes in vivo. Various differentiation factors for human keratinocytes, including high Ca2+ (Todd and Reynolds, 1998), TGFβ (Datto et al., 1995), and TPA (Todd and Reynolds, 1998), are known to induce p21CIP1/WAF1.
In the human epidermis, p21CIP1/WAF1 protein was detected in the lower and middle suprabasal layers, but not in the uppermost layers (Fig. 8 D; Ponten et al., 1995). This seems reasonable because overexpressed p21CIP1/WAF1 was shown to inhibit terminal differentiation of mouse keratinocytes at later stages (Di Cunto et al., 1998). Alternatively, p21CIP1/WAF1 may simply not be needed in irreversibly arrested cells.
Regulation of growth and differentiation in keratinocytes
TGFβ transduces signals via Smad proteins. In HaCaT cells treated with TGFβ, Smad3 and Smad4 were found to cooperate with Sp1 for transcriptional activation of the p21CIP1/WAF1 promoter (Pardali et al., 2000). When we treated HaCaT and NHK cells with TGFβ, S100C/A11 was translocated to nuclei (unpublished data). Thus, S100C/A11 may be a partial converging point of high Ca2+ and TGFβ.
The proposed S100C/A11-mediated pathway for Ca2+-induced growth arrest (Fig. 8 A) has partly revealed the thus-far-unknown link from the initial events triggered by high Ca2+ to the induction of Cdk inhibitors, particularly p21CIP1/WAF1. However, we still have little information on how S100C/A11 is phosphorylated upon stimulation by high Ca2+. The specific phosphorylation observed in the present paper in terms of culture conditions and substrates (Fig. 3, B and D) provides an opportunity for the identification of the involved protein kinase(s). Furthermore, the function of S100C/A11 for liberation of Sp1 and Sp3 in nuclei was born by residues 1–23 of the NH2-terminal peptide. Considering the ubiquitous expressions of nucleolin and Sp1/Sp3 and the large variety of promoters that are regulated by Sp1 and Sp3, further narrowing down of the functional structural domain of the peptide and screening for chemicals that have similar structure may be a promising venture eventually leading to pharmacologically useful substances.
Materials And Methods
NHK cells (Kurabo Industries, Ltd.) were cultured in Defined Keratinocyte-SFM (GIBCO BRL) with Defined Keratinocyte-SFM Growth Supplement (GIBCO BRL). HaCaT cells, a gift from Dr. Fusenig (German Cancer Research Center, Heidelberg, Germany), were cultured in DME with 10% FBS. For monitoring DNA synthesis, 1 μCi/ml tritiated thymidine ([3H]TdR; American Radiolabeled Chemicals) was added to the cultures 1 h before cell harvest. An anti-BrdU antibody (Neomarkers) was used to stain BrdU-incorporated cells. Subfractionation of cells was performed as described previously (Lindeman et al., 1997). Imaging analysis of the intracellular Ca2+ level was performed as reported previously (Yamamoto et al., 2000).
Determination of phosphorylation sites
To determine the phosphorylation sites of endogenous S100C/A11, the protein was collected from HaCaT cells cultivated in the low or high Ca2+ medium using an affinity column with anti-S100C/A11 antibody. After further purification by PAGE, the protein was cleaved with cyanogen bromide and the peptides were fractionated by HPLC. Four peptides (Fig. 3 D) were identified by partial sequencing. pep1 was further cleaved with endoproteinase Asp-N (Roche). The purified p1-1 and p4 were subjected to NH2-terminal sequencing on a peptide sequencer (model 491; Applied Biosystems). Phosphorylated amino acid residues were run through the column, and thus disappeared from the expected position of unphosphorylated residues.
Introduction of proteins into cells
Peptides and S100C/A11 protein with relatively low molecular sizes were introduced into cells by the aid of the protein transduction domain of TAT protein (YGRKKRRQRRR) from human immunodeficiency virus (Schwarze et al., 1999). A Chariot kit (active motif) and cationic PEI-conjugated S100C/A11 antibody were used to introduce antibodies into HaCaT and NHK cells, respectively. To functionally inactivate proteins within the cells, rabbit anti–human S100C/A11, mouse anti–human p16INK4a (Ab-4, clone 16P04; Neomarkers), and mouse anti-p21CIP1/WAF1 (Ab-11, clone CP74: Neomarkers) antibodies were used. The anti-S100C/A11 antibody detects only a single band in Western blot analysis using cells irrespective to Ca2+ levels in the media (Fig. 6 C). Efficacy for neutralization of the target proteins p16INK4a and p21CIP1/WAF1 by the antibodies was confirmed by in vitro phosphorylation assay.
Antibodies and plasmid constructs
Antibodies and plasmid constructs used in the present work are listed in the Online supplemental material section.
An optimal target site of nucleolin mRNA for siRNA was determined as 5′-GACAGTGATGAAGAGGAGG-3′ by a program provided by QIAGEN, and the synthetic siRNA was purchased from the same company. siRNA against GAPDH mRNA (Ambion) was used as a control. The siRNA was transfected to the logarithmically growing HaCaT cells using LipofectAMINE™ 2000 (Invitrogen).
Cdk2 kinase assay
An in vitro cdk2 kinase assay was performed using immunoprecipitates with anti–human cyclin E antibody and histone H1 as a substrate as described previously (Ohashi et al., 1999).
Signal Transduction AntibodyArray™ (Hypromatrix) was used under the conditions indicated by the manufacturer. In brief, the antibody membrane was incubated with 1-mg HaCaT cell extracts, followed by the application of 10-μg full-length recombinant proteins (GST, GST-S100C/A11, GST-nucleolin, or GST-nucleolin + S100C/A11(TD)). After washing, the bound proteins were detected using HRP-conjugated anti-GST antibody (Santa Cruz Biotechnology, Inc.).
Gel shift assay
A gel mobility shift assay was performed as described by Nakano et al. (1997). We used GC-rich regions located between 120 and 94 bp (A), 93 and 74 bp (B), and 73 and 47 bp (C) of the human p21CIP1/WAF1 promoter as probes. The 32P-labeled probes (A + B + C; 104 cpm) were mixed with 4 μg crude nuclear extracts of NHK or HaCaT cells, incubated for 20 min at RT, and electrophoresed in a 5% polyacrylamide gel under nondenaturing conditions. For a super-shift experiment, 1 μg mouse anti–human Sp1 antibody (Santa Cruz Biotechnology, Inc.) or rabbit anti–human Sp3 antibody (Santa Cruz Biotechnology, Inc.) was added to the reaction mixture.
Acquisition and processing of images
For GFP-labeled vital cells, images were acquired by a fluorescent microscope (IX71–22FL/PH; CCD camera, DP50; objective lens, LCPlan F1 40×; Olympus) and processed using Adobe Photoshop® 6.0. For fixed cells labeled with FITC, Alexa® 488, or TRITC, a laser-scanning microscope (Axioplan 2; objective lens, Plan-Apochromat® 63× 1.4 oil DC and Plan-Neofluar® 40×/0.75; Carl Zeiss MicroImaging, Inc.) was used.
Online supplemental material
Fig. S1 shows dose-dependent inhibition of DNA synthesis by TAT-S100C. Fig. S2 shows expression of variant S100C/A11 proteins. Fig. S3 shows effects of high Ca2+ on endogenous S100C/A11 levels. Fig. S4 depicts intracellular localization of nucleolin. Fig. S5 shows necessity of nucleolin for nuclear translocation of S100C/A11. Fig. S6 depicts binding of S100C/A11 pepA with nucleolin. Fig. S7 shows proteins recovered by protein G from HaCaT cells applied with anti-S100C antibody or IgG. Fig. S8 shows Fab of anti-S100C/A11 antibody and reduced phosphorylation level of S100C/A11 induced by high Ca2+. Fig. S9 depicts induction of p21CIP1/WAF1 by variant S100C/A11 proteins. Fig. S10 shows interaction among nucleolin, Sp1, and S100C/A11. Fig. S11 shows competition of S100C/A11 peptides with Sp1 and Sp3 for binding to nucleolin. Fig. S12 shows the dominant-negative effect of nucleolin Seg-3 for the induction of p21CIP1/WAF1 by TAT-S100C/A11. The supplemental material is available.
We are indebted to Dr. T. Sakai (Kyoto Prefectural University of Medicine, Kyoto, Japan) for providing a plasmid.
This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (14370260 to N. Huh) and the Japan Society for the Promotion of Science (0104310 to M. Sakaguchi).
The online version of this article includes supplemental material.
Abbreviations used in this paper: NHK, normal human keratinocyte; PEI, polyethyleneimine; siRNA, small interfering RNA.


Welcome to Luxxery Boutique, a comprehensive plastic surgery and cosmetic enhancement practice designed to help both women and men renew and rejuvenate their appearance with exceptional results. At Luxxery, we invite you to immerse yourself in beauty by taking advantage of the many advanced surgical and non-surgical treatments available that can help you feel great about the way you look. Dr. Ayman Hakki and Dr. Nadya Clarke are experienced plastic surgeons who will customize your treatment based on your distinct needs and goals in order to produce results that not only meet your expectations, but also look natural and not artificial. From surgical body, breast, and facial enhancement procedures to non-surgical cosmetic treatments that can reduce excess fat pockets and rejuvenate the skin, we are committed to providing you with a vast selection of options from a skilled team of professionals who have your best interests at heart.
Dr. Ayman Hakki is certified by the American Board of Plastic Surgery and is a Fellow of the American College of Surgeons. Dr. Hakki is an extensively trained and skilled plastic surgeon who is experienced with surgical aesthetic enhancement procedures for the face, breasts, and body, as well as minimally-invasive procedures designed to diminish signs of age and help patients achieve a more youthful appearance.
Dr. Nadya Clarke is an experienced and immensely talented female plastic surgeon who performs an array of cosmetic enhancement procedures designed to recontour the body and enhance the look and feel of the skin. Dr. Clarke is adept with both surgical and non-surgical treatments that can help women and men renew their appearance and restore their self-confidence.


Bad habits for your heart
Everyone wants to have a healthy heart. Still,cardiovascular disease affects more than 1 in 3 adults in the United States.
The good news is that some simple, everyday habits can make a big difference in your ability to live a healthy lifestyle.
Here are the worst habits for your heart, and how to avoid them.
Sitting for hours on end increases your risk of heart attack and stroke, even if you exercise regularly.
“Intermittent exercise doesn’t compensate for the time you sit,” says Harmony R. Reynolds, MD, associate director of the Cardiovascular Clinical Research Center at NYU Langone Medical Center, in New York City.
Why? The lack of movement may affect blood levels of fats and sugars.
Dr. Reynolds advises walking around periodically and, if you’re at work, standing up to talk on the phone.
Leaving hostility and depression unchecked
Are you feeling stressed, hostile, or depressed? It can take a toll on your heart.
While everyone feels this way some of the time, how you handle these emotions can affect your heart health. “Those likely to internalize stress are in greater danger; research has shown a benefit to laughter and social support,” Dr. Reynolds says.
“And it’s helpful to be able to go to someone and talk about your problems.”
Ignoring the snoring
More than a minor annoyance, snoring can be a sign of something more serious: obstructive sleep apnea. This disorder, marked by breathing that is interrupted during sleep, can cause blood pressure to skyrocket.
More than 18 million Americans adults have sleep apnea, which increases the risk of heart disease. People who are overweight or obese are at higher risk for sleep apnea, but slim people can have it too.
If you snore and often wake up feeling tired, talk with your doctor; there are easy ways to screen for apnea, says Robert Ostfeld, MD, associate professor of clinical medicine at Montefiore Medical Center, in New York City.
While the exact reason is unknown, there is a strong link between gum disease and heart disease, Dr. Ostfeld says.
If you don’t floss, sticky, bacteria-laden plaque build up over time, which can lead to gum disease. One theory is that these bacteria trigger inflammation in the body.
“Inflammation promotes all aspects of atherosclerosis,” Dr. Ostfeld says. Treating gum disease can improve blood vessel function.
Withdrawing from the world
It’s no secret that on some days, other human beings can seem annoying, irritating, and just plain difficult to get along with.
However, it makes sense to strengthen your connections to the ones you actually like. People with stronger connections to family, friends, and society in general tend to live longer, healthier lives.
Everyone needs alone time, but you should still reach out to others and keep in touch whenever you can.
You’re either all or nothing
Call it the Weekend Warrior Syndrome.
“I see so many people in their 40s and 50s dive into exercising with good intentions, hurt themselves, and then stop exercising all together,” says Judith S. Hochman, MD, director of the Cardiovascular Clinical Research Center at NYU’s Langone Medical Center.
With exercise, it’s wise to aim for slow and steady. “It’s more important to have a regular exercise commitment,” says Dr. Reynolds. “Be in it for the long game.”
Drinking (too much) alcohol
Sure, studies suggest a small amount of alcohol may be good for your heart. Alas, too many over-imbibe.
Excess alcohol is linked to a greater risk of high blood pressure, high levels of blood fats, and heart failure. In addition, the extra calories can lead to weight gain, a threat to heart health.
If you drink, stick to no more than two drinks per day for men, and no more than one a day for women. (One drink means a 12-ounce beer or 4-ounce glass of wine).
Being overweight is a major risk factor for heart disease, and 72% of men and 64% of women in the U.S are overweight or obese.
Try to eat less, avoid oversize portions, and replace sugary drinks with water.
Dr. Reynolds and Dr. Hochman also suggest cutting portion sizes for high-calorie carbohydrates (think refined pastas and breads) and watching out for foods labeled “low-fat,” which are often high in calories.
Assuming you’re not at risk
Cardiovascular disease—including stroke, heart disease, and heart failure—claims more lives in the United States than any other illness, including cancer.
“Don’t assume you’re not at risk,” says Dr. Ostfeld.
High blood pressure and cholesterol, diabetes, being overweight, and smoking are all risk factors that should be kept in check.


FOR IMMEDIATE RELEASE:
Thursday, July 18, 2013
First Class of Peace Corps Global Health Service Partnership Volunteers Sworn In at the White House
Volunteers prepare to leave for one-year assignments working as medical or nursing educators in Tanzania, Malawi and Uganda
WASHINGTON, D.C. July 18, 2013 – Thirty U.S. doctors and nurses from across the country were sworn in at the White House today as the first class of Peace Corps Global Health Service Partnership volunteers. The new volunteers will leave this weekend for one-year assignments as medical or nursing educators in Tanzania, Malawi and Uganda, where they will work alongside local faculty to train the next generation of healthcare professionals.
“These volunteers will soon depart for a ground-breaking adventure – an opportunity to make a real difference in communities abroad while enhancing their own skills,” Peace Corps Deputy Director Carrie Hessler-Radelet said. “The Global Health Service Partnership is an exciting continuation of the Peace Corps’ commitment to global health.”
The Global Health Service Partnership – a collaboration of the Peace Corps, the President’s Emergency Plan for AIDS Relief (PEPFAR), and the non-profit Seed Global Health – presents an opportunity for American physicians and nurses to make a real difference in communities abroad by helping to address the known shortage of skilled physicians, nurses and clinical faculty in resource-limited countries. This innovative public-private partnership represents the first organized effort by the Peace Corps to send U.S. healthcare professionals abroad with a focus on teaching and expanding clinical capacity.
“Whether these volunteers serve in Tanzania, Malawi or Uganda, their work will help strengthen the capacity of health professionals and the capacity of health systems,” said Ambassador Eric P. Goosby, M.D., U.S. Global AIDS Coordinator. “Their contributions will help position partner countries to more effectively, efficiently, and sustainably address some of their greatest health challenges, including HIV/AIDS.”
Sub-Saharan Africa has the greatest shortage of physicians and nurses; the region has 24 percent of the global burden of disease but only three percent of the world’s health workforce. While the United States has 280 physicians and 980 nurses for every 100,000 people, countries in Sub-Saharan Africa, like Tanzania, have just one physician and 24 nurses for every 100,000 people.
“We’re proud to be in this partnership that is sending an outstanding group of doctors and nurses abroad to help build a pipeline of medical professionals in the countries that need them most,” said Vanessa Kerry, M.D., Chief Executive Officer of SEED Global Health. “This effort will help ensure that more well-trained doctors and nurses will be walking the wards and caring for patients in hospitals and clinics in Malawi, Tanzania and Uganda for years to come.”
The inaugural group of Global Health Service Partnership volunteers comes from diverse backgrounds, with clinical experience extending from a few years to decades of experience. The doctors and nurses range in age from 26-70, include seven returned Peace Corps volunteers, and have collectively worked in more than 32 developing countries throughout the world.
Fifty-two years ago, President John F. Kennedy hosted a ceremony at the White House to swear in the first-ever class of Peace Corps Volunteers. While honoring that legacy, the Peace Corps takes a step forward in inaugurating the first volunteers of the Global Health Service Partnership program, one of several new and innovative Peace Corps initiatives that expand service opportunities and use 21st century tools to meet development goals.
About the Peace Corps: Since President John F. Kennedy established the Peace Corps by executive order on March 1, 1961, more than 210,000 Americans have served in 139 host countries. Today, 8,073 volunteers are working with local communities in 76 host countries in agriculture, community economic development, education, environment, health and youth in development. Peace Corps volunteers must be U.S. citizens and at least 18 years of age. Peace Corps service is a 27-month commitment and the agency’s mission is to promote world peace and friendship and a better understanding between Americans and people of other countries. Visit www.peacecorps.gov for more information.
About PEPFAR: The U.S. President’s Emergency Plan for AIDS Relief (PEPFAR) is the U.S. Government initiative to help save the lives of people affected by HIV/AIDS around the world. PEPFAR is the largest commitment by any nation to combat a single disease internationally and PEPFAR investments also provide a platform for efforts to address other public health needs. PEPFAR is driven by a shared responsibility among donor and partner nations and others to make smart investments to save lives. For more information, visit www.pepfar.gov.
About Seed Global Health: Seed Global Health is a non-profit whose mission is to strengthen health systems globally by partnering US physicians and nurses with local educators. Seed Global Health believes educational partnerships can rapidly increase the pool of providers and educators in countries where they are most needed. Committed to recruiting the best-qualified candidates, including those who may have financial constraints to service, Seed Global Health raises and disburses loan repayment and other appropriate stipends of support to individuals chosen for assignments abroad. Visit www.seedglobalhealth.org for more information.
# # #


The CIHI report, which covers 21 health system measurements in six
different clinical areas, employs a high and low rate indicator that
takes into consideration the size of the hospital (caseload), then
affixes an "adjusted rate" to equalize the impact since the comparisons
are based on either per 1,000 or per 100.
SMH had a total inpatient load of 2,278 in this study period, staffed
28 beds, and operating expenditures of $28.6 million. It is included
with other "peer group" "small" hospitals for comparison purposes.
Below are some highlights of the adjusted rates at SMH:
C-sections: 32.7 per 100
VBAC: 20 per 100
Coronary Angio following AMI: 57 per 100
28 day readmission - AMI: 20 per 100
28 day readmission - Stroke: 11 per 100
30 day mortality - AMI: 17 per 100
30 day mortality - Stroke: 4 per 100
30 day readmission - Medical: 12 per 100
30 day readmission - Obstetrics: 1 per 100
30 day readmission - Overall: 8 per 100
30 day readmission - Surgical: 5 per 100
30 day readmission - 19 and under: 3 per 100
SMH spends eight per cent of its operating budget on administration
including finance and human resources. And SMH's cost per weighted case
In January, SMH submitted a 368 page Pre Capital Schedule
A report to the Central Local Health Integration Network
(LHIN) laying out the justification for a multi-million expansion to
the Alliston hospital.
Constructed in 1962 to serve about 7,000 people, it handles more than
30,000 cases through its emergency department, which is about 4,000 sq.
ft in space.
"This is well less than half
of documented hospital standard stated in the Hospital Service Needs
and Capacity Planning report and creates heightened risks for infection
control, patient and staff safety, patient privacy, mobility access,
falls prevention and ambulance off-loading protocols," according to the
submission's cover letter. "The fact that the
main lobby, hospital registration desk, coffee shop, gift shop and
entrance to the Hospital are now shared with the Emergency Department
triage area and waiting room is unacceptable. A recent Ministry of
Health review as well as a HIROC review of these facilities found code
violations and issues that constitute ever increasing risk factors for
both the Hospital and the Ministry."


Medical insurance normally supplies very little protection for psychological health problems. Sites Like Betterhelp…
In fact, the overall cost of therapy is generally similar to the co-payment of a lot of insurance coverage strategies.
The cost for counseling ranges from $80 to $100 per week, depending on which plan you choose, and consists of limitless access to your counselor.
FINDING THE RIGHT THERAPIST IS EASIER
With standard therapy, you might discover it difficult to find the ideal therapist, or any therapist, in your area.
Seventy-seven percent (77%) of counties in the US have a serious lack of psychological healthcare workers.
You’ll likely be restricted to therapists in your insurance coverage company’s company network if you have psychological health insurance coverage.
They may decline new patients, may be booked for weeks beforehand, or may not be experienced in handling your problem.
Regardless of how you locate a therapist, there is no assurance of finding a good match.
‘s intake questionnaire matches you with a therapist who has the qualifications and proficiency you choose in a therapist, typically within 24 hr.
In addition, you can mention your choice concerning your therapist’s gender, age, language, sexual orientation, area of know-how, and more.
And, if for any reason you are not pleased with your therapist, makes it easy for you to switch to a new one.
There’s no unpleasant separation with your existing therapist.
Among the first considerations you probably have when signing up for online treatment is how much it will cost you and if the platform will take insurance. It is essential to think about the logistics of taking on a brand-new weekly or monthly expenditure. Now let’s explore what is, how it works, and what you can anticipate it to cost. We’ll talk about whether insurance coverage will play a role in settling the cost of our online treatment sessions and answer other payment-related concerns.
And since lots of standard therapy sessions are partially covered by insurance, it’s definitely logical to question, “Does insurance cover therapy?” or “Does take insurance?” Before diving into the answer to this question, we genuinely think is an economical, life-changing alternative for those who want to transform their mental health for the better.
is the world’s biggest online therapy platform. This therapy service provides a lineup of over 23,000 board-certified and licensed suppliers and therapists offered to you. is designed to be an economical, convenient, and discrete online platform.
People often ask these concerns about seeking services:
Is it difficult to cancel?
Is a genuine therapist?
All our service needs is a quick sign-up questionnaire and a monthly membership strategy.
Can You Get Identified On?
Many people seek out therapy due to the fact that they are concerned that they may be coping with a mental illness, like depression or an anxiety disorder. If this is the case, you may be questioning if you can get diagnosed by a therapist.
While our therapists are equipped to help you with a large selection of mental health disorders (and a lot of them are qualified to detect), they are not allowed by to diagnose mental disorders on the platform. Their restriction from clinically detecting likewise suggests they can not prescribe medications for any mental health or substance utilize disorders.
To numerous, among the most intimidating parts of starting therapy is discovering a therapist. There are thousands of therapists to pick from, but most people don’t understand how to start limiting the list or even producing a list in the first place.
For those searching for a simpler and more personal method to match with a therapist, the online treatment provider might be a good alternative. Online therapy has become more popular than ever since the COVID-19 pandemic, and has been a typical method to find a licensed therapist.
is not covered by insurance coverage, but your insurance provider may still repay you. And even if you are unable to be compensated, is still one of the more budget friendly alternatives for treatment.
is a membership service that costs $60 to $90 a week (billed every 4 weeks), which indicates you can expect to be billed $240 to $360 a month. The specific service price depends upon your place, therapist preference, and therapist accessibility.
You can cancel your subscription at any time.
For those who can not pay this fee, financial aid might be available. You can contact the group to find out more about payment alternatives.
There are a few methods to get more details about psychological health protection.
You can go directly to your health insurance business’s website and log in. On the website, you need to have the ability to gain access to info about what your medical insurance plan covers and what kind of costs you can anticipate for therapy, such as a copay.
If you want to talk to someone straight, you can always call the toll-free number on the back of your insurance card. In this manner, you can get the most precise details about whether you’ll have the ability to have actually therapeutic services covered, what you need to do to have them covered, and what out-of-pocket costs you ought to anticipate.
Lastly, if you’re guaranteed through your company, you can also call your company’s human resources (HR) department straight for more details on what psychological health services your plan uses.
With online therapy platforms, you can find effective options for handy treatment by matching with a certified psychological health professional. Embellished, budget friendly treatment can be right at your fingertips– from the personal privacy of your own house or wherever you feel comfy. While online therapy is often covered by insurance coverage, it isn’t always. You may feel overwhelmed by navigating the world of coverage and online therapy, but you shouldn’t. Continue reading for a summary and suggestions about insurance coverage of therapy insurance coverage of therapy and online treatment.
deals cost effective rates alternatives for therapy with licensed mental health companies.
Though protection for therapy and other mental health services generally differs based upon the health insurance service provider and private strategy, is not covered by a lot of plans. Contacting your health insurance company is the best way to learn about your coverage.
In general, uses inexpensive rates that may be equivalent with the co-payments needed by most insurance coverage plans.


05 Oct Occipital Nerve Blocks | Charleston, SC
Therapeutic pain blocks can be crucial to patients suffering from severe or chronic pain. Occipital nerve blocks are used to treat chronic and cluster headaches and migraines. It involves injecting a steroid and local anesthetic into the occipital nerves at the back of the head.
What to Expect
When you come in for your appointment, expect to spend a short amount of time. The injection itself takes just a few minutes and is done without sedation. A local anesthetic may be applied to prevent any discomfort during the procedure.
You’ll be asked to lie down for the procedure. A fine needle is inserted just above the occipital nerve, which is located at the back of the head, near the neck. A steroid medication is injected. This will help reduce inflammation and block pain signals, reducing or eliminating headache pain. You may feel as if the side of your head has gone “numb.”
Who Shouldn’t have an Occipital Nerve Block?
Patients who are allergic to any of the medications to be injected, who have poorly controlled diabetes or heart conditions, or who have an active infection should not have the procedure. Also, patients who are on blood thinners should discuss the procedure with their doctor before proceeding.
How Long Does the Block Take to Work?
The full degree of pain relief may be apparent within minutes or take up to a full day to develop. The block works differently for everyone, and it’s impossible to tell whether the pain relief will last or how effective the injection will be. If the pain relief is significant, your physician may suggest repeating the procedure to prolong its effectiveness.
Following the procedure, patients should be alert for changes in mood, sensory abnormalities, swelling or discoloration of the injection site, or a reaction to the anesthesia which can include nausea and chest discomfort. Talk to your doctor about any adverse effects you experience.


Urology provides diagnostic, treatment and surgical services to patients with disorders of the urologic and reproductive systems. The urodynamics and incontinence program at Temple Community Hospital provides patients with the most advanced therapies to correct both common and complex problems in men and women.
To learn more about our Incontinence Program, please provide the following information:
***** All fields with an * are required *****
Comments to [email protected]
PH: (213) 382-7252
© 2002 All Rights Reserved.
Temple Community Hospital, 235 N. Hoover St., Los Angeles, CA 90004


Dr. Damper completed his fellowship at the Non-Surgical Orthopedic Institute in Marietta, GA and specializes in Interventional Spine and Pain management. He is passionate about the holistic and multi-system approach to evaluating, diagnosing and treating patients. His enhanced foundations of osteopathic training were focusing on treating the entire person, not just symptoms and he believes focusing on the mind, body and spirit relationship to enable the body to heal itself. Dr. Damper and his practice will construct and communicate clear diagnoses and treatment plans for every patient under his care.
Walter Calvin Damper, DO, is an interventional spine, pain and sport specialist. Dr. Damper has a special interest in the treatment of nonsurgical care for his patients which would incorporate interventional treatments, electro diagnostic studies, osteopathic manipulation therapies and rehabilitations. He enjoys helping patients during their healing process from complex spinal, musculoskeletal and sports injuries. With innovative treatments, he brings a strong passion to help patients get back to enjoying their daily activities.


Study: Vitamin C May Fight Cancer
Lab Tests Show Possible Effect From IV Doses; More Work Needed
Vitamin C Safety Trial Under Way
Vitamin C was only tested on human cells -- not people -- in this study. But vitamin C is being tested in people in a new project.
"Because IV ascorbate is easily available to people who seek it, a phase 1 safety trial in patients with advanced cancer is justified and underway," write Levine and colleagues.
That trial will see if intravenous vitamin C is safe for people, Levine explains. Tests on animals will be done to see if it truly fights cancer.
"If there is something useful, that will be absolutely wonderful for patients. If it turns out that the mechanisms do not hold up in animals in vivo, then maybe we will say that there is no point to do IV ascorbate," says Levine.
He notes that some complementary and alternative medicine practitioners worldwide use intravenous vitamin C for cancer treatment.
New Twist on an Old Theory
The idea that vitamin C might protect against cancer is more than half a century old and has been "colorful and emotionally charged," says Levine.
The theory was discarded 20 years ago by many scientists when other researchers found no benefit in giving vitamin C to patients with advanced cancer. In that study, vitamin C was given orally, not intravenously, notes Levine.
Scientists may not have considered that injected vitamin C might work differently than oral vitamin C, Levine explains.
Levine's team didn't start by studying vitamin C and cancer. Instead, they were probing nutrition and vitamin recommendations in healthy people.
Normally, the body tightly controls vitamin C taken by mouth. It quickly gets rid of too much vitamin C. Injected vitamin C bypasses those controls, says Levine.
"I realized that concentrations given intravenously at higher doses could be much higher than could ever be achieved by mouth," he says. "This is not nutrition use of ascorbic acid. This is potential use of ascorbic acid as a drug, and it must be said, it's not ready for prime time."
Science has a lot to sort out about vitamin C and cancer. Meanwhile, Levine recommends eating your fruits and vegetables. These include orange and grapefruit juice and fruits, raw red peppers, strawberries, and broccoli.
"Independent of this work, our recommendations are to consume vitamin C through foods and eat at least five varied fruits and vegetables a day," he says.


Worried about money??
Are you in pain but do not have any insurance and are afraid how much the doctor will cost???
I completely understand and hope this bit of information will help. Many people will go to their MD due to pain and in 99% of those cases they will leave his/her office with nothing more than a pain killer, muscle relaxer, or anti-inflammatory prescription. They will be told to come back in two weeks if that does not help. In two weeks they will be told to get an MRI or X-ray and to start physical therapy. The cost for these prescriptions, office visits and tests could be easily over $1000 and unfortunately the patient is still in a lot of pain.
HERE IS A BETTER CHOICE.
Talk to your friends, family and/or co-worker and find a chiropractor that they recommend. Call the chiropractic office and ask what their fee is for a patient with no insurance. Most chiropractors will be less than $100 for the first visit which will include consultation, exam, x-ray and if warranted a chiropractic adjustment. 90% of the time depending on the problem the patient will be feeling a little if not a lot better when they leave the office after that first visit. The doctor will want to see the patient again and give them a care plan to fully resolve the condition. In my office as in most I can fully relieve a patients pain for less than $350-500 which includes consultation, exam, x-rays and adjustments. I can do this in less than a month in most cases.
What a savings!!! Not only does the patient feel better, but they have avoided dangerous medication and unnecessary tests. They have not missed much work if any and they are truly healthier and at less risk of re-injuring their body.
My name is Daniel S. Gordon and I am a chiropractor in West Bloomfield, MI. Do not misunderstand my blog. I am not against modern medicine in any way, I just want people to know that not only can chiropractic help them get out of pain and fix their condition better than pills but it can save them a lot of time and money.


Therapists for Personality Disorders in South Africa
We found 233 therapists for you.
Individual, couple and family therapy for various psychological and psychiatric conditions, as well as scholastic assessments and career assessments.
I am a licensed counselling psychologist, registered with the HPCSA. I work in an insight-oriented and solutions-oriented approach to managing difficulties relating to depression,...
I offer a journey, a conversation, to take you to profound personal growth, and sometimes, to a place of healing.
Nicolas van Zyl
Clinical Psychologist Specializing in adult individual, family, couples and divorce mediation.
Registered (Counselling) Psychologist working from a private practice in Malvern, Queensburgh. I presently work with a wide array of problems in my private practice ranging from...
Nicola de Kock
I offer short and long-term therapy to adults and adolescents and aim to provide a confidential, non-judgemental, collaborative and at times gently challenging therapeutic space.
I am a registered Clinical Psychologist and Psychoanalytic Psychotherapist in private practice in Morningside, Sandton.
Garth Newman is a clinical psychologist with years of experience working with children, adolescents and their families at the Child and Family Unit of Lentegeur Hospital. His work...
Caroline Phofi a social worker in private practice runs a psychosocial Services counseling/education platform focusing counselling, trauma debriefing and emotional containment,...
Psychologist in the West Rand and Lenasia areas. Consults with all age groups. Special interest in self development and school assessments.
I am a Counselling Psychologist who enjoys working with teens and adults. I have worked in many different settings and with a wide variety of people.
I am a solution focused therapist who has worked in many areas, I believe in maximising an individuals potential in getting them to where they would like to be.
Dr Ravi Govender
I am a psychiatrist with practices in Tamboerskloof and Durbanville. I have an interest in Anxiety Disorders, Depression, Bipolar Disorder, Adult Attention Deficit Disorder and...
Individual and couples therapy with adults and adolescents. Both short and long-term psychotherapy
I provide short and long term therapy to children, adolescents and adults in a warm, non-judgemental and safe space.
Rina van der Watt
Our assessment and remedial therapy centre assists kids, parents, caretakers and teachers to address special learning needs. We also diagnose disorders such as autism and ADHD....
I am a clinical psychologist offering long- and short-term psychotherapy to adults. I aim to provide a safe, non-judgemental and empathic environment in which to explore and...
Michelle Joanne Andrews
I use an integrative approach tailoring my therapeutic model to you unique needs.
The collaborative therapeutic relationship can be a successful vehicle for change.
I am a Clinical Psychologist practicing in Illovo, Sandton. I specialise in couples’ counselling and life coaching, and work with individuals, couples and families from within a...
Dr Laura Comrie
I am a Sandton -based Clinical Psychologist offering Psychotherapy to individuals and couples. My goal as a therapist is to help you gain understanding of issues and well-being.
Counselling Psychologist focussing on both individual and family counselling, as well Equine Therapy for both individuals (adults, teenagers and children), and families/groups,...
I am a Clinical Psychologist registered with the Health Professions Council of South Africa and the Board of Health Funders. I offer a personalized short or long term treatment...
Northcliff Clinical Psychologist offering short, medium and long term psychotherapy for a range of psychiatric conditions including depression, anxiety, eating disorders etc.
I am a clinical psychologist registered with the Health Professions Counsel of South Africa and work from my private practice in Morningside, Durban. I provide psychological...
Sino Nocwaka C. Klaas
I work with children, adolescents and adults, as well couples and families. I provide a safe and non-judgmental space for my clients to explore their inner worlds.
I am a clinical psychologist practicing in Bedfordview. I offer therapy to children, teens and adults. I also work with parents in order to help them develop closer relationships...
Claudia Correia Marques
Claudia Marques a clinical psychologist with 5 years experience in the field of psychology. She specializes in adult and adolescent individual and couple psychotherapy. Her areas...
Currently I am working at Riverview Manor Specialist Clinic as part of a multidisciplinary team. Riverview Manor offers an evidence based therapeutic program which incorporates the...
Glynne Zackon is a clinical counselling social worker based in Johannesburg Gauteng. Glynne has had an illustrious career in therapy and has been in the industry for 24 years!...
Philip Van Rensburg
I am a registered Clinical Psychologist (HPCSA: PS0128775; PRACTICE NO: 0636908) and offer a wide range of psychological services.
I strive to provide high-quality psychological...
I aim to work collaboratively with my clients to find a good fit between your goals for therapy whilst being mindful of the real-life constraints
Addiction, Alcoholism, Schizophrenia, Bipolar Affective Disorder, Depression, Adult mental health issues.
I provide Psychotherapy and Psychological Assessment Services to Children, Adolescents, and Adults in both Claremont and Plattekloof, Cape Town.
I'm Natasha Reddy, a registered Counselling Psychologist practicing on the East Rand of Johannesburg. I consider myself a generalist and value working with all kinds of clients.
Lydia van Niekerk
Special interests in trauma, mood disorders, anxiety and chronic pain management.
Angela Cotterell has a Masters degree in Clinical Psychology (University of Stellenbosch, 2005) and is registered with the Health Professions Council of South Africa. She currently...
Using Brain Working Recursive therapy to treat Panic, Anxiety, Phobias, Mild Depression, Self esteem issues, Trauma and Sexual Abuse in as little as 2 or 3 sessions (or as...


How many times have you had to tap on your horn to get a senior driver in front of you to go a little faster? And have you ever felt the urge to help out an elderly woman with her shopping at a local grocery store? If your response was positive for both questions, then you have positively contributed to some of the problems facing the senior folk in the society.
Problems that come with age
Aging comes with numerous challenges. Oftentimes, the seniors struggle to get by all sorts of stereotypes every day. You, for instance, may have felt the empathy to help an old woman get her shopping to a parked car nearby because it appeared she was unable to do it. The slow driver on the road was apparently guilty of excessive old age caution according to you.
Certainly, one of the most common challenges faced by the elderly is diminished physical ability. As you grow older you begin to realize you can no longer keep up with some of the activities that used to be mere pieces of cake only a few years ago.
Even worse is the age discrimination and stereotypes levelled against the elderly. Sometimes people assume so much about the aged. While in the past they would be consulted and given high regard as being vastly knowledgeable and full of experience, today’s societies often stress more on the insomnia that is characteristic of old age and regard the old folk as less capable.
Loss of memory is a real challenge affecting people as they grow old and as such they need help. In such conditions, these people lose their independence.
This is ordinarily the process or condition of deterioration that comes with age. The body cells generally lose their power of division and associated growth. As a result, the body begins to experience diminishing conditions. Senescence includes biological, intellectual, emotional, spiritual and social changes associated with age.
This is perhaps one of the most inescapable of old age challenges. It is the medical condition characterized by the bones becoming brittle and fragile as they lose tissue. It results from hormonal changes in the body. Calcium or vitamin D deficiency are the other typical causes of osteoporosis.
Patients with osteoporosis usually suffer from reduced mobility and inability to perform certain basic tasks that do not ordinarily require the help of another person. As patients of osteoporosis grow older, they become not only less mobile but also potentially disabled particularly if they get fractured from a fall or if their vertebral bodies collapse.
In the US, it is estimated by the National Osteoporosis Foundation that 54 million Americans above 50 years of age are affected by the problem of low bone mass. This puts them at the risk of fracturing or breaking from simple accidents.
There’s Light at The End of the Tunnel
It is these kinds of challenges faced by people on a daily basis that have inspired the creation of Sockee, a revolutionary solution that helps everyone with mobility issues. With it, the elderly do not need anyone to help them out with the simple tasks such as putting on their socks particularly in the morning when the back is unusually stiff and sore.
The feeling of having a newborn baby can be both euphoric and overwhelming. For a mommy who has delivered her baby through caesarean operation, it is apparent that you will be recovering from a major abdominal surgery. At the same time, you will be handling typical postnatal issues like mood swings, engorged breasts, and vaginal discharge.
You should also be prepared for the baby blues. These are a mild form of depression that affects as many as 70 to 80 percent of new mums. The blues usually begins on the third or the fourth day after birth. They are as a result of hormonal change reactions the body experiences following the birth.
The period after a caesarean surgery requires a certain level of care to experience a quick and smooth recovery process.
Just like in any surgery, you are likely to experience some physical pain. You may feel for instance that you cannot do anything by yourself. Even small movements may become difficult and cause throbbing spasms. At this time, you may find it hard to shift on bed and moving up may require something hold on to, or someone to help out.
It will hurt when you laugh, and you may feel scissoring sensations when you cough. Some simple acts will make the laugh and cough experience a little more bearable, such as supporting your wound with a pillow or your hands.
What you can do to ease the discomfort and aid your recovery
Wear cotton underwear and loose clothes. Wearing knickers of a size bigger than your normal one will be a good idea. Boxer shorts or special knickers prepared from stretchy gauze that allow quality space over your scar will help ease the pain. The National Childbirth Trust (NCT) have some designed for this purpose.
You will also need a proper, well-balanced diet. A recovering body needs vitamins and enhanced immunity. Eat as soon as you feel hungry and drink immediately you feel thirsty.
Take plenty of water and supply your diet with a lot of fiber-rich foods to help prevent constipation. Be sure to get peppermint water from your nurses to soothe the pain caused by trapped wind associated with surgery.
Monitor your wound for any suspicious signs of infection. Watch out for discharge, extra soreness or redness. Alert your doctor or midwife as soon as you spot any of these signs.
Keep your wound clean by washing and drying as directed daily. You can shower or take a bath and use soap on it as long as you rinse it off. Gently pat dry your wound with a clean towel.
Correct your posture by avoiding hunching or bending as these may cause backache. Try to observe an upright posture when sitting and when standing. Look for help on any incidences that could force you to bend. You may consider wearing slip-ons that do not require a lot of effort to put on the feet. Also, if you want to wear socks, use Sockee to wear and remove them. This revolutionary new solution will enable you to wear your socks with your back in its recommended upright posture.
Hip replacement surgery is a life changing step. And as with any major surgery, you will need adequate preparation both physically and psychologically. You will need the moral and material support of your family.
Such procedures impact not just the person undergoing them but the people around as well. Your friends and family must be concerned about how it will be after you are operated on. They need to be prepared as well before you undergo the actual procedure. They will also be in a better position to support you if they have sufficient preparation.
Most of all, the physical condition of your house and the entire homestead may determine just how fast you recuperate. Hip replacement surgery is hardly ever an emergency. So you will often have adequate time to contemplate the situation as it is to be after your surgery. Keep in mind that you will not be as briskly in motion as you currently are, at least not for a while after the surgery.
Take the time to prepare your house to its optimum before you undertake the procedure. These preparations may help speed your recovery after your surgery. A suitable environment will also reduce your stress levels during your recovery.
Examine the surrounding by taking a walk around the house, looking for potential problems. Be sure to identify risky areas.
Hip replacement operation
The hip replacement surgery is a procedure that involves surgically removing a painful hip joint affected by arthritis and replacing it with an artificial joint. The artificial hip joint is often made from plastic and metal components. The procedure is usually done as a last resort when all other options of treatment have failed to provide sufficient relief from pain.
The surgery can be performed in two different approaches. The doctor can follow the traditional approach or use a minimally-invasive technique. The key difference between these two procedures is the extent of the incision.
Pre-surgery steps to take for safety and convenience after surgery
You will be taking the hip replacement surgery to enjoy a pain-free experience that comes after acquiring your new arthritis free hip.
Begin by the simple things: remove any rugs and small items from the floor. You will need free movement around as you recover. Also, ensure you tape down any electrical cords and wires that might cause injury.
Rearrange your furniture to leave wider pathways that will conveniently accommodate crutches, a wheelchair or walker. Set up your bedroom on the ground floor for stress-free access. You may have to learn the art of climbing up and down any stairs afresh.
Ensure you have all the things you frequently use on lower shelves and within arm’s reach.
Buy and install an elevated toilet seat if you don’t already have one.
Prepare a place where you will spend most of your time. Allocate an appropriate chair with a phone, remote control, TV and reading materials in place.
The idea is to reduce any movements that might expose you to the risk of hurting your recuperating hip. Minimize all movements that involve bending. Find alternatives to necessities that may force you to bend, such as wearing shoes and socks. Buy Sockee to use when putting on taking off your socks without any help. This revolutionary solution is designed to improve the lives of everyone with motion problems.
Our lifestyles today expose us to a number of problems. One common lifestyle related problem today is low back pain. When such pain on the lumbar region is experienced for long, it is then considered chronic. The problem can appear as bone pain, pain in the nerves or muscle pain.
Back pain is prevalent in various continents with the United States stats indicating that at least 31 million Americans experience low back pain at any particular time. And is a major cause of absenteeism from in many countries.
Causes of Chronic back pain
Several factors may cause chronic low back pain. Injuries from sports are a major culprit. But this only affects a small group of people, the sports community across the world. Ironically, this group is only affected during the period of injury, after which they recover perfectly and keep on with their sporting life.
There are however millions of people who are not involved in strenuous physical tasks that are serious victims of chronic back pain. The sedentary life that people live today is a primary cause of the pain. While the human body structure is designed to shift from one thing to another, remaining stationary for a long time at the same position puts a lot of pressure on the back and eventually makes it start to ache.
People who spend their professional lives on the computer are at a greater danger. They are more disposed to slouch over the machines and may not realize that the posture is hurting their spine. With time, they may start complaining of back issues that gradually become serious back pain.
Other factors that increase the risk of chronic back pain include obesity, emotional and psychological stress, inactivity and poor physical conditioning. Hidden diseases such as osteoporosis have also been identified as serious causes of back pain. This problem becomes one way through which the silent diseases become noticeable.
Relief from chronic back pain
The fortunate thing about back pain is that it can always disappear in a couple of days or weeks and may not stay for long. But when it takes longer- say three months or more to heal by itself then it cannot be taken lightly. Prescription drugs are normally administered for such patients of chronic back pain.
These drugs have however received their share of criticisms. They are said to be saddled with severe side effects that may be life-threatening. Nonsteroidal anti-inflammatory drugs (NSAIDs) are argued to put the patient at risk of stroke, heart attack and a good many other dangers, yet they are some of the most commonly prescribed painkillers in the market.
There are a lot of safe and utterly effective alternatives to prescription drugs that you should consider. You may need a little patience with some of them, but they pay off as the results are lasting and dependable.
These alternatives include massage, chiropractic care, yoga, Neuro-Structural Integration Technique (NST). And physical exercises for back pain.
Some healthcare providers will often recommend bed rest as therapy for back pain. But most of the chronic cases are majorly caused by a sedentary lifestyle. The best solution should be one that counters that lifestyle and keeps you on the move.
While at it, ensure you take all precaution and avoid any circumstances and activities that may exert pressure on your back. There are obviously some basics activities that are dependent on the same aching back. A major example is wearing and taking off your socks. There’s just no way to do it without bending over. Not anymore…you can now avoid that too. Just get yourself the new innovative solution, Sockee, and avoid bending while wearing your socks.


Beginning Monday, May 11, the Illinois Department of Public Health (IDPH) recommends:
“…oral health providers resume provision of routine oral and dental care [while] minimizing the risk of transmission of COVID-19.”
We at Creekside Dental are excited that we are now able to care for all our patients, especially those who have had to forego dental care, which may cause their oral health to deteriorate.
The IDPH’s Announcement states, in part, that “…there is a need for a plan to resume safe and routine oral and dental treatment as continued delay in the delivery of time-sensitive, disease treating, oral and dental care may result in an over-reliance on antibiotics and analgesics to manage oral health pathologies, patient complications, poorer prognoses, as well as the need for more complex and costly corrective oral and dental care.”
Toward ensuring everyone’s safety, we have increased our infection control protocols. Upon your next visit, you will notice:
- More plastic barriers
- Personal protective equipment (PPE)
- HEPA air purifiers in each treatment room.
We have also installed a central U.V. light air purifier. According to social distancing guidelines, we are managing the number of patients in our reception area.
“Oral health providers should continue to counsel their patients and communities on the primary prevention of oral disease to lessen the progression of pathologies.
To yield good procedural outcomes, oral health providers should consider their patient’s healthcare needs, assess the risks and benefits of any procedures, and appropriately screen patients for COVID-19.”
If you have questions or concerns, please contact us. We are here to support you during these times, and wish you good oral and overall health!
William L. McCune B.S., D.D.S., General Dentist 135 N. Arlington Heights Rd., Suite 185, Buffalo Grove, IL 60089
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Genetic Counseling Programs
FamGenix Free for Genetic Counseling Programs
Provide your students with our next-generation pedigree tools for free.
Genetic counseling students can access the FamGenix Platform for free.
FamGenix is partnering with genetic counseling programs to empower their students with the latest software tools to gather family history and assess hereditary risk more efficiently. Our platform assists students in learning and engaging with patients or research participants following an easy and time-saving workflow.
Learn by using the latest pedigree and risk assessment tools.
- Fast and easy pedigree maker with built-in medical ontology to enter any condition or gene
- Mobile-friendly patient health history collection with unique family sharing features
- Auto-identifies high-risk patients based on NCCN and ACMG/NSGC guidelines
- Built-in cancer risk assessment models, including BOADICEA, Tyrer-Cuzick v8 (IBIS), BayesMendel, Gail and Claus
- Track additional patient health, demographic or lifestyle data


Perhaps it’s dependency. Perhaps it’s not. We could assist you figure things out. Our personal phone-based or in-person assessment process will determine whether addiction rehab makes good sense for you– as well as, if so, what kind.
Where To Locate The Most Effective Adolescent Medication Rehab Programs That Will Help You in Emmetsburg
If several, complicated issues are entailed, we might advise a domestic assessment so our medical professionals could discover more about your circumstance as well as figure out the most effective strategy. Our approach equips youngsters to recognize the unfavorable consequences of utilizing alcohol and various other drugs. We work carefully in order to help teens discover how you can make better choices, begin a program of recuperation and also maintain sobriety.
What You Must Learn about Inpatient Medicine Rehab For Adolescent Teens
Teens and young grownups may have different treatment needs compared to older substance abusers, and lots of in this population may not see the need for treatment on their own. Privacy regarding drug abuse and denial could be common in young compound abusers, and also commonly therapy might originally be sought for them by their moms and dads or guardians. Age-specific and also specialty programs accommodating young adults and young adults might be beneficial in minimizing problematic medication or alcohol usage along with improving addiction healing around 50536.
Can You Anticipate Chemical Abuse Rehab For Teens To Aid in Emmetsburg
Techniques that have been scientifically examined as well as have proof to support their usage are usually called research-based treatments. These treatments incorporate scientific evidence and also the understanding as well as personal experiences of a treatment company. While a number of the very same therapy models that are made use of for grownups may be utilized during drug rehabilitation for a teenager or young grown-up, a few of the specifics of the program may differ to address the one-of-a-kind scenarios of this populace.
As parents watch the problem unfold when their teenager comes to be self-destructive, they wonder how they will ever reconnect with the kid or daughter that they fear they are losing. It is one of the most powerless, helpless feeling worldwide, as well as it could overwhelm a whole family with unthinkable discomfort, mayhem, as well as misery.
The best ways to Control Your Future and Also Get Rid Of The Manacles of Dependency
Life-controlling troubles of teens are not limited to medications or alcohol. Much of them fight with the particles of separation, have experienced traumas like sex-related assault or physical violence, intimidation, peer stress, and awkwardness that goes along with the challenges of expanding up.
Suitable assessment of teenager behavior is important because the line between regular and undesirable actions could be tough for parents to recognize. Because lots of treatment programs concentrate on obtaining teenagers to approve that they have a drug trouble, it is essential to figure out first whether that is genuinely the situation.
Heroin, long thought about one of one of the most dangerous entertainment medications, is a semi-synthetic opioid, not unlike several of the generally made use of prescription pain relievers. It was manufactured in 1874 as well as made use of for many years in hospital settings. Soon sufficient, medicine dealers started offering unclean dosages to individuals searching for a fix, and also heroin mixed a nationwide problem of addiction as well as overdose.
Search For Drug and Alcohol Rehab Centers For Adolescent Teenagers In Your Area
Roughly 23% of adolescents utilizing heroin will certainly create a dependency, and also 75% of heroin individuals were linkeded on opioid painkillers before switching to heroin. Heroin impacts the brain likewise, as well as is really more affordable than many prescription opioids– it’s even being sold in tablet form currently on the roads and about senior high schools. If your child is hooked on painkillers, it’s crucial to resolve their trouble before they turn to heroin or various other immoral drugs.Iowa


What is Psychotherapy?
Psychotherapy is an invitation offered by a trained psychotherapists to aid clients in reaching their full potential. Psychotherapy includes techniques to increase awareness, within your choices of thought, feeling or action. Psychotherapy can increase the sense of well-being and can better manage discomfort or distress. Psychotherapy can help you to deal with behavioral or emotional problems and psychological reactions to life events.
Types of Psychotherapy
This is the traditional talk therapy, where by gaining insight (understanding) from your past and childhood, you can modify how you feel. Types include Freudian, and Jungian therapy. It is extremely useful in treating depression, anxiety, trauma and self-exploration.
Cognitive Behavioral Therapy (CBT)
Cognitive Behavioral Therapy (CBT) is a type of psychotherapy that helps people change how they think, feel, or act in order to improve their mood, reduce stress, or achieve other important health and life goals. Some goals may be specific, such as reducing worrying or procrastination, whereas others can be more general, such as figuring out why one’s life seems to lack meaning, passion or direction, and figuring out what to do about it.
There are three parts to CBT:
- How you think (cognitive) can and does change your behavior
- The way you think may be monitored and altered
- The desired behavior change may be affected through changes in the way you think.
“Cognitive Behavior Therapy is focused on the present rather than the past,” said Deanna Barch, Ph.D., an associate professor of psychology in Arts & Sciences at Washington University and supervisor of the social anxieties group therapy project.
“It’s focused on the kinds of thoughts, beliefs, ideas that people have currently and how that influences their emotions and behaviors. It also focuses on what behaviors you are actually engaging in and not engaging in at the moment, in the here-and-now.”
The Cognitive Model
This therapy focuses around a cognitive model, which states that individual’s perceptions are where problems lie. The notion of the cognitive model is that these perceptions influence the way people think and behave. According to this model, a patient’s symptoms stem from distorted perceptions about themselves and the world around them. When an individual’s perceptions and basic understanding of the world around them are distorted, it creates distress and anxiety. The cognitive model aims to realign the individuals’ perceptions with reality by helping individuals learn how to recognize distorted thoughts and beliefs and then modify them to better represent reality. When the individuals begin to have more realistic thoughts, their symptoms tend to be become less severe, and, in many cases, go away.
Cognitive Therapy Goals
The goal of cognitive therapy is to identify what exactly is causing the patient distress. Then, working from there, the therapist can create a treatment plan to help alleviate, and ideally ultimately extinguish the symptoms. In treatment, the therapist tends to begin by identifying the patient’s assumptions about themselves and the world around them, and seeing how these thoughts are causing negative emotions and behaviors. The therapist then looks for information that can counteract these negative assumptions, slowly showing the patient how to modify their negative assumptions.
Cognitive Therapy in Practice
An example of a patient and therapist in cognitive therapy: A student receives a D grade on a test, and takes that information and begins to think, “I’m a terrible student”. The student then focuses on that negative idea, which leads to negative emotions, perhaps frustration, hopelessness, and despair. The individual begins to truly believe that they are a terrible student because of this one test grade. Because of these thoughts and feelings, the student may begin to exhibit behaviors that reinforce her perception of being a bad student, such as choosing not to participate in class discussions, answer questions, try for extra credit, etc. The individual will then, inadvertently, start to exhibit behaviors of a “bad student” because of this. However, the irony is that what is making the individual a bad student is her own thoughts and emotions of believing that she is a bad student. A therapist working with this individual will help her realize that it is her own perceptions of herself that is causing her to feel like she is a bad student, rather than her actually being a bad student. The reality of the situation is that this individual received one bad grade, and although she has created a world for herself in which this makes her a bad student, in reality, she is not.
Below is the Chinese character meaning “to listen”. The character embodies the essential aspects necessary to listen completely. You can capture the full spirit of listening when you give your ears, eyes, undivided attention, and heart. Listening is the most important part of therapy.
“The most basic of all human needs is the need to understand and be understood. The best way to understand people is to listen to them.”
— Ralph Nichols