Increased vitamin D serum levels correlate with clinical improvement of rheumatic diseases after Dead Sea climatotherapy.

BACKGROUND: Ultraviolet B (UVB) rays are required by the skin for the production of vitamin D. The intensity of UVB at the Dead Sea area is the lowest in the world. Low vitamin D levels are often associated with musculoskeletal symptoms. OBJECTIVES: To assess the effectiveness of climatotherapy at the Dead Sea on the production of vitamin D in Norwegian patients suffering from various rheumatic diseases and to investigate possible associations between increased vitamin D serum levels, musculoskeletal symptoms and disease severity. METHODS: Sixty Norwegian patients who came to the Dead Sea area for 21 days of medical rehabilitation were divided into three groups according to their diagnosis: chronic pain syndromes, i.e., low back pain or fibromyalgia (Group 1, n=33); rheumatoid arthritis (Group 2, n=16); and osteoarthritis (Group 3, n=11). Serum 25-hydroxyvitamin D (25-OH-D) levels were determined at arrival and prior to departure. The treatment protocol included daily sun exposure (climatotherapy), bathing in the Dead Sea and mineral spring water (balneotherapy), mud applications and fitness classes. RESULTS: 25-OH-D serum levels increased significantly from 71.3 +/- 26.6 nM at arrival to 89.3 +/- 23.2 nM prior to departure (P < 0.001). Adjusted for the initial levels of pain (assessed by a visual analog scale) and disease severity, a direct correlation was observed between increased 25-OH-D serum levels and pain reduction (P = 0.012) and reduction of disease severity (P = 0.02). CONCLUSIONS: Climatotherapy at the Dead Sea induces significant changes in vitamin D. Increased 25-OH-D serum levels are associated with reduced musculoskeletal pain and disease severity.

Nutritional status in relation to balance and falls in the elderly: a preliminary look at serum folate.

BACKGROUND/AIM: In elderly persons, fall-related injury is a serious public health problem. We investigated the impact of essential nutritional elements on falls in the elderly. METHODS: Clinical function, balance, gait and disability tests and health and nutritional status assessments were performed. All subjects were interviewed regarding the occurrence of falls in the last year. Blood tests for serum vitamin D, folate and B(12) were conducted among a randomly selected subsample of 54 participants in the same month. RESULTS: One hundred 65- to 91-year-old volunteers participated in the study, and 29 of them fell at least once during the past year. The depression score was higher (indicating more depressive symptoms) among fallers compared with non-fallers (4.0 +/- 3.2 vs. 2.5 +/- 2.3, respectively). The overall function score (indicating better function) was marginally higher in non-fallers. Subsequent comparisons between fallers and non-fallers were adjusted for overall function and depression scores. Serum folate was significantly lower in fallers (9.5 +/- 7.1 vs. 16.2 +/- 6.7 ng/ml, p = 0.02). Dietary intake was equal in both groups. Correlation analyses indicate a significant association between vitamin D and the functional measurements: timed get up and go (negative), Berg balance test, overall functional score, lower extremity score and limitation score (positive correlation coefficients). Serum folate was highly and negatively associated with the number of falls and with prescribed medications and was the only protective factor against falls in a multivariate analysis. CONCLUSIONS: Vitamin D was related to most functional and balance measurements. Serum folate was protective against falls. For every 1 ng/ml increase in serum folate the occurrence of falls decreased by 19%.

Serum 25-OH vitamin D levels in patients with fibromyalgia.

BACKGROUND: The association between low levels of 25-hydroxyvitamin D and non-specific musculoskeletal pain, including fibromyalgia syndrome, is controversial. Several studies have reported a "positive association" and two others found "no association." OBJECTIVES: To test levels of 25OHD in patients with fibromyalgia syndrome and in matched controls. METHODS: The study population comprised 68 premenopausal women with a diagnosis offibromyalgia and 82 age-matched premenopausal women without. The former were identified from the computerized medical databases of five primary care urban clinics in the south of Israel, and the control subjects were attending the participating clinics for regular periodic blood tests. For each patient, the matched control interview and blood test were performed within a week or two from the patient's interview and blood test, thus controlling for expected seasonal variations. RESULTS: Serum 25OHD was measured using different cutoff levels and compared between the groups (< 30 ng/ml, < 20 ng/ml and <15 ng/ml). No statistically significant differences were found between the groups regardless of the cutoff level used. A logistic regression model for predicting women with 25OHD levels 20 ng/ml showed that all the variables examined in both groups (age, country of birth, education) were not statistically significant. We found the expected seasonal variations of 25OHD levels, though these were not statistically significant. CONCLUSIONS: We found no association between fibromyalgia and low 25OHD levels as previously suggested in other studies.

Teaming Up for Trouble: Cancer Cells, Transforming Growth Factor-ß1 Signaling and the Epigenetic Corruption of Stromal Naïve Fibroblasts.

It is well recognized that cancer cells subvert the phenotype of stromal naïve fibroblasts and instruct the neighboring cells to sustain their growth agenda. The mechanisms underpinning the switch of fibroblasts to cancer-associated fibroblasts (CAFs) are the focus of intense investigation. One of the most significant hallmarks of the biological identity of CAFs is that their tumor-promoting phenotype is stably maintained during in vitro and ex vivo propagation without the continual interaction with the adjacent cancer cells. In this review, we discuss robust evidence showing that the master cytokine Transforming Growth Factor-ß1 (TGFß-1) is a prime mover in reshaping, via epigenetic switches, the phenotype of stromal fibroblasts to a durable state. We also examine, in detail, the pervasive involvement of TGFß-1 signaling from both cancer cells and CAFs in fostering cancer development, taking colorectal cancer (CRC) as a paradigm of human neoplasia. Finally, we review the stroma-centric anticancer therapeutic approach focused on CAFs-the most abundant cell population of the tumor microenvironment (TME)-as target cells.

The anti-inflammatory activity of 1,25-dihydroxyvitamin D3 in macrophages.

In a previous study we demonstrated a down-regulatory effect of vitamin D active metabolite (1,25(OH)(2)D(3)) and its vitamin D(2) analog (1,24(OH)(2)D(2)) on TNFalpha expression in macrophages. We also found an inhibitory effect in the physiological concentration (10(-10)M) of 1,25(OH)(2)D(3) which was dose-dependent. This down-regulation, caused by the decrease in NFkappaB activity by 1,25(OH)(2)D(3) and 1,24(OH)(2)D(2), was demonstrated in P388D1 cells transfected with NFkappaB reporter gene (p NFkappaB-Luc) and by EMSA. In our present study we investigated the processes leading to reduced NFkappaB activity on P388D1 cells. A decrease in nuclei NFkappaB-p65 and an increase in cytosolic NFkappaB-p65, were measured, while no changes in total NFkappaB-p65 mRNA and protein levels were observed. Simultaneously, a significant increase in both mRNA and protein levels of the NFkappaB-cytosolic inhibitor, IkappaBalpha, were determined. The half-life of IkappaBalpha-mRNA increased, with a parallel decrease in the phosphorylation of its protein, as the first step of ubiquitinization and degradation. The present results demonstrate that 1,25(OH)(2)D(3) and 1,24(OH)(2)D(2) inhibit TNFalpha expression in macrophages, by increasing IkappaBalpha and decreasing NFkappaB activity. Since NFkappaB is a major transcription factor for TNFalpha and other inflammatory mediators, these findings suggest that 1,25(OH)(2)D(3) and 1,24(OH)(2)D(2) may be used therapeutically as anti-inflammatory agents.

Efficient dual treatment of the hormone-refractory prostate cancer cell line DU145 with cetuximab and 1,25-dihydroxyvitamin D3.

BACKGROUND: Targeting of the epidermal growth factor receptor (EGFR) pathway is a promising treatment strategy for aggressive androgen-refractory prostate cancer (PCa). The effect of treating the androgen-resistant PCa cell line DU145 with a combination of the anti-EGFR drug cetuximab and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) was evaluated. MATERIALS AND METHODS: DU145 cells were treated with 5 nM cetuximab, 100 nM 1,25(OH)2D3 or a combination of both. The effect of the treatments on cell growth, cell-cycle and apoptosis was evaluated. RESULTS: Single-drug treatments decreased DU145 cell growth by up to 25% and caused a 1.5-to 1.7-fold increase of apoptosis, but did not affect the cell-cycle distribution. However, dual treatment with a combination of cetuximab and 1,25(OH)2D3 inhibited DU145 cell proliferation by 40%, caused considerable cell-cycle arrest in the Go/Gl-phase, and enhanced apoptosis by 2.5-fold (compared to the control, p < 0. 0001, p <0. 006 and p <0. 0001, respectively). CONCLUSION: A combination of cetuximab and 1,25(OH)2D3 efficiently suppresses hormone-resistant PCa cell growth and could provide a basis for its clinical application.

Vitamin D and Myofibroblasts in Fibrosis and Cancer: At Cross-purposes with TGF-ß/SMAD Signaling.

The multifaceted involvement of the active vitamin D metabolite 1,25-dihydroxyvitamin D3 (henceforth referred to by the synonyms 1,25(OH)2D3, calcitriol or vitamin D) in blunting the growth of cancer cells is amply recognized. In this review we focused our attention on the cross-talk between 1,25 (OH)2D3 and the tumor microenvironment (TME), signaling out stromal cancer-associated fibroblasts (CAFs), the most abundant TME population, as a target for calcitriol anticancer action. In view of the commonality of the phenotypic signature in myofibroblasts, resident in the cancer stroma and in non-neoplastic fibrotic loci, we examined modes of action of vitamin D in non-neoplastic chronic diseases and in cancer to assess mechanistic similarities and divergences. A constant observation was that 1,25(OH)2D3 or synthetic ligands via the active vitamin D receptor (VDR) impede transforming growth factor (TGF)-ß/mothers against decapentaplegic homologs (SMADs) signaling in myofibroblasts regardless of the initiating insult. The translational impact of 1,25(OH)2D3 in targetting stromal CAFs is discussed.

The combined treatment of 1,25-dihydroxyvitamin D3 and a non-steroid anti-inflammatory drug is highly effective in suppressing prostate cancer cell line (LNCaP) growth.

BACKGROUND: The active metabolite of vitamin D3, 1,25(OH)2D3, is known to possess anti-proliferative and pro-differentiative activities in prostate cancer (PCa) cells. However, its clinical use is limited because of the risk of hypercalcemia. Concurrent administration of lower doses of 1,25(OH)2D3 together with other anticancer drugs may help to overcome this obstacle and lead to an effective and tolerable therapy. In the present in vitro study, we investigated the combined anti-cancer effect of 1,25(OH)2D3 and ibuprofen, a well-known non-steroidal anti-inflammatory drug (NSAID) that is also recognized for its ability to reduce prostate cancer development. MATERIALS AND METHODS: An androgen-sensitive prostate cancer cell line (LNCaP), grown in medium containing androgen (5alpha-dihydrotestosterone (DHT)) or without it, was treated with 1,25(OH)2D3 or ibuprofen alone or with a combination of both drugs. The effects of the treatments on LNCaP cell proliferation, cell cycle and apoptosis were evaluated by the thymidine incorporation method, the propidium iodide method and ELISA, respectively. The unpaired t-test was used for statistical analysis. RESULTS: Simultaneous treatment of LNCaP cells grown without DHT with 10 nM 1,25(OH)2D3 and 0.2 mM ibuprofen decreased cell growth by 42% (p<0.001, compared to the control cells). This effect was found to be additive since each single drug reduced cell proliferation by only 24%. On the other hand, highly significant synergistic cell growth inhibition (67%, p<0.001) was achieved by combined treatment of 1,25(OH)2D3 and ibuprofen in DHT-stimulated LNCaP cells. This combined treatment was also found to be effective in decreasing the cell transition from G1- to S-phase (p<0.003) and effective in enhancing apoptosis. CONCLUSION: Although both 1,25(OH)2D3 and ibuprofen demonstrate in vitro anti-carcinogenic activities as a single drug treatment, the present results showed that the combined use of 1,25(OH)2D3 with ibuprofen is superior to treatment with a single drug.

Novel role of 1,25(OH)(2)D(3) in induction of erythroid progenitor cell proliferation.

OBJECTIVE: Burst-forming unit erythroid and colony-forming unit erythroid growth in vitro is lower in studies of continuous ambulatory peritoneal dialysis patients than healthy controls. Burst-forming unit erythroid growth was potentiated by addition of 1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] and normalized by erythropoietin (Epo) therapy, suggesting an interaction between Epo and 1,25(OH)(2)D(3) at the stem cell level. The objective of this study was to determine the mechanism by which 1,25(OH)(2)D(3) enhances the stimulatory effect of Epo on the growth of erythroid precursor cells. MATERIALS AND METHODS: We examined the effect of 1,25(OH)(2)D(3) and Epo on stem cell proliferation. Proliferation of TF1 cells of erythroid origin was measured by the XTT method, 3[H] thymidine incorporation, and cell counting by trypan blue exclusion; cord blood (CB) stem cells were counted. Epo receptor (EpoR) quantitation was evaluated by 125I-Epo binding and Scatchard analysis, immunoprecipitation, and Western blotting. Expression of EpoR mRNA was measured by reverse transcriptase polymerase chain reaction. RESULTS: The stem cell factor-dependent CB stem cells and the TF1 cells responded to Epo and 1,25(OH)(2)D(3) by increased proliferation, while their simultaneous addition potentiated cell proliferation in a synergistic manner (25.67% +/- 4.8% of Epo proliferation at day 10 for CB cells; p < 0.005). 1,25(OH)(2)D(3) produced an up-regulation of EpoR number in TF1 cells and increased the expression of EpoR mRNA (p < 0.01). CONCLUSIONS: The increase in EpoR expression induced by 1,25(OH)(2)D(3) might explain the synergistic interaction between Epo and 1,25(OH)(2)D(3) in stem cells.

Usefulness of lipoprotein changes during acute coronary syndromes for predicting postdischarge lipoprotein levels.

The aim of our study was to evaluate the lipoprotein changes that occur during acute coronary events, and to determine the lipoprotein threshold levels that identify patients who require future statin therapy. Lipoprotein levels were measured at admission, at 6 hours, the morning after admission, before discharge, and 3 months after discharge in patients with myocardial infarction and unstable angina. Patients with myocardial infarction on thrombolytic therapy (n = 63) and patients with unstable angina (n = 33) had a decrease in low-density lipoprotein (LDL) cholesterol levels < or = 24 hours after admission (-12 +/- 20% and -6 +/- 23%, respectively), but these levels returned to baseline before discharge. In patients with myocardial infarction who did not receive thrombolytic therapy (n = 37), the decrease was more gradual and peaked before hospital discharge (-7 +/- 19%). There was good correlation between LDL cholesterol levels at admission and after discharge, especially in normotriglyceridemic patients. Over 90% of patients with LDL cholesterol > or = 125 mg/dl on the morning after admission were candidates for statin therapy after discharge. Thus, the need for future statin therapy can be predicted with fair reliability during the initial 24 hours after admission. However, elevated baseline triglyceride levels significantly affect these LDL cholesterol changes and complicate prediction of long-term lipoprotein levels.

Sodium valproate, a histone deacetylase inhibitor, enhances the efficacy of vinorelbine-cisplatin-based chemoradiation in non-small cell lung cancer cells.

AIM: To enhance the anticancer activity of vinorelbine, cisplatin and ionizing radiation (IR) combination against non-small cell lung cancer (NSCLC) cells by co-administration of sodium valproate (VPA), a histone deacetylase inhibitor, and to elucidate molecular events underpinning treatment efficacy. MATERIALS AND METHODS: The NSCLC A549 cell line was treated with cisplatin (0.2 µg/ml), vinorelbine (2 nM), VPA (1 mM) and IR (2.5 Gy) alone, or in combination. Cell proliferation, cell-cycle distribution, apoptosis, and levels of DNA double-strand breaks, activated DNA damage checkpoint kinases pCHK1, pCHK2, cell-cycle inhibitors p21CIP1/WAF1 and p27KIP1 were assessed. RESULTS: VPA markedly enhanced the DNA-damaging effect of the cisplatin-vinorelbine-IR combination and induced increased DSBs, and expression of pCHK2, pCHK1, p21CIP1/WAF1 and p27KIP1. These molecular changes led to cell-cycle arrest and increased apoptosis and consequently markedly curtailed cancer cell growth. CONCLUSION: VPA markedly enhances the anticancer activity of cisplatin-vinorelbine-IR combination. This finding has translational implications for enhancing the efficacy of anticancer treatment and for reducing side-effects by reducing doses of radiation and drugs.

Association between bone mineral density and incidence of breast cancer.

INTRODUCTION: Previous studies have suggested an inverse relationship between bone mineral density (BMD) and breast cancer incidence. The primary objective of this study was to assess whether BMD is associated with risk of subsequent breast cancer occurrence in the female population of southern Israel. METHODS: The electronic medical charts of women who underwent BMD at the Soroka Medical Center (SMC) between February 2003 and March 2011 were screened for subsequent breast cancer diagnoses. Women were divided by tertiles of BMD at 3 skeletal sites: lumbar spine (LS, L1-4), total hip (TH) and femoral neck (FN). The incidence of breast cancer was calculated. RESULTS: Of 15268 women who underwent BMD testing, 86 were subsequently diagnosed with breast cancer. Most women in the study were older than 50 years (94.2% and 92.7%, respectively; p = 0.597). Women who subsequently developed breast cancer had a higher mean body-mass index (BMI) (30.9 ± 5.5 vs. 29.1 ± 5.7 p = 0.004) and the mean BMD Z-score was significantly higher than in those without breast cancer for all 3 skeletal sites (LS: 0.36 ± 1.58 vs. -0.12 ± 1.42, p = 0.002; TH: 0.37 ± 1.08 vs. 0.03 ± 1.02, p = 0.002; FN: 0.04 ± 0.99 vs. -0.18 ± 0.94; p = 0.026). Women in the highest Z-score tertiles at the FN and TH had a higher chance of developing breast cancer compared to the lowest tertile; odds ratio of 2.15, 2.02, respectively (P = 0.004 and 0.01 respectively). No association was found between the BMD Z-score and the stage, histology, grade or survival from breast cancer. CONCLUSIONS: This study provides additional support for an inverse association between BMD and the risk of breast cancer.

A combined pretreatment of 1,25-dihydroxyvitamin D3 and sodium valproate enhances the damaging effect of ionizing radiation on prostate cancer cells.

Radiotherapy is one of the curative treatment options for prostate cancer (PCa). However, effective doses of ionizing radiation (IR) have a high risk of side effects. To increase sensitivity of PCa to IR we pretreated human androgen-refractory DU145 PCa cells with a combination of sodium valproate (VPA), a well-tolerated drug with histone deacetylases inhibiting activity, and 1,25-dihydroxyvitamin D3, 1,25(OH)2D3, the active metabolite of vitamin D, a well known anticancer agent. The results show that irradiation (4Gy) of DU145 PCa cells pretreated with a combination of 1 mM VPA and 100 nM 1,25(OH)2D3 efficiently suppressed (87.9%) PCa cell proliferation. IR after combined pretreatment resulted in increased DNA double-strand breaks expressed as levels of phosphorylated histone H2A.X, compared with non-treated cells the increase was 58.1% in pretreated cells and 11.8% in non-pretreated cells (p<0.002). Combined pretreatment enhanced IR-induced activation of DNA damage checkpoint kinase Chk2, 39.0% in pretreated cells compared to 23.8% in non-pretreated cells (p<0.05). These molecular changes led to DNA replication blockade, S-phase cell-cycle arrest and enhanced apoptosis. Cumulatively, the results indicate that combined pretreatment with VPA and 1,25(OH)2D3 followed by IR is a highly effective treatment for human PCa cells. This observation may have important implications for reducing doses of radiation administered to cancer patients thus limiting the severity of side effects.

Vitamin D decreases NFkappaB activity by increasing IkappaBalpha levels.

BACKGROUND: In a previous study we demonstrated the inhibitory effect of 1,25-dihydroxyvitamin D (1,25(OH)(2)D(3)) and its less calcaemic analog 1,24(OH)(2)D(2) on the production of tumour necrosis factor alpha (TNFalpha) by human peritoneal macrophages. The aim of the present study is to examine whether this vitamin D inhibition of TNFalpha is mediated by its major transcription factor, nuclear factor-kappaB (NFkappaB). METHODS: Murine macrophage cells (P388D1) were incubated with 10(-7) M 1,25(OH)(2)D(3) or 1,24(OH)(2)D(2) and then stimulated with lipopolysaccharide. NFkappaB activity was assayed using a reporter gene and by electrophoretic mobility shift assay (EMSA). In addition, we evaluated mRNA and protein levels of NFkappaB-p65 and of IkappaBalpha, a potent NFkappaB inhibitor, and phosphorylated IkappaBalpha. RESULTS: Both 1,25(OH)(2)D(3) and 1,24(OH)(2)D(2) induced a 60% reduction of TNFalpha secretion. By using a reporter gene and EMSA we found that vitamin D markedly reduced NFkappaB activity. 1,25(OH)(2)D(3) or 1,24(OH)(2)D(2) decreased NFkappaB-p65 levels in the nucleus and increased NFkappaB-p65 levels in the cytosol; no changes were observed in the total levels of NFkappaB-p65 protein and mRNA. Concurrently, vitamin D induced a significant increase in mRNA and protein levels of IkappaBalpha (approximately 6.5- and 4.5-fold, respectively). Elevated levels of IkappaBalpha can be explained by the vitamin D-induced prolongation of IkappaBalpha-mRNA half-life from 110 to 190 min and by the decrease in IkappaBalpha phosphorylation. CONCLUSIONS: Vitamin D up-regulates IkappaBalpha levels by increasing mRNA stability and decreasing IkappaBalpha phosphorylation. The increase in IkappaBalpha levels reduces nuclear translocation of NFkappaB and thereby downgrades its activity. Since NFkappaB is a major transcription factor of inflammatory mediators, these findings suggest that the less-calcaemic analog, 1,24(OH)(2)D(2) may be effective as an anti-inflammatory therapeutic agent.